ABSTRACT Fungal polysaccharides possess an important bioactive potential, including antioxidant and anticarcinogenic activity. The aim of this work was to determine the antioxidant activity and cytotoxicity against tumour and non-tumour cell lines acidic polysaccharides (NAAPs) of the fungus Nothophellinus andinopatagonicus . The effect of NAAPs on tumour cells lines was evaluated by MTT assay and flow cytometry. The analyses determined that glucose was the most abundant monomer and IR spectrum showed the typical peaks of β-glucans in the NAAPs. The cell viability assays revealed significant activity of NAAPs against HL-60, HCT-116 and MCF-7 tumour cell lines (IC50 = 767,16 μg mL-1, 1256 μg mL-1 and 4241,7 μg mL-1, respectively); but a much lower cytotoxicity against the non-tumour cell line HGF-1 (outside the range of the highest concentration tested (>10 mg mL-1)). NAAPs affected the cell cycle of HL-60 tumour cells, increasing the percentage of cells in the sub G1 phase and reducing it in the S/G2/M phases. Moreover, low concentrations of NAAPs also showed an effective cytotoxic activity against tumour cell lines while the non-tumour cell line was unaffected, maintaining a viability close to 100%. The antioxidant activity of the highest NAAPs concentration tested was 6.24% and 4.63%, for DPPH and ABTS method, respectively.

Resumo O presente trabalho estabelece uma análise acerca dos conceitos de ondas eletromagnéticas e sua aplicação no processo de difusão de calor em sólidos com o propósito de compreender e difundir o conceito de ondas térmicas, tão pouco abordado dentro da academia. Com essa análise é possível identificar como a temperatura varia em função das propriedades termofísicas dos materiais quando uma amostra é submetida a uma excitação modulada e periódica por meio de ondas eletromagnéticas. Isso permite a utilização de técnicas fototérmicas, por exemplo, radiometria fototérmica, para determinar as propriedades térmicas de materiais como filmes finos e sólidos homogêneos. O conteúdo aqui desenvolvido pode ser amplamente abordado sob a forma de disciplinas em cursos de graduação e pós-graduação para alunos de física, engenharias e áreas afins.

Abstract The current work establishes an analysis of the electromagnetic waves concepts and its application in the process of heat diffusion in solids in order to understand and spread the concept of thermal waves, so little addressed within the academy. It is possible to identify with this analysis how the temperature can change as a function of the thermophysical properties of the materials when a sample is subjected to a modulated and periodic excitation by means of electromagnetic waves. This allows some photothermal techniques, for instance, photothermal radiometry, to determine the thermal properties of materials such as thin films and homogeneous solids. The content developed in this article can be widely addressed in the form of disciplines in undergraduate and graduate courses intended for physics, engineering and students of related fields.

For proper plant growth, soil substrates must be successful for chemical and physical properties in a way that ensures a high capability to reach the nutritional demand of the plant. A suitable nutritional method is the application of organic compounds, urea and, recently, biochar. Here, using the photoacoustic absorption spectroscopy in the visible range, it was analyzed the nutritional capacity of soil samples, submitted to different treatments. This analysis was carried out through the absorption peaks of A and B chlorophylls and cuticle of papaya leaves planted in soils with addition of biochar and with (without) mycorrhiza. All treated samples showed an increase in the peaks of chlorophyll activity in relation to the base sample. Moreover, the samples containing both mycorrhiza and biochar presented the highest peaks of chlorophyll activity, proving to be the best soil fertilization treatment.

