RESUMEN
We have recently made advancements in a linear electrodynamic quadrupole (LEQ) device for capturing and levitating either single or multiple micro-particles that provides significant improvements in capture efficiency, reliability, and optical measurement access. We have used our LEQ to trap particles ranging from 30 to less than 0.5 µm in size and provide a controlled environment to study particle physical/chemical dependencies on temperature, relative humidity, and gas constituents. To demonstrate this approach, we present data and analysis of liquid-droplet evaporation rates for two materials: glycerol and dibutyl sebacate. Droplet size was monitored as a function of time by two independent optical methods: direct imaging and fixed-angle light scattering. This new approach provides a means to rapidly characterize a wide range of aerosol particle properties and a platform for development of new aerosol optical-diagnostic measurements.
RESUMEN
Plasmodium falciparum is a highly lethal malaria parasite of humans. A major portion of its life cycle is dedicated to invading and multiplying inside erythrocytes. The molecular mechanisms of erythrocyte invasion are incompletely understood. P. falciparum depends heavily on sialic acid present on glycophorins to invade erythrocytes. However, a significant proportion of laboratory and field isolates are also able to invade erythrocytes in a sialic acid-independent manner. The identity of the erythrocyte sialic acid-independent receptor has been a mystery for decades. We report here that the complement receptor 1 (CR1) is a sialic acid-independent receptor for the invasion of erythrocytes by P. falciparum. We show that soluble CR1 (sCR1) as well as polyclonal and monoclonal antibodies against CR1 inhibit sialic acid-independent invasion in a variety of laboratory strains and wild isolates, and that merozoites interact directly with CR1 on the erythrocyte surface and with sCR1-coated microspheres. Also, the invasion of neuraminidase-treated erythrocytes correlates with the level of CR1 expression. Finally, both sialic acid-independent and dependent strains invade CR1 transgenic mouse erythrocytes preferentially over wild-type erythrocytes but invasion by the latter is more sensitive to neuraminidase. These results suggest that both sialic acid-dependent and independent strains interact with CR1 in the normal red cell during the invasion process. However, only sialic acid-independent strains can do so without the presence of glycophorin sialic acid. Our results close a longstanding and important gap in the understanding of the mechanism of erythrocyte invasion by P. falciparum that will eventually make possible the development of an effective blood stage vaccine.
Asunto(s)
Eritrocitos/metabolismo , Eritrocitos/parasitología , Malaria Falciparum/metabolismo , Glicoproteínas de Membrana/metabolismo , Ácido N-Acetilneuramínico/metabolismo , Plasmodium falciparum/fisiología , Receptores de Complemento/metabolismo , Receptores Inmunológicos/metabolismo , Animales , Western Blotting , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Humanos , Inmunoprecipitación , Malaria Falciparum/virología , Merozoítos/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Lectina 1 Similar a Ig de Unión al Ácido SiálicoRESUMEN
Calculations of the depolarization ratio, D(Theta, lambda) = 1 - (S22)/(S11), for light scattered from an ensemble or cloud of single aerosolized spores in air were studied using the discrete dipole approximation (DDA), sometimes also called the coupled-dipole approximation. Here S(ij) is the appropriate Mueller matrix element for scattering angle Theta and wavelength lambda. The effect of modest shape changes on D(Theta, lambda) was determined. The shapes compared were prolate ellipsoids versus right circular cylinders joined smoothly to end caps consisting of hemispheres of the same diameter as the cylinder. Using the same models, the graphs of (S34)/(S11) versus angle were compared with those for D(Theta, lambda). The latter shows sensitivity to length in some cases we examined, while (S34)/(S11) does not. Size parameters and optical constants suggested by measurements of Bacillus cereus endospores were used. An ensemble of spores was modeled with prolate spheroids. The results of this model were compared with results of a model using the same size and optical parameters, but for capped cylinders. The two models produced distinguishably different results for the same parameters. In calculations for all the graphs shown, averaging over random orientations was performed. Averaging over size distributions similar to those from experimental measurements was performed where indicated. The results show that measurements of D(Theta, lambda) could be quite useful in characterizing the shape of particles in an unknown aerosol and for distinguishing between two likely shapes, but not to reconstruct the shapes from the graphs alone without additional information.
Asunto(s)
Microbiología del Aire , Aire/análisis , Algoritmos , Bacillus cereus/aislamiento & purificación , Técnicas Biosensibles/métodos , Monitoreo del Ambiente/métodos , Refractometría/métodos , Esporas Bacterianas/aislamiento & purificación , Luz , Reproducibilidad de los Resultados , Dispersión de Radiación , Sensibilidad y EspecificidadRESUMEN
The application in light scattering of the Mueller matrix ratio (S34)/(S11) for determining average particle size is extended to a large size parameter range for spherical or randomly oriented rod-shaped particles such as micro-organisms. It is shown that combining the graph of this ratio with a Coulter counter measurement of particle volume gives results in agreement with microscopic measurements. Thus this combination provides a method to measure particle diameter and width simultaneously in real time for elongated particles such as bacteria, which are measured in vivo with this method. An approximate empirical formula is developed to estimate the motion of the extrema in the graph of the oscillating matrix ratio as size changes occur. This formula is also shown to be consistent with wavelength changes.