Quantum dots have emerged as an important class of material that offers great promise to a diverse range of applications ranging from energy conversion to biomedicine. Here, we review the potential of using quantum dots and quantum dot conjugates as sensitizers for photodynamic therapy (PDT). The photophysics of singlet oxygen generation in relation to quantum dot-based energy transfer is discussed and the possibility of using quantum dots as photosensitizer in PDT is assessed, including their current limitations to applications in biological systems. The biggest advantage of quantum dots over molecular photosensitizers that comes into perspective is their tunable optical properties and surface chemistries. Recent developments in the preparation and photophysical characterization of quantum dot energy transfer processes are also presented in this review, to provide insights on the future direction of quantum dot-based photosensitization studies from the viewpoint of our ongoing research.

The effects of an additional keto group on absorption wavelength and the corresponding metal complexes Zn(II), Cu(II) In(III) on singlet oxygen production and photodynamic efficacy were examined among the alkyl ether analogs of pyropheophorbide-a. For the preparation of the desired photosensitizers, the methyl 13²-oxo-pyropheophorbide-a obtained by reacting methyl pyropheophorbide-a with aqueous LiOH-THF was converted into a series of alkyl ether analogs. These compounds were evaluated for photophysical properties and in vitro (by means of the MTT assay and intracellular localization in RIF cells) and in vivo (in C3H mice implanted with RIF tumors) photosensitizing efficacy. Among the alkyl ether derivatives, the methyl 3-decyloxyethyl-3-devinyl-13²-oxo-pyropheophorbide-a was found to be most effective and the insertion of In(III) into this analog further enhanced its in vitro and in vivo photosensitizing efficacy. Fluorescence microscopy showed that, in contrast to the hexyl and dodecyl ether derivatives of HPPH (which localize in mitochondria and lysosomes, respectively), the diketo-analogs and their In(III) complexes localized in Golgi bodies. The preliminary in vitro and in vivo results suggest that, in both free-base and metalated analogs, the introduction of an additional keto group at the five-member exocyclic ring in pyropheophorbide-a diminishes its photosensitizing efficacy. This may be due to a shift in subcellular localization from mitochondria to the Golgi bodies. The further introduction of In(III) enhances photoactivity, but not by shifting the localization of the photosensitizer.

Although strong fluorescence makes the R-phycoerythrin (R-PE) proteins increasingly useful in biological and clinical assays, they are subject to nonlinear effects including transitions to collective dark states and photodegradation, which complicate quantitative applications. We report measurements of R-PE fluorescence intensity as a function of incident power, duration of illumination and temperature. Emission intensity in the band at 570 nm is proportional to incident power for low power levels. At higher incident power, the emission at 570 nm is smaller than expected from a linear dependence on power. We propose that R-PE undergoes both reversible emission cessation on a millisecond time scale attributed to transitions to a collective dark state, and irreversible photodegradation on a time scale of minutes. Singlet oxygen scavengers such as dithiothreitol and n-propyl gallate have protective effects against the latter effect but not the former. Electrophoretic analysis of irradiated solutions of R-PE indicates that significant noncovalent aggregation is correlated with photodegradation. A multistate model based on fluorescence measurements and geometric analysis is proposed for the fluorophores in R-PE. The phycobilin fluorophores are partitioned into three groups: the phycourobilins (PUB) absorbing at 490 nm, one group of phycoerythobilins (PEB) absorbing at 530 nm (PEB-530) and another group of PEB absorbing at 560 nm (PEB-560). The two processes that result in the loss of fluorescence intensity are most likely associated with the PEB-560 group.

Thrombospondin 1 (TSP1) is an extracellular glycoprotein and a recognized inhibitor of angiogenesis. Recent studies have demonstrated that UV radiation induces an angiogenic switch, by which it alters the balance between pro- and anti-angiogenic factors in the skin. Here we describe the effects of acute UV exposure on TSP1 expression in human skin epidermis, primary keratinocytes and the epidermal cell line HaCaT. We found that protein and mRNA expressions of TSP1 are significantly reduced in human skin in vivo and in keratinocytes in vitro by a single UV exposure. In human skin and keratinocytes, UV exposure induced the phosphorylation of Akt, a downstream target of the PI3K pathways. Specific inhibitors of PI3K, wortmannin and LY294002, completely blocked Akt activation and UV-induced TSP1 downregulation in keratinocytes. We showed that a specific Akt phosphorylation inhibitor and small interfering RNA–mediated Akt depletion were also blocked by UV-induced TSP1 downregulation in keratinocytes. In conclusion, our findings demonstrate that acute UV exposure downregulates TSP1 expression via PI3K-Akt activation in human keratinocytes. These novel findings may help us understand the regulatory mechanisms of UV-induced skin angiogenesis.

