Spectral monitoring of biologically active solar UVB radiation using an in vitro model of vitamin D synthesis

Vitamin D synthetic capacity of sunlight is the most well known beneficial effect of solar UV radiation. The article describes the new method of biological dosimetry of solar (and artificial) UV radiation based on an in vitro model of vitamin D synthesis (D-dosimeter). Particular attention is given to spectrophotometric analysis of the multicomponent mixture of vitamin D photoisomers that has been developed for the most effective use of D-dosimeter in situ.     

In vitro model of vitamin D synthesis by UV radiation in an Australian urban environment

Vitamin D, an important constituent of human health, is produced through the exposure of human skin to short wave (280-315 nm) ultraviolet radiation (UV). We aimed to establish whether an urbanized environment with tall buildings in close proximity (an "urban canyon") significantly reduced the capacity of sunlight to synthesize vitamin D, when compared with a typical suburban area (～2.5 km away); and to investigate the association of UV and vitamin D production with pollution, temperature, and humidity. Measurements of ambient UV (295-400 nm) (using a portable photometer/radiometer and detector) and synthesized vitamin D (from an in vitro model) were taken regularly at urban and control sites over 3 months in Brisbane, Australia. During a typical 20 min measurement, urban and control sites received 0.26 and 1.03 W m(-2) mean total UV respectively (P < 0.001), and produced 0.12 and 0.53 μg mL(-1) mean vitamin D (P < 0.001). Pollution, temperature and humidity were not associated with UV or vitamin D production. This demonstrates a large difference in vitamin D synthesis between an urban canyon and a nearby control site. Although the results cannot be directly applied to humans, they emphasize the need for further study of human vitamin D production in urban environments.     

Phage T7 in biological UV dose measurement.

An experimental method complete with theoretical considerations is presented for the measurement of different biological UV doses. The method is based on the high sensitivity of phage T7 activity to UV light. A precisely determined T7 inactivation action spectrum is presented over a wide optical range (240-514 nm). Using the T7 spectral sensitivity in relation to the minimal erythema dose (MED) and the effective spectral irradiance from solar radiation for the MED, an example is given to determine the MED value based on the measurement of T7 inactivation for a given case. The advantages and applicability of the method are discussed.     

Calculated ultraviolet exposure levels for a healthy vitamin D status

The dangers of overexposure to sunlight have been well publicized, but less attention has been given to an acknowledged benefit of exposure to UV radiation; that being the cutaneous synthesis of vitamin D3. Here we define a standard vitamin D dose on the basis of recently recommended requirements for vitamin D that take account of its risk reduction role in a variety of diseases, and present a web-based tool that enables the reader to calculate associated exposure times for any time and place using either default values or user-selected conditions. Either it is not possible to synthesize vitamin D3 at high latitudes in winter, or the exposure time required to reach a standard dose is sometimes impractical. Where solar UV is sufficient, a risk-benefit analysis of sunburn vs. vitamin D3 synthesis shows that the best time for brief sun exposure is in the middle of the day. For low solar elevation angles common at high latitudes, a fine line exists between adequate UV exposure for vitamin D3 synthesis and a risk of sun burn.     

Determining an Effective UV Radiation Exposure Time for Vitamin D Synthesis in the Skin Without Risk to Health: Simplified Estimations from UV Observations

UV radiation contains erythemally weighted UV, as well as UV that synthesizes vitamin D₃. Here, we attempted to determine the relationship between these factors by numerical simulation of atmospheric parameters, such as total ozone, using a simplified “SMART2” model for radiative transfer. Both forms of UV were almost linearly correlated with each other for a comparably large UV radiation exposure, larger than UV Index ~1.6. If erythemally weighted UV, which carries a risk of sunburn, is known, the amount of UV exposure needed for vitamin D synthesis in the epidermis can be estimated using this relationship. The production of 10 μg (400 IU) of vitamin D per day takes approximately 1/3 of the time needed to reach the minimal erythemal dose (MED) for an effective skin area of 600 cm² for skin phototype III. For an area of 1200 cm², 1/6 of that exposure time suffices. From a UV Index that is commonly used, the risks and benefits can be evaluated using this linear relationship, which will enable people to effectively manage their UV exposure and consider the risks and benefits to optimize health outcomes.     

