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The role of photosynthetically active radiation (400–700 nm) (PAR) in modifying plant sensitivity and photomorphogenic responses to ultraviolet-B (280–320 nm) (UV-B) radiation has been examined by a number of investigators, but few studies have been conducted on ultraviolet-A (320–400 nm) (UV-A), UV-B and PAR interactions. High ratios of PAR–UV-B and UV-A–UV-B have been found to be important in ameliorating UV-B damage in both terrestrial and aquatic plants. Growth chamber and greenhouse studies conducted at low PAR, low UV-A and high UV-B often show exaggerated UV-B damage. Spectral balance of PAR, UV-A and UV-B has also been shown to be important in determining plant sensitivity in field studies. In general, one observes a reduction in total biomass and plant height with decreasing PAR and increasing UV-B. The protective effects of high PAR against elevated UV-B may also be indirect, by increasing leaf thickness and the concentration of flavonoids and other phenolic compounds known to be important in UV screening. The quality of PAR is also important, with blue light, together with UV-A radiation, playing a key role in photorepair of DNA lesions. Further studies are needed to determine the interactions of UV-A, UV-B and PAR.
Environmental lighting powerfully suppresses the physiologic release of melatonin, which typically peaks in the middle of the night. This decreased melatonin production has been hypothesized to increase the risk of cancer. Evidence from experimental studies supports a link between melatonin and tumor growth. There is also fairly consistent indirect evidence from observational studies for an association between melatonin suppression, using night work as a surrogate, and breast cancer risk.
Singlet oxygen has been detected in single nerve cells by its weak 1270 nm phosphorescence (a1Δg→X3Σg−) upon irradiation of a photosensitizer incorporated in the cell. Thus, one can now consider the application of direct optical imaging techniques to mechanistic studies of singlet oxygen at the single-cell level.
Photosensitizer biodistribution change inside tissue is one of the dominant factors in photodynamic therapy efficacy. In this study, the pharmacokinetics of a benzoporphyrin derivative (BPD), delivered in verteporfin for injection formulation, have been quantified in the rat Dunning prostate tumor MAT-LyLu model, using both subcutaneous and orthotopic sites. Blood plasma sampling indicated that BPD had a bi-exponential metabolic lifetime in vivo, with the two lifetimes being 9.6 min and 8.3 h. The spatial distributions in the tumor were quantified as a function of distance from the perfused blood vessels, using fluorescence histologic images of the tumor. A fluorescent vascular marker was used to obtain locations and shapes of perfused capillaries at a wavelength of emission different from that of BPD and to allow colocalized images to be acquired of vessel and BPD locations. Using the BPD fluorescence images obtained 15 min after intravenous administration, a forward finite-element solution to the diffusion equation was used to predict the drug distribution by matching the fluorescence intensity images observed microscopically. An inverse solver was used to minimize the root mean square error between the image of simulated diffusion and the experimental image, resulting in estimation of the diffusion coefficient of BPD in the tumor models. Effective diffusion coefficients were 0.88 and 1.59 μm2/s for the subcutaneous and orthotopically grown tumors, respectively, indicating that orthotopic tumors have significantly higher vascular extravasation rates as compared with subcutaneous tumors. This analysis supports the hypothesis that leakage rates of the photosensitizer vary considerably. Thus, although varying the time between injection and optical irradiation may be used to vary the targeting between vascular and less vascular areas, the precise time of treatment will depend on the nature of the permeability of the vasculature in the tissue being treated.
In the past few years, there has been an increase in the application of photosensitizers for medical purposes. A good standardized test system for the evaluation of the mutagenic potentials of photosensitizers is therefore an indispensable device. In the standard Ames test, white light itself was proven to be mutagenic and the result influenced by the light source. Lack of a reliable positive control is another problem in many genotoxicity test systems used for the evaluation of mutagenicity of photosensitizers. Based on the validated somatic mutation and recombination test, known as SMART, and using Drosophila melanogaster, we developed the Photo-SMART and demonstrated that methylene blue, known to induce photomutagenicity, can act as a positive control in the presented test system. The SMART scores for the loss of heterozygosity caused predominantly by homologous mitotic recombination. The Photo-SMART can be used to detect photogenotoxicity caused by short-lived photoproducts or by stable photoproducts or both. We demonstrated the Photo-SMART to be a good standardized test system for the evaluation of mutagenic potentials of the photosensitizer 5,10,15-tris(4-methylpyridinium)-20-phenyl-[21H,23H]-porphine trichloride (TPP). We demonstrated that TPP was mutagenic using the Photo-SMART. For hematoporphyrin, the results of the Photo-SMART indicate the absence of mutagenicity.
We studied the spectral characteristics of the larvae of three sympatric Belgian species of fireflies, Lampyris noctiluca, Phosphaenus hemipterus and Lamprohiza splendidula. An in vivo spectral study was performed to compare bioluminescence spectra. The emission spectrum of a laboratory reared female L. noctiluca was recorded by a different, more exact method. The mean peak wavelength (λmax = 546 nm) and shapes of the unimodal emission spectra are visually similar for the larvae of all three species. The emission spectrum of the adult female L. noctiluca peaked in the same range as the larval bioluminescence between 546 and 551 nm. The bandwidth at half-maximum intensity was slightly greater for larval L. noctiluca (77 ± 4 nm) compared with P. hemipterus (70 ± 10 nm). The bandwidth of larval L. splendidula (77 ± 8 nm) was not different compared with the other larvae, whereas the females' bandwidth was somewhat narrower (68 nm). The ecological significance of the color of bioluminescence and conservancy of green emission in larval fireflies and other luminescent beetle larvae is discussed.