Photography has been used in human skin research for some time. With the advent of digital photography in recent years, its use has increased. However, the focus has now turned from documentation to actual analysis and quantification of skin color changes. The advantages of digital photography outweigh any shortcomings as long as consistent, standardized procedures are followed and quality control is implemented. We present a simple procedure to standardize images and discuss a computer-assisted digital image evaluation (CADIE) technique to quantify skin color changes following UV exposure. The CADIE approach is illustrated with examples from two different studies on UV responses in human skin. Using the Commission Internationale de l'Eclairage L*a*b* color coordinate system in combination with a personal computer and image-editing software, we analyzed digital images obtained in these two studies. We demonstrate the feasibility of using digital photography for objective evaluation of UV erythema in different racial/ethnic groups and for measuring pigmentation changes caused by repeated exposures over a period of several weeks. Our results indicate how objective assessment using CADIE can be an adjunct to visual and optical observation in clinical and scientific evaluations.

Endogenous reduced nicotinamide adenine dinucleotide (NADH) fluorescence provides an intrinsic indicator of the cellular metabolic state, but prolonged monitoring is limited by photobleaching and/or phototoxicity. Multiphoton excitation of NADH by ultrashort, 740-nm laser pulses provides a significant improvement over UV excitation by eliminating peripheral photobleaching; however, molecules within the subfemtoliter excitation volume remain susceptible. We have investigated the photophysical mechanisms responsible for multiphoton photobleaching of NADH in living cells to permit the imaging technique to be optimized. The loss of fluorescence because of multiphoton photobleaching was measured by repetitively imaging individual planes within rat basophilic leukemia cells. The photobleaching rate was proportional to the fourth power of the laser intensity. Based on these measurements, we propose a double-biphotonic, four-photon photobleaching mechanism and estimate the quantum yield of photobleaching of intracellular NADH to be 0.0073 ± 0.0002 by this mechanism. In addition to photobleaching, the development of bright, punctate fluorescent lesions can also be observed. The frequency of lesion formation also increased approximately as the fourth power of the laser intensity after an intensity-dependent threshold number of images had been exceeded. The consequences for two-photon metabolic imaging are discussed.

Ultraviolet B (UVB) irradiation exerts hazardous effects such as acute photodamage, skin cancer and photoaging. In this study we evaluated the protective effects of a nonsedative histamine H1-receptor antagonist, mizolastine, on UVB-exposed skin dermal fibroblasts. Therefore, primary human skin fibroblasts were incubated with mizolastine or dexamethasone after 100 mJ/cm² UVB irradiation. Leukotriene B4 (LTB4) in fibroblast supernatants was detected with enzyme immunoassays, expression of 5-lipoxygenase (5-LOX) messenger RNA (mRNA) in skin fibroblasts was examined by reverse transcriptase–polymerase chain reaction and expression of 5-LOX protein was measured by immune blotting and immunofluorescent staining with rabbit anti-human 5-LOX antibody. It was found that 0.01 mM mizolastine inhibited UVB-induced LTB4 production from skin fibroblasts at 12, 24 and 36 h. Meanwhile, mizolastine down-regulated 5-LOX mRNA expression and inhibited 5-LOX translocation from nucleus to cytoplasm in fibroblasts. On the basis of these findings, we propose that mizolastine might play a protective role in the pathogenesis of UV radiation–induced acute photodamage of the skin.

In clinical 5-aminolevulinic acid (ALA)–based photodynamic therapy (PDT) of skin tumors it is desirable to develop vehicles that minimize the penetration of ALA through normal stratum corneum and maximize it through the compromised stratum corneum of the tumors to improve tumor selectivity. We have designed a bioadhesive patch, which may be able to achieve this aim. It induces levels of protoporphyrin IX (PpIX) in skin overlying tumors similar to those induced by the proprietary cream (Porphin®) but at the same time induces less PpIX to form in normal skin and at distant sites. The mechanisms of action of the patch, as compared with that of the cream, were studied by means of Cuprophan® barriers that mimic compromised tumor stratum corneum and in a mouse model with transplanted tumors.

