Alberto Colasanti

Photodynamic therapy with topical δ-aminolaevulinic acid for the treatment of plantar warts

Treatments currently employed for plantar warts are often painful (electrosurgery, cryotherapy) and not always effective (keratolytic agents). In this paper we investigate the effect of photodynamic therapy (PDT) with topical delta-aminolaevulinic acid (ALA) on plantar warts. In order to remove the superficial hyperkeratotic layer of the warts an ointment containing 10% urea and 10% salicylic acid was applied for 7 days. After gentle curettage, a cream containing 20% ALA was applied under an occlusive dressing for 5 h on 64 warts, while 57 warts (controls) received only the vehicle. Both the ALA-treated warts and the controls were irradiated using a visible light lamp (with a range of 400-700 nm, peaking at 630 nm). The light dose was 50 J/cm(2). Patients were followed-up for 22 months. Two months after the last irradiation session 48 (75.0%) out of 64 ALA-PDT treated warts had resolved. By contrast only 13 (22.8%) of the 57 control warts had done so. During the treatment a few patients complained of a mild burning sensation. The absorption of ALA by the verrucous tissue was demonstrated by in vivo fluorescence spectroscopy. This study shows that topical ALA-PDT can be an alternative treatment for plantar warts. Further studies will be necessary in order to optimize the concentration of ALA and duration of treatment.

LASER PHOTOSENSITIZATION OF CELLS BY HYPERICIN

Abstract— Administering a light dose of 90 J/cm2 at 599 nm during incubation with hypericin to a highly differentiated normal epithelial cell line(FRTL–5), derived from Fisher rat thyroid, and to a neoplastic cell line(MPTK–6), derived from the lung metastases of a thyroid carcinoma induced in Fisher rats, produces cell kill at drug doses 1000 times lower than those necessary to cause the same mortality in the dark. The photocytocidal activity of this polycyclic quinone drug on neoplastic cells is superior to that of antitumor anthraquinone drugs, such as daunomycin and mitoxanthrone, and to the photosensitized antiviral activity previously reported for hypericin.

Hypericin photosensitization of tumor and metastatic cell lines of human prostate

We have investigated the photoactivating effect of hypericin on two cancer cell lines: PC-3, a prostatic adenocarcinoma non-responsive to androgen therapy and LNCaP, a lymphonodal metastasis of prostate carcinoma responsive to androgen therapy. The two cell lines are incubated for 24 h with hypericin at concentrations ranging from 0.001 to 0.3 microg/ml in cell culture medium. The cells are irradiated at 599 nm (fluence = 11 J/cm2) using a dye laser pumped by an argon laser. Hypericin exerts phototoxic effects on both cell lines, while it does not produce toxic effects in the absence of irradiation. These results suggest that photodynamic therapy (PDT) with hypericin could be an alternative approach to the treatment of prostatic tumors, and could be beneficial in tumors that are non-responsive to androgen therapy.

Multiple processor version of a Monte Carlo code for photon transport in turbid media

Abstract Although Monte Carlo (MC) simulations represent an accurate and flexible tool to study the photon transport in strongly scattering media with complex geometrical topologies, they are very often infeasible because of their very high computation times. Parallel computing, in principle very suitable for MC approach because it consists in the repeated application of the same calculations to unrelated and superposing events, offers a possible approach to overcome this problem. It was developed an MC multiple processor code for optical and IR photon transport which was run on the parallel processor computer CRAY-T3E (128 DEC Alpha EV5 nodes, 600 Mflops) at CINECA (Bologna, Italy). The comparison between single processor and multiple processor runs for the same tissue models shows that the parallelization reduces the computation time by a factor of about N , where N is the number of used processors. This means a computation time reduction by a factor ranging from about 10 2 (as in our case where 128 processors are available) up to about 10 3 (with the most powerful parallel computers with 1024 processors). This reduction could make feasible MC simulations till now impracticable. The scaling of the execution time of the parallel code, as a function of the values of the main input parameters, is also evaluated.