Resumo Objetivo: Analisar e comparar a função pulmonar de crianças e adolescentes obesos e eutróficos saudáveis, sem asma, pela espirometria e capnografia volumétrica. Métodos: Estudo transversal com 77 indivíduos (38 obesos) entre cinco e 17 anos. Todos fizeram espirometria e capnografia volumétrica. Os obesos repetiram as avaliações após o uso de broncodilatador. Resultados: Na avaliação da espirometria, os indivíduos obesos, quando comparados com o grupo controle, apresentaram menores valores no volume expiratório forçado no primeiro segundo pela capacidade vital forçada (VEF1/CVF) e nos fluxos expiratórios a 75% da CVF e entre 25-75% da mesma (p < 0,05). A capnografia volumétrica demonstrou que os obesos apresentam maior volume produzido de dióxido de carbono e volume corrente alveolar (p < 0,05). Além disso, a relação entre o volume espaço morto e volume corrente, bem como o slope da fase 3 normalizado pelo volume corrente, foi menor nos indivíduos saudáveis (p < 0,05). Esses dados sugerem que a obesidade não altera a homogeneidade da ventilação, mas sim dos fluxos. Ao subdividir os grupos por idade, foi observada maior diferença na função pulmonar entre indivíduos obesos e saudáveis na faixa acima de 11 anos (p < 0,05). Conclusão: Mesmo sem o diagnóstico de asma por critérios clínicos e sem resposta ao uso de broncodilatador, os indivíduos obesos apresentaram menores valores no VEF1/CVF e nos fluxos expiratórios forçados, o que indica a presença de processo obstrutivo. A capnografia volumétrica indicou nos indivíduos obesos maior volume corrente alveolar, sem alterações na homogeneidade da ventilação, o que sugere alteração nos fluxos, sem comprometimento dos volumes pulmonares.

Abstract Objective: To analyze and compare lung function of obese and healthy, normal-weight children and adolescents, without asthma, through spirometry and volumetric capnography. Methods: Cross-sectional study including 77 subjects (38 obese) aged 5-17 years. All subjects underwent spirometry and volumetric capnography. The evaluations were repeated in obese subjects after the use of a bronchodilator. Results: At the spirometry assessment, obese individuals, when compared with the control group, showed lower values of forced expiratory volume in the first second by forced vital capacity (FEV1/FVC) and expiratory flows at 75% and between 25 and 75% of the FVC (p < 0.05). Volumetric capnography showed that obese individuals had a higher volume of produced carbon dioxide and alveolar tidal volume (p < 0.05). Additionally, the associations between dead space volume and tidal volume, as well as phase-3 slope normalized by tidal volume, were lower in healthy subjects (p < 0.05). These data suggest that obesity does not alter ventilation homogeneity, but flow homogeneity. After subdividing the groups by age, a greater difference in lung function was observed in obese and healthy individuals aged >11 years (p < 0.05). Conclusion: Even without the diagnosis of asthma by clinical criteria and without response to bronchodilator use, obese individuals showed lower FEV1/FVC values and forced expiratory flow, indicating the presence of an obstructive process. Volumetric capnography showed that obese individuals had higher alveolar tidal volume, with no alterations in ventilation homogeneity, suggesting flow alterations, without affecting lung volumes.

ABSTRACT Piperaceae species can be found worldwide in tropical and subtropical areas and many of them have been used for centuries in traditional folk medicine and in culinary. In Brazil, species of Piperaceae are commonly used in some communities as local anesthetic and analgesic. Countrified communities have known some species of the genus Ottonia as "anestesia" and it is a common habit of chewing leaves and roots of Ottonia species to relief toothache. The purpose of this study is to report our findings on new molecules entities obtained from the roots of Ottonia anisum Spreng, in which local anesthetic activity (sensory blockage) is demonstrated for the first time in vivo guinea pig model. Phytochemical investigation led to the isolation of three amides (pipercallosidine, piperine and valeramide) and in an enriched mixture of seven amides (valeramide, 4,5-dihydropiperlonguminine, N-isobutil-6-piperonil-2-hexenamide, piperovatine, dihydropipercallosidine, pipercallosidine and pipercallpsine). Our findings demonstrated the anesthetic potential for the methanolic extract from roots, its n-hexane partition and amides from O. anisum and it is in agreement with ethnobotanical survey.

