RESUMO
OBJECTIVE: Cannabis is an illegal drug that has been under the spotlight in recent years, due to its vast array of effects on different biological systems. The role of cannabis has been investigated in the management of pain in acute pancreatitis (AP), even though some studies suggest that it may have a causative effect in this pathology and could be considered the underlying etiology in some cases of idiopathic AP. In this case report, we discuss the case of a young man who presented with three different episodes of AP, with apparently no significant history of alcohol and drug consumption, and with no evidence of a biliary, genetic or, autoimmune etiology. During the third episode, in which he had developed a voluminous pseudocyst, treated trough ultrasound (EUS)-guided drainage, he admitted consumption of cannabis daily. The Naranjo score resulted to be 6 (confirming the possible causality), and it was suggested to the patient to avoid cannabis consumption. Since then, he did not develop any other AP episodes. In summary, cannabis should be considered among the possible AP etiologies, as its causative identification and interruption may significantly improve the course of several idiopathic APs.
Assuntos
Canabinoides/efeitos adversos , Cannabis/efeitos adversos , Pancreatite/induzido quimicamente , Adulto , Humanos , Masculino , Pancreatite/diagnóstico por imagemRESUMO
We have characterized a commercial confocal scanning head for the detection of single molecule fluorescence by two-photon excitation. We have verified that the distribution of the fluorescence emitted by dyes and labeled proteins on glass substrates is discrete with quanta proportional to a common reference signal. We describe and test a simple and quantitative tool to discriminate between single molecules and molecular aggregates on single snapshots based on the analysis of the intensity distribution. We have verified the square dependence of the fluorescence intensity vs. the excitation power, suggesting that no appreciable saturation and fast photo-damage of the chromophores takes place at the excitation power employed here.
Assuntos
Microscopia de Fluorescência , Fluorescência , CinéticaRESUMO
We have combined a confocal laser scanning head modified for TPE (two-photon excitation) microscopy with some spectroscopic modules to study single molecules and molecular aggregates. The behavior of the TPE microscope unit has been characterized by means of point spread function measurements and of the demonstration of its micropatterning abilities. One-photon and two-photon mode can be simply accomplished by switching from a mono-mode optical fiber (one-photon) coupled to conventional laser sources to an optical module that allows IR laser beam (two-photon/TPE) delivery to the confocal laser scanning head. We have then described the characterization of the two-photon microscope for spectroscopic applications: fluorescence correlation, lifetime and fluorescence polarization anisotropy measurements. We describe the measurement of the response of the two-photon microscope to the light polarization and discuss fluorescence polarization anisotropy measurements on Rhodamine 6G as a function of the viscosity and on a globular protein, the Beta-lactoglobulin B labeled with Alexa 532 at very high dilutions. The average rotational and translational diffusion coefficients measured with fluorescence polarization anisotropy and fluorescence correlation methods are in good agreement with the protein size, therefore validating the use of the microscope for two-photon spectroscopy on biomolecules.
Assuntos
Microscopia Confocal , Microscopia de Fluorescência , Animais , Imageamento Tridimensional , Microscopia de Polarização , Modelos Teóricos , Fótons , Análise EspectralRESUMO
Multi-photon driven photo-switching between dark and bright (fluorescent) states of a green fluorescent protein (GFP) mutant is demonstrated. A single-protein investigation shows the existence of two distinct bright states that display sharp two-photon cross-section bands peaked at 780 nm and at 870 nm. Fluorescence of these two species can be independently switched on and off. These results highlight a new photoconversion pathway for photochromic GFPs and can have significant applications in multi-photon confocal microscopy and in optical data-storage architectures.
Assuntos
Proteínas de Fluorescência Verde/análise , Proteínas de Fluorescência Verde/química , Microscopia Confocal/métodos , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Espectrometria de Fluorescência/métodos , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/efeitos da radiação , Mutagênese Sítio-Dirigida , Proteínas Recombinantes/análise , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/efeitos da radiaçãoRESUMO
OBJECTIVES: To describe our single-port experience in gynecologic oncology surgery, and emphasize the feasibility to use the single-port in this surgery. PATIENTS AND METHODS: It is a retrospective, feasibility study, monocentric. All patients who were operated by the single-port, between 1st January 2010 to 1st November 2011, were included. RESULTS: We note that 107 patients were included. We made different interventions: uni- and bilateral salpingo-ovariectomy, hysterectomy, pelvic and para-aortic lymph node sampling or lymphadenectomy in gynecologic malignancies. The median age of the population and the body mass index were respectively 52 and 22.6 kg/m(2). In total, six interventions will be converted. The median hospital stay of patients, all procedures combined, was 2 days. We find low rate of postoperative complications. CONCLUSION: Gynecological cancer surgery appears feasible for single-port. However, we need other studies to confirm a benefit of using the single-port compared to conventional laparoscopy.
