Short communication: Metabolic synthesis of vitamin D2 by the gut microbiome.

The origin of vitamin D2 in herbivorous animals was investigated in vivo in sheep and in bovine as well as mouse gastrointestinal tracts. A high concentration of 25-hydroxyvitamin D2 in blood plasma of sheep both in summer and winter appeared to be incompatible with the undetectable level of vitamin D2 in the pasture on which the sheep were grazing. Studies with bovine rumen contents from a cow grazing the same pasture as the sheep, demonstrated an increased concentration of vitamin D2 on anaerobic incubation in a 'Rusitec' artificial rumen, which was further enhanced when cellulose powder was added as a fermentation substrate. The colon contents of mice that were fed from weaning on a vitamin D-free diet were found to contain vitamin D2. The results of these comparative studies in 3 animal species indicated that vitamin D2 was being generated by microbial anaerobic metabolism in the gastrointestinal tract.

Serum 1,25-dihydroxyvitamin D levels in the diagnosis and pathogenesis of nutritional rickets - a multivariable re-analysis of a case-control study.

BACKGROUND: A multivariable logistic regression model resulting from a case-control study of nutritional rickets in Nigerian children suggested that higher levels of serum 25(OH)D may be required to prevent nutritional rickets in populations with low-calcium intakes. OBJECTIVES: This current study evaluates if adding serum 1,25-dihydroxyvitamin D [1,25(OH)2D] to that model shows that increased levels of serum 1,25(OH)2D are independently associated with risk of children on low-calcium diets having nutritional rickets. METHODS: Multivariable logistic regression analysis was used to model the association between serum 1,25(OH)2D and risk of having nutritional rickets in cases (n = 108) and controls (n = 115) after adjusting for age, sex, weight-for age z-score, religion, phosphorus intake and age began walking and the interaction between serum 25(OH)D and dietary calcium intake (Full Model). RESULTS: Serum 1,25(OH)2D levels were significantly higher (320 pmol/L vs. 280 pmol/L) (P = 0.002), and 25(OH)D levels were lower (33 nmol/L vs. 52 nmol/L) (P < 0.0001) in children with rickets than in control children. Serum calcium levels were lower in children with rickets (1.9 mmol/L) than in control children (2.2 mmol/L) (P < 0.001). Dietary calcium intakes were similarly low in both groups (212 mg/d) (P = 0.973). In the multivariable logistic model, 1,25(OH)2D was independently associated with risk of having rickets [coefficient = 0.007 (95% confidence limits: 0.002-0.011)] after adjusting for all variables in the Full Model. CONCLUSIONS: Results confirmed theoretical models that in children with low dietary calcium intake, 1,25(OH)2D serum concentrations are higher in children with rickets than in children without rickets. The difference in 1,25(OH)2D levels is consistent with the hypothesis that children with rickets have lower serum calcium concentrations which prompt the elevation of PTH levels resulting in an elevation of 1,25(OH)2D levels. These results support the need for additional studies to identify dietary and environmental risks for nutritional rickets.

Differences in career paths of alumni of the Cornell Leadership Program from North American universities compared with those alumni from universities in other countries.

OBJECTIVE: To characterize and compare the careers of alumni of the Cornell Leadership Program for Veterinary Students according to the countries where they studied and obtained their veterinary qualification. The Cornell Leadership Program is a 10-week residential research experience program for veterinary students from around the world who have ambitions for research-related careers. SAMPLE: Data on the career development of all 672 alumni were collected each year over the period of 1990 to 2019. PROCEDURES: The annual career profile of each alumnus was recorded and coded in 1 of 28 different categories. The careers and postveterinary qualifications of alumni from universities in the US and Canada (referred to as North American universities) were compared with those alumni who graduated from universities in other countries. RESULTS: Analysis of this 30-year database revealed that a considerable proportion (45.7% [307/672]) of the total 672 alumni are following the traditional career path of veterinary clinical practice rather than the research-related careers they aspired to as students during the Leadership Program. Furthermore, a higher proportion of the 325 North American alumni (56% [182/325]) were in clinical practice compared with 33.6% (112/333) of the 333 alumni from other countries. CLINICAL RELEVANCE: Many veterinary schools now provide research experience programs to encourage highly talented students who have ambitions for careers in which they can advance knowledge about animal disease and contribute to solving the health problems of animals through hypothesis-based research. Comparison of the careers of the Leadership Program alumni indicates that research experience alone is not sufficient to maintain the career goals of alumni. Follow-up mentoring of alumni of such programs is recommended while they complete their veterinary studies to reinforce their career aspirations and provide advice on how to achieve research-related careers.

