Associations between bone and energy metabolism in cows fed diets differing in level of dietary cation-anion difference and supplemented with cholecalciferol or calcidiol.

Bone-derived hormones play an important role in metabolism. This study examined the hypothesis that interactions between bone and energy metabolism, particularly those involving osteocalcin, are present in dairy cattle and have feedback mechanisms over time. Associations between metabolites in blood were examined in 32 Holstein cows blocked by parity and milk yield and randomly allocated to diets containing either 0.27 mg/kg dry matter (DM) calcidiol or cholecalciferol for an anticipated intake of 3 mg/d (120,000 IU/d) at 11 kg of DM, and positive (+130 mEq/kg DM) or negative (-130 mEq/kg DM) dietary cation-anion difference (DCAD) from 252 d of gestation to calving. Blood was sampled every 3 d, from 9 d prepartum to 30 d postpartum, and plasma concentrations of vitamin D3, 25-hydroxyvitamin D3, adiponectin, C-telopeptide of type 1 collagen (CTX1), glucose, insulin-like growth factor 1 (IGF1), insulin, undercarboxylated osteocalcin (uOC), and carboxylated osteocalcin (cOC) were determined. Feeding calcidiol compared with cholecalciferol increased plasma concentrations of 25-hydroxyvitamin D3 pre- (264.2 ± 8.0 vs. 61.3 ± 8.0 ng/mL) and postpartum (170.8 ± 6.2 vs. 51.3 ± 6.2 ng/mL) but decreased concentrations of vitamin D3 pre- (1.2 ± 0.6 vs. 14.5 ± 0.6 ng/mL) and postpartum (1.9 ± 0.4 vs. 3.2 ± 0.6 ng/mL). Prepartum, cows fed the negative DCAD diet had reduced concentrations of vitamin D3 and glucose compared with cows fed a positive DCAD. The combination of negative DCAD and cholecalciferol reduced IGF1 concentrations prepartum. The DCAD treatment had no effect on postpartum concentrations of metabolites. Nulliparous cows had increased concentrations of OC, CTX1, IGF1, glucose, and insulin compared with parous cows. Time series analysis identified associations between metabolites on the same day and over 3-d lags up to ±9 d that suggest feedback between 25-hydroxyvitamin D3 and vitamin D3 in the negative lags, indicating that 25-hydroxyvitamin D3 may exert feedback on vitamin D3 but not vice versa. We found evidence of a feedback mechanism between vitamin D3 and IGF1, with positive effect size (ES) on the same day and 3 d later, and negative ES 9 d later, that was more evident in cholecalciferol-fed cows. This suggests an important role of IGF1 in integrating bone metabolism with energy and protein metabolic pathways. Evidence of feedback was found between uOC and particularly cOC with IGF1, with positive ES on the same day but negative ES 6 d before and 6 d after. An association between uOC or cOC and IGF1 has not been previously identified in cattle and suggests that both uOC and cOC may have marked biological activity. Associations between OC and insulin identified in mice were not observed herein, although associations between OC and glucose were similar to those between IGF1 and glucose, supporting associations between glucose, OC, and IGF1. We provide further statistical evidence of crosstalk between vitamin D compounds, bone hormones, and energy metabolism in cattle. In particular, associations between uOC or cOC and IGF1 may provide links between prepartum diets and observations of prolonged increases in milk production and allow better control of peripartum metabolism.

Sunlight exposure is just one of the factors which influence vitamin D status.

Studies on the determinants of vitamin D status have tended to concentrate on input - exposure to ultraviolet B radiation and the limited sources in food. Yet, vitamin D status, determined by circulating concentrations of 25-hydroxyvitamin D (25(OH)D), can vary quite markedly in groups of people with apparently similar inputs of vitamin D. There are small effects of polymorphisms in the genes for key proteins involved in vitamin D production and metabolism, including 7-dehydrocholesterol reductase, which converts 7-dehydrocholesterol, the precursor of vitamin D, to cholesterol, CYP2R1, the main 25-hydroxylase of vitamin D, GC, coding for the vitamin D binding protein which transports 25(OH)D and other metabolites in blood and CYP24A1, which 24-hydroxylates both 25(OH)D and the hormone, 1,25-dihydroxyvitamin D. 25(OH)D has a highly variable half-life in blood. There is evidence that the half-life of 25(OH)D is affected by calcium intake and some therapeutic agents. Fat tissue seems to serve as a sink for the parent vitamin D, which is released mainly when there are reductions in adiposity. Some evidence is presented to support the proposal that skeletal muscle provides a substantial site of sequestration of 25(OH)D, protecting this metabolite from degradation by the liver, which may help to explain why exercise, not just outdoors, is usually associated with better vitamin D status.