RESUMO Este trabalho foi conduzido com o objetivo de investigar a aplicabilidade das redes neurais artificiais (RNAs) para estimar a evapotranspiração do cultivo protegido de pimentão. Os dados utilizados compreenderam o segundo ciclo da cultura, cultivado de setembro de 2013 a fevereiro de 2014, constituindo 135 dias de dados meteorológicos diários: temperatura e umidade relativa do ar, velocidade do vento e radiação solar global, utilizadas como variáveis preditoras e evapotranspiração, determinada a partir de dados obtidos por lisímetro de pesagem, usada como variável resposta. Os dados registrados no período foram divididos em três conjuntos para treinamento, teste e validação. O processo de aprendizado da RNA foi estabelecido com precisão aceitável, com erro médio quadrático de 0,005, em que os padrões de evapotranspiração no ambiente protegido estimado pela rede, em comparação àqueles gerados a partir dos dados de lisimetria, apontaram coeficiente de determinação da ordem de 0,87 para uma arquitetura de rede 4-21-1, de múltiplas camadas, algoritmo de aprendizagem de retropropagação do erro e taxa de aprendizagem de 0,1.

ABSTRACT This study aimed to investigate the applicability of artificial neural networks (ANNs) in the prediction of evapotranspiration of sweet pepper cultivated in a greenhouse. The used data encompass the second crop cycle, from September 2013 to February 2014, constituting 135 days of daily meteorological data, referring to the following variables: temperature and relative air humidity, wind speed and solar radiation (input variables), as well as evapotranspiration (output variable), determined using data obtained by load-cell weighing lysimeter. The recorded data were divided into three sets for training, testing and validation. The ANN learning model recognized the evapotranspiration patterns with acceptable accuracy, with mean square error of 0.005, in comparison to the data recorded in the lysimeter, with coefficient of determination of 0.87, demonstrating the best approximation for the 4-21-1 network architecture, with multilayers, error back-propagation learning algorithm and learning rate of 0.01.

The relationship between gross photosynthesis (GPS) and electron transport rate (ETR) in marine algae has been shown to vary as a function of irradiance; however, little is known about the effect of nutrients on the this relationship in seagrasses. The objective of this study was to evaluate the effect of nitrate concentration on the GPS (measured as O2 evolution) vs ETR (estimated by fluorescence quenching analysis) relationship of the seagrass Cymodoceanodosa from the Spanish Mediterranean Sea. Carbon levels in the tissue increased 6.5% when nitrate in the culture medium augmented from 0 to 100 µM. Nitrogen in the tissue, however, increased more than 60% when nitrate concentration in the medium reached 100 µM. Chlorophyll a + b levels increased approximately 30%, while absorptance augmented 15% when nitrate increased from 0 to 100 µM. In general, maximum oxygenic photosynthesis and maximum ETR values increased when nitrate in the medium increased. The relationship between GPS and ETR did not show a linear response at low nitrate levels and high irradiances. In contrast, a linear relationship was observed at nitrate levels above 50 µM, even at high irradiances. The results from this study suggest that the lack of correlation between ETR and GPS is the result of low nitrogen levels in the tissue of marine macrophytes. They also suggest that seasonal fluctuations in nitrate levels or nitrogen pulses, such as those observed during upwelling events, may affect the relationship between GPS and ETR in C. nodosa or other marine macrophytes.