In order to generate renewable and clean fuels, increasing efforts are focused on the exploitation of photosynthetic microorganisms for the production of molecular hydrogen from water and light. In this study we engineered a ‘hard-wired’ protein complex consisting of a hydrogenase and photosystem I (hydrogenase–PSI complex) as a direct light-to-hydrogen conversion system. The key component was an artificial fusion protein composed of the membrane-bound [NiFe] hydrogenase from the β-proteobacterium Ralstonia eutropha H16 and the peripheral PSI subunit PsaE of the cyanobacterium Thermosynechococcus elongatus. The resulting hydrogenase-PsaE fusion protein associated with PsaE-free PSI spontaneously, thereby forming a hydrogenase–PSI complex as confirmed by sucrose-gradient ultracentrifuge and immunoblot analysis. The hydrogenase–PSI complex displayed light-driven hydrogen production at a rate of 0.58 μmol H₂·mg chlorophyll⁻¹·h⁻¹. The complex maintained its accessibility to the native electron acceptor ferredoxin. This study provides the first example of a light-driven enzymatic reaction by an artificial complex between a redox enzyme and photosystem I and represents an important step on the way to design a photosynthetic organism that efficiently converts solar energy and water into hydrogen.

A new tendency in cosmetic formulations is the association of botanical extracts and vitamins to improve skin conditions by synergic effects. The objective of this study was to determine the antioxidant activity of associated bioflavonoids, retinyl palmitate (RP), tocopheryl acetate (TA) and ascorbyl tetraisopalmitate (ATIP), as well as their photoprotective effects in preventing increased erythema, transepidermal water loss (TEWL) and sunburn cell formation in hairless mouse skin. The antioxidant activity of solutions containing the association or each substance separately was evaluated in vitro by a chemiluminescence assay. The photoprotective effect was evaluated by means of in vivo tests. Dorsal skin of hairless mice was treated daily by topical applications for 5 days with formulations containing or not containing (vehicle) the flavonoid-vitamins association (5%). The skin was irradiated (UVA/B) 15 minutes after the last application. The results showed that bioflavonoids had in vitro antioxidant properties and also that when they were associated with vitamins their antioxidant activity was more pronounced. On the other hand, erythema and UV damage to the permeability barrier function (TEWL) was not significantly reduced by previous treatment with the flavonoid-vitamin-association formulations, when compared to the irradiated vehicle-treated area. However, the treatment protected the skin from UV damage because it reduced the number of sunburn cells, when compared to the vehicle-treated area. Finally, the association of vitamins and bioflavonoids added to a dermocosmetic formulation showed a relevant biological activity in terms of photoprotection, because the association of bioflavonoids and vitamins acted by different mechanisms, such as antioxidation and absorption of UV radiation, which suggests its use in antiaging and photoprotective products.

A small and robust dosimeter for determining the biologically effective dose of ambient UV radiation has been developed using UV-sensitive mutant spores of Bacillus subtilis strain TKJ6312. A membrane filter with four spots of the spores was snapped to a slide mount. The slide was wrapped and covered with two or more layers of polyethylene sheet to protect the sample from rain and snow and to reduce monthly-cumulative doses within the measurable range. From 1999, monthly data were collected at 17 sites for more than 1 year, and data for 4 to 6 consecutive years were obtained from 12 sites. Yearly total values of the spore inactivation dose (SID) ranged from 3200 at subarctic Oulu to 96 000 at tropical Denpasar, and the mean yearly values of SID exhibited an exponential dependence on latitude in both hemispheres with a doubling for about every 14 degrees of change. During the observation period, increasing trends of UV doses have been observed at all sites with more than 5 years of data available. Year-to-year variations at high and middle latitude sites are considered due mostly to climatic variation. At three tropical sites, negative correlations between the yearly doses and the column ozone amounts were observed. The results verified the applicability of spore dosimetry for global and long-time monitoring of solar UV radiation, in particular at tropical sites where no monitoring is taking place.

A crucial factor in choosing a porphyrin or analogous photosensitizer for photodynamic therapy (PDT) is its ability to incorporate into the cells. For hydrophobic compounds that partition passively into the cytoplasmic membrane, a partition coefficient between an organic solvent and water, P, is one factor that could be used to predict the molecule's ability to diffuse into biomembranes. We synthesized several porphyrins, modified with two, three or four meso-substituents and studied their spectroscopic and photophysical properties. The octanol–water partitioning coefficients, log P, were calculated as a parameter for hydrophobicity. We found these porphyrins to be very hydrophobic, with log P values in the range of 8.9–11.8. These were correlated with the binding constants of these porphyrins into liposomes, K_b, as well as to their uptake by cells. The correlation between the estimated log P and K_b is nearly linear but negative, indicating, apparently, that there is lesser binding to liposomes with increased hydrophobicity. On the other hand, all of the studied porphyrins are taken up by cells, but there is no clear correlation between cellular uptake and the log P or K_b. Lipinski's pharmacological “rule of 5” predicts poor permeation of drugs into cells when log P is greater than five. This may be relevant for diffusional binding to liposomes, where aqueous aggregation can interfere strongly with cellular uptake. In such extreme conditions, neither liposome binding nor other rules seem to predict porphyrin behavior in vitro.

