Risk of Eye Damage from the Wavelength-Dependent Biologically Effective UVB Spectrum Irradiances

A number of previous studies have discussed the risk of eye damage from broadband ultraviolet (UV) radiation. As the biologically damaging effectiveness of UV irradiation on the human body is known to be wavelength-dependent, it is necessary to study the distribution of the UV spectral irradiance. In order to quantify the ocular biologically effective UV (UVBE) irradiance exposure of different wavelengths and assess the risk of eye damage, UV exposure values were measured at Sanya, China (18.4° N, 109.7°E, altitude 18 m), using a manikin and a dual-detector spectrometer to measure simultaneously the ocular exposure and ambient UV spectral irradiance data and solar elevation angle (SEA) range (approximately 7°–85°). The present study uses the ocular UV spectral irradiance exposure weighted with the action spectra for photokeratitis, photoconjunctivitis and cataracts to calculate the ocular UVBE irradiance exposure for photokeratitis (UVBE_pker), photoconjunctivitis (UVBE_pcon) and cataracts (UVBE_cat). We found that the ocular exposure to UV irradiance is strongest in the 30°–60° SEA range when ∼50% of ocular exposure to UV irradiance on a summer’s day is received. In the 7°–30° SEA range, all the biologically highly effective wavelengths of UVBE_pker, UVBE_pcon and UVBE_cat irradiances are at 300 nm. However, in other SEA ranges the biologically highly effective wavelengths of UVBE_pker, UVBE_pcon and UVBE_cat irradiances are different, corresponding to 311 nm, 300 nm and 307 nm, respectively.     

Toxicity and mutagenicity of radiation from fluorescent lamps and a sunlamp in L5178y mouse lymphoma cells

Unfiltered broad spectrum radiation emitted by black light, cool white, and black light blue fluorescent lamps and a sunlamp, is both toxic and mutagenic to L5178Y mouse lymphoma cells when the cells are irradiated in phosphate-buffered saline. The increase in mutant frequency seen after exposure of the cells is linear throughout the range of exposures tested. The linear increase in mutagenesis is observed even at exposure levels which do not cause significant toxicity. To facilitate comparison of the differing rates of mutagenesis derived from exposure-response curves obtained for each light source, we have defined a parameter, joule-equivalent mutagenesis (jem), equal to mutants per 10(5) survivors per joule per square meter. Jem values are calculated using the integrated irradiance of each lamp. Based on jem values, the relative mutagenicity of the various lamps tested (compared with a germicidal ultraviolet lamp) is 3 x 10(-3) for the sunlamp, 1 x 10(-4) for the black light and cool white lamps, and 3 x 10(-5) for the black light blue lamp. The toxic and mutagenic effects of the lamps are in reasonable agreement with their relative spectral output from 290 to 330 nm.     

Photic sensitivity ranges of hamster pupillary and circadian phase responses do not overlap

Mammalian retinal photoreceptors form an irradiance detection system that drives many nonvisual responses to light such as pupil reflex and resetting of the circadian clock. To understand the role of pupil size in circadian light responses, pupil diameter was pharmacologically manipulated and the effect on behavioral phase shifts at different irradiance levels was studied in the Syrian hamster. Dose-response curves for steady-state pupil size and for behavioral phase shifts were constructed for 3 pupil conditions (dilated, constricted, and control). Retinal irradiance was calculated from corneal irradiance, pupil size, retinal surface area, and absorption of ocular media. The sensitivity of photic responses to retinal irradiance is approximately 1.5 log units higher than to corneal irradiance. When plotted against corneal irradiance, pharmacological pupil constriction reduces the light sensitivity of the circadian system, but pupil dilation has no effect. As expected, when plotted against retinal irradiance all dose-response curves superimposed, confirming that the circadian system responds to photon flux on the retina. Pupil dilation does not increase the circadian response to increasing irradiance, since the response of the circadian system attains saturation at irradiance levels lower than those required to induce pupil constriction. The main finding shows that due to the different response sensitivities, the effect of pupil constriction on the light sensitivity of the circadian system in the hamster under natural conditions is virtually negligible. We further suggest the existence of distinct modulating mechanisms for the differential retinal irradiance sensitivity of the pupil system and the circadian system, which enables the different responses to be tuned to their specific tasks while using similar photoreceptive input.     