La relación entre la fotosíntesis bruta (FB) y la tasa de transporte electrónico (TTE) en algas marinas varía en función de la irradiancia; sin embargo, poco se sabe sobre el efecto de la concentración de nutrientes sobre la relación entre FB y TTE en fanerógamas marinas. El objetivo de este estudio fue evaluar el efecto de la concentración de nitrato sobre la relación FB (determinada como evolución de O2) vs TTE (estimada mediante análisis de fluorescencia) en la fanerógama marina Cymodoceanodosade la costa del Mediterráneo español. Los niveles de carbono en el tejido incrementaron 6.5% cuando el nitrato del medio aumentó de 0 a 100 µM. En contraste, los niveles de nitrógeno en el tejido aumentaron más de 60% cuando los niveles de nitrato alcanzaron 100 µM. Los niveles de clorofila a + b incrementaron aproximadamente 30%, mientras que la absorptancia aumentó 15% cuando el nitrato aumentó de 0 a 100 µM. En general, los valores de fotosíntesis máxima (O2) y los valores de TTE máxima incrementaron cuando los niveles de nitrato aumentaron en el medio. La relación entre FB y TTE no mostró una respuesta lineal a bajas concentraciones de nitrato en el medio y a altas irradiancias. En contraste, una relación lineal fue observada cuando los niveles de nitrato fueron mayores a 50 µM, aun a altas irradiancias. Los resultados de este estudio sugieren que la falta de correlación entre FB y TTE es el resultado de los bajos niveles de nitrógeno en el tejido de macrófitos marinos. Los resultados sugieren también que fluctuaciones estacionales de los niveles de nitrato o pulsos de nitrógeno, como los observados durante eventos de surgencia, podrían afectar la relación entre FB y TTE en C. nodosa y otras macrófitos marinos.

Objetivo: Testamos a hipótese de que a injeção intramiocárdica direta de células mononucleares de medula óssea em pacientes portadores de cardiomiopatia dilatada não-isquêmica pode melhorar a função ventricular e a capacidade física. Métodos: Trinta pacientes com cardiomiopatia dilatada não isquêmica e fração de ejeção 35% foram randomizados na razão 1:2 em grupos controle e tratado. Grupo células mononucleares de medula óssea recebeu 1,06 ± 108 células mononucleares de medula óssea por mini-toracotomia. Grupo controle não recebeu intervenção. Avaliação foi feita clinicamente e por teste de caminhada 6 minutos (T6m), ressonância magnética e ecocardiogramas. Resultados: Grupo células mononucleares de medula óssea mostrou tendência de melhora da Fração de ejeção - ressonância magnética aos 3 meses, 27,80±6,86% para 30,13±9,06% (P=0,08), retornando ao basal aos 9 meses (28,78%, P=0,77). Grupo controle não apresentou variação (28,00±4,32%; 27,42±7,41% e 29,57±4,50%). Ecocardiogramas - fração de ejeção melhorou no grupo células mononucleares de medula óssea aos 3 meses: 25,09±3,98 para 30,94±9,16 (P=0,01) e aos 12 meses (30,07±7,25%, P=0,001), enquanto o controle não variou: 26,1±4,4 vs. 26,5±4,7 e 30,2±7,39%, P=0,25 e 0,10, respectivamente). Células mononucleares de medula óssea melhorou classe funcional New York Heart Association: 3,40±0.50 para 2,41±0,79 (P=0,002); controles não mudaram (3,37±0,51 para 2,71±0,95; P=0,17). T6m melhorou no grupo células mononucleares de medula óssea (348,00±93,51 m inicial para 370,41±91,56 m aos 12 m, P=0,66) e declinou sem significância no controle (361,25±90,78 m para 330,00±123,42 m aos 12 meses, P=0,66). Não houve diferenças significativas entre os grupos. Conclusão: A tendência intragrupo de melhora funcional e subjetiva não se confirmou quando comparado com controle. Portanto, a injeção direta intramiocárdica de células mononucleares de medula óssea não se associou a mudança significativa na função ventricular. As diferenças observadas no grupo tratado poderiam ser devidas a efeito placebo ou a baixo poder estatístico.

