Ontogeny and effect of vitamin D deprivation on rat serum 25-hydroxyvitamin D binding protein.

Specific serum binding of 25-hydroxy-cholecalciferol (25-OHD3) was measured by saturation analysis in rats of various ages, and during vitamin D deprivation. The serum binding capacity for 25-OHD3 was observed to increase until age 6-8 weeks, then decline and remain stable thereafter at 3.7 x 10(-6) M. The 25-hydroxyvitamin D (25-OHD) concentration decreased in serum from rats fed vitamin D-free diet (t1/2 = 7 days). Rats fed 2 IU of vitamin D3/g of diet maintained stable serum levels of 25-OHD at 10-12 ng/ml. Serum binding capacity and affinity for 25-OHD3 was not affected by vitamin D deprivation or hypocalcemia. In addition, the binding affinity did not differ as a function of age (Kd = 3.3 x 10(-9) M). Since normal serum concentrations of 25-OHD in the rat are 2-5 x 10(-8) M, only 1-2% of the serum binding sites for this sterol are occupied under physiological conditions.     

Serum 25-hydroxyvitamin D and vitamin D-binding protein levels and mineral metabolism after partial and total gastrectomy

The effects of gastrectomy, especially total gastrectomy, on the serum levels of 25-hydroxyvitamin D and vitamin D-binding protein and on mineral metabolism were examined. The serum 25-hydroxy-vitamin D levels were markedly decreased in patients with total gastrectomy and Billroth II gastrectomy. Decreased levels of serum vitamin D-binding protein and serum calcium, and increased levels of serum alkaline phosphatase were observed in both patients with partial gastrectomy and patients with total gastrectomy. The results show that vitamin D deficiency could develop in high frequency patients with total gastrectomy and Billroth II gastrectomy, and that deranged mineral metabolism could develop in patients with any type of gastrectomy with or without vitamin D deficiency. The decreased levels of serum vitamin D-binding protein in postgastrectomy patients may be a sensitive reflection of the failure of hepatic protein synthesis.     

25-hydroxyvitamin D levels and hypertension rates

Vitamin D deficiency has been linked to cardiovascular disease and risk factors including hypertension. The authors sought to determine prevalence rates of hypertension in adults tested for 25-hydroxyvitamin D categorized by their levels and evaluate odds ratios for hypertension at lower 25-hydroxyvitamin D levels compared with optimal levels. A cross-sectional study was conducted January 1, 2004, through December 31, 2006, of patients aged 18 years and older within a large ethnically diverse population. Diagnosis of hypertension was determined by International Statistical Classification of Diseases and Related Health Problems codes. Patients were categorized into quartiles according to 25-hydroxyvitamin D levels: ideal (≥40 ng/mL), adequate (30-39 ng/mL), deficient (15-29 ng/mL), and severely deficient (<15 ng/mL). Prevalence rates of hypertension and odds ratios were calculated for each 25-hydroxyvitamin D quartile, adjusting for age, sex, race, and renal insufficiency. A total of 2722 individuals met the inclusion criteria for the study. The overall prevalence of hypertension in the study population was 24%. Hypertension rates were 52%, 41%, 27%, and 20% in 25-hydroxyvitamin D quartiles <15 ng/mL, 15 to 29 ng/mL, 30 to 39 ng/mL, and ≥40 ng/mL, respectively (P<.001). Odds ratios (95% confidence intervals) for hypertension adjusting for age, sex, race, and renal insufficiency were 2.7 (1.4-5.2), 2.0 (1.5-2.6), and 1.3 (1.2-1.6) for 25-hydroxyvitamin D levels <15 ng/mL, 15 to 29 ng/mL, and 30 to 39 ng/mL, respectively, compared with the ≥40 ng/mL group. This study demonstrates increased rates of hypertension in individuals who tested for lower levels of 25-hydroxyvitamin D starting at levels <40 ng/mL. This retrospective analysis raises the question of whether supplementing to optimal vitamin D levels can prevent or improve hypertension.     

