Visible Light Induced Photodegradation and Phototoxicity of Phloxine B and Uranine

Ohjective To determine the visible light-induced photodegradation kinetics of two xanthene photosensitizers, phloxine B and uranine, in solution and on the surface of silica TLC plates, and to examine the phototoxicity of residues of degradation, which could provide valuable safety data on the two photosensitizers and other xanthene chemicals when applied in the environment. Methods UV-Vis absorption during photodegradation was monitored with a Unico 2102 spectrophotometer. Organic content of samples was measured with a Shimadzu TOC 4100. Phototoxicity tests were carried out using Saccharomyces cerevisiae with the methods modified from Daniels. Results When phloxine B and uraniue degraded in solution, their apparent rate constant k was 0.0019 and 0.0027 min^-1, respectively. The total organic carbon （TOC） content decreased by approximately 50% during the 8 h irradiation period, which led to a gradual decrease in phototoxicity of the residues. The photodegradation of photosensitizers on the surface of silica TLC plates was much faster than that in the solution. The apparent rate constant k and the half life of phloxine B were 0.0073 min^-1 and 95 min, respectively. Conclusion Visible light can rapidly induce photodegradation of phloxine B and uranine. The phototoxicity of residues is also decreased. The environmental risk of applications of phloxine B and uranine is minimal.

Connexin 40-formed GJIC increases the phototoxicity of photodynamic therapy through ROS-and calcium-mediated pathways

OBJECTIVE To explore the effect of connexin(Cx)40-formed gap junctional intercellular communication(GJIC)on Photofrin-photodynamic therapy(PDT)phototoxicity in Cx40-transfected He La cells and its potential mechanisms.METHODS He La cell line stably transfected to express Cx40 was seeded at high and low cell density,respectively,to assess in vitro photosensitivity using CCK8 assay.Western blot assay was performed to detect the expression of Cx40.The intracellular ROS and Ca^(2+) concentrations were determined using flow cytometer.4-HNE and ceramide were measured using ELISA assay.RESULTS Cx40-composed GJ formation at high density enhances the phototoxicity of PhotofrinPDT.When the Cx40 is not expressed or Cx40 channels are blocked,the phototoxicity in high-density cultures substantially reduces,indicating that the enhanced PDT phototoxicity at high density is mediated by Cx40-composed GJIC.The GJIC-mediated increase in PDT phototoxicity was associated with ROS and calcium-mediated stress signaling pathways.CONCLUSION The work uniquely presents the ability of Cx40-composed GJIC to enhance the sensitivity of malignant cells to PDT,and indicates that maintenance or increase of Cx40-formed GJIC may be a profitable strategy towards the enhancement of PDT therapeutic efficiency.

Quantitative in vitro assessment of drug phototoxicity by a chemiluminescence method

Objective To establish a test model for phototoxic agents using the method of chemiluminescence Methods The phototoxicity of pipemidic acid, doxycycline, griseofuvin and chlorpromazine was detected These agents and distilled water were irradiated with ultraviolet A (UVA) in the presence of nicotinamide adenine dinucleotide reduced (NADH), and the duplicated samples were incubated in the dark as dark controls Then luminol was added to the test samples, and the chemiluminescent value was counted and calculated Results Chemiluminescent values of pipemidic acid, doxycycline and griseofuvin were significantly higher than those in controls The result of linear regression analysis showed that phototoxic intensity was linear correlated with UVA dosage The regression coefficient for distilled water was 0 56, indicating that the luminscent value (LV) rose slightly after UVA irradiation For pipemidic acid, griseofuvin and doxycycline, the regression coefficients reached 76 96 , 254 33 and 92 61 respectively, significantly increased in comparison with those of negative controls (P<0 01) Conclusion Phototoxicity of pipemidic acid, doxycycline and griseofuvin can be detected with the method of chemiluminescence

Protective effect of saturated hydrogen saline against blue light-induced retinal damage in rats

