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Quantifying circulating nutrient concentrations in sera of free-ranging subjects will help to establish a basis from which we can evaluate the nutritional status and needs of the captive population. We collected serum samples from 26 free-ranging black-and-gold howlers (Alouatta caraya) in San Cayetano forest in northern Argentina. We analyzed them for concentrations of lipoproteins; amino acids; vitamins A, D, and E; carotenoids; and minerals. There are a few significant differences between sexes in concentrations of high-density lipoprotein cholesterol, certain amino acids, vitamin E, lutein + zeaxanthin, and copper. Most nutritional parameters are similar to the ones measured in free-ranging Mexican mantled howlers (Alouatta palliata mexicana) and in captive New World primates (NWPs). Carotenoid, vitamin D, and phosphorus concentrations are the exceptions. Carotenoid concentrations are higher in free-ranging Alouatta caraya than reported for other free-ranging and captive species. Vitamin D concentrations are 14 times greater in the free-ranging black-and-gold howlers than in captive NWPs. Phosphorus concentrations are also higher than expected and higher than typically occur in captive primates, leading to a 1:1.6 calcium:phosphorus ratio. Because we based our study on a small number of free-ranging howlers, additional samples from different regions and throughout the year would better define desirable nutritional parameters for captive howlers. Fil: Schmidt, Debra A.. Lincoln Park Zoo; Estados Unidos Fil: Kowalewski, Miguel Martin. University Of Illinois At Urbana; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; Argentina Fil: Ellersieck, Mark R.. University Of Missouri; Estados Unidos Fil: Zunino, Gabriel Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; Argentina Fil: Stacewicz Sapuntzakis, Maria. University Of Illinois; Estados Unidos Fil: Chen, Tai C.. Boston University; Estados Unidos Fil: Holick, Michael F.. Boston University; Estados Unidos

Calcidiol (25-hydroxyvitamin D) values are reported for 22 wild Ricord's iguanas (Cyclura ricordii) and seven wild rhinoceros iguanas (Cyclura cornuta cornuta). Calcitriol (1,25-hydroxyvitamin D) values are reported for 12 wild Ricord's iguanas and seven wild rhinoceros iguanas. These animals were captured as part of a larger health assessment study being conducted on Ricord's iguanas in Isla Cabritos National Park, Dominican Republic. A total of 13 captive rhinoceros iguanas held outdoors at Parque Zoológico Nacional were also sampled for comparison. Mean concentrations of 25-hydroxyvitamin D were 554 nmol/L (222 ng/ml) with a range of 250-1,118 nmol/L (100-448 ng/ml) for wild Ricord's iguanas, 332 nmol/L (133 ng/ml) with a range of 260-369 nmol/L (104-148 ng/ml) for wild rhinoceros iguanas, and 317 nmol/L (127 ng/ml) with a range of 220-519 nmol/L (88-208 ng/ml) for captive rhinoceros iguanas. On the basis of these results, serum concentrations of at least 325 nmol/L (130 ng/ml) for 25-hydroxyvitamin D should be considered normal for healthy Ricord's and rhinoceros iguanas.

To determine the effect of four vitamin D supplement doses on falls risk in elderly nursing home residents.Secondary data analysis of a previously conducted randomized clinical trial.Seven hundred twenty-five-bed long-term care facility.One hundred twenty-four nursing home residents (average age 89).Participants were randomly assigned to receive one of four vitamin D supplement doses (200 IU, 400 IU, 600 IU, or 800 IU) or placebo daily for 5 months.Number of fallers and number of falls assessed using facility incident tracking database.Over the 5-month study period, the proportion of participants with falls was 44% in the placebo group (11/25), 58% (15/26) in the 200 IU group, 60% (15/25) in the 400 IU group, 60% (15/25) in the 600 IU group, and 20% (5/23) in the 800 IU group. Participants in the 800 IU group had a 72% lower adjusted-incidence rate ratio of falls than those taking placebo over the 5 months (rate ratio=0.28; 95% confidence interval=0.11-0.75). No significant differences were observed for the adjusted fall rates compared to placebo in any of the other supplement groups.Nursing home residents in the highest vitamin D group (800 IU) had a lower number of fallers and a lower incidence rate of falls over 5 months than those taking lower doses. Adequate vitamin D supplementation in elderly nursing home residents could reduce the number of falls experienced by this high falls risk group.

We report new analogs of 1α,25-dihydroxyvitamin D 3 ( 1 ) in three categories. First, design and synthesis of ligands for a mutant vitamin D receptor (VDR)(Arg274Leu), which possess proper functional groups at both C1α and C2α positions of 1 to study the biological activity of the mutant VDR. Among our synthetic analogs, 1α-methyl-2α-(3-hydroxypropyl)-25-hydroxyvitamin D 3 ( 8 ) showed 7.3-fold greater transcriptional activity for the VDR(Arg274Leu) than that of 1 . Next, we examined the antiproliferative activity of 2-substituted 19-norvitamin D 3 analogs on an immortalized normal prostate cell line, PZ-HPV-7, and we found MART 10 ( 14 ) showed the activity even at very low concentration of 10 −10 to 10 −11 M. We also synthesized 25-hydroxy-19-norvitamin D 3 ( 13 ) using Julia-type olefination to connect between the C5 and C6 positions, effectively, to test it as a prohormone type agent for antiprostate diseases. Synthesized compound 13 showed potent antiproliferative activity in PZ-HPV-7, which has high 1α-hydroxylase activity. Finally, we describe design and synthesis of a new TEI-9647 analog, 2α-(3-hydroxypropoxy)-24-propyl-25-dehydro-1α-hydroxyvitamin D 3 -26,23-lactone ( 17 ), which showed the strongest VDR antagonism. Its IC 50 value is 7.4 pM to inhibit differentiation of HL-60 cells induced by 10 nM of 1 .

Laboratory and epidemiological studies have indicated that 1alpha,25-dihydroxyvitamin D(3) [1alpha,25(OH)(2)D(3)] and dietary omega3-polyunsaturated fatty acids (PUFAs) are capable of inhibiting prostate cancer at the initiation and progression stages. The objective of this study is to investigate the influence of 1alpha,25(OH)(2)D(3) and PUFAs in the form of fish oil applied alone or in combination on cell cycle kinetics in the exponentially growing androgen-dependent and -independent prostate cancer cells. Our data indicate that the high passage androgen-independent cell line, LNCaP-c115 had a much greater inhibitory response at the level of the G(1)/S-phase transition in response to fish oil treatment than androgen-dependent low passage LNCaP-c38 cells. When LNCaP-c38 and LNCaP-c115 cells were treated with fish oil (50mug/ml), 1alpha,25(OH)(2)D(3) (10(-8)M) or fish oil (50mug/ml)+1alpha,25(OH)(2)D(3) (10(-8)M), a synergistic growth inhibitory effect was observed with 1alpha,25(OH)(2)D(3)+fish oil group in LNCaP-c115 cell line at the levels of the G(1)/S-phase transition and cell division. This interaction appears to be specific for androgen-independent prostate cancer cell lines. Based on these results, we hypothesize that dietary components, such as omega3PUFAs and Vitamin D, have the potential to delay the progression of prostate cancer cells to an aggressive and un-treatable state.

