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BACKGROUND Low vitamin D status is a global public health issue that vitamin D food fortification and biofortification may help to alleviate. OBJECTIVES We investigated the effect of vitamin D food fortification and biofortification on circulating 25-hydroxyvitamin D (25(OH)D) concentrations. We expanded the scope of earlier reviews to include adults and children, to evaluate effects by vitamin D vitamer, and investigate linear and nonlinear dose-response relations. METHODS We conducted a systematic review and meta-analysis. We searched CINAHL, MEDLINE, PubMed, Embase, the Cochrane Library, and gray and unpublished literature sites for randomized controlled trials, including people of all ages, with the criteria: absence of illness affecting vitamin D absorption, duration ≥4 wk, equivalent placebo food control, dose quantification, dose ≥5 μg/d, baseline and endpoint or absolute change in 25(OH)D concentrations reported, random allocation, and participant blinding. Quality was assessed using the Jadad Scale. RESULTS Data from 34 publications (2398 adults: 1345 intervention, 1053 controls; 1532 children: 970 intervention, 562 controls) were included. Random-effects meta-analysis of all studies combined (mean dose 16.2 μg/d) indicated a pooled treatment effect of 21.2 nmol/L (95% CI: 16.2, 26.2), with a greater effect for studies using cholecalciferol than ergocalciferol. Heterogeneity was high (I2 > 75%). Metaregression analyses for all studies combined suggested positive effect differences for baseline circulating 25(OH)D concentrations

BACKGROUND: Serum 25-hydroxyvitamin D [25(OH)D] concentration is an indicator of vitamin D exposure, but it is also influenced by clinical characteristics that affect 25(OH)D production and clearance. Vitamin D is the precursor to 25(OH)D but is analytically challenging to measure in biological specimens. OBJECTIVES: We aimed to develop and validate a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for quantification of vitamins D(3) and D(2) in serum and to explore the potential of circulating vitamin D as a biomarker of exposure in supplementation trials. METHODS: The method was validated using guideline C62-A from the Clinical and Laboratory Standards Institute and was applied in 2 pilot clinical trials of oral vitamin D(3) supplementation. Pilot study 1 included 22 adults randomly assigned to placebo or 2000 IU/d. Blood was collected at baseline, 1, 3, 6, and 12 mo. Pilot study 2 included 15 adults randomly assigned to 2000 or 4000 IU/d. Blood and subcutaneous (SUBQ) adipose tissue were collected at baseline and 3 mo. RESULTS: In study 1, mean change (baseline to 3 mo) in serum vitamin D(3) was −0.1 ng/mL in the placebo group and 6.8 ng/mL in the 2000 IU/d group (absolute difference: 6.9; 95% CI: 4.5, 9.3 ng/mL). In study 2, mean change (baseline to 3 mo) in serum vitamin D(3) was 10.4 ng/mL in the 2000 IU/d group and 22.2 ng/mL in the 4000 IU/d group (fold difference: 2.15; 95% CI: 1.40, 3.37). Serum and adipose tissue vitamin D(3) concentrations were correlated, and the dose-response of vitamin D(3) in adipose mirrored that in serum. CONCLUSIONS: We validated a sensitive, robust, and high-throughput LC-MS/MS method to quantify vitamins D(3) and D(2) in serum. Serum and SUBQ adipose tissue vitamin D(3) concentrations increased proportionally to dose with 3 mo of daily supplementation. These trials were registered at clinicaltrials.gov as NCT00552409 (pilot study 1) and NCT01477034 (pilot study 2).

Background\ud A large amount of mushroom waste is generated during mushroom production (accounting for up to 20% of total production) and is mainly composed of mushrooms that do not meet the specifications set by retailers because of misshapen caps and/or stalks. Mushrooms are notable for their ergosterol (a precursor of vitamin D2) content which is converted to vitamin D2 after exposure to natural or artificial ultraviolet (UV) irradiation. Therefore, mushroom waste could be used as a source for the recovery of both ergosterol and vitamin D2 which could be valorized by both pharmaceutical and food industries.\ud \ud Scope and approach\ud The current review presents a comprehensive summary of research performed regarding the extraction, purification and determination of ergosterol and vitamin D2 (ergocalciferol) from mushroom matrices. Additionally, studies related to the impact of sample preparation and especially of drying methods on the retention of ergosterol and vitamin D2 are presented. Finally, the potential valorization of mushroom waste sterols by food and pharmaceutical industries is discussed.\ud \ud Key findings and conclusions\ud Ergosterol and vitamin D2 contents vary among different mushroom species. Sample drying is a crucial step that precedes sterol extraction and has a significant impact on the retention of ergosterol and vitamin D2. The extraction of sterols from mushrooms can be conducted by either conventional (e.g., Soxhlet extraction) or non-conventional methods (e.g., ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE), deep eutectic solvents (DES) extraction, supercritical fluid extraction (SFE), and pressurized liquid extraction (PLE)) or their combination. The application of non-conventional methods such as UAE and MAE facilitate in shorter extraction times than the conventional methods. The valorization of mushroom extracts enriched in ergosterol and vitamin D2 by both pharmaceutical and food industries requires further work.