We studied the effects of high-light exposure (500 μmol m⁻² s⁻¹ of photosynthetic active radiation) on the cyanobacteria Nostoc spongiaeforme Agardh, a fresh-water alga, and Phormidium corium Agardh (Gomont), a marine alga, with respect to photosynthesis, pigments, sugar content, lipid peroxidation, fatty acids composition, antioxidant enzymes activity and DNA. It was seen that the ratio of variable fluorescence (Fv) to maximum fluorescence (Fm), which is indicative of photosynthetic efficiency, decreased because of the light treatment. The damage to photosynthesis occurred in the antenna system and the photosynthetic II reaction center. Photobleaching of photosynthetic pigments was also observed. High-light treatment also resulted in decreased sugar content, which was probably due to the effect on photosynthesis. Peroxidation of membrane lipids, indicating oxidative damage to lipids and a high level of unsaturation in the cell membrane, was also observed. The activity of antioxidant enzyme superoxide dismutase and ascorbate peroxidase was increased, probably as a result of oxidative damage observed in the form of lipid peroxidation. Quantitative decreases in phospholipid and glycolipid levels were also observed. The level of unsaturated fatty acids in total lipids and glycolipids remained unchanged in both species; however, the level of saturated fatty acids decreased, which slightly changed the ratio in favor of unsaturated fatty acids. Degradation of DNA was also observed in both species. There was a transient plateau 2–4 h after exposure to high-light treatment in the Fv/Fm ratio and in levels of phycobilisome pigments, sugars and antioxidant enzymes after an initial decrease 1 h after the treatment. These findings may indicate a period of partial adaptation to high light that is due to the efficiency of protective processes operational in the two species, which subsequently failed after a longer exposure duration of 4–6 h.

The photoreduction of oxidized bovine heart cytochrome c oxidase (CcO) by visible and UV radiation was investigated in the absence and presence of external reagents. In the former case, the quantum yields for direct photoreduction of heme A (heme a heme a₃) were 2.6 ± 0.5 × 10⁻³, 4 ± 1 × 10⁻⁴, and 4 ± 2 × 10⁻⁶ with pulsed laser irradiation at 266, 355 and 532 nm, respectively. Within experimental uncertainty, the quantum yields did not depend on pulse energy, implying that the mechanism is monophotonic. Irradiation with 355 nm light resulted in spectral changes similar to those produced independently by reduction with dithionite, whereby the low-spin heme a and Cu_A are reduced first. Extended illumination at 355 and 532 nm yielded substantial amounts of reduced heme a₃. Heme decomposition was noted with 266 nm light. In the presence of formate and cyanide ions, which bind at the binuclear heme a₃/copper center in CcO, irradiation at 355 nm caused selective reduction of only the low-spin heme a and Cu_A. The addition of ferrioxalate ion dramatically increased the efficiency of cytochrome c oxidase photoreduction. The quantum efficiency for heme A reduction was found to be near unity, significantly greater than for other known methods of photoreduction. The active reductant is most likely ferrous iron, and its reduction of the enzyme is thermodynamically driven by the reformation of ferrioxalate in the presence of excess oxalate ion. Other metalloenzymes with redox potentials similar to those of cytochrome c oxidase should be amenable to indirect photoreduction by this method.

A fluorescence imaging technique was used to monitor intracellular localization of protein kinase C (PKC) in U-87 MG human glioma cells in the presence of hypericin (Hyp) and phorbol 12-myristate-13-acetate (PMA). It is shown that PKC localization, which reflects its activity, is influenced by Hyp and this influence is different from that observed for PMA which acts as PKC activator. Fluorescence binding experiments were used to determine the binding constants of Hyp to several isoforms of PKC. The obtained values of K_ds (∼100 nM) suggest that Hyp binds with high affinity to PKC. Finally, molecular modeling was used to compare structural models of the interaction of C1B domain of PKC (PKC isoforms α, δ, γ) with Hyp and our previously published model of the (C1B domain PKCγ)/PMA complex. The influence of Hyp on PKC translocation in U-87 MG cells in comparison with PMA, colocalization fluorescence pattern of Hyp and PKC, the higher binding affinity of Hyp to PKC in comparison with known binding constants of phorbol esters, as well as the binding mode of Hyp and PMA to the C1B domain of PKC suggested by molecular modeling, support the idea that Hyp and PMA might competitively bind to the regulatory domain of PKC.

