Low vitamin D in dark-skinned immigrants is mainly due to clothing habits and low UVR exposure: a Danish observational study.

Low 25-hydroxyvitamin D3 (25(OH)D) among dark-pigmented persons has been observed. To elucidate the reason for this we examined sun behaviour, sun-exposed body area, solar UVR exposure and 25(OH)D levels in immigrants with dark pigmented skin and Danes with light pigmented skin. Clothing, sun behaviour, and diet were recorded daily during a Danish summer season (93 analysed days). Erythema-weighted UVR doses were measured by personal electronic UVR dosimeters (with erythema response, measurement every 5th second) and 25(OH)D was measured in 72 participants (33 dark-skinned and 39 light-skinned). The immigrants exposed 28% less skin area, received 70% less UVR dose, and had 71% less 25(OH)D increase during the summer. The UVR reactivity (Δ25(OH)D per joule accumulated UVR dose) was similar (P = 0.62) among the immigrants (0.53 nmol l-1 J-1) and the Danes (0.63 nmol l-1 J-1). In the groups combined, 25(OH)D levels after summer were mainly influenced by UVR dose to exposed skin (28.8%) and 25(OH)D start level (27.9%). Height and measured constitutive skin pigmentation were of minor influence: 3.5% and 3.2%, respectively. Sun exposure and clothing habits were the main reasons for lower 25(OH)D level after summer in the darker immigrants, as both groups had similar UVR reactivity.

Serum 25(OH)D levels after oral vitamin D3 supplementation and UVB exposure correlate.

BACKGROUND: The inter-individual variation in 25(OH)D3 increase (Δ25(OH)D3 ) after vitamin D3 supplementation was determined and compared with the UVB irradiation response. METHODS: Nineteen Danish participants received 85 µg vitamin D3 (cholecalciferol) daily for nine weeks with regular serum 25(OH)D3 measurements. These participants had three years earlier taken part in a 9-week controlled UVB study. The Δ25(OH)D3 was not confounded by ambient UVB, BMI or ethnicity. RESULTS: Δ25(OH)D3 was 53 nmol L-1 and almost identical to Δ25(OH)D3 (52 nmol L-1 ) after UVB. Δ25(OH)D3 ranged from 17 to 91 nmol L-1 (span 74 nmol L-1 ) and was about half of that observed after UVB irradiation (span 136 nmol L-1 ). The interquartile ranges for vitamin D3 supplementation (38.8-71.4 nmol L-1 , span: 32.6 nmol L-1 ) and UVB irradiation (35.7-65.4 nmol L-1 , span: 29.7 nmol L-1 ) were similar indicating a comparable response of the two interventions. As the 25(OH)D3 start levels (R2  = 0.398, P = 3.8 × 10-3 ), 25(OH)D3 end levels (R2  = 0.457, P = 1.5 × 10-3 ) and Δ25(OH)D3 (R2  = 0.253, P = 0.028) between both interventions were correlated, this suggested a possible common individual background for the variation. Four pigment SNPs influenced the variation in the vitamin D3 -induced and UVB-induced Δ25(OH)D3 . A combined model including the influence of these four SNPs and the 25(OH)D3 start level explained 86.8% (P = 1.6 × 10-35 ) of the individual variation after vitamin D3 supplementation. CONCLUSION: The inter-individual variation in the two interventions was comparable and had no common demographic but a partly common genetic background.

Pigment genes not skin pigmentation affect UVB-induced vitamin D.