Physiological significance of vitamin D produced in skin compared with oral vitamin D.

Since the discovery of vitamin D, it has been accepted that its physiological supply is either from food or by endogenous synthesis in skin exposed to solar UV light. Yet vitamin D is a component of very few foods and its supply as a natural nutrient is unable to maintain good vitamin D status for human populations. One aspect of vitamin D physiology that has been ignored is that the mechanisms for its transport and processing from these two sources are quite different. Excess intake of vitamin D causes hypercalcaemic toxicity. However, experiments with different animal species have shown that long-term supply of oral vitamin D in apparently non-toxic amounts causes atherosclerosis in large arteries. A mechanism for this toxicity is proposed. Alternative strategies for addressing widespread vitamin D deficiency by food fortification should be considered in light of the angiotoxicity caused by oral vitamin D in animal experiments.

Vitamin D toxicity related to its physiological and unphysiological supply.

Vitamin D is defined as a nutrient despite its rare occurrence in food. Vitamin D status is determined mainly by solar UV light action in skin. However, the strategy to combat vitamin D deficiency has been to increase oral intake of vitamin D in greater amounts than could be obtained from food. Persistent large intakes of vitamin D can cause hypercalcaemic toxicity. Although the amounts recommended to prevent deficiency are far less than those causing such toxicity, the possibility of other toxic actions from increased intake of vitamin D has been ignored. Animal experiments have demonstrated that moderate amounts of oral vitamin D over time result in atherosclerosis. Differences in the fate of vitamin D could explain this angiotoxicity.

Skeletal Muscle and the Maintenance of Vitamin D Status.

Vitamin D, unlike the micronutrients, vitamins A, E, and K, is largely obtained not from food, but by the action of solar ultraviolet (UV) light on its precursor, 7-dehydrocholesterol, in skin. With the decline in UV light intensity in winter, most skin production of vitamin D occurs in summer. Since no defined storage organ or tissue has been found for vitamin D, it has been assumed that an adequate vitamin D status in winter can only be maintained by oral supplementation. Skeletal muscle cells have now been shown to incorporate the vitamin D-binding protein (DBP) from blood into the cell cytoplasm where it binds to cytoplasmic actin. This intracellular DBP provides an array of specific binding sites for 25-hydroxyvitamin D (25(OH)D), which diffuses into the cell from the extracellular fluid. When intracellular DBP undergoes proteolytic breakdown, the bound 25(OH)D is then released and diffuses back into the blood. This uptake and release of 25(OH)D by muscle accounts for the very long half-life of this metabolite in the circulation. Since 25(OH)D concentration in the blood declines in winter, its cycling in and out of muscle cells appears to be upregulated. Parathyroid hormone is the most likely factor enhancing the repeated cycling of 25(OH)D between skeletal muscle and blood. This mechanism appears to have evolved to maintain an adequate vitamin D status in winter.

Is it reasonable to ignore vitamin D status for musculoskeletal health?

Severe vitamin D deficiency-25-hydroxyvitamin D (25OHD) concentrations below around 25-30 nmol/L-may lead to growth plate disorganization and mineralization abnormalities in children (rickets) and mineralization defects throughout the skeleton (osteomalacia) and proximal muscle weakness. Both problems are reversed with vitamin D treatment. Apart from this musculoskeletal dysfunction at very low vitamin D levels, there is apparent inconsistency in the available data about whether concentrations of 25OHD below around 50 nmol/L cause muscle function impairment and increase the risk of fracture. This narrative review provides evidence to support the contention that improving vitamin D status, up to around 50 nmol/L, plays a small causal role in optimizing bone and muscle function as well as reducing overall mortality.