The lens eyes of the box jellyfish <Emphasis Type="Italic">Tripedalia cystophora</Emphasis> and <Emphasis Type="Italic">Chiropsalmus sp.</Emphasis> are slow and color-blind

Box jellyfish, or cubomedusae, possess an impressive total of 24 eyes of four morphologically different types. Compared to other cnidarians they also have an elaborate behavioral repertoire, which for a large part seems to be visually guided. Two of the four types of cubomedusean eyes, called the upper and the lower lens eye, are camera type eyes with spherical fish-like lenses. Here we explore the electroretinograms of the lens eyes of the Caribbean species, Tripedalia cystophora, and the Australian species, Chiropsalmus sp. using suction electrodes. We show that the photoreceptors of the lens eyes of both species have dynamic ranges of about 3 log units and slow responses. The spectral sensitivity curves for all eyes peak in the blue-green region, but the lower lens eye of T. cystophora has a small additional peak in the near UV range. All spectral sensitivity curves agree well with the theoretical absorbance curve of a single opsin, strongly suggesting color-blind vision in box jellyfish with a single receptor type. A single opsin is supported by selective adaptation experiments.     

Differences in gas exchange contribute to habitat differentiation in Iberian columbines from contrasting light and water environments

During photosynthesis, respiration and transpiration, gas exchange occurs via the stomata and so plants face a trade‐off between maximising photosynthesis while minimising transpiration (expressed as water use efficiency, WUE). The ability to cope with this trade‐off and regulate photosynthetic rate and stomatal conductance may be related to niche differentiation between closely related species. The present study explored this as a possible mechanism for habitat differentiation in Iberian columbines. The roles of irradiance and water stress were assessed to determine niche differentiation among Iberian columbines via distinct gas exchange processes. Photosynthesis–irradiance curves (P–I curves) were obtained for four taxa, and common garden experiments were conducted to examine plant responses to water and irradiance stress, by measuring instantaneous gas exchange and plant performance. Gas exchange was also measured in ten individuals using two to four field populations per taxon. The taxa had different P–I curves and gas exchange in the field. At the species level, water stress and irradiance explained habitat differentiation. Within each species, a combination of irradiance and water stress explained the between‐subspecies habitat differentiation. Despite differences in stomatal conductance and CO₂ assimilation, taxa did not have different WUE under field conditions, which suggests that the environment equally modifies photosynthesis and transpiration. The P–I curves, gas exchange in the field and plant responses to experimental water and irradiance stresses support the hypothesis that habitat differentiation is associated with differences among taxa in tolerance to abiotic stress mediated by distinct gas exchange responses.     

Flower Colours through the Lens: Quantitative Measurement with Visible and Ultraviolet Digital Photography

Background

The study of the signal-receiver relationship between flowering plants and pollinators requires a capacity to accurately map both the spectral and spatial components of a signal in relation to the perceptual abilities of potential pollinators. Spectrophotometers can typically recover high resolution spectral data, but the spatial component is difficult to record simultaneously. A technique allowing for an accurate measurement of the spatial component in addition to the spectral factor of the signal is highly desirable.

Methodology/Principal findings

Consumer-level digital cameras potentially provide access to both colour and spatial information, but they are constrained by their non-linear response. We present a robust methodology for recovering linear values from two different camera models: one sensitive to ultraviolet (UV) radiation and another to visible wavelengths. We test responses by imaging eight different plant species varying in shape, size and in the amount of energy reflected across the UV and visible regions of the spectrum, and compare the recovery of spectral data to spectrophotometer measurements. There is often a good agreement of spectral data, although when the pattern on a flower surface is complex a spectrophotometer may underestimate the variability of the signal as would be viewed by an animal visual system.