Carryover effects of potassium supplementation on calcium homeostasis in dairy cows at parturition.

The purpose of this study was to test whether supplementation with K improves bone mineral density (BMD) in older cows so that by parturition their bone is better able to mobilize Ca. Twenty-four Holstein Friesian cows (6 mo pregnant, lactating, and in their third or later lactation) were allocated to 2 equal groups and individually fed twice daily a total diet comprising low K oaten hay plus a pelleted concentrate fortified with or without K(2)CO(3) to achieve 3.12% K/kg of DM in the total diet of the K-supplemented (KS) cows compared with 1.50% K/kg of DM for the control cows. The cows were fed their respective diets from the beginning of their sixth month of pregnancy until 2 wk before the expected date of parturition. The strategy was to use K to stimulate a mild increase in extracellular pH to potentially improve BMD well before parturition, when high K contents in the diet are considered safe, but cease supplementing in the few weeks prepartum, when high intakes of K are known to be problematic. The expectation was that the effect of the denser bone would carry through to benefit the cow's plasma Ca, P, and Mg status at parturition. Prior to the period of K supplementation, the cows were part of a commercial pasture-based herd, to which they were returned at the end of the supplementation period and treated as 1 group from at least 11 d prepartum until the end of the study at d 42 of the next lactation. Supplementation with K successfully induced a sustained increase of urinary pH throughout late lactation and into the dry period, as expected. The KS cows consistently averaged a urine pH 0.25+/-0.10 U higher than the controls. However, there was no significant effect of K supplementation on BMD, bone mineral concentrations, plasma osteocalcin, urinary deoxypyridinoline:creatinine plasma Ca, or plasma P concentrations during or immediately after the cessation of supplementation, nor where there any carryover effects during parturition or by d 42 of lactation. Instead, there was an unexpected decrease in the concentration of Mg in plasma of the KS cows compared with the control cows that extended from 0.5 to 2.5 d postpartum. The timing of the decline in plasma Mg was paralleled by declines in plasma concentrations of 1,25 dihydroxy-vitamin D(3) and urinary excretion of Ca and Mg, whereas urinary excretion of P increased; all changes were consistent with a hypomagnesemia that could increase the risk of hypocalcemia. These data suggest that, in addition to the well-documented negative effects of K when fed immediately at parturition, the effects of high dietary K diets can carry over for at least 11 d to trigger a mild hypomagnesemia at parturition. Because K supplementation did not improve BMD prepartum, it was not possible to conclude for or against an ability of denser bone to reduce the risk of hypocalcemia in older cows at parturition.

Relationship between vitamin D status, body composition and physical exercise of adolescent girls in Beijing.

UNLABELLED: Little is known about the prevalence of actual vitamin D deficiency in healthy school-aged adolescents, particularly in China. The aim of this study was to examine the prevalence of hypovitaminosis D and to identify whether there was any association between vitamin D status, body composition and physical exercise in 323 Chinese adolescent girls in Beijing, China (40 degrees N). INTRODUCTION: It is well recognized that persistent severe vitamin D deficiency is associated with the bone abnormalities of rickets and osteomalacia. However, there is now evidence suggesting that low vitamin D status, not previously considered to be a state of deficiency is associated with secondary hyperparathyroidism, increased bone remodelling and other clinical signs thought only to be found in severe vitamin D deficiency. Hypovitaminosis D in healthy children and adolescents has been reported frequently in many countries, especially in winter. METHODS: We performed a cross-sectional analysis of 323 Chinese adolescent girls in Beijing in winter. Mean age of the subjects was 15.0 (+/-0.4) years. About 32.8%, 68.4% and 89.2% of the subjects were at risk of vitamin D deficiency when defined as plasma concentrations of 25OHD of 25, 37.5 or 50 nmol/L, respectively. RESULTS: This cross-sectional analysis of 323 Chinese adolescent girls in Beijing in winter showed that hypovitaminosis D was common in these subjects. In addition, body mass index, milk intake, participation in organized sports and total physical activity were all significant independent determinants of vitamin D status. An inverse association was found between plasma 25OHD and intact-parathyroid hormone (iPTH) concentration. Body mass index (BMI), milk intake, participation in organized sports and total physical activity all emerged as major independent determinants of vitamin D status as assessed by plasma 25OHD concentration. Vitamin D status was positively associated with lean body mass (LBM), but there was no association with the degree of body adiposity. Regardless of the concentration of 25OHD in blood used to define vitamin D deficiency, hypovitaminosis D was common in these subjects. CONCLUSION: It is recommended that policies be developed to prevent vitamin D deficiency in adolescent girls. Further studies are needed to identify the mechanisms whereby vitamin D status is related to exercise and to body composition during growth.