The effects of electron-beam or gamma irradiation (both applied at 1.5 kGy and 2.5 kGy) and subsequent storage under modified atmospheres (35 d at 4 °C) were evaluated on total carbohydrates, chitins, β-glucans, proteins, total phenols, sterols and ergocalciferol concentrations of Tuber melanosporum ascocarps. Irradiation procedures reduced chitin and protein concentrations and modified total phenol levels depending on the dose and irradiation type utilized. Further storage of irradiated truffles maintained the levels of all determined compounds unchanged except for the phenolic compounds and ergosterol levels that increased probably due to their yeast colonization after 21 d storage. Therefore, irradiation plus storage under modified atmosphere packings could be used as a preservation method to extent truffles quality and shelf life, but it is only recommended up to 21 d. The lower irradiation doses are encouraged to diminish protein degradation.

In India, changes in the life style pattern together with low consumption of vitamin D rich foods resulted in 50–90% of population as vitamin D deficient. Fortification is the common practice followed to overcome such micronutrient deficiencies. Vitamin D is fat soluble in nature and simultaneously sensitive to light and heat, however binding to milk protein improves the solubility and stability towards heat and light. Sodium caseinate, succinylated sodium caseinate (SNaCas), reassembled sodium caseinate (RNaCas) and reassembled succinylated sodium caseinate (RSNaCas) were complexed with ergocalciferol (vitamin D2). Milk protein-vitamin D2 complexes (NaCas-VD, SNaCas-VD, RNaCas-VD and RSNaCas-VD) and free vitamin D2 (free-VD) showed highest stability when stored at −20 °C, followed by 4 °C and 37 °C. At the end of storage period of 48 h, the stability (%) of vitamin D2 in milk protein-vitamin D2 complex and free vitamin D2 (free-VD) fortified milk was significantly higher (p

Chronic kidney disease patients have a high prevalence of vitamin D insufficiency/deficiency. Vitamin D deficiency has been associated with a variety of bone, metabolic and cardiovascular disorders. However, the role of native vitamin D supplementation (ergocalciferol, cholecalciferol or calcifediol) remains unclear in chronic kidney disease (CKD), particularly in the pre-dialytic phase. Several international guidelines have been developed on CKD–Mineral and Bone Disorder, but the optimal strategy for native vitamin D supplementation and its clinical benefit remains a subject of debate in the scientific community. This paper aims to review the available literature, including randomized clinical trials that evaluated the effects of native vitamin D supplementation on pre-dialysis CKD on biochemical and clinically relevant outcomes. Resumen: Los pacientes renales crónicos tienen una elevada prevalencia de insuficiencia/deficiencia de vitamina D. El déficit de vitamina D se ha asociado con una serie de cambios óseos, metabólicos y cardiovasculares. Sin embargo, continúa por aclarar el papel de la suplementación con vitamina D nativa (ergocalciferol, colecalciferol o calcifediol) en la enfermedad renal crónica (ERC), especialmente en la fase pre-dialítica. Varias pautas internacionales se han desarrollado sobre la enfermedad mineral y ósea relacionada con la ERC, pero la estrategia ideal de suplementación con vitamina D nativa y su beneficio clínico continúan siendo objeto de debate en la comunidad científica. Este trabajo pretende revisar la literatura disponible, incluyendo ensayos clínicos aleatorizados que evaluaron los efectos de la suplementación con vitamina D nativa en la ERC pre-diálisis en resultados bioquímicos y clínicamente relevantes. Keywords: Chronic renal disease, Native vitamin D, Ergocalciferol, Cholecalciferol, Calcifediol, Secondary hyperparathyroidism, Palabras clave: Enfermedad renal crónica, Vitamina D nativa, Ergocalciferol, Colecalciferol, Calcifediol, Hiperparatiroidismo secundario