Conclusion

Digital imaging presents a significant new opportunity to reliably map flower colours to understand the complexity of these signals as perceived by potential pollinators. Compared to spectrophotometer measurements, digital images can better represent the spatio-chromatic signal variability that would likely be perceived by the visual system of an animal, and should expand the possibilities for data collection in complex, natural conditions. However, and in spite of its advantages, the accuracy of the spectral information recovered from camera responses is subject to variations in the uncertainty levels, with larger uncertainties associated with low radiance levels.     

Testing the consistency of wildlife data types before combining them: the case of camera traps and telemetry

Wildlife data gathered by different monitoring techniques are often combined to estimate animal density. However, methods to check whether different types of data provide consistent information (i.e., can information from one data type be used to predict responses in the other?) before combining them are lacking. We used generalized linear models and generalized linear mixed-effects models to relate camera trap probabilities for marked animals to independent space use from telemetry relocations using 2 years of data for fishers (Pekania pennanti) as a case study. We evaluated (1) camera trap efficacy by estimating how camera detection probabilities are related to nearby telemetry relocations and (2) whether home range utilization density estimated from telemetry data adequately predicts camera detection probabilities, which would indicate consistency of the two data types. The number of telemetry relocations within 250 and 500 m from camera traps predicted detection probability well. For the same number of relocations, females were more likely to be detected during the first year. During the second year, all fishers were more likely to be detected during the fall/winter season. Models predicting camera detection probability and photo counts solely from telemetry utilization density had the best or nearly best Akaike Information Criterion (AIC), suggesting that telemetry and camera traps provide consistent information on space use. Given the same utilization density, males were more likely to be photo-captured due to larger home ranges and higher movement rates. Although methods that combine data types (spatially explicit capture–recapture) make simple assumptions about home range shapes, it is reasonable to conclude that in our case, camera trap data do reflect space use in a manner consistent with telemetry data. However, differences between the 2 years of data suggest that camera efficacy is not fully consistent across ecological conditions and make the case for integrating other sources of space-use data.     

An inexpensive photosynthetic irradiance sensor for ecological field studies

The design and characteristics of inexpensive and simply constructed equal-energy response photosynthetic irradiance sensors is described for use particularly where several cells are required in comparative ecological studies either above or below water. The dimensions of the sensors can be changed proportionally to suit different applications or components. The response of the sensor to irradiance at varying angles corresponds very closely to that required by the cosine law. The sensor is comparatively insensitive to other environmental variables in field use and gave a stable output; the long term drift was proportional to electrical output but in continuous use, drift is regular and could reach -0.08 year^-1 of the total. The spectral range and cosine response is discussed in comparison to other more expensive (x 5–10) commercially available, sensors and to local standards.     

Colour As a Signal for Entraining the Mammalian Circadian Clock

Twilight is characterised by changes in both quantity (“irradiance”) and quality (“colour”) of light. Animals use the variation in irradiance to adjust their internal circadian clocks, aligning their behaviour and physiology with the solar cycle. However, it is currently unknown whether changes in colour also contribute to this entrainment process. Using environmental measurements, we show here that mammalian blue–yellow colour discrimination provides a more reliable method of tracking twilight progression than simply measuring irradiance. We next use electrophysiological recordings to demonstrate that neurons in the mouse suprachiasmatic circadian clock display the cone-dependent spectral opponency required to make use of this information. Thus, our data show that some clock neurons are highly sensitive to changes in spectral composition occurring over twilight and that this input dictates their response to changes in irradiance. Finally, using mice housed under photoperiods with simulated dawn/dusk transitions, we confirm that spectral changes occurring during twilight are required for appropriate circadian alignment under natural conditions. Together, these data reveal a new sensory mechanism for telling time of day that would be available to any mammalian species capable of chromatic vision.     