Why did the dinosaurs become extinct? Could cholecalciferol (vitamin D3) deficiency be the answer?

Palaeontological deductions from the fossil remnants of extinct dinosaurs tell us much about their classification into species as well as about their physiological and behavioural characteristics. Geological evidence indicates that dinosaurs became extinct at the boundary between the Cretaceous and Paleogene eras, about 66 million years ago, at a time when there was worldwide environmental change resulting from the impact of a large celestial object with the Earth and/or from vast volcanic eruptions. However, apart from the presumption that climate change and interference with food supply contributed to their extinction, no biological mechanism has been suggested to explain why such a diverse range of terrestrial vertebrates ceased to exist. One of perhaps several contributing mechanisms comes by extrapolating from the physiology of the avian descendants of dinosaurs. This raises the possibility that cholecalciferol (vitamin D3) deficiency of developing embryos in dinosaur eggs could have caused their death before hatching, thus extinguishing the entire family of dinosaurs through failure to reproduce.

Career paths of alumni of the Cornell Leadership Program for veterinary students.

The Cornell Leadership Program at Cornell University, usa, aims to assist talented veterinary students to embark on careers in research, academia, government agencies or industry. Over 400 students have participated since the Program began in 1990 and their subsequent careers have been followed. In this study, five sources of data were analysed: application documents of the participants; audio recordings of interviews with each participant from 2000 to 2007; annual tracking records of alumni after graduating with a veterinary degree; spontaneous comments from alumni about how the Program influenced their career plans; and a list of published scientific papers by alumni. Analysis revealed that about 50 per cent of veterinary graduates were establishing themselves in careers envisaged by the Program, although many of them experienced conflicts between a vocational commitment to clinical practice and a desire to solve problems through research. Many alumni asserted that the Program had influenced their career plans, but they had difficulty in accepting that rigorous scientific training was more important in acquiring research skills than working directly on a veterinary research problem. One career of great appeal to alumni was that of veterinary translational science, in which disease mechanisms are defined through fundamental research. It is concluded from the data that there are three challenging concepts for recently qualified veterinarians aiming to advance the knowledge of animal disease: research careers are satisfying and rewarding for veterinarians; a deep understanding of the chosen field of research is needed; and a high standard of scientific training is required to become an effective veterinary scientist.

1,25-Dihydroxycholecalciferol (calcitriol) modifies uptake and release of 25-hydroxycholecalciferol in skeletal muscle cells in culture.

The major circulating metabolite of vitamin D3, 25-hydroxycholecalciferol [25(OH)D], has a remarkably long half-life in blood for a (seco)steroid. Data from our studies and others are consistent with the hypothesis that there is a role for skeletal muscle in the maintenance of vitamin D status. Muscle cells internalise vitamin D-binding protein (DBP) from the circulation by means of a megalin/cubilin plasma membrane transport mechanism. The internalised DBP molecules then bind to actin and thus provide an intracellular array of high affinity binding sites for its specific ligand, 25(OH)D. There is evidence that the residence time for DBP in muscle cells is short and that it undergoes proteolytic degradation, releasing bound 25(OH)D. The processes of internalisation of DBP and its intracellular residence time, bound to actin, appear to be regulated. To explore whether 1,25-dihydroxycholecalciferol (calcitriol) has any effect on this process, cell cultures of myotubes and primary skeletal muscle fibers were incubated in a medium containing 10-10M calcitriol but with no added DBP. After 3h pre-incubation with calcitriol, the net uptake of 25(OH)D by these calcitriol-treated cells over a further 4h was significantly greater than that in vehicle-treated control cells. This was accompanied by a significant increase in intracellular DBP protein. However, after 16h of pre-incubation with calcitriol, the muscle cells showed a significantly depressed ability to accumulate 25(OH)D compared to control cells over a further 4 or 16hours. These effects of pre-incubation with calcitriol were abolished in fibers from VDR-knockout mice. The effect was also abolished by the addition of 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid (DIDS), which inhibits chloride channel opening. Incubation of C2 myotubes with calcitriol also significantly reduced retention of previously accumulated 25(OH)D after 4 or 8h. It is concluded from these in vitro studies that calcitriol can modify the DBP-dependent uptake and release of 25(OH)D by skeletal muscle cells in a manner that suggests some inducible change in the function of these cells.