Structural analyses of DNA oligonucleotides indicate the presence of strongly bound water molecules in the major and minor grooves. These water molecules may react with guanosine radical cation to form 8-oxo-7,8-dihydroguanine and other oxidation products, which can result in strand scission or other DNA damage. We probed the reaction of guanosine radical cation with water by incorporating alkyl-substituted cytidines and thymidines into anthraquinone-linked DNA duplexes and irradiating them with UV light at 350 nm. The incorporation of these hydrophobic substituents disrupts the DNA spine of hydration and we have observed that these modifications do not affect the trapping or long-distance hopping of radical cations in DNA.

Free electron laser–photoelectron emission microscopy (FEL-PEEM), femtosecond absorption spectroscopy and electron paramagnetic resonance (EPR) measurements of oxygen photoconsumption were used to probe the threshold potential for ionization of eumelanosomes and pheomelanosomes isolated from human hair. FEL-PEEM data show that both pigments are characterized by an ionization threshold at 282 nm. However, pheomelanosomes exhibit a second ionization threshold at 326 nm, which is interpreted to be reflective of the benzothiazine structural motif present in pheomelanin and absent in eumelanin. The lower ionization threshold for pheomelanin is supported by femtosecond transient absorption spectroscopy. Unlike photolysis at 350 nm, following excitation of solubalized synthetic pheomelanin at 303 nm, the transient spectrum observed between 500 and 700 nm matches that for the solvated electron, indicating the photoionization threshold for the solubalized pigment is between 350 and 303 nm. For the same synthetic pheomelanin, EPR oximetry experiments reveal an increased rate of oxygen uptake between 338 nm and 323 nm, narrowing the threshold for photoionization to sit between these two wavelengths. These results on the solubalized synthetic pigment are consistent with the FEL-PEEM results on the human melanosomes. The lower ionization potential observed for pheomelanin could be an important part of the explanation for the greater incidence rate of UV-induced skin cancers in red-haired individuals.

Lipids extracted from the skin of C57BL/6J mice injected subcutaneously with α-(4-pyridyl-1-oxide)-N-tert-butylnitrone (POBN) and exposed to topical protoporphyrin IX (PPIX) and visible light had significantly higher levels of POBN spin adducts compared with dark PPIX exposed or vehicle-treated controls. Computer analysis of the POBN adduct electron paramagnetic (spin) resonance (EPR) spectra indicated that two radical species were present in each extract, one of which was a lipid-derived carbon-centered adduct (1, a^N = 14.8 G and a^H = 2.6 G), whereas the other (2, a^N = 13.8 G and a^H = 1.8 G) was probably oxygen centered. Adduct 2 was present in greater proportion in lipids extracted from PPIX/light-exposed mice compared with dark or vehicle-treated controls. These findings suggest that PPIX/light generates free radicals in mouse skin, thus providing a radical mechanism for PPIX-induced photosensitivity. Our approach may be useful for the detection of free radicals generated by other skin photosensitizers and may also provide a means for testing putative skin-protecting agents.

Transgenic rats with the P23H mutation in rhodopsin exhibit increased susceptibility to light damage, compared with normal animals. It is known that light-induced retinal damage requires repetitive bleaching of rhodopsin and that photoreceptor cell loss is by apoptosis; however, the underlying molecular mechanism(s) leading to photoreceptor cell death are still unknown. Photoproducts, such as all-trans retinal or other retinoid metabolites, released by the extensive bleaching of rhodopsin could lead to activation of degenerative processes, especially in animals genetically predisposed to retinal degenerations. Using wild-type and transgenic rats carrying the P23H opsin mutation, we evaluated the effects of acute intense visible light on retinoid content, type and distribution in ocular tissues. Rats were exposed to green light (480–590 nm) for 0, 5, 10, 30 and 120 min. Following light treatment, rats were sacrificed and neural retinas were dissected free of the retinal pigment epithelium. Retinoids were extracted from retinal tissues and then subjected to HPLC and mass spectral analysis. We found that the light exposure affected relative levels of retinoids in the neural retina and retinal pigment epithelium of wild-type and P23H rat eyes similarly. In the P23H rat retina but not the wild-type rat retina, we found a retinoic acid–like compound with an absorbance maximum of 357 nm and a mass of 304 daltons. Production of this retinoic acid–like compound in transgenic rats is influenced by the age of the animals and the duration of light exposure. It is possible that this unique retinoid may be involved in the process of light-induced retinal degeneration.