1alpha,25-Dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)) is known to inhibit the proliferation and invasiveness of prostate cancer cells. However, 1alpha,25(OH)(2)D(3) can cause hypercalcemia and is not suitable as a therapeutic agent. 19-Nor-vitamin D derivatives are known to be less calcemic when administered systemically. In order to develop more potent anti-cancer agents with less calcemic side effect, we therefore utilized (3)H-thymidine incorporation as an index for cell proliferation and examined the antiproliferative activities of nine C-2-substituted 19-nor-1alpha,25(OH)(2)D(3) analogs in the immortalized PZ-HPV-7 normal prostate cell line. Among the nine analogs we observed that the substitution with 2alpha- or 2beta-hydroxypropyl group produced two analogs having antiproliferative potency that is approximately 500- to 1000-fold higher than 1alpha,25(OH)(2)D(3). The (3)H-thymidine incorporation data were supported by the cell counting data after cells were treated with 1alpha,25(OH)(2)D(3), 19-nor-2alpha-(3-hydroxypropyl)-1alpha,25(OH)(2)D(3) or 19-nor-2beta-(3-hydroxypropyl)-1alpha,25(OH)(2)D(3) for 7 days. 19-Nor-2alpha-(3-hydroxypropyl)-1alpha,25(OH)(2)D(3) and 19-nor-2beta-(3-hydroxypropyl)-1alpha,25(OH)(2)D(3) were also shown to be about 10-fold more active than 1alpha,25(OH)(2)D(3) in cell invasion studies using prostate cancer cells. In conclusion, a substitution at the C-2 position of 19-nor-1alpha,25(OH)(2)D(3) molecule with a hydroxypropyl group greatly increased the antiproliferative and anti-invasion potencies. Thus, these two analogs could be developed to be effective therapeutic agents for treating early and late stages of prostate cancer.

Four long-term captive adult rhinoceros iguanas (Cyclura cornuta) on display at the Dallas Zoo (Dallas, TX) were maintained for several years in an outdoor enclosure in the summer and in a smaller indoor enclosure in the winter. While fully exposed to sunlight and given the opportunity to supplement their diet with live vegetation each summer, they were deprived of the benefits of full sun exposure, including ultraviolet B (UVB) exposure, and they received no dietary vitamin D for several months each winter. Maintained with this protocol, the animals seemed healthy and they reproduced. However, increasing awareness of the important role of UVB in vitamin D–mediated calcium metabolism and the potential health risk of UVB and vitamin D deprivation stimulated us to focus more on seasonal changes in vitamin D status and possible subtle changes in health status throughout winter. From September 2008 to October 2010, an 85% decline in the serum 25-hydroxyvitamin D3 (calcidiol) concentrations was measur...

It has been suggested that the major source of vitamin D should come from dietary sources and not sun exposure. However, the major fortified dietary source of vitamin D is milk which often does not contain at least 80% of what is stated on the label. Fish has been touted as an excellent source of vitamin D especially oily fish including salmon and mackerel. Little is known about the effect of various cooking conditions on the vitamin D content in fish. We initiated a study and evaluated the vitamin D content in several species of fish and also evaluated the effect of baking and frying on the vitamin D content. Surprisingly, farmed salmon had approximately 25% of the vitamin D content as wild salmon had. The vitamin D content in fish varied widely even within species. These data suggest that the tables that list the vitamin D content are out-of-date and need to be re-evaluated.

The presence of extra-renal 25-hydroxyvitamin D3 [25(OH)D3]-1alpha-hydroxylase (1alpha-OHase) has been reported in several cell types including prostate and colon cancer cells. Additionally, alterations in the local production of 1alpha,25-dihydroxyvitamin D [1alpha,25(OH)2D3] have been implicated in the tumorigenesis of these malignancies. The aim of this study was to analyze whether normal breast tissue or breast cancer cells expressed 1alpha-OHase and to evaluate whether breast tissue possessed the capacity to produce 1alpha,25(OH)2D3 from 25(OH)D3.Total RNA was extracted from normal breast tissue (n = 11), breast carcinomas (n = 12) and cultured MCF-7 breast cancer cells for real-time (LightCycler using specific hybridization probes) and conventional PCR analysis.mRNA for 1alpha-OHase was detected in breast cancer tissue and in MCF-7 breast cancer cells. Interestingly, the mRNA levels for 1alpha-OHase were significantly increased in breast cancer compared to normal breast tissue. When the MCF-7 cells were treated with 1alpha,25(OH)2D3, cell proliferation was inhibited in a dose-dependent manner. Incubation of the MCF-7 cells with [3H]-25(OH)D3 resulted in its conversion to [3H]-1,25(OH)2D3. The 1alpha-OHase activity in the MCF-7 cells was blocked by a specific cytochrome P450 inhibitor, clotrimazole.The data suggest that at least breast cancer cells expressed 1alpha-OHase mRNA and, therefore, might have the ability to synthesize 1alpha,25(OH)2D3 within the cells. The local production of 1alpha,25(OH)2D3 might play an important role in regulating the proliferation and differentiation of breast cells. We hypothesize that alterations in the local production of 1alpha,25(OH)2D3 may be involved in the tumorigenesis of breast cancer. Additionally, breast cancer may be a target for treatment with precursors of biologically-active vitamin D analogs.

Prostate cells can produce 1alpha,25-dihydroxyvitamin D3 (1alpha,25(OH)2D3) from 25-hydroxyvitamin D3 (25(OH)D3) to regulate their own growth. Here, the questions of whether prostate cells express vitamin D-25-hydroxylase (25-OHase) and can convert vitamin D3 to 1alpha,25(OH)2D3 were investigated.Protein and receptor binding assays were used to determine 25(OH)D3 and 1alpha,25(OH)2D3, respectively. Measurements of proliferation by 3H-thymidine incorporation, and 1alpha,25(OH)2D-responsive gene expression by real-time qPCR and by Western blot were used as functional assays for the presence of 25-OHase activity.Prostate cells metabolized vitamin D3 to 1alpha,25(OH)2D3. Vitamin D3 up-regulated 25(OH)D-24R-hydroxylase and IGFBP3, two 1alpha,25(OH)2D-responsive genes, in prostate cells. CYP2R1 was the major form of 25-OHase expressed in normal and cancerous prostate cells as determined by qPCR.The autocrine synthesis of 1alpha,25(OH)2D3 from vitamin D3 suggests that maintaining adequate levels of serum vitamin D could be a safe and effective chemo-preventive measure to decrease the risk of prostate cancer.

Vitamin D deficiency has been associated with increased risk of colon cancer in epidemiologic and prospective clinical studies. In vitro and in vivo studies demonstrated that 1,25-dihydroxycholecalciferol [1,25(OH)2D3] and its analogs inhibit colon cancer cell proliferation. Few studies have evaluated the effect of vitamin D deficiency on the development and growth of colon cancer. To assess the antiproliferative effects of 25-hydroxyvitamin D [25(OH)D] and 1,25(OH)2D3 in vitro, we cultured MC-26 (a colon cancer cell line) in the presence of 25(OH)D3 and 1,25(OH)2D3 and performed [3H]thymidine incorporation. The proliferation of MC-26 was significantly inhibited by both 25(OH)D3 and 1,25(OH)2D3. To determine the effect of vitamin D deficiency on colon cancer proliferation, Balb/c mice were rendered vitamin D deficient by feeding them a vitamin D-deficient diet for 3 mo. A group of vitamin D-sufficient mice was given the same diet with supplemental vitamin D. The mice were injected with MC-26 colon cancer cells and the tumors were measured daily for 20 d. Vitamin D-sufficient mice had 40% smaller tumors than vitamin D-deficient mice. The tumors were evaluated for mRNA expression of the vitamin D receptor (VDR) and 25-hydroxvitamin D-1alpha-hydroxylase (1alpha-OHase) by quantitative RT-PCR. The expression of the mRNA for the VDR and the 1alpha-OHase was 37- and 6-fold higher, respectively, in the vitamin D-sufficient mice compared with the vitamin D-deficient mice. We conclude that vitamin D deficiency enhances the growth of colon cancer in mice. The tumor expression of VDR and 1alpha-OHase indicates possible autocrine/paracrine cell growth regulation by vitamin D.