The visual ecology of Puerto Rican anoline lizards: habitat light and spectral sensitivity

The visual ecology of six closely related species of Puerto Rican anoline lizards was investigated and they were found to occupy four distinct habitat types in terms of light conditions: “full shade”, “partial shade”, “no shade”, and “forest canopy.”The habitats differed substantially in total radiance and irradiance as well as in the shape of the irradiance spectrum. The shape of the radiance spectrum was similar in all of the habitats. We used electroretinogram (ERG) flicker photometry to measure spectral sensitivity and found the curves for all six species to be similar. The spectral sensitivity peaked in the range 550–560 nm, which matched the peak in spectral radiance for all of the habitats. The shape of the spectral-sensitivity curve was similar to those of a number of other terrestrial vertebrates. We suggest that the convergence of the shape of the photopic ERG-determined spectral-sensitivity curve in many terrestrial vertebrates may, in part, be due to the fact that the background radiance of many terrestrial habitats is dominated by the reflectance spectrum of green vegetation which peaks at 550 nm. Accepted: 14 May 1997     

Photocontrol of Anthocyanin Synthesis: VI. Spectral Sensitivity, Irradiance Dependence, and Reciprocity Relationships

The spectral sensitivity and the irradiance dependence of anthocyanin synthesis, a “high irradiance response,” in cabbage (Brassica oleracea, cv. Red Acre) and tomato (Lycopersicon esculentum, cv. Beefsteak) seedlings exposed to continuous irradiation depend upon the length of the exposure. In cabbage, blue and red are more effective than far red when the irradiations are shorter than 12 hours and less effective than far red when the irradiations are longer than 12 hours. The irradiance dependence is negligible under red and becomes evident under blue and far red red only for exposures longer than 12 hours. Anthocyanin synthesis under intermittent light treatments, of efficiency comparable to that of continuous treatments, obeys the Bunsen-Roscoe reciprocity law and is a function of the dose (irradiance × time), rather than of the irradiance alone. The validity of the reciprocity relationships suggests that only one photoreceptor is responsible for the photocontrol of the response in the blue, red, and far red spectral regions. The characteristics of the response suggest that the photoreceptor is phytochrome, at least in cabbage.     

A "melanopic" spectral efficiency function predicts the sensitivity of melanopsin photoreceptors to polychromatic lights

Photoreception in the mammalian retina is not restricted to rods and cones but extends to a small number of intrinsically photosensitive retinal ganglion cells expressing the photopigment melanopsin. These mRGCs are especially important contributors to circadian entrainment, the pupil light reflex, and other so-called nonimage-forming (NIF) responses. The spectral sensitivity of melanopsin phototransduction has been addressed in several species by comparing responses to a range of monochromatic stimuli. The resultant action spectra match the predicted profile of an opsin:vitamin A-based photopigment (nomogram) with a peak sensitivity (λ(max)) around 480 nm. It would be most useful to be able to use this spectral sensitivity function to predict melanopsin's sensitivity to broad-spectrum, including "white," lights. However, evidence that melanopsin is a bistable pigment with an intrinsic light-dependent bleach recovery mechanism raises the possibility of a more complex relationship between spectral quality and photoreceptor response. Here, we set out to empirically determine whether simply weighting optical power at each wavelength according to the 480-nm nomogram and integrating across the spectrum could predict melanopsin sensitivity to a variety of polychromatic stimuli. We show that pupillomotor and circadian responses of mice relying solely on melanopsin for their photosensitivity (rd/rd cl) can indeed be accurately predicted using this methodology. Our data therefore suggest that the 480-nm nomogram may be employed as the basis for a new photometric measure of light intensity (which we term "melanopic") relevant for melanopsin photoreception. They further show that measuring light in these terms predicts the melanopsin response to light of divergent spectral composition much more reliably than other methods for quantifying irradiance or illuminance currently in widespread use.     

