Spectroscopic Characterization of Emulsions Generated with a New Laser-Assisted Device.

This paper presents a spectroscopic study of emulsions generated with a laser-assisted device. Fourier transform infrared (FTIR), Raman and UV-Vis-NIR reflectance spectra of emulsions, recorded before and after exposure to laser radiation were used to characterize the effect of laser irradiation. The paper also presents a comparison between the calculated IR spectra and the experimental FTIR spectra of an emulsion's components. FTIR measurements allowed the identification of absorption bands specific to each of the emulsions' components. Moreover, it enabled the observation of destabilization of the emulsion in real-time. Raman spectroscopy allowed the observation of the modifications at a molecular level, by identifying the vibrations of the representative functional groups and the polymerization of sodium tetradecyl sulfate (STS) molecules by analyzing the evolution of the carbonyl band. UV-Vis-NIR reflectance spectra of emulsions before and after exposure to laser radiation showed that the physical characteristics of the emulsions changed during irradiation-the dimensions of the droplets decreased, leading to an emulsion with a better time stability. These results proved that the employed spectroscopy techniques were powerful tools in emulsion analysis.

Fluorescence and Time-Delayed Lasing during Single Laser Pulse Excitation of a Pendant mm-Sized Dye Droplet.

Fluorescence and lasing emission that are produced separately in time during excitation laser pulse for an mm-sized Rhodamine 6G dye-water droplet are reported. The droplet acts as a quasi-spherical closed optical resonator and due to multiple internal reflections, the resonant amplified emission is delayed with respect to fluorescence emission. Measurements of the temporal evolution of the droplet's emission were performed by varying the signal acquisition gate width and gate delay with respect to the pumping pulse. The droplet emission spectra are structured in two bands which appear one after the other in time: first, the fluorescence emission band which follows pumping laser pulse time shape and then a second band, the lasing band, placed at shorter wavelengths and formed in time after the peak of the pumping laser pulse intensity, on the pulse tail. The lasing threshold pumping intensity is much lower than those for typical dye lasers.

Case series about ex vivo identification of squamous cell carcinomas by laser-induced autofluorescence and Fourier transform infrared spectroscopy.

An ex vivo case series aimed at identification of normal laryngeal tissue from laryngeal epidermoid squamous keratinized carcinoma by measuring laser-induced autofluorescence (LIAF) and Fourier transform infrared-attenuated total reflectance (FTIR-ATR) spectra is presented. The case series results were obtained for paired samples extracted from three patients (exclusion: macroscopic changes of normal vocal cord observed during surgery; surgical intervention on vocal cord, treated only with chemotherapy or radiotherapy for carcinoma; inclusion: men, aged 57-68, non-smokers). For LIAF analysis, a 375-nm picosecond pulsed laser diode with 31 MHz pulse repetition rate, 100 ps full-time width at half-maximum, and average power 0.49 µW was used. LIAF and FTIR-ATR spectra show noticeable differences between normal and malignant tissues. LIAF spectra differed in shape of emitted band, peak position, and band relative intensity of the two kinds of samples, evidencing hypsochromic shift and mean fluorescence intensity decrease of (75.42 ± 3)% in malignant tissue with respect to the normal one. The lack of 1745 cm-1 band in FTIR-ATR spectra for malignant tissues could be considered an important indicative of the presence of this kind of tissue; moreover, it resulted a greater contribution of lipids and proteins in normal tissue and of collagen in malignant tissue. Penetration depth of the evanescent wave was about 2 µm at an angle of 42°. The two spectroscopic methods are complementary, are applicable for real-time measurements, and may enhance cancer detection and diagnostics. Results presented in this study evidence the potential of the two methods for future in vivo studies.

Laser induced autofluorescence lifetime to identify larynx squamous cell carcinoma: Short series ex vivo study.

Laser induced autofluorescence (LIAF) lifetime is useful to distinguish between normal laryngeal tissues and squamous cell carcinoma (SCC) based on variations of their biochemical composition and structure alterations. LIAF was collected from samples constituted by pairs of normal and malignant tissue, which were excised from three patients. Exclusion criteria for samples harvest were: (i) macroscopic changes of normal vocal cord observed during surgery; (ii) previous surgical intervention on vocal cord, (iii) patients treated only with chemotherapy or radiotherapy for carcinoma. Inclusion conditions: men, aged 57-68, non-smokers. A pulsed laser diode excited LIAF at 375 nm and 31 MHz repetition rate; beam full-time width at half-maximum was 87 ps at an average power of 0.49 mW. Mean LIAF lifetime for normal tissues was (3.75 ± 0.49) ns and for malignant (4.37 ± 0.85) ns: it is longer in malignant than in normal tissue. Variance analysis made with Fisher's test has shown no significant difference between patients for normal tissues; the same was true for malignant. Though, when malignant tissue was compared to normal for the same patients as well as between patients, a significant difference (significance level of 5%) was evidenced. Time-resolved LIAF may allow better differentiation between normal and malignant tissues in patients diagnosed with larynx SCC.

Chlorpromazine transformation by exposure to ultraviolet laser beams in droplet and bulk.