Skin pigmentation is believed to contribute to the generally low serum 25-hydroxyvitamin D (25(OH)D) concentrations observed in darker-skinned persons. The influence of measured skin pigmentation on UVB-induced 25(OH)D increase was investigated together with 9 demographic and 13 genetic parameters (pigment SNPs). Forty participants representing a wide range in measured skin pigmentation were exposed to identical UVB doses on identical body areas over nine weeks with weekly measurements of serum 25(OH)D. This study took place in Denmark during winter, a period with negligible ambient UVB, so variation in 25(OH)D synthesis was not influenced by latitude, season, sun and clothing habits. The increase in 25(OH)D concentration displayed considerable variation (range: 2.9 to 139 nmol L-1). Constitutive and facultative skin pigmentation exerted separate influence on the variation of the UVB-induced linear 25(OH)D increase. However, this influence was statistically non-significant in the presence of separate significant pigment SNPs. The variation in the 25(OH)D increase in the combined linear model was not explained by measured skin pigmentation but by sex, height, age and seven SNPs located in the ASIP, MTAP, MIR196A29 and Solute Carrier Family genes. This linear model including individual intercepts and the 10 parameters influencing the slope explained 77.4% of the variation. This study confirmed the influence of sex, age and height on 25(OH)D increase and found that pigment genes provided a better relation to UVB-induced 25(OH)D increase compared to the actual measured skin pigmentation. Therefore, only investigating skin pigmentation obscures other causal parameters for low 25(OH)D.

The half-life of 25(OH)D after UVB exposure depends on gender and vitamin D receptor polymorphism but mainly on the start level.

The 25-hydroxy vitamin D (25(OH)D) production caused by UVB exposure is usually underestimated as the concurrent degradation of 25(OH)D is not considered. Therefore, the decrease in 25(OH)D was investigated during a 7-week period in winter when ambient UVB is negligible. Twenty-two healthy Danish individuals (113 samples) participated and had a mean and steady maximal 25(OH)D start level of 132 nmol l-1 (range of 68-216 nmol l-1) due to long-term UVB treatment prior to this study. In this group with high 25(OH)D start levels, the decrease in 25(OH)D was best described by an exponential model. This suggests a quantitatively larger elimination of 25(OH)D at high 25(OH)D start levels. A linear model (logarithm of 25(OH)D) including personal start levels as intercepts and a slope influenced by gender and the vitamin D receptor gene polymorphism rs2228570 explained 87.8% of the observed variation. The mean half-life was 89 days with a difference in half-life of 120 days between a male with rs2228570 genotype GG (59 days) and a female with rs2228570 genotype AA/AG (179 days). Thus, these two parameters explained a large part of the observed inter-individual variation of 25(OH)D. Furthermore, the decrease was analysed in two groups with medium and low 25(OH)D start levels resulting in longer half-lives of 149 days and 199 days, respectively. The longer half-lives at lower 25(OH)D levels may be caused by storage mobilisation, changed catabolism or increased intestinal absorption.

Major inter-personal variation in the increase and maximal level of 25-hydroxy vitamin D induced by UVB.

Vitamin D influences skeletal health as well as other aspects of human health. Even when the most obvious sources of variation such as solar UVB exposure, latitude, season, clothing habits, skin pigmentation and ethnicity are selected for, variation in the serum 25-hydroxy vitamin D (25(OH)D) response to UVB remains extensive and unexplained. Our study assessed the inter-personal variation in 25(OH)D response to UVR and the maximal obtainable 25(OH)D level in 22 healthy participants (220 samples) with similar skin pigmentation during winter with negligible ambient UVB. Participants received identical UVB doses on identical body areas until a maximal level of 25(OH)D was reached. Major inter-personal variation in both the maximal obtainable UVB-induced 25(OH)D level (range 85-216 nmol l(-1), mean 134 nmol l(-1)) and the total increase in 25(OH)D (range 3-139 nmol l(-1), mean 48 nmol l(-1)) was found. A linear model including measured 25(OH)D baselines as personal intercepts explained 54.9% of the variation. By further including personal slopes in the model, as much as 90.8% of the variation could be explained. The explained variation constituted by personal differences in slopes thus represented 35.9%. Age, vitamin D receptor gene polymorphisms, height and constitutive skin pigmentation (a skin area not exposed to UVB) explained 15.1% of this variation. Despite elimination of most known external sources of variation, our study demonstrated inter-personal variation corresponding to an observed maximal difference of 136 nmol l(-1) in the total increase of 25(OH)D and 131 nmol l(-1) in the maximal level of 25(OH)D.

