Free vitamin D levels in steroid-sensitive nephrotic syndrome and healthy controls.

INTRODUCTION: Body stores of vitamin D are measured as "total" serum 25-hydroxy vitamin D (25(OH)D). Its largest component is protein bound and lost in urine in nephrotic syndrome (NS). Our study investigates whether "free" 25(OH)D levels are a better guide to bone health and need for vitamin D supplementation in patients with steroid-sensitive NS (SSNS). METHODS: A cross-sectional study was performed in children with SSNS and healthy controls. Blood was tested for albumin, creatinine, calcium, phosphate, ALP, total and free (by direct ELISA) 25(OH)D, iPTH, and urine for protein-creatinine ratio. RESULTS: Seventy-nine NS patients (48 in relapse, 31 in remission) and 60 healthy controls were included. The levels of total 25(OH)D were significantly different (lowest in NS relapse and highest in controls) (p < 0.001). Corrected calcium and phosphate levels were normal, and there were no differences in free 25(OH)D, ALP, or iPTH levels between groups. Only total and not free 25(OH)D correlated significantly and negatively with urinary protein creatinine ratios (rs = - 0.42 vs. 0.04). Free 25(OH)D values of 3.75 and 2.85 pg/ml corresponded to total 25(OH)D levels of 20 and 12 ng/ml, respectively, in healthy controls. CONCLUSION: These results confirm that total 25(OH)D levels are low in NS and related to degree of proteinuria. However levels of free 25(OH)D, ALP, and iPTH did not change in relapse or remission in comparison with healthy controls. Our results suggest that in proteinuric renal diseases, free 25(OH)D rather than total 25(OH)D levels should be used to diagnose vitamin D deficiency and guide therapy.

Paricalcitol and Peritoneal Protein Loss in Peritoneal Dialysis: A Double-Center Study.

BACKGROUND/AIMS: Peritoneal protein loss (PPL) is associated with cardiovascular disease and mortality in peritoneal dialysis (PD). Controversial results have been published about the effect of paricalcitol in PPL among PD patients. This study intends to analyze the relationship between paricalcitol and PPL in PD. METHODS: In a retrospective study, prevalent PD patients were divided into 2 groups: "with paricalcitol" and "without paricalcitol". X2-test, Student's t test, Pearson correlation coefficient and Logistic Regression analysis were applied. RESULTS: Eighty-two patients were included. PPL was lower among patients medicated with paricalcitol (5.17 ± 1.71 vs. 6.79 ± 2.10 g/24 h, p = 0.0001). In multivariate analysis, paricalcitol and dialysate/plasma ratio of creatinine (D/P creatinine) were independently related to PPL (OR 4.270 [1.437-12.684], p = 0.009 and OR 0.205 [0.064-0.659], p = 0.008, respectively), adjusted for diabetes. CONCLUSION: Paricalcitol and D/P creatinine were independently related to PPL. Paricalcitol may have an effect on PPL in PD patients.

Bioavailability of vitamin D2 from enriched mushrooms in prediabetic adults: a randomized controlled trial.

BACKGROUND/OBJECTIVES: Based on the growing evidence of risk reduction from fresh fruit and vegetable consumption and an inverse relationship between serum 25-hydroxyvitamin D (25OHD) and the risk of type 2 diabetes (T2D), we determined the benefits of regularly consuming vitamin D-enriched mushrooms in a prediabetic cohort. Exposing edible mushrooms to ultraviolet B (UVB) light increases vitamin D2 (D2) and raises serum 25OHD2 in healthy young adults; however, their benefit to deficient prediabetics and glucose metabolism remains untested. SUBJECTS/METHODS: Forty-three prediabetic, D-deficient adults (25OHD≤20 ng/ml), BMI>25 were randomized to four groups consuming daily entrées containing 100 g fresh sliced cooked mushrooms prepared by a chef for 16 weeks. Two groups were fed UVB-treated mushrooms initially containing: 600 IU D2 or 4000 IU D2; each one also received one capsule of placebo daily. Two control groups were fed untreated mushrooms and D3 dietary supplements at two label doses: 600 IU D3 and 4000 IU D3. D2 and D3 content were analyzed in mushrooms, before and after cooking and in over-the-counter supplements. RESULTS: After 16 weeks, both D2-UVB-mushroom entrée doses, which were significantly lower after cooking, produced modest or no increases in 25OHD2 or total 25OHD relative to the positive control subjects who actually consumed about 1242 and 7320 IU per day of D3 (higher than stated on the label). CONCLUSIONS: Unanticipated D2 cooking loss from fresh UVB mushrooms and probable low absorption and/or hydroxylation may explain the smaller increase in 25OHD2 in our prediabetic overweight/obese cohort compared with past findings in younger, healthy subjects. Moreover, no dose or vitamin D source was associated with modifying T2D risk factors.