Vitamin D supply to the rat fetus and neonate.

The prevention of neonatal rickets by oral supplementation with vitamin D2 (ergocalciferol) has tended to obscure our ignorance of the natural mechanism by which young mammals receive an adequate supply of vitamin D. To investigate the possibility of specific intrauterine transfer and storage of vitamin D in fetal tissues, vitamin D-deficient female rats were given depot injections of 3H- or 14C-labeled vitamin D3 (cholecalciferol) before mating and the 3H-labeled animals were killed at stages during the last third of gestation. Analysis of lipid extracts from whole fetuses revealed a linear increase in the concentration of 25-hydroxyvitamin D3, 24,25-dihydroxyvitamin D3, and D3 itself between days 14 and 19 of gestation. During this period the elimination half-time of 3H-labeled molecules in maternal plasma fell from 27.1 to 4.4 d, suggesting that a specific mechanism was transferring vitamin D molecules into the fetuses. The vitamin was stored predominantly as 25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3, with the highest concentrations in fetal muscle. Immediately after birth, pups from 3H- and 14C-labeled mothers were exchanged and later killed after 1-3 wk of suckling. Analysis of total lipid extracts for 3H and 14C content determined the relative contributions of vitamin D supplied before birth via the placenta and after birth in the maternal milk. The vitamin D content of the rat milk was relatively high, between 1.0 and 3.5 micrograms/liter. Nevertheless, the supply of vitamin D in utero, rather than from milk, was the main determinant of vitamin D status in early neonatal life. This is the first indication in a mammal of a specific transfer mechanism that allows the fetus to accumulate vitamin D from the mother during the last third of gestation.

Vitamin D plasma binding protein. Turnover and fate in the rabbit.

The metabolic disposition of the plasma binding protein (DBP) for vitamin D and its metabolites was studied in adult rabbits. Apo-DBP was purified from rabbit plasma and enzymatically labeled with radioiodine. The radioiodine-labeled protein retained its ability to bind vitamin D sterols and its physicochemical properties. When 125I-labeled DBP and 131I-labeled rabbit albumin were simultaneously injected intravenously, the 125I was cleared from plasma at a faster rate (t 1/2 = 1.7 d) than 131I (t 1/2 = 5 d) and 125I was present in excess of 131I in kidney, liver, skeletal muscle, heart, lung, intestine, testis, and bone 1 h after injection. In contrast to DBP, 25(OH)D3 was cleared more slowly (t 1/2 = 10.7 d). Compared to albumin, DBP radioactivity appeared earlier and in greater quantity in the urine of catheterized rabbits. Gel filtration analyses of plasma revealed most of the 125I to elute in the position of DBP, with only small amounts in the less than 1,000-dalton region. In contrast, almost all of the urine 125I eluted in this small molecular weight fraction. The molar ratio of DBP to 25(OH)D3 in normal rabbit plasma was 138/1. The extravascular pool of DBP was calculated to be 1.5-2.4 times larger than the intravascular DBP pool, and the molar replacement rate of DBP was 1,350-fold higher than that of 25(OH)D3. The plasma disappearance curves of holo-DBP, prepared either by saturating with 25(OH)D3 or by covalently linking 3 beta-bromoacetoxy-25(OH)D3, were very similar to that of apo-DBP. Neuraminidase treatment of DBP did not alter its plasma survival. These studies indicate that DBP or DBP-25(OH)D3 complex is removed from plasma by a variety of tissues, that the DBP moiety is degraded during this process, and that a significant recirculation of 25(OH)D3 probably occurs. The molar excess of DBP to 25(OH)D3 in plasma, and the relatively rapid turnover of DBP indicate that a high capacity, high affinity, and dynamic transport mechanism for vitamin D sterols exists in rabbit plasma.

Effects of diet and exercise on plasma vitamin D (25(OH)D) levels in Vietnamese immigrant elderly in Sydney, Australia.