To determine which vitamin D dose, formulation, and schedule most effectively and safely achieves a 25-hydroxyvitamin D (25[OH]D) level of30 ng/mL (75 nmol/L).In this prospective study, 100 subjects from the NY Harbor HCS Brooklyn Campus, ages 25 to 85 years, with 25(OH)D30 ng/mL (75 nmol/L), were randomized into four groups: cholecalciferol (D3) 2,000 international units (IU) daily; D3 3,000 IU daily; ergocalciferol (D2) 50,000 IU weekly; and D2 50,000 IU twice weekly. All were supplemented with 500 mg calcium carbonate daily. 25(OH)D, parathyroid hormone (PTH), urinary calcium, urinary creatinine, and other variables were measured during 7 visits over 12 months.All groups achieved a mean vitamin D level30 ng/mL (75 nmol/L) by visit 4 (5 months). Those receiving 50,000 IU D2 twice weekly displayed the most rapid and robust response, with 25(OH)D reaching30 ng/mL (75 nmol/L) after only 1 month and plateauing at 60 ng/mL (150 nmol/L) by 7 months. Although no statistically significant difference was seen in mean 25(OH)D levels between groups 1 through 3, subjects on 50,000 IU D2 weekly more consistently showed higher mean levels than either groups 1 or 2. No episodes of significant hypercalcemia occurred. There was a negative correlation in mean PTH levels and mean vitamin D levels in group 4 and all groups combined.All four schedules of vitamin D replacement were effective in safely achieving and maintaining 25(OH)D30 ng/mL (75 nmol/L). D2 50,000 IU twice weekly provided the most rapid attainment and highest mean levels of vitamin D.25(OH)D = 25-hydroxyvitamin D; BMI = body mass index; BUN = blood urea nitrogen; Ca/Cr = calcium/creatinine; D2 = ergocalciferol; D3 = cholecalciferol; IU = international units; PTH = parathyroid hormone.

The Quality by Design (QbD) method was used to develop a new simple, rapid, and sensitive ultra-performance liquid chromatography and time of flight mass spectrometric (UPLC-QToFMS) method for quantitative estimation of ergocalciferol (D 2 ), cholecalciferol (D 3 ), phylloquinone (K 1 ), menaquinone-4 (K 2 -4), menaquinone-7 (K 2 -7), and menadione (K 3 ). Four independent parameters were tested using a full factorial design (FFD): pH of the aqueous component of the mobile phase, flow rate, injection volume, and percentage of the organic modifier. Two factors were discovered to be significant using analysis of variance (ANOVA). To further optimise chromatographic conditions, a central composite design (CCD) was used. At a flow rate of 0.5 mL/min and an injection volume of 5 µL, the analysis was carried out on an Accucore C-18 (50 x 4.6 mm; 2.6 µm) column with a gradient mobile phase containing methanol and 0.1% formic acid buffer, pH 3.5. The proposed method was validated under optimal conditions, with a linear range of 15 to 1500 ng/mL for analytes and correlation coefficient (R 2 ) values greater than 0.99. The percentage of analyte recovery ranged from 97.19 to 100.84 percent. RSD values for both inter-day and intra-day precisions were less than 1%. The proposed method was discovered to be precise, specific, and robust.

Previous studies of the effect of vtamin D on serum levels of fibroblast growth factor- 23 (FGF-23) have yeilded an inconsistent findings. This systematic review and meta-analysis of randomized controlled trials (RCTs) sought to investigate the effect of vitamin D supplementation on serum levels of FGF-23. PubMed, Scopus, ISI Web of Science, and the Cochrane Library were searched, from database inception to November 2020, for RCTs that evaluated the effects of native or active vitamin D supplementation on serum levels of FGF-23 in adults. Weighted mean difference (WMD) were calculated and random effects meta-analysis was used to estimate the overall effects. Twenty-seven trials were included in the meta-analysis. Supplementation with native vitamin D (23 studies, n = 2247 participants; weighted mean difference [WMD] = 0.5 pg/mL, 95 % CI: -0.52 to 1.51, P = 0.33; I2 = 29.9 %), and active vitamin D (5 studies, n = 342 participants, WMD = 29.45 pg/mL, 95 % CI: -3.9 to 62.81, P = 0.08; I2 = 99.3%) had no significant effects on serum FGF-23 concentration. In subgroup analyses, supplementation with ergocalciferol (3 studies, n = 205 participants; WMD = 18.27 pg/mL, 95 % CI: 5.36-31.17, P = 0.006), and daily dosing regimens (9 studies, n = 1374 participants; WMD = 0.41 pg/mL, 95 % CI: 0.22 to 0.59, P < 0.001) increased serum FGF-23 levels compared to control. Overall, our findings revealed no significan effect of vitamin D supplementation on serum FGF-23 concentration. However, further high quality, large-scale studies are needed to better elucidate this relationship.

Vitamin D is essential for bone health and has significant roles in non-skeletal health and organ function. Dermal synthesis through exposure to ultraviolet B light is the major natural source of vitamin D, while only a small portion of the necessary amount can be acquired by a diet without fortified foods. In recent years, vitamin D deficiency as a result of lifestyles with inadequate sun exposure, has received increased attention due to its association with the increased risk of serious chronic diseases. This review summarizes our current understanding of food fortification strategies with vitamin D and the resulting health impact. Conventional and biotechnological approaches can be used for the production of new and novel vitamin D rich or vitamin D fortified foods. The availability of a wider range of every-day consumed fortified foods as part of a "Daily D" public health policy can contribute to the improvement of vitamin D status and to prevention of vitamin D deficiency.