Differences of bone healing in metaphyseal defect fractures between osteoporotic and physiological bone in rats.

Discrepancies in bone healing between osteoporotic and non-osteoporotic bone remain uncertain. The focus of the current work is to evaluate potential healing discrepancies in a metaphyseal defect model in rat femora. Female Sprague-Dawley rats were either ovariectomized (OVX, n=14) and combined with a calcium-, phosphorus- and vitamin D3-, soy- and phytoestrogen-free diet or received SHAM operation with standard diet rat (SHAM, n=14). Three months post-ovariectomy, DEXA measurement showed a reduction of bone mineral density reflecting an osteoporotic bone status in OVX rats. Rats then underwent a 3 mm wedge-shaped osteotomy at the distal metaphyseal area of the left femur stabilized with a T-shaped mini-plate and allowed to heal for 6 weeks. Biomechanical competence by means of a non-destructive three-point bending test showed significant lower flexural rigidity in the OVX rats at 3 mm lever span compared to SHAM animals (p=0.048) but no differences at 10 mm lever span. Microcomputer tomography (µCT) showed bridging cortices and consolidation of the defect in both groups, however, no measurable differences were found in either total ossified tissue or vascular volume fraction. Furthermore, histology showed healing discrepancies that were characterized by cartilaginous remnant and more unmineralized tissue presence in the OVX rats compared to more mature consolidation appearance in the SHAM group. In summary, bone defect healing in metaphyseal bone slightly differs between osteoporotic and non-osteoporotic bone in the current 3 mm defect model in both 3mm lever span biomechanical testing and histology.

[Ovariectomy and calcium/vitamin D2/D3 deficient diet as a model of osteoporosis in the spine of Sprague-Dawley rats]. / Ovarektomie und Kalzium-/Vitamin D2/D3-arme Diät als Osteoporosemodell an der Wirbelsäule von Sprague-Dawley-Ratten.

BACKGROUND: Osteoporosis is a widespread disease characterised by low bone mass and structural deterioration of bone resulting in an increased susceptibility to fractures. Osteoporosis affects women more frequently than men; every second woman older than 50 years suffers from an osteoporotic fracture, frequently a vertebral fracture. The aim of this study was to induce osteoporosis in rats to establish an osteoporotic small-animal model that simulates the human pathology particularly in the spine. Therefore, bone density parameters, which are routinely determined in the spine of osteoporotic patients, were investigated by Dual-Energy X-ray Absorptiometry (DEXA). MATERIALS AND METHODS: Fourteen-week-old female Sprague-Dawley rats (n = 50) were either sham-operated (control group: sham) or ovariectomised (experimental group). Ovariectomised rats were further divided into two groups; one received calcium/vitamin D2/D3 deficient diet (OVX + diet), and the other received subcutaneous steroid injections (dexamethasone 0.3 mg/kg body weight) twice a month (OVX + steroid). Rats were scanned by DEXA at three time points (Month = M, 0 M, 1 M and 3 M). DEXA measurement of the spine delivered T-value, Z-value, bone mineral content (BMC), and the scanned area. Fifteen female patients at an age of 57-72 years were scanned in 8-10 regions of the spine (150 measurements). T-values and Z-values were pre-calculated based on patient databases. Statistical analysis was performed using two-way ANOVA followed by Bonferroni correction, with significance considered at p < 0.05. RESULTS: T-value and Z-value of both rat groups were compared with the patient data as well as with each others. Both treated rat groups revealed significantly lower T- and Z-values than controls. Despite the significant difference, the reference line (-2.5 for T-value and -1.5 for Z-value) was only reached by the OVX + diet group. On the other hand, the sham group showed an increase in BMC over time, while no change was seen in OVX + diet or OVX + steroid. Bone area demonstrated a significant increase up to M3. However, the increase in bone area within the OVX + diet group was notably higher than in both sham and OVX + steroid groups. Patients showed significantly lower T- and Z-values than sham and OVX + steroid but insignificant ones when compared with OVX + diet. CONCLUSION: A reproducible vertebral osteoporosis can be generated in a rat model by combination of ovariectomy with administration of a calcium/vitamin D3 deficient diet. T- and Z-values of this experimental group mimicked values obtained from osteoporotic patients, reflecting a simulation of their pathology. Interestingly, the increase in bone area over time with the steady BMC results in lower mineral density (BMD) of the OVX + diet group. Therefore, this rat model presents a reliable experimental set-up that may serve as a tool to better understand and treat osteoporosis.