Relationships among vitamin D, 25-hydroxyvitamin D, and vitamin D-binding protein concentrations in the plasma and milk of human subjects

We measured plasma and milk concentrations of vitamin D2, vitamin D3, 25-hydroxyvitamin D2 (25OHD2), 25-hydroxyvitamin D3 (25OHD3), and vitamin D-binding protein (DBP) in a group of lactating women. All vitamin D compounds were quantitated using competitive protein binding assay, while DBP concentrations were determined by rocket electrophoresis. Vitamin D3 was the most abundant vitamin D compound in human milk, followed by vitamin D2, 25OHD3, and, finally, 25OHD2. The average vitamin D activity in milk was between 33-68 IU/liter, depending on the criterion of biological activity used. DBP concentrations in milk were approximately 3% of those in plasma. Significant relationships were found between plasma and milk levels for all vitamin D compounds. The milk to blood concentration ratio was greatest for vitamin D2, followed by vitamin D3, 25OHD2, and 25OHD3. (Thus, the parent compounds gained access into milk in a much more efficient fashion than their 25-hydroxy metabolites. It is postulated that this differential translocation is controlled by the DBP in the circulation.) There was no significant correlation between plasma and milk DBP concentrations, nor were milk DBP concentrations related to the vitamin D content of milk. This investigation supports the concept that the nutritional status of lactating mothers affects the vitamin D sterol potential of her milk which, in turn, would likely have an effect on the vitamin D status of her nursing infant.     

Clinical significance of competitive protein binding assay of 25-hydroxyvitamin D concentration

The level of serum 25-hydroxyvitamin D (25-OH-D3) was determined in 94 cases of healthy volunteers and 98 patients with various diseases. The mean level of the controls was 16.2 +/- 4.6 ng/ml (x /- S). The patients were divided into 5 groups, 1) Osteomalacia and rickets 42 cases, showing typical changes of bone in X-ray films. In 12 of them the disease was caused by vitamin D deficiency. The mean value of 25-OH-D3 was 3.3 +/- 2.0 ng/ml, being much lower than that of controls (P less than 0.001). In 14 cases of hypophosphatemia, 8 cases of renal tubular acidosis and 8 cases of renal insufficiency all complicated with osteomalacia the mean value of 25-OH-D3 was 17.9 +/- 11.4, 18.2 +/- 9.6 and 19.2 +/- 5.6 ng/ml (P greater than 0.05) respectively. 2) Intestinal malabsorption 18 cases. The mean level of 25-OH-D3 was 4.5 +/- 3.2 ng/ml (P less than 0.001). Some of them had hypocalcemia and/or secondary hyperparathyroidism. 2 showed osteoporosis and 1 osteomalacia. 3) Chronic liver disease 24 cases, the mean value of 25-OH-D3 was 6.2 +/- 5.6 ng/ml, being much lower than that of the controls (P less than 0.001). 4) 10 cases after taking anti-epileptic drugs had a mean level of 25-OH-C3 6.9 +/- 5.8 ng/ml (P less than 0.001). 1 had X-ray evidence of osteomalacia. 5) 4 cases of overdosage of vitamin D had a mean value of 110.3 +/- 22.5 ng/ml, being much higher than that of the controls (P less than 0.001). Competitive protein binding assay for measuring 25-OH-D3 is a simple and economic method. It is sensitive and specific as it provides distinct discrimination between healthy controls and patients with vitamin D deficiency or overdosage.     

Serum 25-hydroxyvitamin D levels are associated with prognosis in hematological malignancies