Objective: We tested the hypothesis that direct intramyocardial injection of bone marrow mononuclear cells in patients with non-ischemic dilated cardiomyopathy can improve left ventricular function and physical capacity. Methods: Thirty non-ischemic dilated cardiomyopathy patients with left ventricular ejection fraction <35% were randomized at a 1:2 ratio into two groups, control and treated. The bone marrow mononuclear cells group received 1.06±108 bone marrow mononuclear cells through mini-thoracotomy. There was no intervention in the control group. Assessment was carried out through clinical evaluations as well as a 6-min walk test, nuclear magnectic resonance imaging and echocardiogram. Results: The bone marrow mononuclear cells group showed a trend toward left ventricular ejection fraction improvement, with magnectic resonance imaging - at 3 months, showing an increase from 27.80±6.86% to 30.13±9.06% (P=0.08) and returning to baseline at 9 months (28.78%, P=0.77). Magnectic resonance imaging showed no changes in left ventricular ejection fraction during follow-up of the control group (28.00±4.32%, 27.42±7.41%, and 29.57±4.50%). Echocardiogram showed left ventricular ejection fraction improved in the bone marrow mononuclear cells group at 3 months, 25.09±3.98 to 30.94±9.16 (P=0.01), and one year, 30.07±7.25% (P=0.001). The control group showed no change (26.1±4.4 vs 26.5±4.7 and 30.2±7.39%, P=0.25 and 0.10, respectively). Bone marrow mononuclear cells group showed improvement in New York Heart Association functional class, from 3.40±0.50 to 2.41±0.79 (P=0.002); patients in the control group showed no change (3.37±0.51 to 2.71±0.95; P=0.17). Six-minute walk test improved in the bone marrow mononuclear cells group (348.00±93.51m at baseline to 370.41±91.56m at 12 months, P=0.66) and there was a non-significant decline in the control group (361.25±90.78m to 330.00±123.42m after 12 months, P=0.66). Group comparisons were non-significant. Conclusion: The trend of intragroup functional and subjective improvement was not confirmed when compared to the control group. Direct intramyocardial application of bone marrow mononuclear cells in non-ischemic dilated cardiomyopathy was not associated with significant changes in left ventricular function. Differences observed within the bone marrow mononuclear cells group could be due to placebo effect or low statistical power.

OBJECTIVES: Proper assessment of dyspnea is important in patients with heart failure. Our aim was to evaluate the use of the 5-point Likert scale for dyspnea to assess the degree of pulmonary congestion and to determine the prognostic value of this scale for predicting adverse events in heart failure outpatients. METHODS: We undertook a prospective study of outpatients with moderate to severe heart failure. The 5-point Likert scale was applied during regular outpatient visits, along with clinical assessments. Lung ultrasound with ≥15 B-lines and an amino-terminal portion of pro-B-type natriuretic peptide (NT-proBNP) level >1000 pg/mL were used as a reference for pulmonary congestion. The patients were then assessed every 30 days during follow-up to identify adverse clinical outcomes. RESULTS: We included 58 patients (65.5% male, age 43.5±11 years) with a mean left ventricular ejection fraction of 27±6%. In total, 29.3% of these patients had heart failure with ischemic etiology. Additionally, pulmonary congestion, as diagnosed by lung ultrasound, was present in 58% of patients. A higher degree of dyspnea (3 or 4 points on the 5-point Likert scale) was significantly correlated with a higher number of B-lines (p = 0.016). Patients stratified into Likert = 3-4 were at increased risk of admission compared with those in class 1-2 after adjusting for age, left ventricular ejection fraction, New York Heart Association functional class and levels of NT-proBNP >1000 pg/mL (HR = 4.9, 95% CI 1.33-18.64, p = 0.017). CONCLUSION: In our series, higher baseline scores on the 5-point Likert scale were related to pulmonary congestion and were independently associated with adverse events during follow-up. This simple clinical tool can help to identify patients who are more likely to decompensate and whose treatment should be intensified.