Low 25OH vitamin D2 levels found in untreated Alzheimer's patients, compared to acetylcholinesterase-inhibitor treated and controls.

Following contradictory reports, the aim of this study was to apply our highly specific novel assay to delineate the relationship between vitamin D forms and Alzheimer's disease. The study incorporated patients, both untreated and treated with acetylcholinesterase inhibitors, along with controls. Patients were grouped as A: untreated (n=26) and B: treated with donepezil, rivastigmine or galantamine (n=44). The study included a control Group (C, n=35) with no cognitive impairment. Cognitive function was assessed using the MMSE. Levels of vitamin D forms were measured using liquid chromatography-mass spectrometry (LC-MS/MS) and calcium measurements were conducted using inductively coupled plasma-mass spectrometry (ICP-MS). In the cohort studied, no relationship was observed between MMSE score, calcium and any form of vitamin D. The indisputable finding is that the level of 25hydroxyvitamin D2 (25OHD2) (3.165 ± 6.352 nmol/L, p < 0.001) was significantly lower in the untreated Group (A) compared to the control and treated groups (7.932 ± 9.196 and 12.138 ± 15.682 nmol/L, respectively). In contrast, the levels of the primary forms, vitamin D2 and total vitamin D were the highest for the untreated group. Vitamin D levels, assessed as 25OHD are significantly lower in patients suffering from Alzheimer's disease arising from extremely low levels of 25OHD2 along with low levels of 25OHD3. Treatment with acetylcholinesterase inhibitors reverses this deficit. Further research is warranted to delineate the mode of action of acetylcholinesterase inhibitors with respect to normalising 25OHD2 levels. These observations resulted in the hypothesis that along with the common functions of vitamin D, different forms have distinct roles in health and disease.

Treatment of vitamin D deficiency: divergence between clinical practice and expert advice.

BACKGROUND: Current recommendations for the treatment of vitamin D deficiency vary from calciferol 800 IU per day to loading doses of vitamin D followed by maintenance therapy of up to 2000 IU per day. OBJECTIVE: To assess the preparations and doses of vitamin D used to load and maintain patients with serum 25-hydroxyvitamin D (25OHD) <25 nmol/l. METHODS: We examined all requests for serum 25OHD over a 12-month period, from September 2009 to 2010 in southwest Scotland. We wrote to all 33 general practices asking whether they usually started replacement therapy with a loading dose and/or recommended over-the-counter maintenance preparations. We accessed the Emergency Care Summary for all patients with serum 25OHD <25 nmol/l to determine whether they had been prescribed maintenance therapy. RESULTS: Serum 25OHD was requested in 1162 patients. Levels were <25 nmol/l in 282 (24%) patients, only 173 (61%) of whom were receiving vitamin D replacement therapy 3-15 months after diagnosis. Only four (1.4%) were prescribed a loading dose. One hundred and fifty-three (54%) were treated with cholecalciferol or ergocalciferol and 19 (7%) with alfacalcidol or calcitriol. The median dose of chole/ergocalciferol was 800 IU per day, usually in combination with 1200 mg calcium per day. CONCLUSIONS: We have shown a divergence between clinical practice and even the most conservative expert advice for vitamin D replacement therapy. Possible explanations are conflicting advice on treatment and difficulty obtaining suitable vitamin D preparations, particularly high dose vitamin D and vitamin D without calcium, in the UK.

Differences in outcomes between cholecalciferol and ergocalciferol supplementation in veterans with inflammatory bowel disease.