AIM: To explore the effect of saturated hydrogen saline on blue light-induced retinal damage in rats. METHODS: The retinal damage of rats was induced by blue light exposure for 6 hours and examined 8 hours, 16 hours and 24 hours after the exposure. One hundred female Sprague-Dawley rats were randomly divided into four groups. Group 1 included 30 rats received light exposure without any other treatment. Group 2 included 30 rats received light exposure with intraperitoneal injection of normal saline. Group 3 included 30 rats received light exposure with intraperitoneal injection of saturated hydrogen saline. And Group 4 included the other 10 rats which did not receive any treatment. The amount of intraperitoneal injection of saturated hydrogen saline and normal saline was calculated in the ratio of 1ml/100g of rat weight. Specimens were collected and processed by H-E staining, ultrastructure observation, biochemical measurement. Morphological changes were observed by light microscope and transmission electron microscope (TEM) and the retinal outer nuclear layer (ONL) thickness was measured by IPP 6.0, while the malondialdehyde (MDA) was measured by colorimetric determination at 532nm. RESULTS: Although the structure of retina in Group 1 and Group 2 was injured heavily, the injury in Group 3 was mild. The differences between Group 1 and Group 2 were not significant. Compared with the rats in Group 1 and Group 2, the ones in Group 3 had more clearly demarcated retina structure and more ordered cells by light microscope and TEM observation. The ONL thicknesses (400 times) of four groups at each time point except between Group 1 and Group 2 were significantly different (P<0.05). The thicknesses of the ONL in Group 1 at three time points were 30.41 +/- 4.04 mu m, 26.11 +/- 2.82 mu m and 20.63 +/- 1.06 mu m, in Group 2 were 31.62 +/- 4.54 mu m, 25.08 +/- 3.63 mu m and 19.07 +/- 3.86 mu m, in Group 3 were 29.75 +/- 3.62 mu m, 28.83 +/- 1.97 mu m and 27.61 +/- 1.83 mu m. In Group 4 the mean of the thickness was 37.35 +/- 1.37 mu m. As time went by, the damage grew more severely. At 24h point, the differences were most significant. Compared with Group 4, the thickness was 46.23% thinner in Group 1, 50.29% thinner in Group 2 and 28.04% thinner in Group 3. The stack structures of membranous disc in Group 3 were injured slightly, but in Group 1 and Group 2 the damage was more obvious by TEM. Compared with Group 4 at each time point, the content of MDA in Group 1 was higher (P<0.05). The content of MDA in Group 3 was significantly lower than those of Group 1 ( P<0.05) and Group 2 (P<0.05). Between the Group 1 and Group 2, the MDA concentration at each time point was no significant difference (P>0.05). CONCLUSION: Saturated hydrogen saline could protect the retina from light-induced damage by attenuating oxidative stress.

Photoinduced toxicity single and binary mixtures of four polycyclic aromatic hydrocarbons to the marine diatom Skeletonema costatum

Diatom Skeletonema costatum Cleve is one of the main predominant phytoplankton species in the Changjiang Estuary in China. In order to provide some basic information for future assessment of the potential risk on phytoplankton communities in this estuary caused by polycyclic aromatic hydrocarbons（ PAHs）, this alga was selected as a representative to investigate the photoinduced toxicity of PAHs, in single and mixture. Four PAHs including three-ring phenanthrene and anthracene, four-ring fluoranthene and pyrene were tested in the laboratory. The single toxicity of each PAH on this microalga was compared with and without the simulated solar UV radiation. The results showed that this microalga was sensitive to PAH＇ s photoinduced toxicity. Ratios of the 72 h median effect concentration obtained for fluorescent and UV light tests were about 8.4 for phenanthrene, 13.0 for anthracene, 6. 5 for fluoranthene, and 5.7 for pyrene, indicating that UV light enhanced the PAH toxicity to this alga significantly. Under the fluorescent radiation （lacking UV ）, the dose-response curves based on chemical concentrations revealed that the order of toxic strength was fluoranthene greater than pyrene greater than anthracene greater than phenanthrene; while under the UV radiation （476 μW/cm^2 for UVA, 6.5 μW/cm^2 for UVB） it became fluoranthene approximately equaling anthracene greater than pyrene greater than phenanthrene, indicating that the UV light also changed its relative toxicity to this alga. The photoinduced toxicity of PAHs to the marine diatom S. costatum might be a synergistic effect of photosensitization reactions （ e. g. , generation of singlestate oxygen） and photomodification （photooxidation and/or photolysis）. The combined effects of six binary mixtures on the marine diatom S. costatum were investigated using the additive-index method. Four binary-mixtures （ phenanthrene plus anthracene ; phenanthrene plus pyrene ; anthracene plus fluoranthene ; anthracene plus pyrene） were found to be synergistic joint action mode, while two binary-mixtures （phenanthrene plus fluoranthene; fluoranthene plus pyrene） displayed antagonist, revealing a complex pattern of possible interactions. The combined action mode of PAHs might be related to various factors such as the mixture compounds, mixture ratios and test conditions, etc.