The objective was to determine the seasonal fluctuations in serum 25-hydroxyvitamin D (25-OHD) in a group of healthy adolescents living in a northern climate. Twenty-three 9- to 11-year-old girls participated in the study from September 2000 to March 2003. Serum 25-OHD and parathyroid hormone levels were measured each September and March. Dietary intake of vitamin D was assessed each summer and winter. Summer-sun exposure was evaluated using reports of time spent outdoors. The mean decrease in serum 25-OHD from September to March was 28%. Vitamin D insufficiency (at least one serum 25-OHD level

We compared the natural ultraviolet B (UV-B) exposure, dietary vitamin D, and skin-generated vitamin D synthesis for adult males of two species of Jamaican anoles. The more shade-tolerant and thermal-conforming Anolis lineotopus merope, rarely exposed to full sun, experienced less UV-B irradiation in its shady environment than the more heliophilic and thermophilic Anolis sagrei, which frequently basked in full sun during the morning hours (0800-1100 hours). Both species obtained detectable levels of vitamin D(3) in their diet, but the heliophilic A. sagrei obtained more. To compensate for less availability of UV-B and dietary vitamin D, the skin of A. lineotopus merope seems to have acquired a greater sensitivity than that of A. sagrei regarding UV-B-induced vitamin D(3) photobiosynthesis. We assessed this by observing a greater conversion of provitamin D to photoproducts in skin exposed to UV-B from a sunlamp. The reduced skin sensitivity of A. sagrei regarding vitamin D photobiosynthesis may reflect a correlated response associated with less need for vitamin D photobiosynthesis and greater need for UV-B screening capacity as an adaptation to a more damaging UV-B environment. However, the possibility that adaptations for photobiosynthesis of vitamin D and for protection from skin damage could involve independent mechanisms needs investigation. Also, the ability to behaviorally regulate UV-B exposure, as shown for the panther chameleon, would benefit both species of Anolis and should be investigated.

Severe burn injury is associated with vitamin D deficiency, low bone turnover, and abnormalities in calcium homoeostasis. Patients do not routinely receive vitamin D supplementation and sun exposure is currently not controlled. By analysis of skin biopsy samples for vitamin D3 precursors after exposure to ultraviolet B light we found that the conversion of 7-dehydrocholesterol to previtamin D3 was reduced in children a mean of 14 months after the burn. Low serum 25-hydroxyvitamin D concentrations were also found. We conclude that vitamin D supplementation is necessary after burn injury.

The steroid hormone 1,25-dihydroxyvitamin D(3), [1,25(OH)(2)D(3), calcitriol], the active metabolite of vitamin D, exerts pleiotropic antitumor effects against several malignancies. However, the clinical use of this hormone is limited by hypercalcemia. 25-Hydroxyvitamin D(3), the prohormone of 1,25(OH)(2)D(3), is hydroxylated to the active hormone by the enzyme 25-hydroxyvitamin-1-alpha-hydroxylase [1 alpha(OH)ase]. 1 alpha(OH)ase is found primarily in the kidney, but also is expressed in the prostate, colon and other tissues. Using immunohistochemistry, we report that 1 alpha(OH)ase is highly expressed in both normal and malignant pancreatic tissue. Expression of this enzyme and enzymatic activity was also detected in four pancreatic tumor cell lines. 25(OH)D(3) inhibited the growth of three of four pancreatic cell lines in a manner that correlated with the level of induction of the cyclin-dependent kinase inhibitors p21 and p27 and with the induction of cell cycle arrest at the G(1)/S checkpoint. The growth of a cell line stably transfected with a mutant Ki-ras allele and of a second cell line with an endogenous Ki-ras activating mutation was also inhibited by 25(OH)D(3), indicating that activating Ki-Ras mutations, which occur in almost 90% of pancreatic adenocarcinomas, do not interfere with the growth-inhibitory effects of 25(OH)D(3). The expression of 1 alpha(OH)ase in normal and malignant pancreatic tissue and the antiproliferative effects of the prohormone in these cells, suggest that 25(OH)D(3) may offer possible therapeutic and chemopreventive options for pancreatic cancer.

Human prostate cells contain receptors for 1alpha,25-dihydroxyvitamin D, the active form of vitamin D. Prostate cancer cells respond to vitamin D(3) with increases in differentiation and apoptosis, and decreases in proliferation, invasiveness and metastasis. These findings strongly support the use of vitamin D-based therapies for prostate cancer and/or as a second-line therapy if androgen deprivation fails. The association between either decreased sun exposure or vitamin D deficiency and the increased risk of prostate cancer at an earlier age, and with a more aggressive progression, indicates that adequate vitamin D nutrition should be a priority for men of all ages. Here we summarize recent advances in epidemiological and biochemical studies of the endocrine and autocrine systems associated with vitamin D and their implications for prostate cancer and in the evaluation of vitamin D(3) and its analogs in preventing and/or treating prostate cancer.

25-Hydroxyvitamin D-1alpha-hydroxylase (lalpha-OHase) is expressed in prostate cells. The expression suggests that local production of 1,25-dihydroxyvitamin D could provide an important cell growth regulatory mechanism. However, there is differential expression of 1alpha-OHase activity among the primary cultures of prostate cells derived from cancerous, benign prostatic hypertrophy and normal tissue, and among noncancerous (PZHPV-7) and various cancer cell lines (PC-3, DU145). No activity was found in cancer cell line LNCaP. The observed marked decrease in 1alpha-OHase activity in prostate cancer cells suggests some defect of the 1alpha-OHase in these cells. Using luciferase reporter gene assay, we observed a step-wise decrease in the basal promoter activity in two truncated promoter fragments, AN2 (-1,100 bp) and AN5 (-394 bp), with the highest basal activities found in PZHPV-7 and with loss of promoter activity in LNCaP. In order to understand the mechanism underlying the differential promoter activities among different prostate cells, we investigated the possible role of phosphorylation of cyclic AMP response element binding protein (CREB) on the regulation of 1alpha-OHase promoter activity in the four prostate cell lines. First we compared the levels of CREB phosphorylation among PZHPV-7, DU145, PC-3 and LNCaP cells by Western blot analysis using antibody against phosphorylated CREB. We observed that CREB was phosphorylated to a greater extent in PZHPV-7 than in DU145 cells. No significant phosphorylation of CREB was found in PC-3 and LNCaP cells. Next, we utilized activators and inhibitors of protein kinase A (PKA), protein kinase C (PKC), mitogen-activated protein kinase kinase (MAPKK) and calcium/calmodulin-dependent protein kinase II (CaMKII) to determine which kinases might be involved in phosphorylating the CREB in PZHPV-7 cells. We demonstrated that forskolin (an activator of PKA) increased the AN2 basal promoter activity 50%, whereas H-89 (an inhibitor of PKA) inhibited the basal and forskolin-stimulated AN2 promoter activity 40% and 70%, respectively. We also showed that PD98059 (an inhibitor of MAPKK) decreased the AN2 promoter activity 70%. Phorbol 12-myristate 13-acetate (an activator of PKC), GF109203 (an inhibitor of PKC) and KN-93 (an inhibitor of CaMKII) had no effect on AN2 promoter activity in PZHPV-7 cells. Thus, our results suggest that differential phosphorylation of CREB through PKA and MAPK pathways may be involved in the regulation of 1alpha-OHase promoter activity.

