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Biomedical subjects

F Docchio

Publications and source records attributed to F Docchio.

24 records · Page 2Linked to original sources

Time-resolved fluorescence spectroscopy of hematoporphyrin-derivative in human lymphocytes.

This paper reports on time-resolved microfluorimetric measurements on hematoporphyrin-derivative (HpD)-treated lymphocytes. HpD is at present widely used as a tumor-locating and photosensitizing drug. It is therefore of great importance to study the extent to which the HpD uptake process depends on cell functional and structural properties. Time-resolved fluorescence measurements in single cells are very useful in this respect, since they give information on the content of fluorescent molecules through fluorescence peak-intensity, and, indirectly, on the binding properties through the fluorescence decay times. In particular, we studied the dependence of HpD fluorescence on the cellular functional state. To this end, we performed in-cell fluorescence measurements on human lymphocytes, both in quiescent conditions and in the pre-replicative phase, after stimulation with phytohemagglutinin (PHA). We found a higher HpD content in stimulated lymphocytes. Moreover, we found a spectral band around 575 nm, corresponding to a particular porphyrin species, in which the differences between normal and stimulated lymphocytes are more striking. The porphyrin species emitting in this band seems to play a role in the specific interaction of HpD with tumors, since a similar emission band has also been found in tumor cells containing HpD.

Hematoporphyrin Derivative↗

Spectroscopic studies of hematoporphyrin-derivative in culture medium.

This work reports on studies of hematoporphyrin-derivative (HpD) behaviour in culture medium. Absorption, excitation and emission spectra, together with time-resolved fluorescence measurements, were performed. In previous works, similar studies had been carried out on HpD in saline and in lymphocytes: a new porphyrin species (NPS) and the environmental conditions for its formation in saline were studied. A fluorescent emission similar to that presented by the NPS is reported to be more likely in tumor rather than in normal HpD-treated cells, it was also found in greater amounts in lymphocytes in the pre-replicative phase, as compared with quiescent ones. The higher NPS content in stimulated rather than in quiescent lymphocytes may be due either to a differential uptake, as compared with other HpD components, or to a differential formation rate in cells, because of different microenvironmental conditions. To distinguish between these two main assumptions, the formation of NPS in culture medium was studied. The process was very slow: no NPS appeared within the first 40 h. The incubation time of lymphocytes in culture medium added with HpD in the experiments performed was only 1 h and therefore a differential formation rate of NPS may explain the higher content found in stimulated lymphocytes.

Culture Media↗

An automatic pulsed laser microfluorometer with high spatial and temporal resolution.

The paper describes an automatic pulsed laser microfluorometer with high spatial and temporal resolution, developed in our laboratories. The instrument consists of: (i) a nitrogen-laser-pumped dye-laser for the excitation of the fluorescence, (ii) a microscope with additional optics to focus the excitation beam on the sample and to collect the fluorescence, (iii) filters or monochromators to select the output wavelength, (iv) a fast photomultiplier tube to detect the signal, and (v) a dual time-scale microprocessor-controlled signal averager for the acquisition and processing of the signal. Examples are given that show the potential of the time-resolved fluorescence microscopy in studying, quantitatively and qualitatively, the properties of fluorescent molecules.

Animals↗

Measurement of radiation from photochemotherapy equipment using thermoluminescence detector elements (BeO TLD).

The results of experimental dosimetry carried out with beryllium oxide thermoluminiscent material (BeO TLD) are presented. In particular, this material shows a good linearity of response to UV radiation at 365 nm, up to 200 mJ/cm2, and a spectral sensitivity with a peak at 340 nm. The advantages and disadvantages of BeO TLD in comparison with solid state detectors are discussed and suggested for personal and environmental dosimetry of UVA radiation in photochemotherapy.

Beryllium↗

Time-resolved fluorescence microscopy of hematoporphyrin-derivative in cells.

This work presents measurements of time-resolved fluorescence microscopy of hematoporphyrin-derivative (HpD) in single cells of mice tissue (both tumor and normal cells), in HeLa Cells, and in solution. The measurements were performed using a pulsed-laser microfluorometer with high spatial and temporal resolution. In agreement with the results obtained with other techniques, it has been found that the tumor cells examined present an HpD uptake about five times higher than that of the normal cells of the corresponding tissue and that, within a cell, HpD become localized mainly in the cytoplasm. It has also been found that the fluorescence decay time is different in cells as compared with solution, and that the presence of HpD stabilizes cell auto-fluorescence. These results are discussed.

Animals↗

Corneal autofluorescence in diabetic and normal eyes.

Corneal autofluorescence has been lately studied as a predictor of retinopathy severity in diabetic patients. We measured corneal autofluorescence in 138 eyes of 69 diabetic patients and 64 eyes of 32 healthy controls. Diabetic patients were subdivided by the severity of retinopathy according to the Modified Airlie House Classification (stage 1: no or minimal retinopathy; stage 2: minimal background retinopathy; stage 3: background retinopathy; stage 4: (pre-) proliferative retinopathy. The fluorescence peak value and the area underlying the corneal autofluorescence curve were measured with a scanning fluorophotometer (Fluorotron Master, Coherent Radiation Palo Alto CA) Healthy controls' values of corneal autofluorescence (peak value: mean 11.03 +/- 3.77 ng. equivalent fluorescein/ml; area: mean 163.85 +/- 61.40 scan-point x ng. equivalent fluorescein/ml) resulted similar (peak value: p = 0.83; area: p = 0.61) to those of diabetic patients without retinopathy (peak value: mean 11.2 +/- 3.4 ng.eq/ml; area: 170.07 +/- 57.23 scan-pnt.ng.eq/ml). A statistically significant difference was found between diabetic patients without retinopathy and patients with stage 2, 3, 4 retinopathy. No statistically significant difference was found both for the peak value (p = 0.50) and for the area (p = 0.61) between stage 3 and stage 4 retinopathy. The sensitivity and specificity of corneal autofluorescence as a screening test for diabetic retinopathy were 82% and 62% for the peak value, 87% and 60% for the area; the positive predictive value for the presence of diabetic retinopathy was 65% for the peak and 63% for the area.

Adult↗