It has been proposed that Vitamin D has a significant influence on disease progression in malignancy. This study aims to investigate whether serum levels of 25-hydroxyvitamin D [25(OH)D] are associated with prognosis in patients with hematological malignancies. This study is based on 105 patients with hematological disease (acute and chronic leukemias, myelodysplastic syndromes, monoclonal gammapathies, and chronic lymphoid disorders), seen over a 6-months period. 25(OH)D deficiency (<20 ng/ml) appeared very common and an inverse relationship was observed between 25(OH)D levels and the response to therapy: lower levels being related to poorer response. In acute leukemias, a significant difference was noted between patients with long-term disease-free survival in those tested at diagnosis (P=0·001) or in those tested at the time of relapse (P=0·05). Similarly in patients with Philadelphia-positive leukemias, there was a correlation between molecular response and levels of 25(OH)D (P=0·01). Previously identified factors, such as age, season, gender, or nutritional index, were not related to circulating 25(OH)D levels. Lower levels of circulating 25(OH)D appeared related to a progressive stage of the disease and poor response to therapy, and, therefore, to the aggressiveness of the disease. It is a potential marker of prognosis in patients with leukemia.     

Serum 25-hydroxyvitamin D levels are associated with prognosis in hematological malignancies

Abstract

It has been proposed that Vitamin D has a significant influence on disease progression in malignancy. This study aims to investigate whether serum levels of 25-hydroxyvitamin D [25(OH)D] are associated with prognosis in patients with hematological malignancies. This study is based on 105 patients with hematological disease (acute and chronic leukemias, myelodysplastic syndromes, monoclonal gammapathies, and chronic lymphoid disorders), seen over a 6-months period. 25(OH)D deficiency (<20 ng/ml) appeared very common and an inverse relationship was observed between 25(OH)D levels and the response to therapy: lower levels being related to poorer response. In acute leukemias, a significant difference was noted between patients with long-term disease-free survival in those tested at diagnosis (P?=?0·001) or in those tested at the time of relapse (P?=?0·05). Similarly in patients with Philadelphia-positive leukemias, there was a correlation between molecular response and levels of 25(OH)D (P?=?0·01). Previously identified factors, such as age, season, gender, or nutritional index, were not related to circulating 25(OH)D levels. Lower levels of circulating 25(OH)D appeared related to a progressive stage of the disease and poor response to therapy, and, therefore, to the aggressiveness of the disease. It is a potential marker of prognosis in patients with leukemia.     

Free, and not total, 1,25-dihydroxyvitamin D regulates 25-hydroxyvitamin D metabolism by keratinocytes

Greater than 99% of the total circulating 1,25-dihydroxyvitamin D [1,25-(OH)2D] is bound to proteins such as the vitamin D-binding protein (DBP) and albumin; in the normal human only 0.4% of the circulating 1,25-(OH)2D is free. Although it is often assumed that only the free concentration of 1,25-(OH)2D is available to cells, this has not been demonstrated. In particular, it is not clear whether the DBPs facilitate 1,25-(OH)2D entry into target cells or serve only to transport these metabolites within the circulation. To address this question, we evaluated one of the best characterized target tissue responses to 1,25-(OH)2D, namely its ability to inhibit its own production and induce that of 24,25-(OH)2D, using one of the most sensitive cells, the human foreskin keratinocyte. We incubated keratinocytes in the presence of 1,25-(OH)2D (from 10(-11)-10(-8) M) in medium containing albumin (from 0.1-10%) or serum (from 0.1-10%) for 4 h [to inhibit the 25-hydroxyvitamin D (250HD) 1 alpha-hydroxylase] or 16 h (to induce the 250HD 24-hydroxylase) before evaluating [3H]250HD metabolism by these cells during a 1-h incubation in serum- and albumin-free medium. The free fraction of 1,25-(OH)2D was determined in each medium by centrifugal ultrafiltration and varied from 36% to 0.57% in direct proportion to the albumin or serum in the medium. Increasing the serum or albumin concentration in the medium increased the concentration of total 1,25-(OH)2D needed to inhibit its own production or stimulate that of 24,25-(OH)2D. In contrast, the concentration of free 1,25-(OH)2D needed to half-maximally inhibit its own production or induce 24,25-(OH)2D production remained constant at approximately 10(-11) M. We conclude that the free, not the total, 1,25-(OH)2D concentration regulates 250HD metabolism by keratinocytes, that DBPs do not facilitate 1,25-(OH)2D entry into the cell, and that these cells sense only free 1,25-(OH)2D.     