Tracking Veterinary Students Who Aspire to Careers in Science.

The Cornell Leadership Program for Veterinary Students is an intensive 10-week learning experience intended to guide competitively selected scholars into careers in science and public health. It features independent research, vocational counseling, and student-directed learning modules. Program scholars are encouraged to objectively evaluate graduate training as preparation for careers promoted by the program. Prominence is given to experiential learning through research, participation in program enrichment modules, and inspirational experiences achieved through group meetings and individual interactions with established scientists. Program alumni are monitored to determine how the careers they pursue relate to their earlier-stated ambitions. In addition, subjective assessments are made of the quality of graduate training and its impact on alumni career paths. The influence of mentors, vocational counseling, and inspirational experiences on subsequent training is also subjectively assessed. Information is obtained from students' anonymous responses to questionnaires and recorded interviews. Program alumni are contacted annually to determine their current activities and career aspirations. The Leadership Program encourages program graduates to undertake careers in science and public health, yet an unanticipated number of alumni enter private veterinary practice. A factor relevant to that outcome is that many students destined for practice lack a definitive career plan. Persuading veterinary students to consider careers in research or public service is challenging but worth the effort. Critical to that connection is the need for veterinary students to objectively evaluate graduate training options because the vocations they follow appear to be strongly influenced by the experiences they choose.

The Role of Skeletal Muscle in Maintaining Vitamin D Status in Winter.

The status of vitamin D is determined mainly by its formation in skin by the photochemical action of solar UVB light (wavelength 290-320 nm) on the precursor 7-dehydrocholesterol. Because of seasonal variation in intensity of solar UV light, vitamin D status falls in winter and rises in summer. It has been presumed that there is no functional store of vitamin D. Thus, to avoid deficiency, a nutritional supply would be required in winter. However, there is now evidence that the main circulating metabolite of vitamin D, 25-hydroxyvitamin D, accumulates in skeletal muscle cells, which provide a functional store during the winter months. The mechanism is mediated by muscle cell uptake of circulating vitamin D-binding protein (DBP) through a megalin-cubilin membrane transport process. DBP then binds to cytoplasmic actin to provide an array of high-affinity binding sites for 25-hydroxyvitamin D [25(OH)D]. The repeated passage of 25(OH)D into and out of muscle cells would account for its long residence time in blood.

In vivo measurement of strontium absorption from the rumen of dairy cows as an index of calcium absorption capacity.

Absorption of dietary calcium from the rumen is a quantitatively important process in calcium homeostasis of ruminants. In 3 separate experiments in dairy cows, we applied a technique developed in sheep to measure the rate of strontium (Sr) absorption from the rumen as an indicator of calcium absorption capacity. Absorption from the rumen after an oral dose of SrCl2 resulted in a maximum plasma concentration of Sr after 1 h, whereas absorption from the small intestine after injection of SrCl2 into the abomasum through a cannula occurred more slowly. The second experiment demonstrated that the calcium absorption capacity index of the rumen was significantly greater in 21 lactating Friesian cows (230 ± 66, mean ± SEM) than in 6 mature, nonlactating, nonpregnant heifers (101 ± 21, mean ± SEM). In a third experiment, we compared clinically normal cows at the onset of lactation with those that developed parturient paresis. In cows that developed severe hypocalcemia, plasma concentrations of 1,25(OH)2D were significantly elevated (144 ± 60 pg/mL vs. 90 ± 54 pg/mL; means ± SEM) and their rumen calcium absorption index was significantly decreased compared with that of clinically normal cows. Evidence suggested that mobilization of calcium from bone as lactation commenced was significantly depressed in paretic cows compared with those that did not show clinical signs of hypocalcemia. Moreover, ruminal stasis suppressed the absorption of calcium from the rumen. We conclude that measurement of Sr concentration in blood plasma after an oral dose of SrCl2 into the rumen can be used as an index of rumen calcium absorption capacity under different states of calcium homeostasis.

Vocational choices made by alumni of the Leadership Program for Veterinary Students at Cornell University.