FUNDAMENTO: O fator de crescimento endotelial vascular (VEGF - vascular endothelial growth factor) induz a mobilização de células progenitoras endoteliais (CPEs) com capacidade de proliferação e diferenciação em células endoteliais, contribuindo, dessa forma, para o processo angiogênico. OBJETIVO: Buscamos avaliar o comportamento de CPEs em pacientes com doença cardíaca isquêmica e angina refratária que receberam injeções intramiocardicas de 2000 µg de VEGF165 como terapia única. MÉTODOS: O estudo foi uma subanálise de um ensaio clínico. Pacientes com doença cardíaca isquêmica avançada e angina refratária foram avaliados para inclusão no estudo. Os critérios de inclusão foram: sinais e sintomas de angina e/ou insuficiência cardíaca apesar de tratamento medicamentoso máximo e área de isquemia miocárdica de, no mínimo, 5% conforme avaliado por uma tomografia computadorizada por emissão de fóton único (TCEFU). Os critérios de exclusão foram: idade > 65 anos, fração de ejeção do ventrículo esquerdo < 25% e cancer diagnosticado. Os pacientes cujos níveis de CPE foram avaliados foram incluídos. A intervenção consistiu na administração de 2000 µg de VEGF 165 de plasmídeo injetado no miocárdio isquêmico. A frequência de células CD34+/KDR+ foi analisada por citometria de fluxo antes e 3, 9, e 27 dias após a intervenção. RESULTADOS: Um total de 9 pacientes foram incluídos, 8 homens, média de idade de 59,4 anos, fração de ejeção ventricular esquerda de 59,3%, e classe de angina predominante III. Observou-se um aumento significativo dos níveis de CPEs no terceiro dia após a intervenção. Todavia, 9 e 27 dias após a intervenção, os níveis de CPEs foram similares aos basais. CONCLUSÃO: Identificamos uma mobilização transitória de CPE, com pico no terceiro dia após a intervenção com VEGF 165 em pacientes com angina refratária. Todavia, os níveis de CPEs apresentaram-se semelhantes aos basais 9 e 27 dias após a intervenção.

BACKGROUND: Vascular endothelial growth factor (VEGF) induces mobilization of endothelial progenitor cells (EPCs) with the capacity for proliferation and differentiation into mature endothelial cells, thus contributing to the angiogenic process. OBJECTIVE: We sought to assess the behavior of EPCs in patients with ischemic heart disease and refractory angina who received an intramyocardial injections of 2000 µg of VEGF 165 as the sole therapy. METHODS: The study was a subanalysis of a clinical trial. Patients with advanced ischemic heart disease and refractory angina were assessed for eligibility. Inclusion criteria were as follows: signs and symptoms of angina and/or heart failure despite maximum medical treatment and a myocardial ischemic area of at least 5% as assessed by single-photon emission computed tomography (SPECT). Exclusion criteria were as follows: age > 65 years, left ventricular ejection fraction < 25%, and a diagnosis of cancer. Patients whose EPC levels were assessed were included. The intervention was 2000 µg of VEGF 165 plasmid injected into the ischemic myocardium. The frequency of CD34+/KDR+ cells was analyzed by flow cytometry before and 3, 9, and 27 days after the intervention. RESULTS: A total of 9 patients were included, 8 males, mean age 59.4 years, mean left ventricular ejection fraction of 59.3% and predominant class III angina. The number of EPCs on day 3 was significantly higher than that at baseline (p = 0.03); however, that on days 9 and 27 was comparable to that at baseline. CONCLUSION: We identified a transient mobilization of EPCs, which peaked on the 3th day after VEGF 165 gene therapy in patients with refractory angina and returned to near baseline levels on days 9 and 27.

Nanopartículas mesoporosas de sílica são conhecidas por induzirem hemólise de células vermelhas do sangue (RBCs) humano quando ensaios de citotoxicidade são feitos em solução-tampão de fosfato (PBS). Entretanto, em uma abordagem mais realista, a presença de biomoléculas do plasma sanguíneo precisa ser considerada em qualquer avaliação nanotoxicológica de nanopartículas porosas de SiO2 quando se objetiva a sua utilização em aplicações biomédicas através de administração intravenosa. Nesse contexto, demonstrou-se neste trabalho que nanopartículas porosas de sílica não induzem nenhum efeito citotóxico em células vermelhas do sangue quando ensaios de hemólise são feitos na presença de plasma sanguíneo, independentemente da carga superficial (positiva ou negativa) da nanopartícula. A ausência de hemólise está principalmente associada à adsorção de proteínas do plasma sobre a superfície das nanopartículas, levando à formação de um recobrimento proteico estável (denominado protein corona ou PC) que blinda o ambiente microquímico original das nanopartículas.