Vitamin D bioavailability: serum 25-hydroxyvitamin D levels in man after oral, subcutaneous, intramuscular, and intravenous vitamin D administration.

Bioavailability of vitamin D (D) was assessed by competitive protein-binding assay of serum 25-hydroxyvitamin D (25OHD) levels in normal adult volunteers after a single oral, sc, or im dose of commercially available D pharmaceuticals or after a single iv injection of ethanol-propylene glycol solution containing D. Similar increases in serum 25OHD levels were noted after either iv D2 or D3 (100 micrograms/kg), suggesting that the 25-hydroxylation of D2 and D3 is comparable in man. Absolute increases in serum 25OHD levels were similar in subjects receiving D in iv doses of 100 and 250 micrograms/kg; however, subjects receiving the larger dose had higher basal 25OHD levels. This finding suggests an inverse relationship in man between basal serum 25OHD concentrations and relative serum 25OHD increments after administration of pharmacological doses of D. Oil depot sc and im injection of D (200 micrograms/kg) resulted in delayed serum 25OHD increases compared to oral and iv dosing (100 micrograms/kg). Studies after im oil depot injection of [3H]D3 into rats showed that D administered in this manner had delayed bioavailability but remained unaltered in situ. Differences in D pharmaceutical bioavailability should be considered in treatment or prophylaxis with this sterol.     

Plasma 25-hydroxyvitamin D levels and risk of incident hypertension

Hydroxylation of 25(OH)D to 1,25-dihydroxyvitamin D and signaling through the vitamin D receptor occur in various tissues not traditionally involved in calcium homeostasis. Laboratory studies indicate that 1,25-dihydroxyvitamin D suppresses renin expression and vascular smooth muscle cell proliferation; clinical studies demonstrate an inverse association between ultraviolet radiation, a surrogate marker for vitamin D synthesis, and blood pressure. We prospectively studied the independent association between measured plasma 25-hydroxyvitamin D [25(OH)D] levels and risk of incident hypertension and also the association between predicted plasma 25(OH)D levels and risk of incident hypertension. Two prospective cohort studies including 613 men from the Health Professionals' Follow-Up Study and 1198 women from the Nurses' Health Study with measured 25(OH)D levels were followed for 4 to 8 years. In addition, 2 prospective cohort studies including 38 388 men and 77 531 women with predicted 25(OH)D levels were followed for 16 to 18 years. During 4 years of follow-up, the multivariable relative risk of incident hypertension among men whose measured plasma 25(OH)D levels were <15 ng/mL (ie, vitamin D deficiency) compared with those whose levels were >or=30 ng/mL was 6.13 (95% confidence interval [CI]: 1.00 to 37.8). Among women, the same comparison yielded a relative risk of 2.67 (95% CI: 1.05 to 6.79). The pooled relative risk combining men and women with measured 25(OH)D levels using the random-effects model was 3.18 (95% CI: 1.39 to 7.29). Using predicted 25(OH)D levels in the larger cohorts, the multivariable relative risks comparing the lowest to highest deciles were 2.31 (95% CI: 2.03 to 2.63) in men and 1.57 (95% CI: 1.44 to 1.72) in women. Plasma 25(OH)D levels are inversely associated with risk of incident hypertension.     

Vitamin D intake and blood 25-hydroxyvitamin D levels of adult Japanese

This paper is a review of studies on the vitamin D nutritional status of adult Japanese. The average dietary intake of vitamin D by adult Japanese, according to the National Nutrition Survey, is 352 IU/day. Older people have a higher intake than the average value and younger people have a lower intake than the average. Epidemiologic studies using blood 25-hydroxyvitamin D (25 [OH] D) as an index of vitamin D nutritional status have shown that elderly people with low activities-of-daily-living levels and young women have lower 25 (OH) D levels and a higher prevalence of vitamin D insufficiency than the rest of the population. The vitamin D nutritional status of these two high-risk groups should be improved by increasing their vitamin D intake in food or by means of supplements.     