Multiple drug resistance requires a flexible approach to find medicines able to overcome it. One method could be the exposure of existing medicines to ultraviolet laser beams to generate photoproducts that are efficient against bacteria and/or malignant tumors. This can be done in droplets or bulk volumes. In the present work are reported results about the interaction of 266nm and 355nm pulsed laser radiation with microdroplets and bulk containing solutions of 10mg/ml Chlorpromazine Hydrochloride (CPZ) in ultrapure water. The irradiation effects on CPZ solution at larger time intervals (more than 30min) are similar in terms of generated photoproducts if the two ultraviolet wavelengths are utilized. The understanding of the CPZ parent compound transformation may be better evidenced, as shown in this paper, if studies at shorter than 30minute exposure times are made coupled with properly chosen volumes to irradiate. We show that at exposure to a 355nm laser beam faster molecular modifications of CPZ in ultrapure water solution are produced than at irradiation with 266nm, for both microdroplet and bulk volume samples. These effects are evidenced by thin layer chromatography technique and laser induced fluorescence measurements.

Toxicity study in blood and tumor cells of laser produced medicines for application in fabrics.

Phenothiazine derivatives are non-antibiotics with antimicrobial, fungistatic and fungicidal effects. We exposed to a high energy UV laser beam phenothiazines solutions in water at 20mg/mL concentration to increase antibacterial activity of resulting mixtures. Compared to previous results obtained on bacteria, more research is needed about UV laser irradiated phenothiazines applications on cancer cell cultures to evidence possible anticancerous properties. Evaluation of the safety of the newly obtained photoproducts in view of use on humans is also needed. Due to expensive animal testing in toxicology and pressure from general public and governments to develop alternatives to in vivo testing, in vitro cell-based models are attractive for preliminary testing of new materials. Cytotoxicity screening reported here shows that laser irradiated (4h exposure time length) chlorpromazine and promazine are more efficient against some cell cultures. Interaction of laser irradiated phenothiazines with fabrics show that promethazine and chlorpromazine have improved wetting properties. Correlation of these two groups of properties shows that chlorpromazine appears to be more recommended for applications on tissues using fabrics as transport vectors. The reported results concern stability study of phenothiazines water solutions to know the time limits within which they are stable and may be used.

Exposure of chlorpromazine to 266 nm laser beam generates new species with antibacterial properties: contributions to development of a new process for drug discovery.

INTRODUCTION: Phenothiazines when exposed to white light or to UV radiation undergo a variety of reactions that result in degradation of parental compound and formation of new species. This process is slow and may be sped up with exposure to high energy light such as that produced by a laser. METHODS: Varying concentrations of Chlorpromazine Hydrochloride (CPZ) (2-20 mg/mL in distilled water) were exposed to 266 nm laser beam (time intervals: 1-24 hrs). At distinct intervals the irradiation products were evaluated by spectrophotometry between 200-1500 nm, Thin Layer Chromatography, High Pressure Liquid Chromatography (HPLC)-Diode Array Detection, HPLC tandem mass spectrometry, and for activity against the CPZ sensitive test organism Staphylococcus aureus ATCC 25923. RESULTS: CPZ exposure to 266 nm laser beam of given energy levels yielded species, whose number increased with duration of exposure. Although the major species produced were Promazine (PZ), hydroxypromazine or PZ sulfoxide, and CPZ sulfoxide, over 200 compounds were generated with exposure of 20 mg/mL of CPZ for 24 hrs. Evaluation of the irradiation products indicated that the bioactivity against the test organism increased despite the total disappearance of CPZ, that is due, most probably, to one or more new species that remain yet unidentified. CONCLUSIONS: Exposure of CPZ to a high energy (6.5 mJ) 266 nm laser beam yields rapidly a large number of new and stable species. For biological grade phenothiazines (in other words knowing the impurities in the samples: solvent and solute) this process may be reproducible because one can control within reasonably low experimental errors: the concentration of the parent compound, the laser beam wavelength and average energy, as well as the duration of the exposure time. Because the process is "clean" and rapid, it may offer advantages over the pyrogenically based methods for the production of derivatives.

Direct modification of bioactive phenothiazines by exposure to laser radiation.

Whereas exposure of combinations of a phenothiazine and bacterium to incoherent UV increases the activity of the phenothiazine, exposure of the phenothiazine alone does not yield an increase of its activity. Because the laser beam energy is greater than that produced by the incoherent UV sources, exposure of phenothiazines to specific lasers may yield molecules with altered activities over that of the unexposed parent. Chlorpromazine, thioridazine and promethazine active against bacteria were exposed to two distinct lasers for varying periods of time. Absorption and fluorescence spectra were conducted prior to and post-exposure and the products of laser exposure evaluated for activity against a Staphylococcus aureus ATCC strain via a disk susceptibility assay. Exposure to lasers alters the absorption/fluorescence spectra of the phenothiazines; reduces the activity of thioridazine against the test bacterium; produces a highly active chlorpromazine compound against the test organism. Exposure of phenothiazines to lasers alters their structure that results in altered activity against a bacterium. This is the first report that lasers can alter the physico-chemico characteristics to the extent that altered bioactivity results. Exposure to lasers is expected to yield compounds that are difficult to make via chemical manipulation methods. A survey of selected patents of interest, even co-lateral for the subject of this article is shortly made.