A novel modelling and experimental technique to predict and measure tissue temperature during CO2 laser stimuli for human pain studies.

Laser nerve stimulation is now accepted as one of the preferred methods for applying painful stimuli to human skin during pain studies. One of the main concerns, however, is thermal damage to the skin. We present recent work based on using a CO2 laser with a remote infrared (IR) temperature sensor as a feedback system. A model for predicting the subcutaneous skin temperature derived from the signal from the IR detector allows us to accurately predict the laser parameters, thus maintaining an optimum pain stimulus whilst avoiding dangerous temperature levels, which could result in thermal damage. Another aim is to relate the modelling of the CO2 fibre laser interaction to the pain response and compare these results with practical measurements of the pain threshold for various stimulus parameters. The system will also allow us to maintain a constant skin temperature during the stimulus. Another aim of the experiments underway is to review the psychophysics for pain in human subjects, permitting an investigation of the relationship between temperature and perceived pain.

Microemulsion and diafiltration approaches: an attempt to maximize the global yield of DNA-loaded nanospheres.

The yield of DNA-loaded nanospheres in its widest definition includes encapsulation efficiency and the integrity of the loaded molecules plus the production yield of fabricated nanospheres. The former aspect could be considerably improved by adopting the microemulsion concept to enhance the stability of the primary emulsion during the preparation of nanospheres by the double emulsion solvent-removal method. The droplet size of the mentioned emulsion was monitored by means of photon electron correlation spectroscopy and could serve as an index for emulsion fineness and stability. DNA stability as a function of applied mechanical stress was monitored by horizontal agarose gel electrophoresis. The impact of the primary emulsion on nanosphere porosity was assessed as well. Regarding the second aspect of the global yield of nanospheres, i.e. production yield, a modified diafiltration technique was adopted for the washing and recovery processes in comparison with the traditional and for the conservation of particle size characteristics of the recovered nanospheres.

The application of a compact multispectral imaging system with integrated excitation source to in vivo monitoring of fluorescence during topical photodynamic therapy of superficial skin cancers.

A novel, compact and low-cost multispectral fluorescence imaging system with an integrated excitation light source is described. Data are presented demonstrating the application of this method to in vivo monitoring of fluorescence before, during and after topical 5-aminolevulinic acid photodynamic therapy of superficial skin cancers. The excitation source comprised a fluorescent tube with the phosphor selected to emit broadband violet light centered at 394 nm. The camera system simultaneously captured spectrally specific images of the fluorescence of the photosensitizer, protoporphyrin IX, the illumination profile and the skin autofluorescence. Real-time processing enabled images to be manipulated to create a composite image of high contrast. The application and validation of this method will allow further detailed studies of the characteristics and time-course of protoporphyrin IX fluorescence, during topical photodynamic therapy in human skin in vivo.