Meso-substituted cationic 3-and 4-N-Pyridylporphyrins and their Zn(II)derivatives for antibacterial photodynamic therapy

Photodynamic inactivation of microorganisms known as antibacterial photodynamic therapy(APDT)is one of the most promising and innovative approaches for the destruction of pathogenic microorganisms.Among the photosensitizers(PSs),compounds based on cationic porphyrins/metalloporphyrins are most successfully used to inactivate microorganisms.Series of meso-substituted cationic pyridylporphyrins and metalloporphyrins with various peripheral groups in the third and fourth positions of the pyrrole ring have been synthesized in Armenia.The aim of this work was to determine and test the most e®ective cationic porphyrins and metalloporphyrins with high photoactivity against Gram negative and Gram positive microorganisms.It was shown that the synthesized cationic pyridylporphyrins/metalloporphyrins exhibit a high degree of phototoxicity towards both types of bacteria,including the methicillinresistant S.aureus strain.Zinc complexes of porphyrins are more phototoxic than metal-free porphyrin analogs.The e®ectiveness of these Zn–metalloporphyrins on bacteria is consistent with the level of singlet oxygen generation.It was found that the high antibacterial activity of the studied cationic porphyrins/metalloporphyrins depends on four factors:the presence in the porphyrin macrocycle of a positive charge(+4),a central metal atom(Zn2þÞand hydrophobic peripheral functional groups as well as high values of quantum yields of singlet oxygen.The results indicate that meso-substituted cationic pyridylporphyrins/metalloporphyrins cannd wider application in photoinactivation of bacteria than anionic or neutral PSs usually used in APDT.

Photosafety testing of dermally-applied chemicals based on photochemical and cassette-dosing pharmacokinetic data

Drug-induced phototoxicity is elicited by exposure of skin and/or eyes to topical or systemic administration of chemicals,followed by exposure to sunlight.For photosafety testing,a new screening strategy,employing in vitro photochemical/photobiological and in vivo cassette-dosing pharmacokinetic(PK)studies,was proposed[1,2].The screening system provided reliable phototoxic predictions of tested chemicals at least within the same chemical series;however,the applicability of the proposed strategy is not fully understood.

Photolysis of chlortetracycline in aqueous solution:Kinetics,toxicity and products

The aqueous photodegradation of the widely used antibiotic chlortetracycline（CTC） was investigated under simulated sunlight.The quantum yield of photodegradation increased from 3.3 × 10-4 to 8.5 × 10-3 within the pH range of 6.0 to 9.0.The presence of Ca2＋,Fe3＋,and NO3-enhanced the photodegradation of CTC,whereas Mg2＋,Mn2＋,and Zn2＋ inhibited the degradation with the order Mn2＋ Zn2＋ Mg2＋ at pH 7.3.The monovalent cations（Na＋ and K＋） had negligible effect on the photolysis of CTC.Fulvic acid（FA） decreased the photodegradation of CTC due to light screening effect.Hydrogen peroxide（H2O2） was formed concurrently with direct photodegradation of CTC.The generation rate of H2O2 increased from 0.027 to 0.086 μmol/（L.min） when the pH ranged from 6.0 to 9.0.The CTC solution was about three-fold more toxic to the Photobacterium phosphoreum bacteria after irradiation,suggesting that the photoproducts and H2O2 formed in the CTC solution exhibited high risk on the bacteria.By LC-ESI（＋）-MS,the photoproducts of CTC were identified.The direct photodegradation of CTC was involved in hydroxylation and N-demethyl/dedismethyl processes.The main photoproducts included the iso-CTC analog containing hydroxyl groups（m/z 511.4 and 495.4）,and the N-demethyl/dedismethyl products of the photoproduct m/z 495.4（m/z 481.3 and 467.4）.In addition,the photochemical dechlorination of CTC led to tetracycline（m/z 445.5）.