ENHANCEMENT OF ANTITUMOR DRUG CYTOTOXICITY via LASER PHOTOACTIVATION

Abstract— —We investigate the efficacy of daunomycin, some imino‐ and amino‐substituted daunomycin analogues and the disubstituted aminoanthracenedione, mitoxantrone, in photosensitizing short‐term cell kill upon irradiation in the long wavelength visible range, during incubation of Fisher rat thyroid cells with the drugs. While all compounds exhibit similar cytocidal effects on our cell line, in the absence of irradiation, administering 86 J/cm2 at wavelengths either coincident or close to drug absorption peaks causes greater enhancement in cell mortality for the 4‐demethoxydaunomycin analogues than either the parent drug or its 5‐imino‐derivative. A lower enhancement is observed with mitoxantrone. In particular, C50 doses (i.e. concentrations that would kill 50% cells) as low as ∼10−9 M are found for both 6‐ and 11‐amino 4‐demethoxydaunomycin, compared with the values obtained in the absence of light, which are 2.59 × 10−4 and 0.43 × 10−4 M, respectively. Our previous studies of the photophysical and photochemical properties of the excited states of these drugs, and ESR and spin trapping studies of photosensitized generation of singlet oxygen, which were extended in this work to include mitoxantrone, indicate that the cytocidal effects proceed via type I rather than type II mechanisms.

Enhanced response to daunomycin of normal, tumor and metastatic cell lines via drug photoactivation.

Abstract— We have compared the cytotoxicity of daunomycin in vitro to highly differentiated normal epithelial cells (Fisher rat thyroid cells, FRTL‐5) and to two neoplastic cell lines, a thyroid carcinoma (TK‐6) and its lung metastasis (MPTK‐6). Whereas the cell lines are equally sensitive to the drug in the dark, if irradiated during incubation with daunomycin (86 J/cm2 at 488 nm), they become more and differently sensitive. Namely, the drug doses producing 50% mortality decrease by factors of about 22, 28 and 16 for FRTL‐5, TK‐6 and MPTK‐6 cell lines, respectively. This result correlates with differences in drug uptake and resistance observed in the normal and neoplastic cell lines.

Cell photosensitization by 5-iminodaunomycin activated with red light

5-Iminodaunomycin, an anthracycline antitumor drug exhibiting an absorption peak at 595 nm, is shown to photosensitize in vitro cell kill. The photoactivation is performed irradiating the culture dishes during the incubation with the drug for 2 h with 34 mW/cm2 intensity, that is with light doses of up to 245 J/cm2. Long-term effects of administering 50 ng/ml and light for 2 h are studied in terms of growth curves. We show that photoactivation enhances the dark toxicity by a factor of about 10. Immediate cell death is produced by irradiating the cells in the presence of higher drug concentrations (e.g., 1000 ng/ml) which, however, are not toxic in the short term if administered in the dark. The viable cell percentage decreases at increasing light doses, being about 0.6% at the maximum dosage. Administering lower light doses, such as 30 J/cm2, which corresponds to an exposure duration of 15 min, has a short-term effect on the cell survival that strongly depends on the timing of the exposures within the incubation period.

MS‐2 fibrosarcoma characterization by laser induced autofluorescence

MS‐2 fibrosarcoma implanted in BALB‐CDF1 mice was investigated by frequency and time domain measurements of the autofluorescence (AF) radiation emitted upon excitation by a N2 laser beam (337.1 nm).

Excited singlet state properties of anthracenedione photosensitizers.

The absorption, fluorescence and S1 state kinetics of anthracycline antitumour drugs (e.g. daunomycin, adriamycin) and several imino- and/or amino-substituted derivatives are investigated. The study, which includes all anthracyclines which possess photocytocidal activity, is extended to the disubstituted aminoanthracenedione, mitoxantrone, a red-light-absorbing antitumour drug whose activity, both in vitro and in vivo, is enhanced by photoactivation. The S1 state of the anthracycline imino and amino derivatives, in aqueous buffer at pH 7.4, is characterized by bi-exponential decay kinetics which indicates the presence of two ground state populations differing in the extent of hydrogen bonding. The ammonium group of the sugar moiety of anthracyclines contributes to the quenching of the S1 state population through a prototropic mechanism.

Noninvasive spectroscopic analysis of dermatological lesions excited with N2 laser

A system for the excitation and detection of autofluorescence induced by N2 laser light (337.1 nm, 1 divided by 1.5 mJ/pulse) on biological tissues in vivo is described. Spectra obtained with medium spectral resolution spectrograph were detected with a 512 by 512 CCD array. Spectral measurements performed on patients bearing different cutaneous diseases show that, in the wavelength range 420 - 480 nm, pathological skin tissues fluoresce less than healthy ones and these differences might be useful for diagnostic purposes.