AIM: Vitamin D deficiency is a global health issue associated with increased health-care costs, and could play a role in the pathogenesis and management of inflammatory bowel disease. Prior studies show a high prevalence of vitamin D deficiency in veterans with inflammatory bowel disease. We aimed to examine the outcome differences in patients with inflammatory bowel disease, comparing treatment with ergocalciferol to cholecalciferol. METHODS: A retrospective review of electronic medical records of patients with inflammatory bowel disease at a Veterans Affairs Medical Facility in the Southeastern United States was carried out. Those with at least one serum 25(OH) vitamin D level were included. Initial and follow-up vitamin D values were recorded. The type of vitamin D supplementation, whether cholecalciferol or ergocalciferol, was documented. Costs in the year after measurement of vitamin D were divided into separate inpatient and outpatient categories. RESULTS: Veterans (n = 108) with ulcerative colitis or Crohn's disease and an available 25(OH) vitamin D level were studied. There were differences in follow-up vitamin D levels; those who received weekly ergocalciferol had higher subsequent levels than those who received cholecalciferol, especially at a second follow up, although differences did not achieve statistical significance. However, those who received vitamin D3 were less likely to use laboratory, pharmacy, radiology and fee-based services, and had lower laboratory and pharmacy costs. CONCLUSIONS: Our data suggest that cholecalciferol replacement might improve outcomes to a greater extent than ergocalciferol, and might be better in limiting health-care costs and expenses in patients with inflammatory bowel disease.

Vitamin D status and bone health in healthy Thai elderly women.

OBJECTIVE: The prevalence of hypovitaminosis D varies in different countries. Therefore, the current study was designed to assess vitamin D status and bone health in elderly women in Thailand, which is situated near the equator. METHODS: This cross-sectional study was performed in 446 healthy women aged 60-97 y. RESULTS: Serum 25-hydroxyvitamin D (25(OH)D) was 67.6 ± 15.7 (mean ± SD) nmol/L. Daily calcium intake was 309.5 ± 147.2 mg/d. Serum 25(OH)D levels tended to decline with bone mineral density (BMD) status. Based on functional health-based reference values, plasma-intact parathyroid hormone began to rise below serum 25(OH)D level 70 nmol/L and increase significantly when serum 25(OH)D was ≤ 60 nmol/L. Thirty-two percent of elderly women had 25(OH)D insufficiency (≤ 60 nmol/L). There was no trend toward a decrease in the concentration of serum 25(OH)D with age (r = -0.078, P = 0.10) and no significant inverse relationship with plasma intact parathyroid hormone values (r = -0.079, P = 0.097). However, a positive relationship was observed between serum 25(OH)D level and femoral neck BMD (r = 0.156, P = 0.001) but not lumbar spine L(2)-L(4) BMD (r = 0.093, P = 0.050). In addition, BMD at the femoral neck but not lumbar spine of the vitamin D insufficiency group was significantly lower than that of the vitamin D sufficiency group. CONCLUSION: The optimum level of serum 25(OH) value in Thai elderly women should be higher than 70 nmol/L. Vitamin D insufficiency is observed in one-third of elderly women in Bangkok.

Micronutrients at the interface between inflammation and infection--ascorbic acid and calciferol. Part 2: calciferol and the significance of nutrient supplements.

As explained in the first part of the article, vitamins and trace elements influence various metabolic functions that are directly related to immune function. In this context, secosteroid vitamin D has met with growing interest. The discussion has focused on whether and, if so, to what extent, vitamin D might contribute to the prevention and possibly the treatment of infections and autoimmune diseases. We know, for instance, that immune cells are capable of synthesizing calcitriol from its precursor calcidiol, whereby the former enhances the synthesis of antibacterial peptides by macrophages while simultaneously inhibiting the (auto)immune response mediated by T helper cells (Th1). Numerous observational studies support the hypothesis that a vitamin D deficit increases the risk of autoimmune diseases such as type 1 diabetes mellitus, multiple sclerosis, psoriasis and rheumatoid arthritis; however, there are few reliable interventional studies to date. In general, immune status represents a sensitive indicator of micronutrient supply. Conversely, the activity of the immune system has an effect on the status of and requirements for nutrients.

Significant vitamin D deficiency in youth with type 1 diabetes mellitus.

Serum 25-hydroxyvitamin D was measured in 128 youth with type 1 diabetes mellitus. Less than 25% of the patients were vitamin D sufficient. Because individuals with type 1 diabetes mellitus possess multiple risk factors for skeletal fragility, ensuring vitamin D sufficiency throughout childhood and adolescence in this population seems especially warranted.

Simplified method for the determination of 25-hydroxy and 1alpha,25-dihydroxy metabolites of vitamins D2 and D3 in human plasma. Application to nutritional studies.