Vitamin D deficiency may be associated with osteoporosis and fractures in the elderly. In Australia where there is a sizeable Vietnamese population, research has not yet clarified the roles of diet, exercise and sun exposure in determining vitamin D status. Plasma samples for 25-hydroxy-vitamin D (25(OH)D); dietary intake of vitamin D and calcium; muscle strength and sun exposure were measured and weekly dairy intake, exercise levels and smoking habits were surveyed in free-living elderly of Vietnamese and Australian/British origin. There was marginal vitamin D deficiency (<37 nmol/L 25(OH)D) in 63% of Vietnamese but only in 37% of Australian/British born. Low dairy intake and no vigorous exercise were best predictors of vitamin D deficiency in Vietnamese, taking into account age, gender, dietary intake and sun exposure. Since these migrant elderly may not get adequate sun exposure due to either clothing customs or cultural norms that encourage fair (untanned) skin, it is important to encourage increased exercise and dairy intake.

The effect of parathyroid hormone on the uptake and retention of 25-hydroxyvitamin D in skeletal muscle cells.

Data from our studies, and those of others, support the proposal that there is a role for skeletal muscle in the maintenance of vitamin D status. We demonstrated that skeletal muscle is able to internalise extracellular vitamin D binding protein, which then binds to actin in the cytoplasm, to provide high affinity binding sites which accumulate 25-hydroxyvitamin D3 (25(OH)D3) [1]. This study investigated the concentration- and time-dependent effects of parathyroid hormone (PTH) on the capacity of muscle cells to take up and release 3H-25(OH)D3. Uptake and retention studies for 3H-25(OH)D3 were carried out with C2C12 cells differentiated into myotubes and with primary mouse muscle fibers as described [1]. The presence of PTH receptors on mouse muscle fibers was demonstrated by immunohistochemistry and PTH receptors were detected in differentiated myotubes, but not myoblasts, and on muscle fibers by Western blot. Addition of low concentrations of vitamin D binding protein to the incubation media did not alter uptake of 25(OH)D3. Pre-incubation of C2 myotubes or primary mouse muscle fibers with PTH (0.1 to 100 pM) for 3h resulted in a concentration-dependent decrease in 25(OH)D3 uptake after 4 or 16h. These effects were significant at 0.1 or 1pM PTH (p<0.001) and plateaued at 10pM, with 25(OH)D3 uptake reduced by over 60% (p<0.001) in both cell types. In C2 myotubes, retention of 25(OH)D3 was decreased after addition of PTH (0.1 to 100pM) in a concentration-dependent manner by up to 80% (p<0.001) compared to non-PTH treated-C2 myotubes. These data show that muscle uptake and retention of 25(OH)D3 are modulated by PTH, a physiological regulator of mineral homeostasis, but the cell culture model may not be a comprehensive reflection of vitamin D homeostatic mechanisms in whole animals.

The origin and metabolism of vitamin D in rainbow trout.

An explanation for the origin and the high concentration of vitamin D (cholecalciferol) in some species of fish is still not apparent. Because fish may live in deep water and may, thus, not be exposed to solar ultraviolet (UV) light, it is commonly assumed that vitamin D found in their livers and adipose tissue has been derived from a food chain, originating in zooplankton exposed to UV light at the water surface. To investigate the metabolism and possible origin of vitamin D in fish, rainbow trout were reared from eggs, in the absence of light, and were fed a vitamin D-free diet. When small quantities of radioactively-labelled vitamin D were injected or fed to these trout, much of the radioactivity was found as excreted metabolites in bile. Hence, even when they are vitamin D deficient, trout vigorously catabolise and excrete exogenous vitamin D. The main vitamin D metabolite found in plasma of non-deficient trout was 1,25-dihydroxycholecalciferol [1,25(OH)2D3]. This was produced in the liver by an enzyme process that was strongly stimulated in vitamin D deficiency. When vitamin D was fed for several weeks to vitamin D-deficient trout, plasma 1,25(OH)2D3 levels rose to 180 pg/ml and the fish became hypercacemic. When vitamin D-deficient fish were inadvertently exposed to 60 W incandescent light for 24h, they became moribund and died. It was subsequently found that vitamin D-deficient trout can produce vitamin D in skin when exposed to blue light at wavelengths between 380 and 480 nm. It is concluded that trout, like terrestrial vertebrates, produce 1,25(OH)2D3 as the functional form of vitamin D and that this has an effect on calcium homeostasis. Furthermore, vitamin D is formed in the skin of these fish by the photochemical action of visible light on 7-dehydrocholesterol. Elucidation of the physicochemical mechanism of this process requires further research.