OBJECTIVE To compare vocational aspirations and outcomes of participants in the 10-week Leadership Program for Veterinary Students at Cornell University. DESIGN Survey. SAMPLE Veterinary students who participated in the program between 1990 and 2013. PROCEDURES Questionnaires that sought information about the career aspirations of participants at the beginning and end of the program were reviewed, along with records documenting the career progression of participants, audio recordings of interviews conducted with students, and notes of vocation-oriented counseling sessions held during each year's program. RESULTS At the conclusion of the program, 143 of 174 (82%) participants indicated they were more likely than not to undertake research training after completing their veterinary degree, compared with 106 of 174 (61%) at the beginning. Participation also stimulated interest in residency training and industry, but did little to promote interest in careers in government or the military. The percentage of participants who indicated they were more likely than not to pursue additional training in private practice decreased from 97 of 174 (56%) at the beginning of the program to 75 of 174 (43%) at the end. Information on career progression was available for 391 individuals, of whom 177 (45%) were pursuing careers of the kind envisioned by the program. However, 189 (48%) participants had a career in general or specialty clinical practice. CONCLUSIONS AND CLINICAL RELEVANCE The Leadership Program appeared to have a short-term influence on careers anticipated by program participants. However, a substantial proportion pursued careers in clinical practice after graduation.

Calcium transport in bovine rumen epithelium as affected by luminal Ca concentrations and Ca sources.

The quantitative role of different segments of the gastrointestinal tract for Ca absorption, the respective mechanisms, and their regulation are not fully identified for ruminants, that is, cattle. In different in vitro experiments the forestomach wall has been demonstrated to be a major site for active Ca absorption in sheep and goats. In order to further clarify the role of the bovine rumen for Ca transport with special attention to luminal Ca concentrations, its ionic form, and pH, electrophysiological and unidirectional flux rate measurements were performed with isolated bovine rumen epithelial tissues. For Ca flux studies (Jms, Jsm) in vitro Ussing chamber technique was applied. Standard RT-PCR method was used to characterize TRPV6 and PMCA1 as potential contributors to transepithelial active Ca transport. At Ca concentrations of 1.2 mmol L(-1) on both sides of the tissues, Jms were higher than Jsm resulting under some conditions in significant Ca net flux rates (Jnet), indicating the presence of active Ca transport. In the absence of an electrical gradient, Jnet could significantly be stimulated in the presence of luminal short-chain fatty acids (SCFAs). Increasing the luminal Ca concentrations up to 11.2 mmol L(-1) resulted in significant increases in Jms without influencing Jsm. Providing Ca in its form as respective chloride, formate, or propionate salts there was no significant effect on Jms. No transcripts specific for Ca channel TRPV6 could be demonstrated. Our results indicate different mechanisms for Ca absorption in bovine rumen as compared with those usually described for the small intestines.

New data for vitamin D in Australian foods of animal origin: impact on estimates of national adult vitamin D intakes in 1995 and 2011-13.

OBJECTIVES: To assess the potential dietary supply of vitamin D to Australian adults by application of new data for Australian primary foods of animal origin. METHODS: New published analytical data on the vitamin D contents of Australian primary foods from animal products were obtained and assessed for reliability. Using food consumption data from Australian population dietary surveys for 1995 and 2011-2013, estimates were made of the likely average daily intakes of vitamin D equivalents from these sources by Australian adults. RESULTS: Meats, chicken, fish, eggs and dairy produce may alone have contributed about 4.2 µg vitamin D equivalents per day to average Australian diets of adults >18 years in 1995 and 4.3 µg in 2011-2013. CONCLUSIONS: Dietary vitamin D intake in Australia is likely to be higher than previously estimated because new data from improved analytical methods reveal the contributions to vitamin D supply from foods of animal origin. Absence of reliable vitamin D data for milk and milk products, and the gaps in vitamin D data for many commonly consumed seafood, poultry, eggs and processed animal products greatly limit estimation of dietary vitamin D intakes by Australians.

In vivo measurement of the absorption of strontium in the rumen and small intestine of sheep as an index of calcium absorption capacity.