The role of the spectral sensitivity curve in the selection of relevant biological dosimeters for solar UV monitoring

To estimate the risk of enhanced UV-B radiation due to stratospheric ozone depletion, phage T7 and uracil thin-layer biological dosimeters have been developed, which weight the UV irradiance according to induced DNA damage. To study the molecular basis of the biological effects observed after UV irradiation, the spectral sensitivity curves of the two dosimeters and induction of the two major DNA photoproducts, cyclobutane pyrimidine dimers (CPDs) and 6-4 photoproducts ((6-4)PDs), in phage T7 have been determined for polychromatic UV sources. CPDs and (6-4)PDs are determined by lesion-specific monoclonal antibodies in an immunodotblot assay. Phage T7 and uracil biological dosimeters together with a Robertson-Berger (RB) meter have been used for monitoring environmental radiation from the polar region to the equator. The biologically effective dose (BED) established with the three different dosimeters increases according to the changes in the solar angle and ozone column, but the degree of the change differs significantly. The results can be explained based on the different spectral sensitivities of the dosimeters. A possible method for determining the trend of the increase in the biological risk due to ozone depletion is suggested.     

In vitro model of vitamin D3 (cholecalciferol) synthesis by UV radiation: dose-response relationships

Vitamin D deficiency is a major health concern worldwide. Very little is understood regarding its production in the human body by exposure to UV radiation. In particular, we have no means of predicting how much vitamin D (cholecalciferol) will be produced in the skin after exposure to sunlight. Using a refined in vitro model, we found that there is a nonlinear relationship between UV dose and cholecalciferol synthesis. Two minimal erythemal doses (MED) of UV radiation produced 1.84 microg/mL of cholecalciferol whereas 4 MED produced 2.81 microg/mL. We also found that the production of cholecalciferol is restricted by the initial concentration of its precursor (7-dehydrocholesterol, 7-DHC). For example, using an initial concentration of 7-DHC of 102 microg/mL, the resultant cholecalciferol production was 1.05 microg/mL after receiving 4 MED exposure. Under the same exposure conditions, an initial concentration of 305 microg/mL yielded 2.81 g/mL of cholecalciferol. The data presented in this paper has important implications for humans, including: (1) increasing UV exposure does not result in a proportionate increase in the amount of cholecalciferol that is produced; and (2) the initial concentration of 7-DHC in the skin may impact the amount of cholecalciferol that can be synthesized. When translating these results to population groups, we will discuss how the sun exposure message needs to be carefully formulated to account for such considerations.     

Daily duration of vitamin D synthesis in human skin with relation to latitude, total ozone, altitude, ground cover, aerosols and cloud thickness

Vitamin D production in human skin occurs only when incident UV radiation exceeds a certain threshold. From simulations of UV irradiances worldwide and throughout the year, we have studied the dependency of the extent and duration of cutaneous vitamin D production in terms of latitude, time, total ozone, clouds, aerosols, surface reflectivity and altitude. For clear atmospheric conditions, no cutaneous vitamin D production occurs at 51 degrees latitude and higher during some periods of the year. At 70 degrees latitude, vitamin D synthesis can be absent for 5 months. Clouds, aerosols and thick ozone events reduce the duration of vitamin D synthesis considerably, and can suppress vitamin D synthesis completely even at the equator. A web page allowing the computation of the duration of cutaneous vitamin D production worldwide throughout the year, for various atmospheric and surface conditions, is available on the Internet at http://zardoz.nilu.no/~olaeng/fastrt/VitD.html and http://zardoz.nilu.no/~olaeng/fastrt/VitD-ez.html. The computational methodology is outlined here.     

UV index forecasts and measurements of health-effective radiation

While erythemal irradiance as a potentially damaging effect to the skin has been extensively studied and short-term forecasts have been issued to the public to reduce detrimental immediate and long-term effects such as sunburn and skin cancer by overexposure, beneficial effects to human health such as vitamin D(3) production by UV radiation and melatonin suppression by blue visible light have attained more and more attention, though both of them have not become part of forecasting yet. Using 4years of solar radiation data measured at the mid-latitude site Lindenberg (52°N), and forecast daily maximum UV index values, an overall good correspondence has been found. The data base of solar UV radiation and illuminance has also been used to analyze effects of clouds and aerosols on the effective irradiance. Optically thick clouds can strongly modify the ratios between erythemal and vitamin D(3) effective irradiance such that direct radiative transfer modeling of the latter in future UV forecasts should be preferably used. If parameterizations of vitamin D(3) effective irradiance from erythemal irradiance are used instead, the optical cloud depth would have to be taken into account to avoid an overestimation of vitamin D(3) with parameterizations neglecting cloud optical depth. Particular emphasis for the beneficial effects has been laid in our study on low exposure. Daily doses of solar irradiation for both vitamin D(3) and melatonin suppression do not reach minimum threshold doses even with clear sky and unobstructed horizon during the winter months.     