Reprint of "The origin and metabolism of vitamin D in rainbow trout".

An explanation for the origin and the high concentration of vitamin D (cholecalciferol) in some species of fish is still not apparent. Because fish may live in deep water and may, thus, not be exposed to solar ultraviolet (UV) light, it is commonly assumed that vitamin D found in their livers and adipose tissue has been derived from a food chain, originating in zooplankton exposed to UV light at the water surface. To investigate the metabolism and possible origin of vitamin D in fish, rainbow trout were reared from eggs, in the absence of light, and were fed a vitamin D-free diet. When small quantities of radioactively-labelled vitamin D were injected or fed to these trout, much of the radioactivity was found as excreted metabolites in bile. Hence, even when they are vitamin D deficient, trout vigorously catabolise and excrete exogenous vitamin D. The main vitamin D metabolite found in plasma of non-deficient trout was 1,25-dihydroxycholecalciferol [1,25(OH)2D3]. This was produced in the liver by an enzyme process that was strongly stimulated in vitamin D deficiency. When vitamin D was fed for several weeks to vitamin D-deficient trout, plasma 1,25(OH)2D3 levels rose to 180pg/ml and the fish became hypercalcemic. When vitamin D-deficient fish were inadvertently exposed to 60W incandescent light for 24h, they became moribund and died. It was subsequently found that vitamin D-deficient trout can produce vitamin D in skin when exposed to blue light at wavelengths between 380 and 480nm. It is concluded that trout, like terrestrial vertebrates, produce 1,25(OH)2D3 as the functional form of vitamin D and that this has an effect on calcium homeostasis. Furthermore, vitamin D is formed in the skin of these fish by the photochemical action of visible light on 7-dehydrocholesterol. Elucidation of the physicochemical mechanism of this process requires further research.

Vitamin D deficiency and associated factors in adolescent girls in Beijing.

BACKGROUND: Several locally published reports indicate a high prevalence of vitamin D deficiency among adolescents in China, but no systematic population-based survey has been conducted. OBJECTIVE: The objective was to determine the prevalence of vitamin D deficiency and to study associated factors in adolescent girls in Beijing. DESIGN: A cross-sectional study was conducted in a random sample of 1248 Beijing girls aged 12-14 y. Nutrient intakes, ultraviolet light exposure, anthropometric characteristics, physical activity, signs and symptoms of rickets, and plasma concentrations of 25-hydroxyvitamin D, 1,25-dihydroxyvitamin D, and calcium were measured and X-rays of the hand and wrist were taken. RESULTS: The prevalence of clinical vitamin D and calcium deficiency (plasma 25-hydroxyvitamin D <12.5 nmol/L, plasma calcium <2.25 mmol/L, and muscle spasm at least once per week) was 9.4% in winter. The prevalence of subclinical vitamin D deficiency (25-hydroxyvitamin D <12.5 nmol/L) was 45.2% in winter and 6.7% in summer (P < 0.0005). Logistic regression analysis showed that subclinical and clinical vitamin D deficiency in winter were associated with low plasma 25-hydroxyvitamin D concentrations (<12.5 nmol/L) in summer, low calcium intake ( x +/- SD: 280 +/- 48 compared with 440 +/- 61 mg/d), and low plasma calcium concentrations (<2.25 mmol/L) in winter. The odds ratios for these associations were 3.1, 1.5, and 1.5, respectively. CONCLUSIONS: Subclinical vitamin D deficiency was widespread among Beijing adolescent girls in winter. Low plasma 25-hydroxyvitamin D concentrations in summer, low calcium intake, and low plasma calcium concentrations in winter were the main risk factors for vitamin D deficiency in winter.

Milk consumption and bone mineral content in Chinese adolescent girls.