Black light visualized solar lentigines on the shoulders and upper back are associated with objectively measured UVR exposure and cutaneous malignant melanoma.

Previous studies on the association of solar lentigines with ultraviolet radiation (UVR) exposure have been based on retrospective questionnaires about UVR exposure. We aimed to investigate the association between solar lentigines and UVR exposure in healthy individuals using objective measurements, and to investigate the association between solar lentigines and cutaneous malignant melanoma (CMM). Forty-eight patients with CMM and 48 controls that matched the patients individually by age, sex, constitutive skin type and occupation participated. Solar lentigines on the shoulders and upper back were counted and graded into 3 categories using black light photographs to show sun damage. Current UVR exposure in healthy controls was assessed by personal electronic UVR dosimeters that measured time-related UVR and by corresponding exposure diaries during a summer season. Sunburn history was assessed by interviews. Among controls, the number of solar lentigines was positively associated with daily hours spent outdoors between noon and 3 pm on holidays (P = 0.027), days at the beach (P = 0.048) and reported number of life sunburns (P < 0.001). Compared with matched controls CMM patients had a higher number of solar lentigines (P = 0.044). There was a positive association between CMM and higher solar lentigines grade; Category III versus Category I (P = 0.002) and Category II versus Category I (P = 0.014). Our findings indicate that solar lentigines in healthy individuals are associated with number of life sunburns, as well as time spent outdoors around noon on holidays and beach trips during a summer season, most likely reflecting past UVR exposure, and that solar lentigines are a risk factor for CMM.

A 3-year follow-up of sun behavior in patients with cutaneous malignant melanoma.

IMPORTANCE UV radiation (UVR) exposure is the primary environmental risk factor for developing cutaneous malignant melanoma (CMM). OBJECTIVE To measure changes in sun behavior from the first until the third summer after the diagnosis of CMM using matched controls as a reference. DESIGN, SETTING, AND PARTICIPANTS Three-year follow-up, observational, case-control study performed from May 7 to September 22, 2009, April 17 to September 15, 2010, and May 6 to July 31, 2011, at a university hospital in Denmark of 21 patients with CMM and 21 controls matched to patients by sex, age, occupation, and constitutive skin type participated in the study. Exposure to UVR was assessed the first and second summers (n=20) and the first and third summers (n=22) after diagnosis. Data from 40 participants were analyzed. MAIN OUTCOMES AND MEASURES Exposure to UVR was assessed by personal electronic UVR dosimeters that measured time-related UVR in standard erythema dose (SED) and corresponding sun diaries (mean, 74 days per participant each participation year). RESULTS Patients' daily UVR dose and UVR dose in connection with various behaviors increased during follow-up (quantified as an increase in daily UVR dose each year; all days: mean, 0.3 SED; 95% CI, 0.05-0.5 SED; days with body exposure: mean, 0.6 SED; 95% CI, 0.07-1.2 SED; holidays: mean, 1.2 SED; 95% CI, 0.3-2.1 SED; days abroad: 1.9 SED; 95% CI, 0.4-3.4 SED; and holidays with body exposure: mean, 2.3 SED; 95% CI, 1.1-3.4 SED). After the second year of follow-up, patients' UVR dose was higher than that of controls, who maintained a stable UVR dose. No difference was found between groups in the number of days with body exposure or the number of days using sunscreen in the second and third years of follow-up. CONCLUSIONS AND RELEVANCE Our findings suggest that patients with CMM do not maintain a cautious sun behavior in connection with an increase in UVR exposure, especially on days with body exposure, when abroad, and on holidays.

Sunscreen use and failures--on site observations on a sun-holiday.