A mathematical model for seasonal variability of vitamin D due to solar radiation

It is widely recognized that vitamin D deficiency has detrimental health consequences. The ultraviolet (UV) B radiation increases the serum vitamin D level, expressed by 25-hydroxyvitamin-D(3) [25(OH)D]. An analytical model is presented to calculate the serum 25(OH)D changes throughout a year, caused by the solar exposure variability due to geophysical and habitual factors. The model is tuned by taking into account recent experimental results of serum 25(OH)D changes, after a series of artificial (by fluorescent tubes) UV exposures. The model uses the erythemal and vitamin D weighted irradiances, inferred from the Brewer spectrophotometer and the Kipp and Zonen broad-band meter measurements, carried out in Belsk (52°N, 21°E), Poland, in 2010. The modeled seasonal pattern of the serum 25(OH)D concentration in Polish indoor workers is only slightly different, than in subjects with typical outdoor activity habits, and in those with sun-seeking behavior. A deep minimum in the serum 25(OH)D concentration appears in late winter, regardless of outdoor activity habits. An extra sunbathing to boost the vitamin D level is not worth taking, because of a minor improvement of the vitamin D status, and because of a greater erythema risk. It would be much safer and more effective to maintain an adequate vitamin D level through diet supplements, even in summer, for non sun-seeking subjects.     

How does the skin sense sun light? An integrative view of light sensing molecules

The consensus on the effects of excessive sun exposure on human health has long emphasized the negative effects of solar UV radiation. Nevertheless, although UV radiation has been demonized, less is known about the consequences of sun exposure while using sunscreen, which can lead to high visible light exposure. UV and visible light play key roles in vitamin D synthesis, reduction of blood pressure, among other beneficial effects. In this review, we aim to provide a comprehensive view of the wide range of responses of the human skin to sunlight by revisiting data on the beneficial and harmful effects of UV and visible light. We start by exploring the interaction of photons in the skin at several levels including physical (depth of photon penetration), chemical (light absorption and subsequent photochemical events), and biological (how cells and tissues respond). Skin responses to sun exposure can only be comprehensively understood through a consideration of the light-absorbing molecules present in the skin, especially the light-sensing proteins called opsins. Indeed, many of the cellular responses to sun exposure are modulated by opsins, which act as the “eyes of the skin”.     

Analysis of vitamin D and its metabolites using thermospray liquid chromatography/mass spectrometry.

A new method is described for the analysis of vitamin D and its metabolites utilizing thermospray (TSP) mass spectrometry as an on-line detector for high performance liquid chromatography. Ionization conditions were optimized for use with isocratic reversed phase chromatography. TSP mass spectrometry was employed in series with a UV absorbance detector to facilitate comparisons between the two methods of detection. Positive ion TSP mass spectra were recorded for vitamin D2, vitamin D3, 25-hydroxyvitamin D3 (25(OH)D3), 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and 24,25-dihydroxyvitamin D3 (24,25(OH)2D3). The spectra contained protonated molecular ions, ammonium adduct ions and fragment ions due to the loss of one or more molecules of water. A comparison of quantitative precision was made by determining UV absorbance and TSP standard curves for vitamin D3 using two different methods: (1) External standard method with post-column (post UV detector) addition of ammonium acetate. (2) As (1) but using the method of internal standards with a closely eluting internal standard (vitamin D2). In each case the quantitative precision (correlation coefficient) for UV absorbance detection was superior owing to intrinsic instability of the TSP ion beam. A stable isotopically labelled internal standard was employed in the development of an assay for 1,25(OH)2D3. The assay was used to quantify in vitro enzymic conversion of 25(OH)D3 to 1,25(OH)2D3 in guinea pig and sheep renal mitochondrial incubations. TSP LC/MS was also applied to analysis of an extract of human blood plasma in which D3 and each of its principal metabolites were identified in a single analysis.     

