Genetic and seasonal determinants of vitamin D status in Confederated Salish and Kootenai Tribes (CSKT) participants

Background: Vitamin D is a hormone produced in the skin upon ultraviolet B (UVB) radiation. Vitamin D is a crucial regulator of calcium and phosphate levels for bone mineralization and other physiological roles. Vitamin D levels vary globally in human populations due to genetics, geography, and other demographic factors. It is estimated that 20-85 % of the variability in vitamin D levels is driven by genetic variation. To improve our understanding of contributors to vitamin D levels, we conducted a candidate-gene study in partnership with the Confederated Salish and Kootenai Tribes (CSKT). Methods: We recruited 472 CSKT study participants on the Flathead Reservation in Montana. Demographic factors included age, BMI, and gender (185 male and 287 female; ≥ 18 years old). Genomic DNA and plasma were isolated from whole blood. We sequenced 14 vitamin D regulatory candidate genes: CASR, CUBN, CYP2R1, CYP3A4,CYP24A1, CYP27B1, DHCR7, GC, RXRA, RXRB, RXRG, SULT2A1, UGT1A4, and VDR. We also measured plasma levels of vitamin D and vitamin D metabolites by liquid chromatography/mass-spectrometry (LC/MS), including the clinical marker of vitamin D status, 25-hydroxyvitamin D3 [25(OH)D3]. We tested demographic factors as well as common and rare genetic variants for statistical associations with vitamin D levels using bioinformatics software and R statistical programming language code. Results: We identified 7,370 total genetic variants with 8% (n = 585) of them being novel. We identified 60 genetic variants that may be of clinical significance (disease associated or predicted to influence medication response). Vitamin D levels were below sufficiency [25(OH)D3 + 25(OH)D2 levels \u3c 20 ng/mL] in 56 % of CSKT participants across the year. We observed seasonal vitamin D and metabolite level fluctuations in a seasonal, sinusoidal statistical model with peak concentrations in June – August and trough concentrations in December – February. In linear regression analysis, we found that age, BMI, season, and 5 variants in CUBN and CYP3A4 were significantly associated with 25(OH)D3 concentration (p-value\u3c 0.05). In logistic regression, we found that 4 variants in CUBN, CYP3A4, and UGT1A4 were associated with 25(OH)D sufficiency status [25(OH)D3 + 25(OH)D2 levels of 20 ng/mL] (p-value\u3c 0.05). Multivariate linear regression analysis revealed that genetic variation alone explained ~13% of the variability in 25(OH)D3 concentration in CSKT participants. Genetic variation and environmental factors together explained ~23 % of the variability in 25(OH)D3 concentration in CSKT participants. It is likely that genetic variation in additional genes and other environmental factors (e.g., dietary vitamin D intake) that were not included in this study explain the remaining variability in 25(OH)D3 concentration. Conclusion: This research addresses the need for increased inclusion of American Indian and Alaska Natives in precision medicine health research. We are the first to describe the contribution of season and genetics to vitamin D levels in an American Indian population. Our next steps will be to use these findings to perform mechanistic studies and develop interventional strategies for the CSKT people

Effect of high root temperature and excessive insolation upon growth

Summary. 1 1. Under ordinary environmental conditions of temperature and sunlight the growth of peas, as of barley, is seriously hindered by overcrowding, even when each plant receives a similar supply of food and water. Not only is less dry weight produced, but the pods become thin and distorted and fail to develop their seeds properly. 2 Growth tends to be depressed in hot sunny weather when no protection is afforded. The chief detrimental factors concerned appear to be high temperatures at the roots associated with strong and prolonged sunshine, though the two factors acting individually are much less potent for harm. Under these conditions crowding shelters the roots from overheating and the leaves from too much sunlight, and up to a certain point crowded plants make better growth than those spaced well apart. Overcrowding, however, still depresses growth, probably because the light and root temperature reductions are too great. 3 Provided insolation is not excessive the amount of daily fluctuation of root temperature over a total range of about 22oC. (6?7-28-9oC.) has comparatively little influence upon growth; high maxima and low minima give similar results to low maxima and relatively high minima, provided the average mean temperatures are not too dissimilar. 4 With high root temperatures a difference in the degree of insolation or in the angle of incidence of the sun's rays may have a considerable influence on growth, a slight easing off of the solar conditions enabling much better growth to be made. 5 With very strong sunshine reduction of high maximum root temperatures (from 29oC. upwards) allows of satisfactory growth, when unprotected plants are rapidly killed. The inhibitory action of too high temperatures at the roots is thus clearly shown. Nevertheless, the growth so made is less good than under more normal conditions of insolation, thus demonstrating the harmful action of too powerful sunlight, when all the root temperatures rule high. 6 Boot temperatures appear to be of greater importance than atmospheric temperatures, as good growth can be made in hot atmospheres provided the roots are kept relatively cool. 7 There is some reason to believe that the minima are of as much importance as the maxima, i.e. that plants can withstand very high maximum temperatures provided there is a considerable drop to the minima, but cannot put up with the constant conditions of heat induced by fairly high maxima, and high minim