Evaluation of the different levels of variability in the underwater light field of a shallow estuary

The underwater light climate of a shallow estuary located at the southern coast of the Baltic Sea has been investigated, with special emphasis on the spectral irradiance composition and on short-term irradiance fluctuations caused by vertical mixing and wave focussing. The inherent optical properties of the water body were dominated by phytoplankton pigment absorption in the long-wavelength range and by coloured, dissolved organic matter (cDOM) absorption in the wavelength range <500 nm, including ultraviolet-A (UV-A) and ultraviolet-B radiation (UV-B). Pronounced particulate scattering combined with the absorption values to give very high attenuation coefficients, especially for the shorter wavelengths of UV-B radiation. Photosynthetically active radiation (PAR) was found to be reduced to 1% of the surface value within 0.8 m in the inner, hypertrophic end of the estuary and within 1.9 m in the outer, eutrophic parts of this system, with corresponding 1% penetration depths for UV-B of 0.13 and 0.31 m. During late winter and early spring, the period when reduced atmospheric ozone concentrations and enhanced UV-B have been reported over northern Europe, the irradiance levels in the water column were greatly reduced, due to strong attenuation by ice cover and overlying snow. cDOM concentration of the water was also found to remain at a high level during these periods, and indeed throughout the year, thus reducing the exposure of organisms to UV-R and PAR still further. A complex irradiance regime was found in this system, with irregular and high amplitude fluctuations caused by wind-induced vertical mixing and wave focussing being superimposed upon the solar-angle-dependent seasonal and daily cycles. The methods used to quantify the short-term fluctuations are described, and their relevance to phytoplankton physiology is discussed. The wave-focussing effect is unique to the aquatic environment, and measurements showed that average subsurface irradiances could be increased by up to 5 times for periods lasting for <1 s. The highest irradiances recorded during wave-focussing events reached over 9,000 μmol photons m^–2 s^–1. Received in revised form: 7 April 2000 Electronic Publication     

Ship-borne measurements of erythemal UV irradiance and ozone content in various climate zones.

Ship-borne measurements of spectral as well as biologically effective UV irradiance have been performed on the German research vessel Polarstern during the Atlantic transect from Bremerhaven, Germany (53.5 degrees N, 8.5 degrees E), to Cape Town, South Africa (33.6 degrees S, 18.3 degrees E), from 13 October to 17 November 2005. Such measurements are required to study UV effects on marine organisms. They are also necessary to validate satellite-derived surface UV irradiance. Cloud free radiative transfer calculations support the investigation of this latitudinal dependence. Input parameters, such as total ozone column and aerosol optical depth have been measured on board as well. Using these measured parameters, the modelled cloudless noontime UVA irradiance (320-400 nm) shows the expected dependence on varying minimum solar zenith angles (SZA) at different latitudes. The modelled cloudless noontime UVB irradiance (290-320 nm) does not show this clear dependence on SZA due to the strong influence of ozone absorption in this spectral range. The maximum daily dose of erythemal irradiance of 5420 J m(-1) was observed on 14 November 2005, when the ship was in the tropical Atlantic south of the equator. The expected UV maximum should have been observed with the sun in the zenith during local noon (11 November). Stratiform clouds reduced the dose to 3835 J m(-1). In comparison, the daily erythemal doses in the mid-latitudinal Bay of Biscay only reached values between 410 and 980 J m(-1) depending on cloud conditions. The deviation in daily erythemal dose derived from different instruments is around 5%. The feasibility to perform ship-borne measurements of spectral UV irradiance is demonstrated.     

Growth responses to irradiance regime along an ecological gradient of <Emphasis Type="Italic">Quercus</Emphasis> <Emphasis Type="Italic">castaneifolia</Emphasis> seedlings of different provenance

Understory light is known as one of the most important environmental predictors of growth response of woody species. Hence, the primary objective of most forest management practices is optimizing that resource for understory seedlings. In this study, growth responses of Quercus castaneifolia seedlings from five different provenances from west to east of the Hyrcanian forest were investigated along their ecological gradients (from high to low precipitation). An experimental design was executed under controlled conditions at eight different irradiance levels (10, 20, 30, 40, 50, 60, 70 and 100 % full irradiance). Results showed that the irradiance is probably the most important determinant of variation in seedling characteristics. Among all investigated variables, variability in seedling size was affected significantly by provenance, while seedling morphology and their architectural response was affected by different levels of irradiance in a curvilinear manner. The biggest changes were observed at lowest irradiance levels (10–20 %) while at higher irradiance (70–100 %) the curves flatten. It was shown that, unlike at low irradiance levels, there is little capacity in seedling morphology to acclimatize with high irradiance intensity. Attaining maximal biomass varies across provenances and irradiance gradient. The highest biomass for the five provenances could be ranked as follows: 20–60 % and 50–60 % for the wetter and drier provenances, respectively. These results demonstrated that the light requirement increases from wetter to drier provenances, with a negative relationship between light requirement and precipitation regime. Different responses to irradiance levels may be the result of genetic adaptation to the ecological conditions prevailing in native habitat, especially precipitation regime.     