Two boron complexes of 5-phenyldipyrromethenes bearing isothiocyanate groups on the phenyl ring have been synthesized for the first time. The utility of these new fluorescence probes for labeling biologically relevant proteins is demonstrated on two monoclonal antibodies that bind to antigens overexpressed on cancer cells. Spectral comparison of the two structures reveals significant photophysical differences, including bathochromically shifted excitation and emission bands, increased molar absorptivity and a large increase in fluorescence quantum yield of approximately 10 times. Differences in photophysical parameters are linked to hindered rotation of the phenyl ring in one of the probes.

The importance of protection against solar ultraviolet radiation (UVR) in childhood has lead to SunSmart policies at Australian schools, in particular primary schools, where children are encouraged and in many cases required to wear hats at school. Hat styles change regularly and the UVR protection provided by some of the hat types currently used and recommended for sun protection by the various Australian state cancer councils had not been previously evaluated. The UVR protection of the hats was measured using UVR sensitive polysulphone film badges attached to different facial sites on rotating headforms. The sun protection type hats included in this study were broad-brimmed hats, “bucket hats” and legionnaires hats. Baseball caps, which are very popular, were also included. The broad-brimmed hats and bucket hats provided the most UVR protection for the six different sites about the face and head. Legionnaires hats also provided satisfactory UVR protection, but the caps did not provide UVR protection to many of the facial sites. The highest measured UVR protection factors for facial sites other than the forehead were 8 to 10, indicating that, while some hats can be effective, they need to be used in combination with other forms of UVR protection.

We have studied the association reaction of the CF₂Cl radicals with O₂ in presence of N₂. The infrared multiple photon dissociation (IRMPD) technique with a homemade TEA CO₂ laser was used for the CF₂Cl radical generation and the vibrational chemiluminiscence technique was set up for the study of the reaction kinetics. The time-resolved IR fluorescence of the vibrationally excited CF₂O photoproduct was used to measure the disappearance rate of these radicals. A kinetic mechanism is presented to account for the rate of production of CF₂O^*. The CF₂Cl radical association reaction rate with O₂, evidence of a direct channel of photoproduct formation and its reaction rate, and the CF₂O^* collisional deactivation rate have been obtained.

Polyunsaturated fatty acids located in leukemia cell membranes are excellent targets for peroxidation. They can significantly enhance the effectiveness of Photofrin-mediated photodynamic therapy (PDT)-induced cell killing. In this study, the peroxidizability of conjugated fatty acid isomers (9c,11t-linoleic acid and 9c,11c-linoleic acid) and polyunsaturated fatty acids (PUFAs; linoleic acid, γ-linolenic acid and arachidonic acid) with 2,2′-azo-bis(2-amidinpropane)dihydrochloride, soybean lipoxygenase and photomediated peroxidation are compared with each other. Peroxidation was determined using different methods: by means of gas chromatography to estimate the fatty acid (FA) consumption, by photometry for the level of FA peroxides or phospholipid peroxides and by definition of the content of malondialdehyde for thiobarbituric acid reactive substances (TBARS). The results suggest that the generation of oxidation products from individual FAs indicate a different formation rate of oxidation products. Radical FA peroxides were produced most by polyunsaturated arachidonic acid, followed by linoleic acid and γ-linolenic acid, whereas conjugated FA isomers did not generate peroxides. Accordingly, the levels of lipid peroxides and TBARS were substantially increased after incorporation and oxidation of polyunsaturated FAs into U937 cells and could significantly enhance the effectiveness of Photofrin-PDT–induced cytotoxicity. The results showed that PUFA, but not conjugated FA supplementation of U937 cells, can act as a PDT amplification factor.

The photostabilities of bacteriochlorophyll a and several of its derivatives, which are of interest as potential sensitizers in photodynamic tumor therapy, were investigated. The pigments were irradiated with light >630 nm in organic solvents (acetone, tetrahydrofuran, pyridine, methanol, ethanol, n-propanol, 2-propanol and toluene) and in aqueous detergent solutions (cetyl-trimethyl-ammonium bromide [CTAB], lauryldimethyl-aminoxide [LDAO] or sodium dodecyl-sulfate [SDS] and Triton X-100 [TX100]). Their stabilities in these different solvents were determined in the presence and absence of an external sensitizer (pyromethyl–pheophorbide a), oxygen, sodium ascorbate and inert gas (Ar) or vacuum. The photodegradation products of bacteriochlorophyll a in acetone solution were isolated, purified by HPLC and analyzed by their absorption spectra and mass spectroscopy. Besides the well-known dehydrogenation products, such as [3-acetyl]-chlorophyll a, which were obtained as by-products, the major products had low absorption in the visible–near infrared spectral range. The spectral signature of the major component of these products was characteristic of linear open-chain tetrapyrroles, but they lacked the characteristic protonation–deprotonation behavior and reactivity of bilins with Zn .