Mesoporous silica nanoparticles are known to induce the hemolysis of human red blood cells (RBCs) when citotoxicity assays are performed in a phosphate buffer solution (PBS). However, in a more realistic approach, the presence of blood plasma biomolecules must be considered in any nanotoxicological evaluation of porous SiO2 nanoparticles when biomedical applications through intravenous administration are aimed. In this context, it is demonstrated in this work that porous silica nanoparticles do not induce any cytotoxic effect on RBCs when hemolysis assay is done in the presence of blood plasma, regardless the surface charge (positive or negative) of the nanoparticle. The absence of hemolysis is mainly associated with the adsorption of plasma proteins on the nanoparticle surface, which leads to the formation of a stable protein coating (called protein corona or PC) that shields the original microchemical environment of bare nanoparticles.

This paper proposes a strategy to achieve robust optimization of structures against high-cycle fatigue when a potentially large number of uncertain load cases are considered. The strategy is heavily based on a convexity property of some of the most commonly used high-cycle design criteria. The convexity property is rigorously proven for the Crossland fatigue criterion. The proof uses a perturbation technique and involves the principal stress components and analytical expressions for the applicable fatigue criteria. The multiplicity of load cases is treated using load ratios which are bounded but are otherwise free to vary within certain limits. The strategy is applied to a notched plate subject to traditional normal and shear loadings that possess uncertain or unspecified components.

Cardiopatia isquêmica grave com angina refratária a formas convencionais de tratamento apresenta-se em uma crescente incidência. Para tratar angina refratária, terapias alternativas na tentativa de redução da isquemia miocárdica e alívio de sintomas têm sido estudadas. Neste contexto, a terapia gênica representa uma opção, pela possibilidade de induzir angiogênese, estabelecer circulação colateral e reperfundir miocárdio isquêmico. Diversos ensaios clínicos têm sido conduzidos e, com exceção de casos isolados e específicos de efeitos adversos, há indicação de segurança, viabilidade e potencial eficácia da terapia. O benefício clínico não está bem definido. Neste artigo, revisamos os ensaios clínicos que utilizaram terapia gênica para tratamento de pacientes cardiopatas isquêmicos. A abordagem inclui: (1) isquemia miocárdica e angiogênese, sobre os aspectos fisiopatológicos envolvidos; (2) fatores de crescimento, tratando sobre aspectos específicos e justificando a utilização em pacientes cardiopatas isquêmicos sem opções pela terapêutica convencional; (3) ensaios clínicos controlados, onde é apresentado um resumo dos principais estudos envolvendo terapia gênica para tratamento da cardiopatia isquêmica grave; (4) nossa experiência, especialmente sobre resultados preliminares do primeiro ensaio clínico de terapia gênica do Brasil e (5) perspectivas.

Severe ischemic heart disease with refractory angina, occurs in increasing incidence. Alternative forms of treatment, in an attempt to reduce myocardial ischemia and relief of symptoms has been studied. In this context, gene therapy is an option, for the possibility of inducing angiogenesis, establish collateral circulation and reperfuse ischemic myocardium. Several clinical trials have been conducted and, except for specific cases of adverse effects, there is indication of safety, feasibility and potential effectiveness of therapy. The clinical benefit, however, is not yet well established. In this article we review the clinical trials of gene therapy for patients with ischemic heart disease. The approach includes: (1) myocardial ischemia and angiogenesis on the pathophysiological aspects involved, (2) growth factors, dealing with specific aspects and justifying the use in cardiac patients with no option for conventional therapy, (3) controlled clinical trials, where a summary of the main studies involving gene therapy for severe ischemic heart disease is presented, (4) our experience, especially on preliminary results of the first gene therapy clinical trial in Brazil and (5) future prospects.