A cross-sectional study of a random sample of 649 girls, aged 12-14 years (mean +/- SD: 12.9 +/- 0.6 years), in the Beijing area examined the relationship between diet and bone mineral status. Food and nutrient intakes over the past year were estimated by means of a semiquantitative food frequency questionnaire. Bone mineral content (BMC) and bone width (BW) at the distal one-third and one-tenth radius and ulna were measured by single-photon absorptiometry. Results showed Beijing pubertal girls had a low mean milk consumption (fresh and powdered milk, vitamin D-fortified milk, and yogurt) at 50 g/day (95% confidence interval [CI] 44-55 g/day whereas one-third consumed no milk at all. Mean calcium intake was 356 +/- 97 mg/day of which only 21% was provided by milk and milk products. Milk intake varied by region (rural, suburban, and urban: 9, 36, and 83 g/day, respectively, p < 0.0005) as did the proportion of milk consumers in the three areas (30%, 64%, and 91%, p < 0.0005). Bone mineral density (BMD) at the distal one-third and one-tenth radius and one-tenth ulna was positively associated with milk consumption (p < 0.05). Multiple regression analysis of BMC on foods and nutrients as well as confounding factors, including weight, bone age, Tanner stage, and School Physical Activity Score (SPAS), showed that milk intake was the only dietary factor included in the models for BMC at the four bone sites measured. The model explained 54%-65% of the variation in BMC, and milk alone accounted for up to 3.2% of the variation. Milk was the only food group with significant partial correlation with BMC. SPAS, weight, bone age, and Tanner stage each accounted for a smaller variation in BMC (<1.8%). The results indicate that milk (presumably as an integrated source of nutrients) had a beneficial effect on bone mass of Beijing pubertal girls and was a better nutritional determinant of BMC than intake of any milk nutrient alone. Promotion of milk consumption should be considered for achieving optimal bone mass in this population group.

Low body weight and its association with bone health and pubertal maturation in Chinese girls.

OBJECTIVE: To investigate the prevalence of low body weight in Beijing pubertal girls and to establish the cut-off for body mass index (BMI) for underweight for Chinese pubertal girls. DESIGN: Cross-sectional study. SETTING: Three socioeconomic areas (rural, suburban and urban) in Beijing, China. SUBJECTS: Random sample of 1214 adolescent girls aged 12-14 y from 13 middle schools. RESULTS: Using a modified Chinese reference, the rate of low body weight (BMI<18) was 32.2% (95% CI 29.6-34.8%). Compared with desirable weight girls (BMI=18-21), girls with low body weight had a lower bone age, delayed breast and pubic hair development, a lower rate of menarche, lower distal one-third radius and ulna bone mineral content (BMC), bone mineral density and bone width. Logistic regression showed that BMI was one of the predictors of one-third ulna BMC after adjustment for confounding variables. When comparing BMI<18 vs BMI=18-21, the risk of BMC being less than the median increased by 82% (odds ratio 1.82, 95% CI 1.06-3.13). Thinness and stunting rates assessed by WHO recommended cut-offs are also reported. CONCLUSIONS: High prevalence of low body weight (BMI<18) was found to be a major health problem among Beijing pubertal girls. BMI<18 is confirmed as the cut-off for delayed general growth and development for Chinese girls and for screening girls at risk of lower bone mineral status.

Improvement of bone health in childhood and adolescence.

Osteoporosis as a worldwide problem is discussed in the present review and the question of improving peak bone mass to reduce the risk of osteoporosis and osteoporotic fracture is addressed. The available evidence points to pre-puberty and puberty as the most opportune periods for intervention, but the potential for achievable increments in bone mass is shown to be small compared with the overwhelming influence of heredity, body composition and hormonal factors on bone. Lean body mass appears to be positively correlated with bone mass, while black-white racial differences in bone mass appear to be related to greater lean mass and lower bone turnover rate in blacks. Within races, twin and parent-offspring models have suggested that 46-80 % of the variance in bone mineral density can be explained by inherited factors; however, the mechanism of the genetic influence on bone density remains poorly understood. Moderate regular exercise seems to maintain bone mass while more vigorous regular exercise increases it in children and young adults. Ca intake has been found to be positively associated with bone mass in many but not all studies, possibly because of a ceiling at about 1300-1500 mg/d for young people. Other nutritional variables, including vitamin D, have been little investigated in relation to childhood and adolescent bone mass. The influence of milk as a source of highly bioavailable Ca and other nutrients has also been less frequently investigated, which is of concern given the cessation of school milk programmes in Western countries over the last three decades. Intervention studies to improve bone health in young people have mainly been based on Ca milk or exercise. The evidence points to the benefits to bone of such interventions, particularly when commenced pre-puberty, and it seems that daily consumption of 200-300 ml milk/d by children and adolescents has no adverse side effects. The benefits to bone are almost universally shown to be lost fairly rapidly after Ca or exercise intervention ceases; there is therefore no justification in terms of bone health for short-term interventions of this nature. The question of withdrawal of milk supplementation has undergone very little examination. Further, very little evidence is available on the effects of long-term interventions of any sort on bone health. Nevertheless, the data obtained so far permit the suggestion that promotion of Ca intake (e.g. at the higher level of current recommendations) and exercise commencing in the pre-pubertal period should be adopted as policy now.