In the present study, a method was developed for determining the alimentary tract Ca absorption capacity of ruminant animals by measuring the absorption rate of Sr after the administration of an oral dose of strontium chloride acting as a tracer analogue of Ca. A close correlation between the absorption rates of the two tracers was observed upon simultaneous administration of an oral dose of stable Sr and radioactive calcium (r 0·98). The Ca absorption capacity of the rumen and small intestine was determined separately by either directing the solution into the rumen or by diverting it into the post-ruminal tract by vasopressin-induced closure of the ruminoreticular groove. The animals were treated with 1α-hydroxyvitamin D3 administered via subcutaneously implanted mini-osmotic pumps. The effect of elevated plasma 1,25-dihydroxycholecalciferol concentrations on the Ca absorption capacity of the alimentary tract was then determined. An increased rate of Sr absorption was observed in both the rumen and small intestine of sheep after treatment, although it is unclear whether the rumen possesses the same vitamin D-dependent Ca absorption pathway as the small intestine.

The vitamin D receptor (VDR) is expressed in skeletal muscle of male mice and modulates 25-hydroxyvitamin D (25OHD) uptake in myofibers.

Vitamin D deficiency is associated with a range of muscle disorders, including myalgia, muscle weakness, and falls. In humans, polymorphisms of the vitamin D receptor (VDR) gene are associated with variations in muscle strength, and in mice, genetic ablation of VDR results in muscle fiber atrophy and motor deficits. However, mechanisms by which VDR regulates muscle function and morphology remain unclear. A crucial question is whether VDR is expressed in skeletal muscle and directly alters muscle physiology. Using PCR, Western blotting, and immunohistochemistry (VDR-D6 antibody), we detected VDR in murine quadriceps muscle. Detection by Western blotting was dependent on the use of hyperosmolar lysis buffer. Levels of VDR in muscle were low compared with duodenum and dropped progressively with age. Two in vitro models, C2C12 and primary myotubes, displayed dose- and time-dependent increases in expression of both VDR and its target gene CYP24A1 after 1,25(OH)2D (1,25 dihydroxyvitamin D) treatment. Primary myotubes also expressed functional CYP27B1 as demonstrated by luciferase reporter studies, supporting an autoregulatory vitamin D-endocrine system in muscle. Myofibers isolated from mice retained tritiated 25-hydroxyvitamin D3, and this increased after 3 hours of pretreatment with 1,25(OH)2D (0.1 nM). No such response was seen in myofibers from VDR knockout mice. In summary, VDR is expressed in skeletal muscle, and vitamin D regulates gene expression and modulates ligand-dependent uptake of 25-hydroxyvitamin D3 in primary myofibers.

Differences in peripartal plasma parameters related to calcium homeostasis of dairy sheep and goats in comparison with cows.

Recently it has been demonstrated that there are differences between sheep and goats in respect to adaptation to a calcium-restricted diet. It was the aim of the present study to evaluate whether species-specific peculiarities also occur when calcium homoeostasis is challenged by lactation. Therefore, we investigated the time courses of plasma parameters related to calcium homoeostasis (calcium, phosphate, calcitriol, the bone resorption marker CrossLaps and the bone formation marker osteocalcin) during the transition period in multiparous animals of both species and compared the results to data from a former study carried out with dairy cows. As in cows, plasma calcium and the ratio of bone formation to bone resorption decreased at parturition in goats while plasma calcitriol increased. On day 10 post partum the bone parameters of goats reached prepartum values again, which was not the case in cows. Sheep were found to experience a challenge of calcium homoeostasis already 10 d before parturition, reflected by a very low ratio of bone formation to bone resorption, which was not accompanied by an increase in plasma calcitriol. Additionally, sheep and goats which had been in milk for 3 months were sampled, dried-off and sampled again 6 weeks later. In dried-off animals there were no detectable differences in parameters of bone metabolism. In conclusion we could show that the contribution of bone mobilisation to the compensation for the enhanced calcium demand due to lactation differs between the three ruminant species.

Uptake of 25-hydroxyvitamin D by muscle and fat cells.