Low serum 25-hydroxyvitamin D levels are associated with left ventricular hypertrophy in essential hypertension

Background and aimLow serum 25-hydroxyvitamin D [25(OH)D] levels may have an important role in predisposing to hypertension and myocardial disease. We investigated the relationship between 25(OH)D and left ventricular (LV) structure and function, assessed by echocardiography, in a series of patients with essential hypertension (EH).Methods and resultsSixty-two newly diagnosed never-treated patients (32 male/30 female), aged 18–65 years, with grade 1–2 hypertension, no diabetes, no obesity, no hyperlipidemia, and no cardiopulmonary, renal, or hepatic disease, were studied. Twenty-four healthy normotensive sex-, age-, BMI-matched subjects served as controls. Hypertensive patients with 25(OH)D deficiency, defined as serum 25(OH)D levels <50 nmol/L, had higher prevalence of LV hypertrophy (LVH) than their 25(OH)D-sufficient counterparts (57.1 vs 17.6%, P = 0.02); no differences between the two groups were found in blood pressure levels as well as in other biochemical and hormone parameters. There was an inverse correlation between LV mass index and 25(OH)D levels (r = –0.366, P < 0.003) and a direct correlation between LV mass index and BMI (r = 0.333, P < 0.006) in the entire hypertensive population. The two variables remained independently associated with LVH at multivariable logistic regression analysis (OR 1.05, P < 0.005 and OR 1.25, P = 0.03, respectively). Prevalence of 25(OH)D deficiency was similar in EH patients and controls (45.1 vs 41.6%, P = 0.89), whereas no correlation between echocardiographic parameters and hormone levels was found.ConclusionsIn the absence of major cardiovascular risk factors, 25(OH)D deficiency is a frequent finding in EH patients and is independently associated with LVH.     

Serum 25-hydroxyvitamin D levels are inversely associated with systemic inflammation in severe obese subjects

Obesity is frequently characterized by a reduced vitamin D bioavailability, as well as insulin-resistance and a chronic inflammatory response. We tested the hypothesis of an independent relationship between serum concentrations of 25-hydroxyvitamin D (25[OH]D) and several circulating inflammatory markers in a cohort of severely obese individuals. Cross-sectional study was carried out among obese patients undergoing a clinical evaluation before bariatric surgery in our University Hospital. Serum 25(OH)D, fasting and post load glucose and insulin, high-sensitive C-reactive protein (hs CRP), fibrinogen, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), leptin, adiponectin and lipid profile were collected. Insulin-resistance was assessed by insulin sensitivity index (ISI). Total body fat (FAT kg), total percent body fat (FAT%) and truncal fat mass (TrFAT) were assessed with dual-energy X-ray absorptiometry. A total of 147 obese subjects (89 women, 37.8 ± 7.1 years) with mean body mass index (BMI) of 43.6 ± 4.3 kg/m² were enrolled. Patients in the lowest tertile of 25(OH)D were significantly more obese with a higher amount of TrFAT, more insulin-resistant, and had higher levels of fasting and post-challenge glucose (p < 0.05 for all). In a multivariate regression analysis, serum 25(OH)D was inversely related to significant levels of hs CRP, IL-6 and TNF-α after accounting for age, gender, season of recruitment, BMI, FAT kg and TrFAT (p < 0.01 for all). In extremely obese subjects, 25(OH)D serum concentrations are inversely associated with several biomarkers of systemic inflammation, regardless of the total quantity of fat mass.     

Study on the relationship between serum 25-hydroxyvitamin D levels and vascular calcification in hemodialysis patients with consideration of seasonal variation in vitamin D levels