ULTRAVIOLET DOSIMETRY IN OUTDOOR MEASUREMENTS BASED ON BACTERIOPHAGE T7 AS A BIOSENSOR

A simple method has been worked out for measuring the biologically effective dose (BED) of solar radiation The method uses phage T7 as a biosensor and it includes field measurements of global and direct UV radiation from the sun in the air; it has been applied to underwater measurements as well. Results of field measurements are presented with discussion of the angle-dependent sensitivity of the biosensor. A model of spectral irradiance based on the measured values is presented. Relevance of the H^T7 unit—derived earlier by us from T7 phage inactivation upon UV radiation—as a measure of the BED is also discussed.     

PHOTOAGING EFFECTS ON SPECTRAL TRANSMITTANCE OF PLASTIC FILTERS

We examined the effects of ultraviolet (UV) radiation in combination with high levels of infrared (IR) radiation on the spectral transmittance of plastic filters. The biological action spectrum for damage to the human eye and skin changes dramatically in the 300-400 nm wavelength range. Cut-off filters used in this region to shape the spectrum of exposure sources are thus critical to the design of experiments which use broadband light sources. The changes in transmittance of three types of plastic filters were observed over an exposure period of 1000 h. One set of three filters was exposed mainly to UV radiation, while the other set was exposed to UV radiation plus IR radiation. Filters exposed to both UV and IR radiation showed spectral changes in their transmittance, while the filters exposed to UV only showed no measurable changes.     

Location and vitamin D synthesis: is the hypothesis validated by geophysical data?

The literature reports strong correlations between UV exposure and latitude gradients of diseases. Evidence is emerging about the protective effects of UV exposure for cancer (breast, colo-rectal, prostate), autoimmune diseases (multiple sclerosis, type II diabetes) and even mental disorders, such as schizophrenia. For the first time, the available levels of vitamin D producing UV or "vitamin D UV" (determined from the previtamin D action spectrum) and erythemal (sunburning) UV from throughout the USA are measured and compared, using measurements from seven locations in the USA are measured and compared, using measurements from seven locations in the US EPA's high accuracy Brewer Spectrophotometer network. The data contest longstanding beliefs on the location-dependence and latitude gradients of vitamin D UV. During eight months of the year centered around summer (March-October), for all sites (from 18 degrees N to 44 degrees N latitude) the level of vitamin D UV relative to erythemal UV was equal (within the 95% confidence interval of the mean level). Therefore, there was no measured latitude gradient of vitamin D UV during the majority of the year across the USA. During the four cooler months (November-February), latitude strongly determines vitamin D UV. As latitude increases, the amount of vitamin D UV decreases dramatically, which may inhibit vitamin D synthesis in humans. Therefore, a larger dose of UV relative to erythemal UV is required to produce the same amount of vitamin D in a high latitude location. However, the data shows that at lower latitude locations (<25 degrees N), wintertime vitamin D UV levels are equal to summertime levels, and the message of increasing UV exposure during winter is irrelevant and may lead to excessive exposure. All results were confirmed by computer modeling, which was also used to generalize the conclusions for latitudes from 0 degrees to 70 degrees N. The results of this paper will impact on research into latitudinal gradients of diseases. In particular, it may no longer be correct to assume vitamin D levels in populations follow significant latitude gradients for a large proportion of the year.     

UV radiation: balancing risks and benefits

We use action spectra published by the International Commission on Illumination to examine diurnal, seasonal and latitudinal variations in erythemally weighted (sunburning) UV-a health risk, and vitamin D-weighted UV-a health benefit. Vitamin D-weighted UV is more strongly dependent on ozone and solar zenith angle. Consequently, its diurnal, seasonal and geographic variability is more pronounced than for erythemally weighted UV. We then investigate relationships between the two quantities. An algorithm is developed and used to relate vitamin D production to the widely used UV index, to help the public to optimize their exposure to UV radiation. In the summer at noon, there should at mid-latitudes be sufficient UV to photosynthesize optimal vitamin D in approximately 1 min for full body exposure, whereas skin damage occurs after approximately 15 min. Further, while it should be possible to photosynthesize vitamin D in the winter at mid-latitudes, the amount of skin that must be exposed is larger than from the hands and face alone. This raises the question of whether the action spectrum for vitamin D production is correct, since studies have reported that production of vitamin D is not possible in the winter at mid-latitudes.     

