Stability Studies of UV Laser Irradiated Promethazine and Thioridazine after Exposure to Hypergravity Conditions.

Pharmaceuticals carried into space are subjected to different gravitational conditions. Hypergravity is encountered in the first stage, during spacecraft launching. The stability of medicines represents a critical element of space missions, especially long-duration ones. Therefore, stability studies should be envisaged before the implementation of drugs for future deep space travel, where the available pharmaceuticals would be limited and restocking from Earth would be impossible. Multipurpose drugs should be proposed for this reason, such as phenothiazine derivatives that can be transformed by optical methods into antimicrobial agents. Within this preliminary study, promethazine and thioridazine aqueous solutions were exposed to UV laser radiation that modified their structures and generated a mixture of photoproducts efficient against particular bacteria. Subsequently, they were subjected to 20 g in the European Space Agency's Large Diameter Centrifuge. The aim was to evaluate the impact of hypergravity on the physico-chemical and spectral properties of unirradiated and laser-irradiated medicine solutions through pH assay, UV-Vis/FTIR absorption spectroscopy, and thin-layer chromatography. The results revealed no substantial alterations in centrifuged samples when compared to uncentrifuged ones. Due to their stability after high-g episodes, laser-exposed phenothiazines could be considered for future space missions.

Spectral Properties of Foams and Emulsions.

The optical and spectral properties of foams and emulsions provide information about their micro-/nanostructures, chemical and time stability and molecular data of their components. Foams and emulsions are collections of different kinds of bubbles or drops with particular properties. A summary of various surfactant and emulsifier types is performed here, as well as an overview of methods for producing foams and emulsions. Absorption, reflectance, and vibrational spectroscopy (Fourier Transform Infrared spectroscopy-FTIR, Raman spectroscopy) studies are detailed in connection with the spectral characterization techniques of colloidal systems. Diffusing Wave Spectroscopy (DWS) data for foams and emulsions are likewise introduced. The utility of spectroscopic approaches has grown as processing power and analysis capabilities have improved. In addition, lasers offer advantages due to the specific properties of the emitted beams which allow focusing on very small volumes and enable accurate, fast, and high spatial resolution sample characterization. Emulsions and foams provide exceptional sensitive bases for measuring low concentrations of molecules down to the level of traces using spectroscopy techniques, thus opening new horizons in microfluidics.

High performance thin layer chromatography-densitometry method based on picosecond laser-induced fluorescence for the analysis of thioridazine and its photoproducts.

A densitometry method based on steady-state and time-resolved fluorescence assessments for thioridazine and its photoproducts applied on HPTLC plates has been developed. The excitation source was a picosecond diode laser emitting at 375 nm. This method was used for the analysis of the photoproducts resulted from thioridazine irradiation with 266 nm nanosecond-pulsed laser. The validation of the developed method was performed for thioridazine in terms of linearity, precision, limits of detection and quantification. Furthermore, analysis of the photoproducts of irradiated thioridazine was performed by steady-state and time-resolved fluorescence. The fluorescence spectra and fluorescence lifetime of each photoproduct were obtained and the horizontal chromatograms of fluorescence maxima were generated.

Editorial: Trends in Optical/Laser Spectroscopy and Applications.

Optics and optical spectroscopy are dynamic fields that are developing very fast nowadays, triggered by (i) the need to go deeper in the scientific approach to nature's processes and phenomena, (ii) the evolution of applications in technological and industrial processes, art conservation, environment protection and cosmic space, and (iii) the sometimes hard to predict evolutions of knowledge in science, life sciences, artistic culture, technology and industrial processes [...].

Stability of Antimicrobial Drug Molecules in Different Gravitational and Radiation Conditions in View of Applications during Outer Space Missions.