The 25-hydroxyvitamin D-1alpha-OHase (1alpha-OHase) is responsible for producing the active form of vitamin D, 1alpha,25-dihydroxyvitamin D. The enzyme not only is expressed in kidneys, but also is expressed in many nonrenal tissues, including skin. In this study, we compared the regulation of the 1alpha-OHase expression in kidney cells and keratinocytes. Using transfected luciferase reporter gene constructs, we compared the activity and regulatory features of the human 1alpha-OHase gene promoter in C-21 human kidney cells (PTH/PTHrP receptor positive) and cultured human keratinocytes (NHKs). We found that two regions, -1,100 bp and -396 bp from the ATG, were highly sensitive to parathyroid hormone (PTH) in C-21 cells but not in NHK. Furthermore, three CRE-like sequences (CLS) were identified within this PTH-sensitive area of the 1alpha-OHase promoter and when deleted they reduced induction of PTH by 50%-95% in C-21 cells. To further investigate the differential regulation profile, we examined the protein products of 1alpha-OHase in kidney and skin. Western blot analysis of whole cell extracts from these tissues with a 1alpha-OHase-specific antibody revealed the predicted 1alpha-OHase protein product of 56 kDa in kidney and a larger protein product of 59 kDa in skin. Using RT-PCR for the 1alpha-OHase in skin and kidney, we detected an insertion between exons 2 and 3 in skin but not in kidney. These results suggest that the regulation of renal and skin 1alpha-OHase gene expression may be tissue specific and possibly produce different splice variants, and that this specificity is likely conferred by differential expression of CRE-binding proteins in different cell types. In conclusion, the differential tissue expression of 1alpha-OHase gene variants and the tissue-specific regulation profile open up a new paradigm in the understanding of the role of 25-hydroxyvitamin D3 1alpha-hydroxylase gene in the regulation of vitamin D physiology.

Prostate cancer cells contain specific receptors (VDR) for la,25-dihydroxyvitamin D (1alpha,25(OH)2D), which is known to inhibit the proliferation and invasiveness of these cells. These findings support the use of 1alph,25(OH)2D for prostate cancer therapy. However, because 1alpha,25(OH)2D can cause hypercalcemia, analogs of 1alpha,25(OH)2D that are less calcemic but which exhibit potent antiproliferative activity would be attractive as therapeutic agents. We studied four vitamin D compounds: 25-hydroxyvitaminD3 [25(OH)D3], which is converted to 1alpha,25(OH)2D3 in prostate cells, and three analogs of 1alpha,25(OH)2D3: EB1089, 19-nor-1alpha,25(OH)2D2 and hexafluoro-1alpha,25(OH)2D3 (F6-1alpha,25(OH)2D3). 19-nor-1alpha,25(OH)2D2 has been shown to be less calcemic than 1alpha,25(OH)2D3 in clinical trials. F6-1alpha,25(OH)2D3 has been shown to be 100-fold more active than 1alpha,25(OH)2D3 and to be longer-lasting in inhibiting keratinocyte proliferation in vitro. EB1089 has been shown to be less calcemic than 1alpha,25(OH)2D3 in rats implanted with Leydig cell tumors. For 25(OH)D3, 19-nor-1alpha,25(OH)2D2 and F6-1alpha,25(OH)2D3, we studied the in vitro effects and compared their activity to 1alpha,25(OH)2D3 on cellular proliferation by 3H-thymidine incorporation assay. In addition, we studied transactivation of the VDR in the presence of 25(OH)D3 and 19-nor-1alpha,25(OH)2D2 in prostate cells. For EB1089, we compared its inhibition of prostate cancer metastasis to that induced by 1alpha,25(OH)2D3 in vivo in the rat Dunning MAT LyLu prostate cancer model. We found that 1alpha,25(OH)2D3 and 19-nor-1alpha,25(OH)2D2 caused similar dose-dependent inhibition in 3H-thymidine incorporation into DNA in prostate cells and behaved similarly in the CAT reporter gene transactivation assay in PC-3/VDR cells. F6-1alpha,25(OH)2D3 is 10- to 50-fold more active than 1alpha,25(OH)2D3 in 3H-thymidine incorporation into DNA in the primary cultured prostate cells. Likewise, 25(OH)D3 had comparable antiproliferative activity to la,25(OH)2D3. In the rat model, tumor volumes and the number of metastases in the lungs were significantly reduced by both 1alpha,25(OH)2D3 (10.4 +/- 2.81 tumor foci) and EB1089 (7.7+/-1.29 tumor foci) compared to controls (22.7 +/- 1.98 tumor foci). Although serum calcium levels were significantly elevated in both 1alph,25(OH)2D3- and EB1089-treated rats, EB1089 was significantly less calcemic than 1alpha,25(OH)2D3 (12.59+/-0.21 mg/dl versus 14.47+/-.46 mg/dL; 1 microg/kg; p0.001). In conclusion, our data indicate that 25(OH)D3 and the three 1alpha,25(OH)2D analogs represent two different solutions to the problem of hypercalcemia associated with vitamin D-based prostate cancer therapies: 25(OH)D3 requires the presence of 25-hydroxyvitaminD-1alpha-hydroxylase, whereas 19-nor-1alpha,25(OH)2D2, F6-1alpha,25(OH)2D3 and EB1089 do not. These compounds may be good candidates for human clinical trials in prostate cancer.

In this report we describe that 1,25(OH)2D3-3-BE, a VDR-affinity labeling analog of 1,25(OH)2D3, showed strong and dose-dependent growth-inhibitory effect in several epithelial cells, i.e., keratinocytes (primary cells), MCF-7 breast cancer, PC-3, and LNCaP prostate cancer and PZ-HPV-7 immortalized normal prostate cell-lines. Furthermore, 10−6 M of 1,25(OH)2D3-3-BE induced apoptosis specifically in LNCaP and PC-3 cells; and the effect was much less pronounced at lower doses. We also showed that the effect (of 1,25(OH)2D3-3-BE) was not due to probable degradation (hydrolysis) of 1,25(OH)2D3-3-BE or random interaction of this molecule with cellular proteins. Tissue- or cell-specific action of 1,25(OH)2D3 and its mimics is not common due to the ubiquitous nature of VDR. Furthermore, variable effects of 1,25(OH)2D3 and its analogs in various cell-lines potentially limits their application as anticancer agents. We showed that 1,25(OH)2D3-3-BE displayed similar growth-inhibitory and cytotoxic activities towards androgen sensitive LNCaP and androgen-independent PC-3 cell-lines. Therefore, these results raise the possibility that 1,25(OH)2D3-3-BE or similar VDR-cross linking analogs of 1,25(OH)2D3 might be considered for further development as potential candidates for prostate cancer. J. Cell. Biochem. 89: 909–916, 2003. © 2003 Wiley-Liss, Inc.