We examined UV-B radiation flux and its environmental control within and among streams of northern Michigan. UV-B flux was estimated in streams by plastic dosimetry strips, which allow for the simultaneous and repeated in situ measurement of solar radiation. During the summer of 2004, UV-B flux was measured across depth gradients and along longitudinal transects in seven streams, which were chosen to encompass a range of dissolved organic carbon (DOC) concentrations and canopy cover. Attenuation coefficients of UV-B (K_{d UV-B}) were estimated using plastic dosimeters placed along a depth gradient. K_{d UV-B} were positively correlated with DOC concentration and similar to values obtained with laboratory and in situ spectrometry. Along 100 m longitudinal transects, UV-B flux varied along all streams regardless of their canopy cover and DOC concentration. Within-stream fluxes of UV-B were correlated to canopy cover in the only two streams that both had relatively low DOC concentration and variable canopy cover. Large differences were found among streams in the average UV-B flux (corrected for incident solar flux) reaching the dosimeters at 5 cm depth. These among-stream differences were largely accounted for by the stream width, canopy cover, and DOC concentration. Our results illustrate an inherent variability in UV-B flux within and among streams of northern Michigan that is strongly tied to the interactions of DOC concentration, stream size and riparian vegetation.

Ultraviolet light (UV) induces aggregation of Fas-receptor through a Fas-ligand–independent pathway. However, the mechanism of ultraviolet light-induced Fas-receptor aggregation is not known. In this report, we show that lipid rafts mediate ultraviolet light–induced aggregation of Fas. Our data show that UV induces a redistribution of Fas-receptor in a 25–5% Optiprep continuous gradient. The amount of Fas-receptorS is significantly increased in a gradient fraction that contain lipid rafts and is associated with an increase of FADD and caspase-8. Our data also show that the active dimeric form of caspase-8 (p44/p41) is increased in the lipid raft fraction. In addition, our data show that cholesterol, a major component of lipid rafts, is significantly reduced in only the lipid raft fractions after UV-irradiation. However, ceramide, another major lipid raft component, is increased evenly in all gradient fractions after UV-irradiation. These results suggest that UV alters the composition of major lipid raft components, which leads to the recruitment of Fas-receptor and FADD, with subsequent activation of caspase-8. Based on our results, we propose a novel mechanism by which UV induces apoptosis through a membrane lipid raft–mediated signaling pathway.

The aim of our study was to evaluate the changes of interstitial pH and flavin adenine dinucleotide (FAD)/reduced nicotinamide adenine dinucleotide (NADH) ratio in healing skin wounds using fluorescence spectroscopy in Sprague Dawley rats. In the experiment, excisional and incisional models of wound healing were used. The florescein as the pH-sensitive probe using excitation spectra (λ_Em = 535 nm) was used for the measurement of pH changes, and synchronous fluorescence spectra (Δλ = 60 nm) for the monitoring of FAD/NADH ratio changes were measured from the surfaces of healing wounds. Increase of interstitial pH and FAD/NADH ratio was recorded during the time interval from the 15th to the 65th minute after surgery. The decrease of pH between the 48th and the 72nd hour after surgery as well as the increase of FAD/NADH ratio between the 72nd and the 96th hour of wound healing were recorded. The results indicate that the use of fluorescence spectroscopy may be considered as a valuable tool for noninvasive in vivo monitoring of selected redox parameters in the early phases of wound healing.

In this paper, the solvent-dependent photosensitive behaviors of fullerene (C₆₀) were investigated in polar and nonpolar solvents by time-dependent density functional theory (TD-DFT) calculation. Based on the calculated physicochemical parameters on triplet state, it is revealed that excited-state C₆₀ only generates ¹O₂ via energy transfer in benzene, but can give birth to O₂^·− and ¹O₂ in water via energy transfer and electron transfer, respectively. Considering the fact that electron transfer is more favorable compared with energy transfer in polar biological systems, especially with the presence of electron donors, the O₂^·−-generating process will get predominant in physiological systems. These results account well for the experimental observations that O₂^·− and ^·OH are primarily responsible for the photoinduced DNA cleavage by C₆₀ under physiological conditions, whereas ¹O₂ plays a critical role in nonpolar solvents.