The evolution of different antimicrobial drugs in terrestrial, microgravity and hypergravity conditions is presented within this review, in connection with their implementation during human space exploration. Drug stability is of utmost importance for applications in outer space. Instabilities may be radiation-induced or micro-/hypergravity produced. The antimicrobial agents used in space may have diminished effects not only due to the microgravity-induced weakened immune response of astronauts, but also due to the gravity and radiation-altered pathogens. In this context, the paper provides schemes and procedures to find reliable ways of fighting multiple drug resistance acquired by microorganisms. It shows that the role of multipurpose medicines modified at the molecular scale by optical methods in long-term space missions should be considered in more detail. Solutions to maintain drug stability, even in extreme environmental conditions, are also discussed, such as those that would be encountered during long-duration space exploratory missions. While the microgravity conditions may not be avoided in space, the suggested approaches deal with the radiation-induced modifications in humans, bacteria and medicines onboard, which may be fought by novel pharmaceutical formulation strategies along with radioprotective packaging and storage.

Optical Characterization of Ciprofloxacin Photolytic Degradation by UV-Pulsed Laser Radiation.

Ciprofloxacin is one of the most prescribed antibiotics in treating bacterial infections, becoming an important pollutant of the wastewaters. Moreover, ciprofloxacin is hard to be destroyed by conventional water treatment processes; therefore, efficient treatments to destroy it are needed in water decontamination. This study offers insights into the performance of 266 nm laser beams on the photodegradation of ciprofloxacin. An Nd:YAG laser was used that emitted 266 nm at an energy of 6.5 mJ (power of 65 mW) and ciprofloxacin water solutions were irradiated up to 240 min. The irradiated solutions were investigated by UV-Vis and FTIR absorption spectroscopy, pH assay, and laser-induced fluorescence. An HPTLC densitometer was used to characterize the laser-induced fluorescence and fluorescence lifetime of photodegradation products. The UV-Vis absorption, FTIR, and laser-induced fluorescence spectra showed the degradation of ciprofloxacin. Moreover, HPTLC densitometry offered the fluorescence and fluorescence lifetime of ciprofloxacin and its three photoproducts as well as their relative quantification. From the FTIR spectra, the molecular structure of two out of three photoproducts was proposed. In conclusion, the laser irradiation method provided the efficient photodegradation of ciprofloxacin, whereas the analytical techniques offered the proper means to monitor the process and detect the obtained photoproducts.

Anti-staphylococcal activity and mode of action of thioridazine photoproducts.

Antibiotic resistance became an increasing risk for population health threatening our ability to fight infectious diseases. The objective of this study was to evaluate the activity of laser irradiated thioridazine (TZ) against clinically-relevant bacteria in view to fight antibiotic resistance. TZ in ultrapure water solutions was irradiated (1-240 min) with 266 nm pulsed laser radiation. Irradiated solutions were characterized by UV-Vis and FTIR absorption spectroscopy, thin layer chromatography, laser-induced fluorescence, and dynamic surface tension measurements. Molecular docking studies were made to evaluate the molecular mechanisms of photoproducts action against Staphylococcus aureus and MRSA. More general, solutions were evaluated for their antimicrobial and efflux inhibitory activity against a panel of bacteria of clinical relevance. We observed an enhanced antimicrobial activity of TZ photoproducts against Gram-positive bacteria. This was higher than ciprofloxacin effects for methicillin- and ciprofloxacin-resistant Staphylococcus aureus. Molecular docking showed the Penicillin-binding proteins PBP3 and PBP2a inhibition by sulforidazine as a possible mechanism of action against Staphylococcus aureus and MRSA strains, respectively. Irradiated TZ reveals possible advantages in the treatment of infectious diseases produced by antibiotic-resistant Gram-positive bacteria. TZ repurposing and its photoproducts, obtained by laser irradiation, show accelerated and low-costs of development if compared to chemical synthesis.

Laser irradiated phenothiazines: New potential treatment for COVID-19 explored by molecular docking.