FUNDAMENTO: O tramadol é um analgésico de ação central cujo mecanismo de ação envolve a ativação de um receptor opioide. Anteriormente, mostramos que o tramadol e seus enantiômeros apresentavam um efeito inotrópico negativo sobre o músculo papilar no qual o (+)-enantiômero era mais potente que (-)- e (±)-tramadol. OBJETIVO: No presente trabalho, investigamos os efeitos do tramadol e seus enantiômeros na corrente de cálcio tipo L (I Ca-L). MÉTODOS: Os experimentos foram realizados em miócitos ventriculares isolados de ratos Wistar utilizando a técnica de patch-clamp com configuração de célula inteira. RESULTADOS: O tramadol (200 µM) reduziu a amplitude de pico do I Ca-L em potenciais de 0 a +50 mV. Em 0 mV, a I Ca-L foi reduzida em 33,7 ± 7,2%. (+)- e (-)-tramadol (200 µM) produziram uma inibição semelhante da I Ca-L, na qual a amplitude do pico foi reduzida em 64,4 ± 2,8% e 68,9 ± 5,8%, respectivamente a 0 mV (P > 0,05). O tramadol, (+)- e (-)-tramadol mudaram a inativação de estado estacionário de I Ca-L para potenciais de membrana mais negativos. Além disso, tramadol e (+)-tramadol alteraram significativamente a curva de recuperação dependente de tempo da I Ca-L para a direita e reduziram a recuperação de I Ca-L da inativação. A constante de tempo foi aumentada de 175,6 ± 18,6 a 305,0 ± 32,9 ms (P < 0,01) para o tramadol e de 248,1 ± 28,1 ms para 359,0 ± 23,8 ms (P < 0,05) para o (+)-tramadol. O agonista do receptor µ-opioide (DAMGO) não tem nenhum efeito na I Ca-L. CONCLUSÃO: A inibição da I Ca-L induzida por tramadol e seus enantiômeros não teve relação com a ativação de receptores opioides e poderia explicar, pelo menos em parte, seu efeito inotrópico negativo cardíaco.

BACKGROUND: Tramadol is a centrally acting analgesic, whose mechanism of action involves opioid-receptor activation. Previously, we have shown that tramadol and its enantiomers had a negative inotropic effect on the papillary muscle in which the (+)-enantiomer is more potent than (-)- and (±)-tramadol. OBJECTIVE: In this study, we investigated the effects of tramadol and its enantiomers on L-type calcium current (I Ca-L). RESULTS: Tramadol (200 µM) reduced the peak amplitude of I Ca-L at potentials from 0 to +50 mV. At 0 mV, I Ca-L was reduced by 33.7 ± 7.2%. (+)- and (-)-tramadol (200 µM) produced a similar inhibition of I Ca-L, in which the peak amplitude was reduced by 64.4 ± 2.8% and 68.9 ± 5.8%, respectively at 0 mV (p > 0.05). Tramadol, (+)- and (-)-tramadol shifted the steady-state inactivation of I Ca-L to more negative membrane potentials. Also, tramadol and (+)-tramadol markedly shifted the time-dependent recovery curve of I Ca-L to the right and slowed down the recovery of I Ca-L from inactivation. The time constant was increased from 175.6 ± 18.6 to 305.0 ± 32.9 ms (p < 0.01) for tramadol and from 248.1 ± 28.1 ms to 359.0 ± 23.8 ms (p < 0.05) for (+)-tramadol. The agonist of µ-opioid receptor DAMGO had no effect on the I Ca-L. CONCLUSION: The inhibition of I Ca-L induced by tramadol and its enantiomers was unrelated to the activation of opioid receptors and could explain, at least in part, their negative cardiac inotropic effect.