Assuntos
Neoplasias dos Genitais Femininos/cirurgia , Procedimentos Cirúrgicos em Ginecologia/métodos , Adulto , Idoso , Estudos de Viabilidade , Feminino , Humanos , Histerectomia/métodos , Tempo de Internação , Excisão de Linfonodo/métodos , Pessoa de Meia-Idade , Ovariectomia/métodos , Complicações Pós-Operatórias/epidemiologia , Salpingectomia/métodosRESUMO
An organic dye, SAMSA, bound to gold nanoparticles, displays random photoactivated fluorescence blinking whose rate depends on the size of the nanoparticles. We report experiments indicating that (1) the dye emission wavelength is red-shifted (10-30 nm) by applying an external low voltage (1-10 V) and that (2) the fluorescence emission of single dyes can be resonantly driven by tuning the alternating external bias frequency from 1 to 3 Hz, depending on the nanoparticle size. These properties appear highly valuable and promising for devising light emitting nanostructures.
Assuntos
Corantes Fluorescentes/química , Corantes Fluorescentes/efeitos da radiação , Ouro/química , Nanoestruturas/química , Nanoestruturas/efeitos da radiação , Nanotecnologia/métodos , Espectrometria de Fluorescência/métodos , Sítios de Ligação , Cristalização/métodos , Campos Eletromagnéticos , Substâncias Macromoleculares/química , Teste de Materiais , Conformação Molecular , Nanoestruturas/ultraestrutura , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
We have studied the unfolding of single molecules of GFP-mut2 mutant trapped in wet silica gels in a wide range of GuHCl concentration. After the addition of denaturant, the number of fluorescent molecules decreases with unfolding rates (of the order of 0.01 min(-1)) that are in very good agreement with bulk fluorescence and circular dichroism data. Unexpectedly, single molecule experiments show rare fluctuations in the number of fluorescent proteins at equilibrium. On the other hand, although a first approximate description of the number decays can be reasonably performed by single exponential functions, the distributions of the single molecule unfolding times show a maximum at times congruent with 50-100 min up to the denaturation midpoint concentration of [GuHCl] congruent with 2.5 M. A theoretical analysis of the distributions indicates that this feature is a fingerprint of the competition between unfolding and refolding processes when the protein is very far from the midpoint denaturant concentration.
Assuntos
Proteínas de Fluorescência Verde/química , Géis , Proteínas de Fluorescência Verde/genética , Cinética , Mutação , Conformação Proteica , Desnaturação Proteica , Dobramento de Proteína , Sílica Gel , Dióxido de Silício , Espectrometria de FluorescênciaRESUMO
We report the two-photon excitation and emission or a recently developed green fluorescent protein (GFP) mutant, E(2)GFP. Two main excitation bands are found at 780 and 870 nm. Blinking and irreversible and reversible bleaching were observed. Fluorescence blinking occurs in the millisecond range and has been ascribed to conversions between the neutral, anionic and dark zwitterionic states. Bleaching is observed after approximately 10 to 400 ms depending on the excitation power, and it is probably due to a conversion to a dark state. The striking feature of this GFP mutant is that the fluorescence can be recovered with very high efficiency only upon irradiation at 720 +/- 10 nm. This GFP mutant therefore seems promising as an almost permanent chromophore for two-photon excitation (TPE) microscopy or for applications in single-molecule memory arrays.
Assuntos
Recuperação de Fluorescência Após Fotodegradação/métodos , Proteínas de Fluorescência Verde/química , Animais , Ânions , Elétrons , Recuperação de Fluorescência Após Fotodegradação/instrumentação , Proteínas de Fluorescência Verde/metabolismo , Concentração de Íons de Hidrogênio , Hidrozoários , Cinética , Proteínas Luminescentes/química , Microscopia de Fluorescência/métodos , Mutação , Distribuição Normal , Fotoquímica , Fótons , Prótons , Espectrometria de Fluorescência , Temperatura , TermodinâmicaRESUMO
We report on the construction details of a compact autocorrelator set-up for the measurement of the width of infrared laser pulses at the focal plane of a microscope for two-photon excitation fluorescence imaging. One of the novelties of the set-up, which leads to an improved measurement accuracy, is the use of a modulation technique that is achieved by mounting one of the interferometer mirrors on a loudspeaker driven by a sinusoidal bias at low frequency. A non-linear least-square routine selects only that part of the fluorescence signal that is modulated at the same frequency as the loudspeaker bias. To further increase the accuracy, the laser pulse width is obtained from a series of measurements at different values of the modulation bias. The autocorrelator is a compact single bread-board (10 x 20 cm); it is PC-controlled both for the acquisition and the analysis of the data and can be coupled to different ports of the microscope. The increase in the pulse width measured for three different ports of the microscope is well accounted for by the group velocity dispersion and the glass thickness of the optics found along these paths.