The worldwide infection with the new Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) demands urgently new potent treatment(s). In this study we predict, using molecular docking, the binding affinity of 15 phenothiazines (antihistaminic and antipsychotic drugs) when interacting with the main protease (Mpro) of SARS-CoV-2. Additionally, we tested the binding affinity of photoproducts identified after irradiation of phenothiazines with Nd:YAG laser beam at 266 nm respectively 355 nm. Our results reveal that thioridazine and its identified photoproducts (mesoridazine and sulforidazine) have high biological activity on the virus Mpro. This shows that thioridazine and its two photoproducts might represent new potent medicines to be used for treatment in this outbreak. Such results recommend these medicines for further tests on cell cultures infected with SARS-CoV-2 or animal model. The transition to human subjects of the suggested treatment will be smooth due to the fact that the drugs are already available on the market.

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.

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.

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.

In vitro antimicrobial efficacy of laser exposed chlorpromazine against Gram-positive bacteria in planktonic and biofilm growth state.

Aqueous chlorpromazine solutions exposed to 266 nm generated as fourth harmonic of Nd:YAG pulsed laser along time intervals from 1 min to 240 min were investigated for their antimicrobial activity against planktonic and adherent Gram-positive bacterial strains. Qualitative and quantitative assays based on microbiological methods and flow cytometry assays were performed to establish the minimum inhibitory and minimum biofilm eradication concentrations and to reveal some of the possible mechanisms of antimicrobial activity. Optimal irradiation conditions and combinations of photoproducts for achieving the best antimicrobial and antibiofilm effects are suggested. It was confirmed that chlorpromazine solutions irradiated for 15 min and 30 min have the best antimicrobial and antibiofilm activity against Staphylococcus aureus ATCC 6538, methicillin susceptible Staphylococcus aureus, methicillin resistant Staphylococcus aureus, Enterococcus faecium 17-VAR, Enterococcus faecalis 2921, and Bacillus subtilis 6633. Flow cytometry revealed that two of the possible mechanisms of the antimicrobial activity of irradiated chlorpromazine are the inhibition of efflux pumps activity and induction of cellular membrane lesions.

Effects of 1064-nm Nd:YAG long-pulse laser on polidocanol microfoam injected for varicose vein treatment: a controlled observational study of 404 legs, after 5-year-long treatment.

Sclerotherapy continues to be the treatment of choice for varicose veins in the legs. However, isolated treatment using microfoam or lasers requires a high number of sessions to eliminate them. In 2013, we published results about the efficacy and safety 3 years after the combined treatment with microfoam injections and subsequent application of Nd:YAG laser. The aim of this paper is to clinically evaluate the treatment of varices in a control visit after 5 years, when polidocanol microfoam is used and is immediately irradiated in the tissue with 1064-nm Nd:YAG laser beam. The outcome persistence after 5 years was studied in the legs that had received combined treatment and had been studied 3 years after treatment. Patients were contacted by phone, interviewed, and examined with echo-Doppler. Out of the 259 patients who were contacted, 221 agreed to make the appointment, although in the end, only 202 came, which meant analysing 404 legs. At 5 years, the clearance rates were very high: patients were included in class CEAP C1 showing vessels of from 0.5 to 3 mm diameter. The patients showed a high level of satisfaction. Regarding adverse effects, only 4 cases of hypopigmentation described in the previous publication persisted. Although the action mechanisms between the microfoam and the Nd:YAG laser must still be elucidated, it is notable that combining microfoam with laser exposure obtained a complete, effective treatment of legs in only 2 sessions, with high clearance rates and high level of satisfaction among patients.

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.

Protein reactivity with singlet oxygen: Influence of the solvent exposure of the reactive amino acid residues.