Seasonal vitamin D status of Greyhounds in Sydney.

OBJECTIVES: To survey the vitamin D status of a population of Greyhounds in New South Wales, and to establish a reference range for plasma 25(OH)D. To investigate whether any seasonal fluctuation in vitamin D status is detectable in these animals. DESIGN: Vitamin D status was assessed in Greyhounds and crossbred dogs presented to the University of Sydney for teaching purposes over a 24 month period. PROCEDURES: Plasma 25(OH)D concentration was measured as an estimate of vitamin D status. Physical examination and plasma calcium concentration were used to verify the health of the animals, particularly with respect to metabolic bone disease. RESULTS: A plasma 25(OH)D concentration range of 10 to 76 nmol/L was found in healthy adult Greyhounds. There was no sex- or season-dependent variation in vitamin D status in Greyhounds. Concentrations in crossbred dogs did not differ significantly from those in Greyhounds. CONCLUSION: The reference range for plasma 25(OH)D concentration in Greyhound dogs is similar to that previously reported for humans. It would seem that healthy dogs in the Sydney region do not exhibit a seasonal fluctuation in their vitamin D status.

The requirement for natural sunlight to prevent vitamin D deficiency in iguanian lizards.

To investigate possible causes of embryonic and neonatal mortality in a group of captive Fijian iguanas (Brachylophus fasciatus and Brachylophus vitiensis), the vitamin D status of adults in the colony was compared with that of agamid and iguanid lizards either housed in indoor enclosures under artificial ultraviolet light or exposed to natural sunlight (wild-caught or captive animals housed outdoors). Those under artificial lighting had a significantly lower vitamin D status than those housed exclusively outdoors, whereas the vitamin D status of Fijian iguanas that had received intermittent exposure to natural sunlight was intermediate and not significantly different from that of animals housed exclusively outdoors. However, eggs from some of these Fijian iguanas had substantially lower vitamin D content than eggs from outdoor iguanid and agamid animals. Artificial ultraviolet light, therefore, might not be an adequate substitute for natural sunlight to maintain vitamin D status of lizards. This possible inadequacy may be because either artificial ultraviolet light has a lower intensity of the wavelengths that induce vitamin D than does sunlight or the intensity of the artificial lighting is not sufficient.

Evidence for a specific uptake and retention mechanism for 25-hydroxyvitamin D (25OHD) in skeletal muscle cells.

Little is known about the mechanism for the prolonged residence time of 25-hydroxyvitamin D (25OHD) in blood. Several lines of evidence led us to propose that skeletal muscle could function as the site of an extravascular pool of 25OHD. In vitro studies investigated the capacity of differentiated C2 murine muscle cells to take up and release 25OHD, in comparison with other cell types and the involvement of the membrane protein megalin in these mechanisms. When C2 cells are differentiated into myotubes, the time-dependent uptake of labeled 25OHD is 2-3 times higher than in undifferentiated myoblasts or nonmuscle osteoblastic MG63 cells (P < .001). During in vitro release experiments (after 25OHD uptake), myotubes released only 32% ± 6% stored 25OHD after 4 hours, whereas this figure was 60% ± 2% for osteoblasts (P < .01). Using immunofluorescence, C2 myotubes and primary rat muscle fibers were, for the first time, shown to express megalin and cubilin, endocytotic receptors for the vitamin D binding protein (DBP), which binds nearly all 25OHD in the blood. DBP has a high affinity for actin in skeletal muscle. A time-dependent uptake of Alexafluor-488-labeled DBP into mature muscle cells was observed by confocal microscopy. Incubation of C2 myotubes (for 24 hours) with receptor-associated protein, a megalin inhibitor, led to a 40% decrease in 25OHD uptake (P < .01). These data support the proposal that 25OHD, after uptake into mature muscle cells, is held there by DBP, which has been internalized via membrane megalin and is retained by binding to actin.