Previously, we demonstrated stimulation of epidermal proliferation and hair growth in triiodothyronine (T(3)) treated mice. To distinguish skin effects of directly applied T(3) from those of systemic hyperthyroidism, we treated CD-1 mice with either intraperitoneally (IP) or topically administered T(3). Relative to controls, mice receiving T(3) IP had 10% thinner epidermis (p < 0.01) and 48% fewer hairs (p < 0.001). By contrast, mice receiving T(3) topically had 78% thicker epidermis (p < 0.01) and 160% more hairs (p < 0.01). To gain insight into factors responsible for the diverging effects, we contrasted T(3) effect on proliferation of isolated keratinocyte cultures versus keratinocytes cocultured with dermal fibroblasts. For keratinocytes grown in the absence of fibroblasts, T(3) stimulated proliferation in a dose-dependent, biphasic pattern with the peak at 0.5 nM T(3) (84 +/- 30%, p < 0.05). Paradoxically, T(3) inhibited proliferation of keratinocytes cocultured with fibroblasts, the nadir at 0.1 nM T(3) (34% +/- 4%, p < 0.001). These studies are the first describing divergent effects of IP and topically administered thyroid hormone. The data suggest that while T(3) stimulated keratinocyte proliferation, T(3) also stimulated proliferation inhibitory factor(s) from skin fibroblasts. Insight into the interplay among the competing factors will be important in understanding thyroid hormone regulation of skin physiology.

Basking by ectothermic vertebrates is thought to have evolved for thermoregulation. However, another beneficial effect of sunlight exposure, specifically the ultraviolet B (UV-B) component, includes endogenous production of vitamin D(3). In the laboratory, panther chameleons exhibited a positive phototaxis to greater visible, ultraviolet A (UV-A) and UV-B light. However, with equivalent high irradiances of UV-A or UV-B, their response to UV-B was significantly greater than it was to UV-A. Exposure of in vitro skin patches of panther chameleons to high UV-B (90 microW/cm(2)) for 1 h significantly enhanced vitamin D(3) concentration. Voluntary exposure to higher UV-B irradiance (70 vs. 1 microW/cm(2)) resulted in greater circulating 25-hydroxyvitamin D(3) in female panther chameleons (604 vs. 92 ng/mL). Depending on dietary intake of vitamin D(3), chameleons adjusted their exposure time to UV-B irradiation as if regulating their endogenous production of this vital hormone. When dietary intake was low (1-3 IU/g), they exposed themselves to significantly more UV-producing light; when intake was high (9-129 IU/g), they exposed themselves to less. Vitamin D(3) photoregulation seems to be an important additional component of the function of basking.

Evidence suggests that vitamin D may have a protective role for prostate cancer. 1alpha,25-Dihydroxyvitamin D [1alpha,25(OH)(2)D] inhibits growth and induces differentiation of prostate cells. 25-Hydroxyvitamin D-1alpha-hydroxylase [1alpha-OHase], the enzyme that is responsible for the synthesis of 1alpha,25(OH)(2)D, is expressed in cultured prostate cells. We observed a marked decrease in 1alpha-OHase activity in prostate cancer cells, suggesting some defect of the 1alpha-OHase in these cells. To investigate whether the defect was due to dysregulation of the enzyme at the promoter level, a series of deletion constructs of the promoter was synthesized and incorporated upstream into the luciferase reporter gene. Two regions were identified with high basal activity in transfected normal prostate cell line (PZHPV-7), -1100 bp (AN2), and -394 bp (AN5) upstream of ATG start site of the 1alpha-OHase gene. When the reporter gene with either AN2 or AN5 was transfected into prostate cancer cell lines, we observed a lower basal promoter activity in PC-3 cells and DU145 cells than that found in PZHPV-7 cells for both constructs, and a loss of promoter activity in LNCaP cells. Thus, the results suggest that the defect in enzyme activity may result from the decreased promoter activity in prostate cancer cells.

The 25-hydroxyvitamin D-1α-OHase (1α-OHase) is responsible for producing the active form of vitamin D, 1α,25-dihydroxyvitamin D. The enzyme not only is expressed in kidneys, but also is expressed in many nonrenal tissues, including skin. In this study, we compared the regulation of the 1α-OHase expression in kidney cells and keratinocytes. Using transfected luciferase reporter gene constructs, we compared the activity and regulatory features of the human 1α-OHase gene promoter in C-21 human kidney cells (PTH/PTHrP receptor positive) and cultured human keratinocytes (NHKs). We found that two regions, -1,100 bp and -396 bp from the ATG, were highly sensitive to parathyroid hormone (PTH) in C-21 cells but not in NHK. Furthermore, three CRE-like sequences (CLS) were identified within this PTH-sensitive area of the 1α-OHase promoter and when deleted they reduced induction of PTH by 50%-95% in C-21 cells. To further investigate the differential regulation profile, we examined the protein products of 1α-OHase in kidney and skin. Western blot analysis of whole cell extracts from these tissues with a 1α-OHasespecific antibody revealed the predicted 1α-OHase protein product of 56 kDa in kidney and a larger protein product of 59 kDa in skin. Using RT-PCR for the 1α-OHase in skin and kidney, we detected an insertion between exons 2 and 3 in skin but not in kidney. These results suggest that the regulation of renal and skin 1α-OHase gene expression may be tissue specific and possibly produce different splice variants, and that this specificity is likely conferred by differential expression of CRE-binding proteins in different cell types. In conclusion, the differential tissue expression of 1α-OHase gene variants and the tissue-specific regulation profile open up a new paradigm in the understanding of the role of 25-hydroxyvitamin D3 1α-hydroxylase gene in the regulation of vitamin D physiology.

Prostate cancer cells contain specific receptors (VDR) for la,25-dihydroxyvitamin D (1alpha,25(OH)2D), which is known to inhibit the proliferation and invasiveness of these cells. These findings support the use of 1alph,25(OH)2D for prostate cancer therapy. However, because 1alpha,25(OH)2D can cause hypercalcemia, analogs of 1alpha,25(OH)2D that are less calcemic but which exhibit potent antiproliferative activity would be attractive as therapeutic agents. We studied four vitamin D compounds: 25-hydroxyvitaminD3 [25(OH)D3], which is converted to 1alpha,25(OH)2D3 in prostate cells, and three analogs of 1alpha,25(OH)2D3: EB1089, 19-nor-1alpha,25(OH)2D2 and hexafluoro-1alpha,25(OH)2D3 (F6-1alpha,25(OH)2D3). 19-nor-1alpha,25(OH)2D2 has been shown to be less calcemic than 1alpha,25(OH)2D3 in clinical trials. F6-1alpha,25(OH)2D3 has been shown to be 100-fold more active than 1alpha,25(OH)2D3 and to be longer-lasting in inhibiting keratinocyte proliferation in vitro. EB1089 has been shown to be less calcemic than 1alpha,25(OH)2D3 in rats implanted with Leydig cell tumors. For 25(OH)D3, 19-nor-1alpha,25(OH)2D2 and F6-1alpha,25(OH)2D3, we studied the in vitro effects and compared their activity to 1alpha,25(OH)2D3 on cellular proliferation by 3H-thymidine incorporation assay. In addition, we studied transactivation of the VDR in the presence of 25(OH)D3 and 19-nor-1alpha,25(OH)2D2 in prostate cells. For EB1089, we compared its inhibition of prostate cancer metastasis to that induced by 1alpha,25(OH)2D3 in vivo in the rat Dunning MAT LyLu prostate cancer model. We found that 1alpha,25(OH)2D3 and 19-nor-1alpha,25(OH)2D2 caused similar dose-dependent inhibition in 3H-thymidine incorporation into DNA in prostate cells and behaved similarly in the CAT reporter gene transactivation assay in PC-3/VDR cells. F6-1alpha,25(OH)2D3 is 10- to 50-fold more active than 1alpha,25(OH)2D3 in 3H-thymidine incorporation into DNA in the primary cultured prostate cells. Likewise, 25(OH)D3 had comparable antiproliferative activity to la,25(OH)2D3. In the rat model, tumor volumes and the number of metastases in the lungs were significantly reduced by both 1alpha,25(OH)2D3 (10.4 +/- 2.81 tumor foci) and EB1089 (7.7+/-1.29 tumor foci) compared to controls (22.7 +/- 1.98 tumor foci). Although serum calcium levels were significantly elevated in both 1alph,25(OH)2D3- and EB1089-treated rats, EB1089 was significantly less calcemic than 1alpha,25(OH)2D3 (12.59+/-0.21 mg/dl versus 14.47+/-.46 mg/dL; 1 microg/kg; p < 0.001). In conclusion, our data indicate that 25(OH)D3 and the three 1alpha,25(OH)2D analogs represent two different solutions to the problem of hypercalcemia associated with vitamin D-based prostate cancer therapies: 25(OH)D3 requires the presence of 25-hydroxyvitaminD-1alpha-hydroxylase, whereas 19-nor-1alpha,25(OH)2D2, F6-1alpha,25(OH)2D3 and EB1089 do not. These compounds may be good candidates for human clinical trials in prostate cancer.