The singlet oxygen quenching rate constants were measured for three model proteins, bovine serum albumin, ß-lactoglobulin and lysozyme. The results were analyzed by comparing them with the corresponding singlet oxygen quenching rate constants for a series of tripeptides with the basic formula GlyAAGly where the central amino acid (AA) was the oxidizable amino acid, tryptophan, tyrosine, methionine and histidine. It was found that the reaction rate constant in proteins can be satisfactorily modelled by the sum of the individual contributions of the oxidizable AA residues corrected for the solvent accessible surface area (SASA) effects. The best results were obtained when the SASA of the AA residues were determined by averaging over molecular dynamics simulated trajectories of the proteins. The limits of this geometrical correction of the AA residue reactivity are also discussed.

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.

Characterization of mixtures of compounds produced in chlorpromazine aqueous solutions by ultraviolet laser irradiation: their applications in antimicrobial assays.

The study reports an investigation of the photoproducts obtained by exposure of chlorpromazine hydrochloride in ultrapure water (concentration 2 mg/mL) to a 266-nm laser beam obtained by fourth harmonic generation from a Nd:YAG laser (6-ns full time width at half maximum, 10-Hz pulse repetition rate). The photoproducts were analyzed by steady-state UV-Vis absorption, laser-induced fluorescence, Fourier transform infrared spectroscopy, and liquid chromatography-tandem time-of-flight mass spectroscopy. Two figures showing pathways that take place during irradiation for obtaining the final products are shown. The quantum yield of singlet oxygen generation by chlorpromazine (CPZ) was determined relative to standard Zn-phthalocyanine in dimethyl sulfoxide. To outline the role of fluorescence in photoproducts formation rates, fluorescence quantum yield of CPZ during exposure to 355-nm radiation (third harmonic of the fundamental beam of Nd:YAG laser) was investigated relative to standard Coumarin 1 in ethanol. The CPZ solutions exposed 60 and 240 min to 266-nm laser beam, respectively, were tested against Staphylococcus aureus ATCC 25923 strain. For 25 µL of CPZ samples irradiated 240 min, a higher diameter of inhibition has obtained against the tested strain than for the 60-min exposed ones.

Rapid, laser-induced conversion of 20-hydroxyecdysone - a follow-up study on the products obtained.

We have recently reported the set-up of an experimental system for the laser-induced photochemical modification of bioactive substances, where two ecdysteroids, 20-hydroxyecdysone (20E) and its diacetonide derivative served as probes. As a direct continuation of our previous work, three new compounds together with five other ecdysteroid derivatives, have been identified from the novel, laser-induced photo-transformation reaction of 20E. The structures and NMR signal assignment were established by comprehensive one- and two-dimensional NMR spectroscopy supported by mass spectroscopy. Possible ways for the formation of each species is also discussed. Similar to their parental compound, the products obtained are potentially bioactive and worthy for further investigations; due to the low yields, however, a different approach for their higher scale production is suggested.

Interaction of solutions containing phenothiazines exposed to laser radiation with materials surfaces, in view of biomedical applications.

Phenothiazine drugs - chlorpromazine (CPZ), promazine (PZ) and promethazine (PMZ) - were exposed to 266 nm (fourth harmonic of the Nd:YAG pulsed laser radiation) in order to be modified at molecular level and to produce an enhancement of their antibacterial activity. The irradiated samples were analysed by several methods: pH and surface tension measurements, UV-vis-NIR absorption spectroscopy, laser induced fluorescence and thin layer chromatography. The purpose of these investigations was to study and describe the modified properties of the medicines to further investigate their specific interactions with materials such as cotton, polyester and Parafilm M as a model smooth surface. The textile materials may be impregnated with phenothiazines drug solutions exposed to laser radiation in order to be used in treatments applied on the surface of the organism. Some of the phenothiazines solutions exposed prolonged time intervals to laser radiation have much better activity against several bacteria. Therefore, in the paper, it is reported the wetting behaviour of CPZ, PZ and PMZ solutions, irradiated for time intervals between 1 and 240 min, on the surfaces of the three textures in order to draw a conclusion about their wettability as a function of time.