Growth rates of phytoplankton under fluctuating light

• 1 The effect of light fluctuations on the growth rates of four species of freshwater phytoplankton was investigated. Experimental light regimes included constant irradiance and fluctuations of a step function form, with equal proportion of high (maximum of 240 µmol photons m^‐2 s^‐1) and low light (minimum of 5 µmol photons m^‐2 s^‐1) (or dark) in a period. Fluctuations of 1, 8 and 24‐h periods were imposed over several average irradiances (25, 50, 100 and 120 µmol photons m^‐2 s^‐1).
• 2 Growth rate responses to fluctuations were species‐specific and depended on both the average irradiance and the period of fluctuations. Fluctuations at low average irradiances slightly increased growth rate of the diatom Nitzschia sp. and depressed growth of the cyanobacterium Phormidium luridum and the green alga Sphaerocystis schroeteri compared to a constant irradiance.
• 3 Fluctuations at higher average irradiance did not have a significant effect on the growth rates of Nitzschia sp. and Sphaerocystis schroeteri (fluctuations around saturating irradiances) and slightly increased the growth rates of the cyanobacteria Anabaena flos‐aquae and Phormidium luridum (when irradiance fluctuated between limiting and inhibiting levels).
• 4 In general, the effect of fluctuations tended to be greater when irradiance fluctuated between limiting and saturating or inhibiting levels of a species growth‐irradiance curve compared to fluctuations within a single region of the curve.
• 5 The growth rates of species under fluctuating light could not always be predicted from their growth‐irradiance curves obtained under constant irradiance. When fluctuations occur between limiting and saturating or inhibiting irradiances for the alga and when the period of fluctuations is long (greater than 8 h), steady‐state growth‐irradiance curves may be insufficient to predict growth rates adequately. Consequently, additional data on physiological acclimation, such as changes in photosynthetic parameters, may be required for predictions under non‐constant light supply in comparison to constant conditions.

     

Bayesian methods for comparing species physiological and ecological response curves

Many ecological questions require information on species' optimal conditions or critical limits along environmental gradients. These attributes can be compared to answer questions on niche partitioning, species coexistence and niche conservatism. However, these comparisons are unconvincing when existing methods do not quantify the uncertainty in the attributes or rely on assumptions about the shape of species' responses to the environmental gradient. The aim of this study was to develop a model to quantify the uncertainty in the attributes of species response curves and allow them to be tested for substantive differences without making assumptions about the shape of the responses. We developed a model that used Bayesian penalised splines to produce and compare response curves for any two given species. These splines allow the data to determine the shape of the response curves rather than making a priori assumptions. The models were implemented using the R2OpenBUGS package for R, which uses Markov Chain Monte Carlo simulation to repetitively fit alternative response curves to the data. As each iteration produces a different curve that varies in optima, niche breadth and limits, the model estimates the uncertainty in each of these attributes and the probability that the two curves are different. The models were tested using two datasets of mosses from Antarctica. Both datasets had a high degree of scatter, which is typical of ecological research. This noise resulted in considerable uncertainty in the optima and limits of species response curves, but substantive differences were found. Schistidium antarctici was found to inhabit wetter habitats than Ceratodon purpureus, and Polytrichastrum alpinum had a lower optimal temperature for photosynthesis than Chorisodontium aciphyllum under high light conditions. Our study highlights the importance of considering uncertainty in physiological optima and other attributes of species response curves. We found that apparent differences in optima of 7.5 °C were not necessarily substantive when dealing with noisy ecological data, and it is necessary to consider the uncertainty in attributes when comparing the curves for different species. The model introduced here could increase the robustness of research on niche partitioning, species coexistence and niche conservatism.     