25-Hydroxyvitamin D-1alpha-hydroxylase (lalpha-OHase) is expressed in prostate cells. The expression suggests that local production of 1,25-dihydroxyvitamin D could provide an important cell growth regulatory mechanism. However, there is differential expression of 1alpha-OHase activity among the primary cultures of prostate cells derived from cancerous, benign prostatic hypertrophy and normal tissue, and among noncancerous (PZHPV-7) and various cancer cell lines (PC-3, DU145). No activity was found in cancer cell line LNCaP. The observed marked decrease in 1alpha-OHase activity in prostate cancer cells suggests some defect of the 1alpha-OHase in these cells. Using luciferase reporter gene assay, we observed a step-wise decrease in the basal promoter activity in two truncated promoter fragments, AN2 (-1,100 bp) and AN5 (-394 bp), with the highest basal activities found in PZHPV-7 and with loss of promoter activity in LNCaP. In order to understand the mechanism underlying the differential promoter activities among different prostate cells, we investigated the possible role of phosphorylation of cyclic AMP response element binding protein (CREB) on the regulation of 1alpha-OHase promoter activity in the four prostate cell lines. First we compared the levels of CREB phosphorylation among PZHPV-7, DU145, PC-3 and LNCaP cells by Western blot analysis using antibody against phosphorylated CREB. We observed that CREB was phosphorylated to a greater extent in PZHPV-7 than in DU145 cells. No significant phosphorylation of CREB was found in PC-3 and LNCaP cells. Next, we utilized activators and inhibitors of protein kinase A (PKA), protein kinase C (PKC), mitogen-activated protein kinase kinase (MAPKK) and calcium/calmodulin-dependent protein kinase II (CaMKII) to determine which kinases might be involved in phosphorylating the CREB in PZHPV-7 cells. We demonstrated that forskolin (an activator of PKA) increased the AN2 basal promoter activity 50%, whereas H-89 (an inhibitor of PKA) inhibited the basal and forskolin-stimulated AN2 promoter activity 40% and 70%, respectively. We also showed that PD98059 (an inhibitor of MAPKK) decreased the AN2 promoter activity 70%. Phorbol 12-myristate 13-acetate (an activator of PKC), GF109203 (an inhibitor of PKC) and KN-93 (an inhibitor of CaMKII) had no effect on AN2 promoter activity in PZHPV-7 cells. Thus, our results suggest that differential phosphorylation of CREB through PKA and MAPK pathways may be involved in the regulation of 1alpha-OHase promoter activity.

The biologic action of parathyroid hormone (PTH)-related peptide (PTHrP) in normal skin was investigated in cultured human keratinocytes and in SKH-1 hairless mice. The results indicate that the PTHrP agonists human PTHrP-(1-34) and PTH(1-34) are potent inhibitors of epidermal cell proliferation. [Nle8,18,Tyr34]bovine PTH-(7-34)-amide, an antagonist of the PTH/PTHrP receptor, blocked the inhibitory effect of PTH-(1-34) in cultured keratinocytes. In the SKH-1 mice, PTH-(7-34) caused a 244% increase of [3H]thymidine incorporation into isolated epidermal DNA and 246% and 180% increases in the number and length of hair shafts, respectively. Thus, PTH and PTHrP may play an important role in the normal physiology of skin, and their agonists and antagonists have potentially wide therapeutic applications in the treatment of hyperproliferative skin disorders and aging skin and could also be effective in stimulating and maintaining hair growth.

The hormone 1alpha,25-dihydroxyvitamin D (1alpha,25(OH)(2)D) inhibits growth and induces differentiation of prostate cells. The enzyme responsible for 1alpha,25(OH)(2)D synthesis, 25-hydroxyvitamin D (25(OH)D)-1alpha-hydroxylase (1alpha-OHase), has been demonstrated in human prostate cells. We compared the levels of 1alpha-OHase activity in prostate cancer cell lines, LNCaP, DU145 and PC-3 and in primary cultures of normal, cancerous and benign prostatic hyperplasia (BPH) prostate cells. We observed a marked decrease in 1alpha-OHase activity in prostate cancer cells, including an undetectable level of activity in LNCaP cells. Transient or stable transfection of 1alpha-OHase cDNA into LNCaP cells increased 1alpha-OHase activity from undetectable to 4.95pmole/mg+/-0.69pmole/mg and 5.8pmole/mg+/-0.7pmole/mg protein per hour, respectively. In response to 25(OH)D, the prohormone of 1alpha,25(OH)(2)D, the transfected LNCaP cells showed a significant inhibition of 3H-thymidine incorporation (37%+/-6% and 56%+/-4% at 10(-8)M for transiently and stably transfected cells, respectively). These findings support an important autocrine role for 1alpha,25(OH)(2)D in the prostate and suggest that the re-introduction of the 1alpha-OHase gene to prostate cancer cells, in conjunction with the systemic administration of 25(OH)D, constitutes an endocrine form of gene therapy that may be less toxic than the systemic administration of 1alpha,25(OH)(2)D.

Most vertebrates including reptiles and amphibians need a source of ultraviolet B radiation in order to promote an adequate production of vitamin D to satisfy their body’s requirement (1). Reptiles routinely sun themselves, not only to warm their bodies, but also to produce vitamin D3 in their skin. Amphibians exposed to sunlight also have the ability to produce vitamin D3 in their skin (1). It is estimated that there are over ten million households in the United States that have a reptile or amphibian as a pet. Often these animals are housed in a glass enclosure, and are exposed to incandescent lighting. Since incandescent lighting does not emit any ultraviolet B radiation, these animals depend solely on their diet for their vitamin D requirement. Frequently, diets including vegetable matter or live or dead animals do not contain an adequate amount of vitamin D to satisfy their requirements resulting in these animals developing severe vitamin D deficiency. Vitamin D deficiency causes rickets and osteomalacia that can lead to fractures, muscle weakness and ultimately, death. Many zoos also house reptiles and amphibians indoors, in glass enclosures and often experience vitamin D deficiency in these prized animals. The appreciation that these animals require a source of ultraviolet B radiation has prompted the lamp manufactures and distributors to produce florescent lamps that emit ultraviolet B radiation that is similar to lamps used in tanning salons.