A simplified method for the determination of 25-hydroxy and 1alpha,25-dihydroxy metabolites of vitamins D2 and D3 in human plasma was developed. Plasma samples were deproteinizated and applied to a Bond Elut C18OH cartridge to separate 25-hydroxyvitamin D (25-OH-D) and 1alpha,25-dihydroxyvitamin D [1,25(OH)2D] fractions. The 25-OH-D fraction was purified by a Bond Elut C18 cartridge and 25-OH-D2 and 25-OH-D3 were assayed by HPLC using a Zorbax SIL column. The 1,25(OH)2D fraction obtained above was subsequently applied to HPLC using a Zorbax SIL column to separate 1,25(OH)2D2 and 1,25(OH)2D3 fractions which were determined by a radioreceptor assay (RRA) using calf thymus receptor. The method was applied to nutritional studies.

Effect of iron on serum 25-hydroxyvitamin D and 24,25-dihydroxyvitamin D concentrations.

In 13 of 17 infants (aged 10.5 +/- 4.3; mean +/- SD mo) with iron-deficiency anemia, the serum 24,25-dihydroxyvitamin D concentration was below the normal range and in 9 of these 13 the serum 25-hydroxyvitamin D concentration was below the normal range despite the fact that these infants received 10 micrograms vitamin D/d from the age of 1 mo. The infants were treated with intramuscular iron dextran (Imferon). The iron-dextran treatment increased the hemoglobin and serum iron concentrations as well as 25-hydroxyvitamin D and 24,25-dihydroxyvitamin D concentrations. It is known that iron deficiency impairs fat and vitamin A intestinal absorption. Therefore, it is suggested that absorption of vitamin D may also be impaired. This may contribute to the development of vitamin D deficiency. Iron supplementation may have improved the absorption of vitamin D in the small intestine and hence increased the vitamin D concentration in the plasma.

Hearing-loss and calciferol deficiency.

Vitamin D (calciferol) deficiency has recently been claimed to cause cochlear hearing-loss (Brookes and Morrison, 1981; Brookes, 1983, 1984). In view of the therapeutic and pathophysiological implications of this finding, a confirmatory study was undertaken in the Department of Neuro-otology at The Royal National Throat, Nose and Ear Hospital. Screening of 112 consecutive new referrals revealed 32 patients with biochemical abnormalities compatible with vitamin D deficiency. Of these, 26 agreed to serum vitamin D assay and normal values were obtained in all of these cases. This study does not support the claim that vitamin D deficiency is a cause of hearing-loss.

[Effect of calciferol-protein deficiency on the metabolism of bone collagen]. / Vliianie kal'tsiferol-belkovoi nedostatochnosti na obmen kostnogo kollagena.

Growing rats received rations with separate and combined deficiencies of vitamin D, protein and certain essential amino acids, during 60 days. An increment was recorded in the total amount of collagen in the tubular bones at the expense of its mature, insoluble fraction, in the presence of a decreased content of salt- and acid-soluble fractions of this protein. These shifts were more manifest in combined calciferol-protein deficiency. Disorders in the correlation of collagen biosynthesis, maturation and degradation processes were responsible for these shifts, which was confirmed by the corresponding changes in the rate of 4C-glycine-1 build up in proteins of the bone tissue organic matrix and in the intensity of hydroxyproline urinary excretion.

Combined vitamin D parathyroid defect in thalassemia major.

A 17-year-old girl with thalassemia major experienced tetany. The serum calcium level was 5.5 mg/dL, and the phosphorus level was 6.3 mg/dL. Serum levels of parathyroid hormone (PTH) and 25-hydroxyvitamin D (25-OHD) were subnormal at 125 pg/mL and 8.1 ng/mL, respectively, As a result of these findings, serum 25-OHD and PTH levels were measured in an additional 12 patients with thalassemia major. Low levels of both 25-OHD and PTH were found frequently. An increase in serum 25-OHD levels was noted in each of four patients who were examined after iron chelation therapy.

Osteomalacia and disorders of vitamin D metabolism.

A rapidly growing understanding of the biochemical and physiological processes that underlie the metabolism of vitamin D has provided new insights into the pathogenesis of oestomalacia. Many of the vitamin D--resistant osteomalacia syndromes can now be explained on the basis of defects in the metabolic conversion of vitamin D to the biologically active dihydroxylated metabolite 1,25(OH)2D and perhaps, in some instances, to impairement of the actions of 1,25(OH)2D on target tissues. The availability of this new information has made possible the synthesis of 1-hydroxylated forms of the vitamin for therapeutic use in states of vitamin D resistance. Although many questions regarding the pathogenesis and most effective approaches in the management of osteomalacia remain unanswered, considerable progress has been made in this direction as a result of continued research on the subject.