With this observation study we aimed to determine how and when sunscreen was used. 20 sun seekers were observed during a one-week sun holiday in Hurghada, Egypt. The sunscreen application thickness was related to part of body, time outdoors, exposure to ultraviolet radiation and to sunburning. Skin sites with sunscreen were exposed to UVR significantly longer and received significantly higher UVR doses than skin sites without sunscreen. They received an average of 0.62 SED [0.0-9.3 SED] (13% of their MED) before the first sunscreen application of the day. The average sunscreen used was SPF15 and the sunscreen application thickness was in average 0.79 mg cm(-)2 giving an approximated effect of SPF3. For different body parts either the total UVR exposure dose or the UVR exposure time and UVR exposure dose before the first sunscreen application were higher for sunburned than non-sunburned skin sites. In the final model gender, skin type and UVR to skin (adjusted for SPF and sunscreen application thickness) were significant predictors of sunburning. The sunscreen application thickness of 0.79 mg cm(-)2 was less than the 2 mg cm(-2) used for testing SPF. The late start of sunscreen use and improper application thickness was ineffective in preventing sunburn, and therefore could not compensate for the risk of prolonged UVR exposure and high UVR doses. Our results lead us to suspect that the protective effect of sunscreen use against DNA-damage, and thereby skin cancer, is minimal the way sunscreen is used under real sun holiday conditions.

Increase in serum 25-hydroxyvitamin-D3 in humans after solar exposure under natural conditions compared to artificial UVB exposure of hands and face.

Vitamin D studies are often performed under controlled laboratory conditions and the findings may be difficult to translate to natural conditions. We aimed to determine and compare the doses of natural solar ultraviolet radiation (UVR) with doses of artificial UVB radiation of hands and face needed to increase serum 25-hydroxyvitamin-D(3) (25(OH)D). Furthermore, we aimed to investigate the natural course of 25(OH)D due to solar exposure from April to September. 46 Caucasian volunteers were included. 17 volunteers received solar UVR (Group 1) in their natural Danish environment. Individual daily solar UVR doses in standard erythema doses (SEDs) were determined with personal wristwatch UV-dosimeters. 29 volunteers (Group 2) received artificial UVB doses of 6 SEDs (N = 14) and 3 SEDs (N = 15) on hands and face during late-winter/early-spring when outdoor UVB is negligible. 25(OH)D-levels were determined around every second week during study periods. Solar-UVR doses and sun-exposure diaries with information of sun-exposed areas were available from 8 volunteers and used for comparison with artificial UVB doses. However no significant solar-induced Δ25(OH)D was observed when sun-exposed areas were limited to hands and face. Instead the earliest period (week 17-19) with significant Δ25(OH)D, occurring after a mean of 2 days of sun-exposing more than hands and face, was used to estimate an approximate UVR dose required to increase 25(OH)D. This estimate resulted in a dose of 4.1 solar SEDs required to increase 25(OH)D by 1 nmol l(-1). The artificial dose of 6 SEDs of only hands and face significantly increased 25(OH)D and resulted in a dose of 0.52 SEDs required to increase 25(OH)D significantly by 1 nmol l(-1). Artificial UVB was thus at least 8 times more efficient in increasing 25(OH)D than solar UVR at a UV-exposed area consisting of approximately hands and face. Solar UVR exposure of larger areas may lead to enhanced efficacy but was not relevant for this comparison. Significant solar-induced Δ25(OH)D was present earliest at April 8, maximal by early August and decreased by late August.

Association analysis of the LAG3 and CD4 genes in multiple sclerosis in two independent populations.

We have investigated the genetic involvement of the CD4 and the LAG3 genes, two appealing candidates for MS due to their suggested role in MS pathology. We genotyped a Swedish case-control material consisting of 920 MS patients and 778 controls in an initial study of CD4, three SNPs showed a significant association with MS. An independent material consisting of 1720 Nordic MS patients and 1416 controls were used for confirmation of associated markers in CD4 and to do a confirmative study of the LAG3 gene from previous findings. The result, including a total of 2640 MS patients and 2194 controls shows no significant association with CD4 and LAG3 and MS. We conclude that these genes are of minor importance in regard of genetic predisposition to the MS.