Either absorbed in the intestine from the diet or synthesized in the skin through exposure to ultraviolet (UV) radiation, vitamin D undergoes two sequential enzymatic hydroxylations, first in the liver at the 25th carbon position, producing 25-hydroxyvitamin D3 (25(OH)D3), and then in the kidney, at the 1α position, yielding the active hormone, 1α,25-dihydroxyvitamin D3 (1α,25(OH)2D3)1. Since the discovery of extra-renal 25(OH)D-1α-hydroxylase (1α-OHase) in keratinocytes2, this enzyme, responsible for activating vitamin D, has been demonstrated in various tissues, including several prostate cancer cell lines and primary cultures of prostate cells as shown by our group in 19983. In that report, we observed that a primary culture of prostate cells from a patient with benign prostatic hypertrophy (BPH) had a lower 1α-OHase activity level than that obtained from a normal donor, and that prostate cancer cell lines had an even lower 1α-OHase activity level than that found in BPH cells. In fact, no detectable 1α-OHase activity was found in the LNCaP cancer cell line. In fact, no detectable 1α-OHase activity was found in the LNCaP cancer cell line.

Epidemiological and biochemical evidence strongly implicates a role for the active form of vitamin D, 1α,25-dihydroxyvitamin D [1α,25(OH)2D], in the growth and differentiation of human breast, colon and prostate cells. Recently, our laboratory demonstrated the presence of mRNA and enzyme activity for 25-hydroxyvitamin D-1α-hydroxylase (1α-OHase), the enzyme responsible for the synthesis of 1α,25(OH)2D, in human cancer cells derived from those tissues. We further demonstrated a marked decrease in 1α-OHase activity in the primary cultures and cell lines of prostate cancer cells, compared to those found in the primary cultures of normal prostate cells. Transient or stable transfection of LNCaP prostate cancer cells with 1α-OHase cDNA greatly increased the 1α-OHase enzyme activity, and thereby their cellular response to the antiproliferative activity of 25(OH)D. These findings suggest that, by synthesizing la,25(OH)2D within the cell, the enzyme may play an autocrine role in modulating the proliferation and differentiation of breast, colon and prostate cells. Thus, higher circulating concentrations of 25(OH)D may be necessary for the extra-renal 1α-OHase to work at its maximum capacity to produce an adequate amount of 1α,25(OH)2D in tissues such as the breast, colon and prostate to regulate cell growth and protect them from becoming malignant. This may explain why people who live closer to the equator are less likely to die from cancers of the breast, colon and prostate.

As a first step toward documenting efficacy of the Mark IV (Synaptic Tan, Inc., Louisville, KY, synaptictan.com) for clinical vitamin D3 augmentation we sought to determine whether in vitro photoactivation of 7-dehydrocholesterol (7-DHC) occurs. The Mark IV provides uniform illumination (± 15%) over 0.5 m2 at 60 cm utilizing non-spatial collimation with 20–100 µ pulses of UVB/UVA. The optically coated reflectors serve as highly effective bandpass filters with output ranging over 270–370 nm with little light measured above and below this range. The device delivers 1 FDA corrected MED within 1-4 minutes depending on pulse rate and distance. Glass ampoules (total 27) containing 7-DHC in methanol were exposed to 0.5, 1, and 2 MED at 30, 45, and 60 cm. Assay for preVitamin D3 (preD3), Lumisterol (L3), and Tachysterol (T3) identified a dose-dependent linear relationship for the formation of preD3 independent of distance. This confirms that “reciprocity” is preserved with 20–100 µ pulses.

Vertebrates require vitamin D for several bodily processes including the regulation of calcium and phosphorus levels1. Vitamin D is obtained in two ways. First, vitamin D can be obtained from the diet2. Second, vitamin D can be produced in the skin upon exposure to ultraviolet B irradiation (UVB, 280–315nm)2.

Vitamin D deficiency has been implicated as a possible factor in the development of many cancers including cancer of the prostate, breast and colon 1, 2. Garland & Garland reported that mortality rates due to colorectal cancer in the United States are highest in areas with the least amount of solar radiation 3. Countries with the lowest prevalence of colon cancer are located within 20 degrees of the equator 4. Increasing latitude increases the risk of developing colon cancer with the exception of Japan where the diet is high in vitamin D from ingestion of fatty fish 5. A prospective cohort study of 1954 men determined that men who ingested more than 150 IU of vitamin D had a 50% lower incidence of colon cancer than those who consumed less than 150 IU of vitamin D 6. A twelve-year prospective study of over 89,000 female nurses found that the highest quintile of vitamin D intake was associated with a relative risk of 0.42 (95% CI 0.19 – 0.91) of developing colon cancer when compared to the lowest quintile of vitamin D intake 7. A 6 year prospective study of almost 48,000 men revealed that men in the highest quintile of vitamin D intake had half the incidence rate of colon cancer than men in the lowest quintile of vitamin D intake 8. A case-control study of 25,620 adults revealed that individuals with 25(OH)D levels greater than 20 ng/ml had one-third the risk of colon cancer compared to subjects who were vitamin D insufficient9.

Humans have developed many mechanisms for detecting and responding to solar ultraviolet (UV) and visible radiation. The primary UV responses are localized to the skin because of its direct interface with sunlight. There is a complex underlying network of response pathways initiated through hormonal and other biologic systems in the skin after UV irradiation which augment gene expression. It is well established that, during exposure to sunlight, the UVB portion (290–315nm) of the solar spectrum is responsible for the photolysis of epidermal and dermal stores of 7–dehydrocholesterol [provitamin D3] to previtamin D3 (1). Once formed, it is then isomerized by a thermally induced mechanism to vitamin D3 (1). Vitamin D3 a biologically inert form, enters the circulation and requires two successive hydroxylations before becoming biologically active. It is first hydroxylated in the liver by the vitamin D–25–hydroxylase [25–OHase] to form 25–hydroxyvitamin D3 [25(OH)D3] and then in the kidney to its biologically active form 1α,25–dihydroxyvitamin D3 [1α,25(OH)2D3] by the 25–hydroxyvitamin D3 1α–hydroxylase [1α–OHase] or to 24,25–dihydroxyvitamin D3 [24,25(OH)2D3], a catabolic metabolite, by 25–hydroxyvitamin D3 24–hydroxylase [24OHase] (2). Once formed, 1α,25(OH)2D3 acts on its target tissues by interacting with a specific nuclear receptor, the vitamin D receptor (VDR), to bring about its biological effects on gene transcription (3).

Sunlight is a major contributor to an individual’s vitamin D status. Determinants of sun exposure have been used in conjunction with dietary data to study the effect of sunlight on bone health of elderly populations. In order to assess the impact of sun exposure on vitamin D status or bone mass of children, a sun-exposure questionnaire must be devised for children and tested. Because sun exposure is not equivalent over the course of a day, a factor that affects cutaneous previtamin D3synthesis, the questionnaire should account for the varying strengths of ultraviolet B (UVB) radiation throughout a day. This study attempts to improve upon previous methods of quantifying sun exposure and to further investigate in vitro synthesis of previtamin D3at varying UVB levels.

The benefit of the exposure to sunlight as a treatment for cardiocirculatory and cardiovascular disorders has been reported for more than 6,000 years, since the times of the Egyptian emperors Ramses and Nofretete. The medical use of natural heliotherapy has also been reported from Hippokrates (”Corpus Hippocraticum”), from health care habits of ancient Rome and from the physicians of the Islamic culture in ancient Iran [29].