Background/aimsThe aim of this study was to determine the prevalence of vitamin D deficiency in hemodialysis (HD) patients and the relationship between seasonal variations in vitamin D levels and vascular calcification.MethodsAs a prospective observational study, we analyzed 289 HD patients. We have assessed serum 25-hydroxyvitamin D (25D) levels at the end of the summer (September) and winter (March) and analyzed the data to reveal the association of serum 25D level with vascular calcification scores (VCS) at the end of the summer, when vitamin D levels were found to peak. Plan X-ray images of lateral lumbar spine from all subjects were studied for calculation of semiquantitative VCS as described by Kauppila.ResultsThe prevalence of 25D deficiency was 86.2% at the end of the summer and increased to 96.2% at the end of the winter. Female gender and diabetes were associated with vitamin D deficiency. According to univariate analysis, 25D levels were inversely related to vascular calcification. However, after correcting for confounding factors, this relationship lost statistical significance. Multivariate analysis showed that age, systolic blood pressure, and LDL-cholesterol levels were directly associated with a higher VCS.ConclusionVitamin D deficiency was highly prevalent in HD patients with marked seasonal variation. However, low 25D levels could not be identified as an independent predictor of vascular calcification in these patients.     

Group-specific component (Gc) proteins bind vitamin D and 25-hydroxyvitamin D.

Group-specific component (Gc) proteins are human plasma proteins for which a worldwide polymorphism exists. As yet no functional role has been assigned this protein. We show that the products of both Gc alleles, proteins Gc 1 and Gc 2 (distinguished electrophoretically), bind substantial quantities of vitamin D and 25-hydroxyvitamin D. Three lines of evidence are reported: (1) Polyacrylamide gel electrophoresis and autoradiography of serum labeled with (14-C)vitamin D3 revealed patterns of radioactive bands identical to those expected of the two Gc alleles. Population gene frequencies for these proteins binding vitamin D were in the range of those reported for Gc, and individuals of known Gc phenotype were found to have the corresponding vitamin-D-binding phenotype. (2) Immunoelectrophoresis and autoradiography of labeled serum reacted against antiserum to human Gc revealed labeling by (14-C)vitamin D3 of Gc-antibody precipitation ares. (3) (14-C)vitamin D3 or 25-hydroxy(3-H)vitamin D3 was found to coprecipitate specifically with Gc in serum incubated with Gc antiserum. Use of these techniques demonstrated further that plasma proteins that bind vitamin D and that are immunologically similar to human Gc are found in mouse, rat, cow, horse, dog, rhesus monkey, and chimpanzee. We propose that Gc and "vitamin-D-binding alpha-globulin" are in fact the same portein, and that the ability of Gc to bind vitamin D may be directly related to the action of selection on this locus. These techniques may also find application in the study of other plasms transport proteins.     

Vitamin D binding protein,but not vitamin D or vitamin D-related peptides,is associated with septic shock mortality

Background

The aim of this study was to assess the prognostic value of vitamin D, vitamin D binding protein (VDBP) and vitamin D-related peptides in septic shock patients in relation to hospital mortality.

Assessment of the free fraction of 25-hydroxyvitamin D in serum and its regulation by albumin and the vitamin D-binding protein

We measured the free fraction of 25-hydroxyvitamin D (25OHD) in human serum and determined that 25OHD bound to a component with an affinity constant of 7 X 10(8) M-1 and a concentration of 4.5 X 10(-6) M. This concentration was equal to that of the vitamin D-binding protein (DBP) in the same serum sample. We removed DBP from the serum using actin affinity columns and found that the affinity for 25OHD of the remaining serum components was equivalent to that of human serum albumin (6 X 10(5) M-1). We then measured the free fractions of 25OHD, DBP, and albumin in normal and cirrhotic subjects. We calculated that 88 +/- 3% (+/- SD) and 83 +/- 8% of the 25OHD were bound to DBP in the serum of normal and cirrhotic subjects, respectively. We compared previously reported data for the free fraction and the free concentration of 1,25-dihydroxyvitamin D in these subjects with the current data for the free fraction and free concentration of 25OHD. The total concentrations and free fractions of both metabolites correlated to each other and to the DBP and albumin concentrations in these subjects, but the free concentrations of these metabolites did not. We conclude that 25OHD, like 1,25-dihydroxyvitamin D, is transported in blood bound primarily to DBP and albumin. Changes in the concentrations of DBP and albumin affected the total and free fractions of 25OHD in serum, but the actual free concentration of 25OHD was independent of such changes.