A high-density screen for linkage in multiple sclerosis.

To provide a definitive linkage map for multiple sclerosis, we have genotyped the Illumina BeadArray linkage mapping panel (version 4) in a data set of 730 multiplex families of Northern European descent. After the application of stringent quality thresholds, data from 4,506 markers in 2,692 individuals were included in the analysis. Multipoint nonparametric linkage analysis revealed highly significant linkage in the major histocompatibility complex (MHC) on chromosome 6p21 (maximum LOD score [MLS] 11.66) and suggestive linkage on chromosomes 17q23 (MLS 2.45) and 5q33 (MLS 2.18). This set of markers achieved a mean information extraction of 79.3% across the genome, with a Mendelian inconsistency rate of only 0.002%. Stratification based on carriage of the multiple sclerosis-associated DRB1*1501 allele failed to identify any other region of linkage with genomewide significance. However, ordered-subset analysis suggested that there may be an additional locus on chromosome 19p13 that acts independent of the main MHC locus. These data illustrate the substantial increase in power that can be achieved with use of the latest tools emerging from the Human Genome Project and indicate that future attempts to systematically identify susceptibility genes for multiple sclerosis will have to involve large sample sizes and an association-based methodology.

Significantly increased fractions of transformed to total alpha2-macroglobulin concentrations in plasma from patients with multiple sclerosis.

We examined the proteinase inhibitor alpha2-macroglobulin (alpha2M) in plasma from patients with multiple sclerosis (MS); a neurological disease of the central nervous system. The plasma concentrations of native and transformed alpha2M were measured in 90 patients with clinically definite MS, 73 with relapsing-remitting and 17 with secondary progressive MS, and 132 healthy individuals. Significantly lower concentrations of native alpha2M and significantly higher concentrations of transformed alpha2M were found in MS patients. A significant correlation between the concentrations of native and transformed alpha2M was found. The fraction of transformed to total alpha2M in the MS patients was 36% higher than in the healthy individuals. The results suggest an important involvement of alpha2M in regulation of increased proteolytic activity occurring in MS disease.

Concordance for disease course and age of onset in Scandinavian multiple sclerosis coaffected sib pairs.

BACKGROUND: Investigation of coaffected sib pairs is one method to determine the genetic influence on the clinical presentation of many complex diseases, such as multiple sclerosis (MS). Investigation of the clinical concordance in coaffected sib pairs may be a prerequisite to identify genes that modify the clinical outcome. The aim of this study was to investigate a possible genetic influence on selected demographic and clinical variables among familial Scandinavian MS cases. MATERIAL AND METHODS: We identified 136 Caucasian Scandinavian families with MS coaffected sib pairs from Denmark, Norway and Sweden. Cohen's kappa coefficient and the intraclass correlation coefficient were used to assess concordances in sib pairs. Furthermore, clinical features and HLA-DR2 carrier status were compared among the probands of sib pairs. RESULTS: We found significant concordance of the disease course (kappa = 0.28, P < 0.001) and adjusted age of onset (r = 0.23, P = 0.028). Among probands of sib pairs, HLA-DR2 carrier patients had a younger age of onset (P = 0.024). CONCLUSION: Analyses of Scandinavian coaffected sib pairs suggest that disease course and age of onset are partly under genetic control. Furthermore, HLA-DR2 in probands of sib pairs suggests importance for age of onset.

Two genome-wide linkage disequilibrium screens in Scandinavian multiple sclerosis patients.

We report the first two genome-wide screens for linkage disequilibrium between putative multiple sclerosis (MS) susceptibility genes and genetic markers performed in the genetically homogenous Scandinavian population, using 6000 microsatellite markers and DNA pools of approximately 200 MS cases and 200 controls in each screen. Usable data were achieved from the same 3331 markers in both screens. Nine markers from eight genomic regions (1p33, 3q13, 6p21, 6q14, 7p22, 9p21, 9q21 and Xq22) were identified as potentially associated with MS in both screens.