Importance In animal studies, brain-derived neurotrophic factor (BDNF) has been shown to impact neuronal survival and function and improve synaptic plasticity and long-term memory. Circulating BDNF levels increase with physical activity and caloric restriction, thus BDNF may mediate some of the observed associations between lifestyle and the risk for dementia. Some prior studies showed lower circulating BDNF in persons with Alzheimer disease (AD) compared with control participants; however, it remains uncertain whether reduced levels precede dementia onset. Objective To examine whether higher serum BDNF levels in cognitively healthy adults protect against the future risk for dementia and AD and to identify potential modifiers of this association. Design, Setting, and Participants Framingham Study original and offspring participants were followed up from 1992 and 1998, respectively, for up to 10 years. We used Cox models to relate BDNF levels to the risk for dementia and AD and adjusted for potential confounders. We also ran sensitivity analyses stratified by sex, age, and education, as well as related BDNF genetic variants to AD risk. This community-based, prospective cohort study involved 2131 dementia-free participants aged 60 years and older (mean [SD] age, 72 [7] years; 56% women). Main Outcomes and Measures Ten-year incidence of dementia and AD. Results During follow-up, 140 participants developed dementia, 117 of whom had AD. Controlling for age and sex, each standard-deviation increment in BDNF was associated with a 33% lower risk for dementia and AD ( P = .006 and P = .01, respectively) and these associations persisted after additional adjustments. Compared with the bottom quintile, BDNF levels in the top quintile were associated with less than half the risk for dementia and AD (hazard ratio, 0.49; 95% CI, 0.28-0.85; P = .01; and hazard ratio, 0.46; 95% CI, 0.24-0.86; P = .02, respectively). These associations were apparent only among women, persons aged 80 years and older, and those with college degrees (hazard ratios for AD: 0.65, [95% CI, 0.50-0.85], P = .001; 0.63 [95% CI, 0.47-0.85], P = .002; and 0.27 [95% CI, 0.11-0.65], P = .003, respectively). Brain-derived neurotrophic factor genetic variants were not associated with AD risk. Conclusions and Relevance Higher serum BDNF levels may protect against future occurrence of dementia and AD. Our findings suggest a role for BDNF in the biology and possibly in the prevention of dementia and AD, especially in select subgroups of women and older and more highly educated persons.

Generalized atrichia with papules is a rare disorder characterized by loss of hair shortly after birth and development of cutaneous cysts. Mutations in the hairless gene (HR) cause this phenotype in both mouse and human. Here we present a case of atrichia with papules in a patient with a normal HAIRLESS gene but with mutations in both alleles of the VITAMIN D RECEPTOR. The patient exhibited vitamin D resistant rickets, which was confirmed by an absent response of her fibroblasts to 1,25-dihydroxyvitamin D3 in vitro. Similar to individuals with HAIRLESS mutations, her skin showed an absence of normal hair follicles and the presence of follicular remnants and cysts. The cyst epithelium contained keratin-15- and keratin-17-positive cells suggesting derivation from the hair follicle bulge and the presence of epithelial stem cells. Although hair loss has been reported in association with hereditary vitamin D resistant rickets, we now characterize this alopecia as clinically and pathologically indistinguishable from generalized atrichia with papules, which was previously thought to be caused only by mutations in HAIRLESS. These findings suggest that VDR and HR, which are both zinc finger proteins, may be in the same genetic pathway that controls postnatal cycling of the hair follicle.

Background: Obesity is associated with vitamin D insufficiency and secondary hyperparathyroidism. Objective: This study assessed whether obesity alters the cutaneous production of vitamin D3 (cholecalciferol) or the intestinal absorption of vitamin D2 (ergocalciferol). Design: Healthy, white, obese [body mass index (BMI; in kg/m 2 ) ≥ 30] and matched lean control subjects (BMI ≤ 25) received either whole-body ultraviolet radiation or a pharmacologic dose of vitamin D2 orally. Results: Obese subjects had significantly lower basal 25hydroxyvitamin D concentrations and higher parathyroid hormone concentrations than did age-matched control subjects. Evaluation of blood vitamin D3 concentrations 24 h after whole-body irradiation showed that the incremental increase in vitamin D3 was 57% lower in obese than in nonobese subjects. The content of the vitamin D 3 precursor 7-dehydrocholesterol in the skin of obese and nonobese subjects did not differ significantly between groups nor did its conversion to previtamin D3 after irradiation in vitro. The obese and nonobese subjects received an oral dose of 50 000 IU (1.25 mg) vitamin D2. BMI was inversely correlated with serum vitamin D3 concentrations after irradiation (r = � 0.55, P = 0.003) and with peak serum vitamin D2 concentrations after vitamin D2 intake (r = � 0.56, P = 0.007). Conclusions: Obesity-associated vitamin D insufficiency is likely due to the decreased bioavailability of vitamin D 3 from cutaneous and dietary sources because of its deposition in body fat compartments. Am J Clin Nutr 2000;72:690‐3.

Rickets (rachitic pelvis) was a common cause of death in childbirth in industrialized nations at the turn of the 20th century and associated with a steep rise in cesarean section rates. The addition of vitamin D to milk caused the virtual disappearance of rickets, but recent reports suggest that vitamin D deficiency is now widespread in industrialized nations. In 2 previous studies, a high incidence of vitamin D deficiency was found in pregnant and lactating women. Another study reported that 50% of mothers and 65% of newboms infants were severely vitamin D deficient (25-hydroxyvitamin D [25(OH)D

Vitamin metabolism and its regulation is controlled by a complex feedback loop. The biologically active metabolite calcitriol [1,25(OH)2D3] is a hormone that promotes its actions via a specific receptor (VDR) which belongs to the superfamily of the steroid-retinoid-vitamin D transcription regulatory factors.

During exposure to sunlight, the UVB portion of the solar spectrum (295-315 nm) photolyzes 7-dehydrocholesterol (7-DHC) in epidermis to previtamin D3. Previtamin D3 then thermoisomerizes to vitamin D3 (1). Vitamin D3 is hydroxylated in the liver at C-25 to form 25-hydroxyvitamin D3 [25(OH)D3], followed by hydroxylation at the C-lin kidneys to form 1,25dihydroxyvitamin D3 [1,25(OH)2D3], the active form of vitamin D. However, further exposure to sunlight can photoisomerize previtamin D3 and vitamin D3 to a variety of photoproducts, including 5,6-trans-vitamin D3, tachysterol, and lumisterol (2,3). Since they are produced locally in the skin, their concentration could be very high. Among these photoproducts, tachysterol and 5,6-trans-vitamin D3 have a pseudo-1-a-hydroxyl structure due to the 180 degree rotation of the 3-hydroxyl group during the isomerization (Figure 1).

The use of sunlight to treat infections is one of the oldest healing methods. Today most of the attention is paid to the immunosuppressive and cancerogenic effects of UV-light. The positive aspects of ultraviolet radiation have become of little importance to research and clinical practice.

Since Cowles and Bogert’s [1] classic study on thermoregulation, numerous studies have established the importance of basking behavior as a means of thermoregulation of many reptiles [2,3,4]. However, the sun provides not only heat produced by long-wave infrared radiation but also biologically important radiation at the other end of the spectrum, specifically ultraviolet-B (UVB) radiation (290-320 nm). These wavelengths penetrate the skin of vertebrates and promote photobiosynthesis of previtamin D, from provitamin D3 [5,6], which is then isomerized to vitamin D3 by a temperature-dependant process. Despite its potentially important role, basking behavior has never been seriously considered an important means of UV photoregulation for vitamin D photobiosynthesis in reptiles.

Vitamin D deficiency is a major complication in chronic renal failure, leading to secondary hyperparathyroidism and renal osteodystrophy. The usual treatment is the substitution of calcitrol or alphacalcidol orally or intravenously.

In the state of vitamin D deficiency parathyroid hormone (PTH) increases in healthy people during winter time as well as in patients with chronic renal failure, that lead to the state of secondary hyperparathyroidism (1).