Operation of the xanthophyll cycle in the seagrass Zostera marina in response to variable irradiance

Changes in the photobiology and photosynthetic pigments of the seagrass Zostera marina from Chesapeake Bay (USA) were examined under a range of natural and manipulated irradiance regimes. Photosynthetic activity was assessed using chlorophyll-a fluorescence, and photosynthetic pigments were measured by HPLC. Large changes in the violaxanthin, zeaxanthin, and antheraxanthin content were concomitant with the modulation of non-photochemical quenching (NPQ). Photokinetics (F_v/F_m, rapid light curves (RLC), and non-photochemical quenching) varied as a result of oscillating irradiance and were highly correlated to xanthophyll pigment content. Zeaxanthin and antheraxanthin concentrations increased under elevated light conditions, while violaxanthin increased in darkened conditions. Unusually high concentrations of antheraxanthin were found in Z. marina under a wide range of light conditions, and this was associated with the partial conversion of violaxanthin to zeaxanthin. These results support the idea that xanthophyll intermediate pigments induce a photoprotective response during exposure to high irradiances in this seagrass.     

Effects of water-filtered infrared A irradiation on human fibroblasts

Infrared radiation is a substantial part of the solar energy output reaching the earth surface. Therefore, exposure of humans to infrared radiation is common. However, whether and how infrared (IR) or infrared A acts on human skin cells is still under debate. Recently the generation of reactive oxygen species by water-filtered infrared A (wIRA) irradiation was postulated. wIRA shows a spectral distribution similar to that of solar irradiation at the earth's surface. Thus, the need for protection of human skin from both solar- and artificially generated infrared A irradiation was concluded. Here we demonstrate that in human dermal fibroblasts this reactive oxygen species generation is dependent on heat formation by infrared A and can be reproduced by thermal exposure. On the other hand wIRA irradiation had no detectable effect if the temperature in the cells was kept constant, even if irradiance exceeded the extraterrestrial solar irradiance in the IR range by a factor of about 4 and the maximum at noontime in the tropics by a factor up to about 6. This could be demonstrated by the measurement of oxidant formation using H₂DCFDA and the determination of protein carbonyls. In additional experiments we could show that during thermal exposure the mitochondria contribute significantly to oxidant production. Further experiments revealed that the major absorbance of infrared is due to absorption of the energy by cellular water.     

Mechanistic investigation of doxycycline photosensitization by picosecond-pulsed and continuous wave laser irradiation of cells in culture

In order to elucidate the photophysical mechanisms of cellular phototoxicity sensitized by doxycycline, MGH-U1 human bladder carcinoma cells in vitro were treated with 20.7 microM doxycycline and irradiated with either a pulsed (lambda = 355 nm, pulse duration = 24 ps) or a continuous wave (lambda = 351 nm) laser. Cumulative radiant exposure and irradiance were systematically varied in experiments with both lasers. Phototoxicity was assessed by epifluorescence microscopy of unfixed cells using rhodamine 123 labeling of mitochondria. With the continuous wave source, the cumulative radiant exposure required for induction of phototoxic injury was independent of irradiance. With the 24-ps-pulsed source, a significantly lower cumulative radiant exposure was required to induce the phototoxicity when the peak irradiance was 5.8 x 10(7) or 1.3 x 10(8) watts cm-2 compared with when peak irradiance was either lower (6.0 x 10(6) watts cm-2) or higher (7.6 x 10(8) watts cm-2). The measured fluorescence lifetimes of doxycycline in buffered saline solution were longer than the laser pulse duration of 24 ps. The increased efficiency of photosensitization at the optimal peak irradiance in the ps domain appears to result from sequential multiphoton absorption involving higher excited states of the singlet manifold. At the highest irradiance studied, on the other hand, reduced efficiency of photosensitization is attributed to increased photodegradation of doxycycline from higher excited states by processes such as photoionization. A model consistent with these observations is presented along with calculations, based on simple rate equations, that fit the essentials of the proposed model.