Vitamin D status, measured as serum 25-hydroxyvitamin D (25OHD) concentration, is determined by rates of input and of degradation. The half-life of 25OHD is surprisingly long for a steroid and much longer than its blood transporter, vitamin D binding protein. There is some evidence to suggest that vitamin D itself is stored in fat, whereas 25OHD concentrations are usually related to muscle-related parameters such as lean body mass and exercise. Both muscle and fat cells come from the mesenchymal cell lineage. We recently published evidence for net uptake of 25OHD into differentiated muscle cells, in a process that was megalin dependent, and speculated that this uptake might contribute to the extended half-life of 25OHD. Whether 25OHD is also taken up into cells of the adipocyte lineage is not clear. In the current study, we used the C2 muscle cell line as a source of myoblasts that were differentiated in culture to myotubes and 3T3-L1 pre-adipocytes that were differentiated into adipocytes in culture. We incubated the cells with trititated 25OHD and measured net uptake 4 and 16h afterwards. Differentiated myotubes took up labeled 25OHD in a time-dependent process to a far greater extent than myoblasts. In contrast, pre-adipocytes, but not differentiated adipocytes, accumulated labeled 25OHD in a time-dependent manner, though to a lesser extent than myotubes. Myotubes, but not myoblasts, showed megalin expression by immunohistochemistry. Pre-adipocytes, but not adipocytes, also showed expression of megalin. Since skeletal muscle consists mainly of differentiated muscle cells, while adipose tissue is mainly differentiated fat cells, it seems likely that muscle, but not fat tissue, provides a large extravascular pool through which 25OHD circulates and that this protects 25OHD from degradation. This article is part of a Special Issue entitled '16th Vitamin D Workshop'.

Culture and sun exposure in immigrant East Asian women living in Australia.

In this qualitative study, researchers examined cultural and attitudinal factors that might be related to sun-exposure behaviors among East Asian women living in Australia. Researchers asked Chinese (n = 20) and Korean (n = 16) immigrant women who participated in a larger cross-sectional quantitative study of vitamin D blood levels to volunteer to participate in an in-depth interview in 2010. These women reported a number of cultural factors related to their attitudes and behaviors with regard to sun exposure. They expressed preference for fair skin, a tradition of covering skin when outdoors, and no sunbathing culture. They believed that fair skin was more beautiful than tanned skin. They reported that beauty was the reason for active avoidance of sunlight exposure. Although they reported knowledge of the need for sun avoidance due to skin cancer risk, few reported knowledge about the benefits of sun exposure for adequate vitamin D levels. These findings may provide some reasons for vitamin D deficiency previously reported in these populations. Thus, researchers recommend that these attitudes of excessive sun protection and limiting sun exposure be further investigated as they may have implications for planning and delivery of health promotion programs to this growing population of immigrants in Australia.

Hypertension, pulse, and other cardiovascular risk factors and vitamin D status in Finnish men.

BACKGROUND: Debate exists about the relationship between hypovitaminosis D and cardiovascular (CVD) risk. METHODS: This study investigated baseline (n=2,271) 25-hydroxyvitamin D (25(OH)D) and baseline and 4 year (n=1,957) CVD risk in a cohort of Finnish middle-aged male smokers. RESULTS: The prevalences of measured hypertension, high pulse rate, diabetes, and coronary heart disease were 63%, 16%, 5%, and 10% at baseline and were 64%, 20%, 6%, and 16% at 4 years after baseline. The mean 25(OH)D was 41±18 nmol/L. At baseline, systolic blood pressure (ß = -0.048; P = 0.02), and pulse rate (ß = -0.043; P = 0.04) were both associated with lower 25(OH)D levels but not coronary heart disease or diabetes prevalence. On remeasuring CVD risk 4 years after baseline, the only significant association with baseline 25(OH)D levels was high pulse rate (ß = -0.077; P = 0.001). In addition, a higher 25(OH)D level at baseline was associated with a change in pulse rate (ß = -0.055; P = 0.01). These trends for hypertension (baseline) and high pulse rate (baseline and 4 years after baseline) were also seen on adjusted categorical analysis (P trend < 0.05). CONCLUSIONS: Vitamin D deficiency at baseline was associated with hypertension in Finnish male smokers, but not after 4 years. These results are consistent with recent findings in other large cohort studies with measured blood pressure. Change in pulse rate over time continued to be significantly associated with lower 25(OH)D baseline levels; this new finding should be investigated further.