The photolysis of acetophenone, benzophenone, 4-carboxybenzophenone and benzil was studied in air-saturated aqueous solution in the presence of alcohols. The overall reaction is an oxidation of 2-propanol to acetone. The quantum yield of oxygen uptake (Φ_−O2) increases with increasing 2-propanol concentration up to 0.9. The photoreaction can also be initiated by quenching of the ketone triplet state by ascorbic acid, formate or an amine e.g. triethylamine. Subsequent reactions of the involved radicals with oxygen yield the superoxide radical and eventually hydrogen peroxide. For the ketones in the presence of 3–30 mM ascorbic acid or triethylamine Φ_−O2 = 0.3–0.9. The specific properties of ketones, including 4-methoxyacetophenone and 2-acetonaphthone, the radicals involved and the pH and concentration dependences of Φ_−O2 are discussed.

Photochemical internalization (PCI) is a targeting technique that facilitates endosomal escape of macromolecules, such as transgenes, in response to photochemical treatment with endosome/lysosome-localized photosensitizers, such as disulfonated meso-tetraphenylporphine (TPPS_2a). In gene therapy this leads to enhanced transgene expression. Moreover, photochemical treatment generally activates transcription of stress-response genes, such as heat-shock proteins (HSPs), via stimulation of corresponding promoters. Therefore, we used HSP70 (HSPp; a promoter from the HSP family gene) and investigated whether the PCI stimulus could also activate HSPp and thereby stimulate transcription (expression) of the HSPp-controlled transgene internalized via PCI. Using human colorectal carcinoma and hepatoma cell lines in vitro, we showed that TPPS_2a-based photochemical treatment enhances expression of cellular HSP70, which correlated with a photochemically enhanced expression (approximately 2-fold, at PCI-optimal doses) of the HSPp-controlled transgene integrated in the genome. Furthermore, PCI enhanced expression of the HSPp-controlled episomal transgene delivered as a plasmid. However, in plasmid-based transfection, PCI-mediated enhancement with HSPp did not exceed the enhancement achieved with the constitutive active CMV promoter. In conclusion, we demonstrated that the PCI-relevant treatment initiates HSP70 response and that the HSP70 promoter can be used in combination with PCI, leading to PCI-enhanced expression of the HSPp-controlled transgene.

It has been proposed that photodegradation of folates may be the reason for the pigmentation of races living under high fluence rates of ultraviolet radiation. The photodegradation of folic acid (FA) induced by ultraviolet-A (UV-A) radiation, in solution and in the presence of human serum albumin (HSA), was studied with absorption and fluorescence spectroscopy. FA photodegradation, with formation of p-aminobenzoyl-l-glutamic acid, 6-formylpterin and pterin-6-carboxylic acid, was found to follow an exponential trend. A scheme of FA photodegradation, which involves photosensitization of FA degradation by its photoproducts, was proposed. The rate of FA photodegradation decreased drastically in the presence of HSA, whereas the spectral characteristics of the photoproducts remained constant. The reduction of the FA photodegradation rate by HSA was accompanied by degradation of tryptophan in HSA. Tryptophan, when added to solutions of FA, had a similar effect as HSA. In solutions of FA and HSA the FA photoproducts cause photodamage mainly to HSA rather than to FA itself. The oxygen dependence of FA photodegradation and the inhibition of this process by sodium azide indicate that singlet oxygen may participate in the photosensitizing activity of FA photoproducts.

Di-2,2′-diethylhexyl-3,5-dimethoxy-4-hydroxy-benzylidenemalonate (INCI name diethylhexyl syringylidene malonate, DESM), the target photostabilizer, was synthesized in one step by condensation of 3,5-dimethoxy-4-hydroxy benzaldehyde (Syringaldehyde) with di-2,2′-diethylhexyl malonate. Photostability data in sunscreen formulations showed that DESM is photostable and improves the photostability of avobenzone significantly when compared to control (without a photostabilizer). Photostable broad-spectrum sunscreen formulations with high SPF (>30) have been achieved by combining avobenzone, DESM and UV-B sunscreens, such as homosalate, octisalate or other UV-B sunscreens. It seems that (a) triplet-state energy transfer from avobenzone to DESM and (b) scavenging of reactive species are responsible for the observed stabilization of avobenzone. In vitro study of the two formulations containing DESM clearly showed critical wavelength of well over 370 nm and can thus be categorized as broad-spectrum sunscreens. DESM does not have any contribution to in vivo SPF; instead it boosts SPF by about 5 units in high-SPF products. DESM was found to be an excellent singlet-oxygen quencher, thereby reducing photodegradation of avobenzone caused by singlet oxygen. In short, the multiplicity of effects and formulation benefits seen with DESM makes it an ideal choice as a unique antioxidant photostabilizer for a variety of cosmetic products targeting young and mature skin alike.