BACTERIAL PHOTOPROTECTION THROUGH EXTRACELLULAR CADMIUM SULFIDE CRYSTALLITES

Ultraviolet light and the heavy metal, cadmium, both have toxic effects on many microorganisms. In this communication we describe a method by which the bacterium Klebsiella aerogenes surmounts both problems using one biological process: the synthesis of cadmium sulfide (CdS) crystallites. These semiconductor particles absorb radiation in the UV spectral region and therefore, when K. aerogenes produces extracellular CdS material in response to environments containing cadmium ions, a photoprotective layer is formed. The effect of UVA radiation on cultures of Klebsiella aerogenes was monitored using electron microscopy, energy-dispersive X-ray analysis and electronic spectroscopy. The results show that at wavelengths 320 nm Λ 400 nm, a photoprotection period of between 4 and 6 h is induced, which eventually fails due to photodegradation of the semiconductor layer to metallic cadmium and elemental sulfur.     

UV doses worldwide

UV radiation affects human health. Human exposure to UV radiation causes a few beneficial health effects like vitamin D3 formation but it causes many detrimental health effects: sunburn, ocular damage, photoaging, immune suppression, DNA damage and skin cancer. In countries with fair-skinned populations, skin cancer is the most diagnosed of all cancers. In the United States in 2002, there were over one million new skin cancer cases. That means one out of every 285 people got skin cancer. Skin cancer of fair-skinned individuals is increasing at an alarming rate (4-6% per year) around the world and has now reached so-called "pandemic" proportions. Thus, it is important to know what UV doses people around the world get throughout their lives. This review covers how the outdoor UV doses are weighted for different biological effects, the most commonly used measuring devices for terrestrial and personal UV doses, the natural and other effects on terrestrial and personal UV doses, the time people spend outside, their ambient exposures and the terrestrial and personal UV doses of adult outdoor and indoor workers as well as children and adolescents around the world. Overall, outdoor-working adults get about 10%, while indoor-working adults and children get about 3% (2-4%) of the total available annual UV (on a horizontal plane). People's UV doses increase with increasing altitude and decreasing latitude; most indoor-working adult Europeans get 10,000-20,000 J/m2 per year, Americans get 20,000-30,000 J/m2 per year and Australians are estimated to get 20,000-50,000 J/m2 per year (excluding vacation, which can increase the dose by 30% or more).     

Standard ultraviolet daylight for nonextreme exposure conditions

The skin is exposed to ultraviolet radiation (UVR) from natural or artificial sources on a daily basis. The effects of chronic low dose exposure merit investigation, even when these effects are neither conspicuous nor clinically assessable. The purpose of the present study was to define a relative spectral UV irradiance that is representative of frequent nonextreme sun exposure conditions and therefore more appropriate for studies of the long-term and daily effects of solar UV on the skin. Solar spectral UV irradiance values were calculated for different dates and locations by using a radiative transfer model. The spectral irradiance values obtained when the solar elevation is lower than 45 degrees were averaged. An important feature is the dUVA (320-400 nm) to dUVB (290-320 nm) irradiance values ratio, which was found to be 27.3 for the overall average. When the months corresponding to extreme irradiance values (low or high) were excluded from the calculations, the dUVA to dUVB ratio ranged from 27.2 to 27.5. The mean spectral irradiance of the model presented here represents environmental UV exposure conditions and can be used both as a standard to investigate the biological effects of a nonextreme UVR and to assess the effectiveness of products for daily skin protection.     

The vitamin D debate: translating controlled experiments into reality for human sun exposure times

Exposure to sunlight, specifically the ultraviolet radiation, has both positive and negative health effects. Maximizing the benefits (vitamin D synthesis) while minimizing the damage is a multifaceted problem in which many of the elements are poorly quantified. Here we show how rigorously conducted large sample size laboratory studies of the effect of ultraviolet radiation dose on vitamin D status can be applied to real-life situations. This was achieved by modeling the radiation incident on different surfaces for different solar locations, and equating with the controlled exposures in the laboratory studies. Results from both model and experimental data show that relatively short exposures of a modest amount of unprotected skin to summer sunlight in northern climes, on a regular basis during lunchtime hours, increases vitamin D to sufficiency status (≥20 ng mL(-1) ) in the white Caucasian population. While both sun exposure conditions and human skin responses are variable in real life, these quantitative findings provide a guide for authorities devising sunlight exposure recommendations.