Chloroplast reorientations within mesophyll cells are among the most rapid physiological responses of higher plants to blue light. At light intensities below the saturation point of photosynthesis, chloroplasts move to the cell walls perpendicular to the direction of light and maximize light absorption (low–fluence rate response [LFR]). At light intensities above the saturation point of photosynthesis, chloroplasts redistribute to cell walls parallel to the direction of light (high–fluence rate response [HFR]). The actin-based mechanism is responsible for the light-induced chloroplast movements. We have found that an inhibitor of phosphoinositide-3-kinases, wortmannin, potently and irreversibly inhibited LFR and HFR chloroplast responses to blue light in Lemna trisulca L. mesophyll cells. Microscopic observations and photometric measurement indicated that 100 nM wortmannin specifically inhibited LFR in Lemna, whereas HFR displayed no sensitivity to the inhibitor at this concentration. A complete inhibition of the HFR could be obtained by 1 μM wortmannin. These data indicate that LFR is more sensitive to wortmannin than HFR and suggest that these two responses may be under the control of different cellular mechanisms. Our results suggest that phosphoinositide kinases and other phosphoinositide cycle enzymes may play a role in the transduction of the light signal to the actin cytoskeleton in Lemna as factors specifying the direction of chloroplast movements. A hypothetical model assuming three signaling pathways regulating light-induced chloroplast reorientations in mesophyll cells is proposed.
Cell survival, synergistic interaction, liquid-holding recovery (LHR) kinetics and inactivation forms after the simultaneous treatment with UV light (254 nm) and various high temperatures were studied in diploid yeast cells Saccharomyces cerevisiae. The synergistic interaction was observed within a certain temperature range in which there was a temperature that maximizes the synergistic effect. The LHR study revealed that both the extent and the rate of recovery greatly decreased with the increase in exposure temperature. A quantitative approach describing the LHR process as a decrease in the effective radiation dose was used to estimate the probability of recovery per unit time and the irreversible component of damage. Using the experimental data obtained and the mathematical model described, it was shown that the irreversible component, i.e. the fraction of cells incapable of recovery, increased with the exposure temperature, whereas the recovery constant, i.e. the probability of recovery per unit time, was independent of the exposure temperature. The increase in the irreversible component was accompanied by an increase in cell death without postirradiation division. It is concluded based on this that the synergistic interaction of UV light radiation and hyperthermia in yeast cells is not related to the impairment of the recovery process itself and that it may be attributed to an increased yield of the irreversible damage.
Under 254 nm irradiation, [Pt(bpy)Cl2] is converted to [Pt(bpy)Cl4] in a solvent-initiated process. The reaction is very nearly zero order throughout. The rate decreases slightly with increasing starting concentration. These characteristics can be rationalized by a rate law of the form afS, where fS is the fraction of light absorbed by chloroform. The species that reacts with [Pt(bpy)Cl2] is believed to be CCl3OO.
Many responses of the zygomycete fungus Phycomyces blakesleeanus are mediated by blue light, e.g. the stimulation of β-carotene synthesis (photocarotenogenesis) and the formation of fruiting bodies (photomorphogenesis). Even though both responses have been described in detail genetically and biophysically, the underlying molecular events remain unknown. Applying a pharmacological approach in developing mycelia, we investigated the possible involvement of heterotrimeric G proteins in the blue-light transduction chains of both responses. G protein agonists (guanosine triphosphate analogues, cholera toxin, pertussis toxin) mimicked in darkness the effect of blue light for both responses, except for cholera toxin, which was ineffective in increasing the β-carotene content of dark-grown mycelia. Experiments combining the two toxins indicated that photocarotenogenesis could involve an inhibitory G protein (Gi) type, whereas photomorphogenesis may depend on a transducin (Gt type)–like heterotrimer. The determination of the carB (phytoene dehydrogenase) and chs1 (chitin synthase 1) gene expression under various conditions of exogenous challenge supports the G protein participation. The fluctuations of the time course measurements of the carB and chs1 transcripts are discussed.
Photodynamic therapy (PDT) relies on three main ingredients, oxygen, light and photoactivating compounds, although the PDT response is definitively contingent on the site and level of reactive oxygen species (ROS) generation. This study describes the development of a novel, fluorescent-based actinometer microsphere system as a means of discerning spatially resolved dosimetry of total fluence and ROS production. Providing a high resolution, localized, in situ measurement of fluence and ROS generation is critical for developing in vivo PDT protocols. Alginate-poly-l-lysine-alginate microspheres were produced using ionotropic gelation of sodium alginate droplets, ranging from 80 to 200 μm in diameter, incorporating two dyes, ADS680WS (ADS) and Rhodophyta-phycoerythrin (RPE), attached to the spheres' inside and outside layers, respectively. To test the responsivity and dynamic range of RPE for ROS detection, the production of ROS was initiated either chemically using increasing concentrations of potassium perchromate or photochemically using aluminum tetrasulphonated phthalocyanine. The generation of singlet oxygen was confirmed by phosphorescence at 1270 nm. The resulting photodegradation and decrease in fluorescence of RPE was found to correlate with increased perchromate or PDT treatment fluence, respectively. This effect was independent of pH (6.5–8) and could be inhibited using sodium azide. RPE was not susceptible to photobleaching with light alone (670 nm; 150 Jcm−2). ADS, which absorbs light between 600 and 750 nm, showed a direct correlation between radiant exposure (670 nm; 0–100 Jcm−2) and diminished fluorescence. Photobleaching was independent of irradiance (10–40 mW cm−2). We propose that actinometer microspheres may provide a means for obtaining high spatial resolution information regarding delivered PDT dose within model systems during investigational PDT development and dosimetric information for clinical extracorporeal PDT as in the case of ex vivo bone marrow purging.