This paper investigates the effect of direct sunlight during natural sun drying and solar drying in a forced convection type solar dryer on vitamin D 2 content of brown button mushrooms ( Agaricus bisporus ). Commercial UVB light, sample processing and drying methods on vitamin D 2 content were also studied via different UVB-drying strategies. Mushrooms of the same flush were divided in different groups: i) slices were dried under the sun and in an innovative solar dryer, ii) slices were UVB-treated and dried either by freeze- or convective drying, iii) whole fruit bodies were UVB-treated, sliced and freeze-dried, iv) freeze dried samples were UVB-treated in the form of slices or powder. UVB-treatment of whole mushrooms led to remarkable vitamin D 2 enrichment (45 μg/g dry matter), while slicing of mushrooms prior to UVB exposure resulted in a ten-fold increase in D 2 content (406 μg/g). UVB light exposure of already dried slices and powder also induced high vitamin D 2 generation. Sun-dried and solar-dried mushrooms contained 36 and 39 μg/g dry matter vitamin D 2 , respectively. Although commercial UVB light boost D 2 synthesis in mushrooms, solar drying represents a viable method for the natural formation of ergocalciferol in mushrooms and an excellent vitamin D source for vegans.

There is substantial evidence that the prevalence of vitamin D deficiency is unacceptably high in the population, and this requires action from a public health perspective. Circulating 25-hydroxyvitamin D [25(OH)D] is a robust and reliable marker of vitamin D status and has been used by numerous agencies in the establishment of vitamin D dietary requirements and for population surveillance of vitamin D deficiency or inadequacy. In a wider context, modeling of serum 25(OH)D data and its contributory sources, namely dietary vitamin D supply and UVB availability, can inform our understanding of population vitamin D status. The aim of this review is to provide the current status of knowledge in relation to modeling of such vitamin D–relevant data. We begin by highlighting the importance of the measurement of 25(OH)D and its standardization, both of which have led to new key data on the prevalence of vitamin D deficiency and inadequacy in North America and Europe. We then overview how state-of-the-art modeling can be used to inform our understanding of the potential effect of ergocalciferol and 25(OH)D on vitamin D intake estimates and how meteorological data on UVB availability, when coupled with other key data, can help predict population serum 25(OH)D concentration, even accounting for seasonal fluctuations, and lastly, how these in silico approaches can help inform policymakers on strategic options on addressing low vitamin D status through food-based approaches and supplementation. The potential of exemplar food-based solutions will be highlighted, as will the possibility of synergies between vitamin D and other dairy food–based micronutrients, in relation to vitamin D status and bone health. Lastly, we will briefly consider the interactions between season and vitamin D supplements on vitamin D status and health.

Vitamin D has been found have various biological effects that may be potent in preventing bovine mastitis. Two forms of vitamin D, vitamin D2 (D2) and vitamin D3 (D3), can be hydroxylated to functional metabolites in cattle. The objectives of the present study were to investigate the effects of D2 and D3 compounds on bovine mammary epithelial cell proliferation and Staphylococcus aureus (S. aureus) invasion.. Results showed that 1,25-dihydroxyvitamin D2 have an anti-proliferation activity comparable to 1,25-dihydroxyvitamin D3, while D2 and 25-hydroxyvitamin D2 (25(OH)D2) was slightly more potent than D3 and 25-hydroxyvitamin D3 (25(OH)D3) in inhibiting MAC-T cell viability in vitro. S. aureus growth was inhibited by high concentrations of D2, D3, 25(OH)D2 and 25(OH)D3. 25(OH)D2 and 25(OH)D3 induced CYP24A1 expression but reduced VDR mRNA expression, whereas the expression of CYP27B1, occludin, and E-cadherin did not change. Additionally, the induction of CYP24A1 expression by 25(OH)D3 was higher than that of 25(OH)D2, which may contribute to the differences in anti-proliferation activity. S. aureus invaded into MAC-T cells and universally inhibited gene expressions. MAC-T cells pre-treated with 25(OH)D2 reduced S. aureus adhesion while pre-treatment with 25(OH)D3 inhibited S. aureus invasion, but neither of the compounds attenuated the S. aureus-induced gene expression reduction. In conclusion, the present study showed that D2 compounds have comparable effects on cell proliferation and bacterial invasion to their D3 analogues in vitro, suggesting that D2 and its metabolites may also be effective in the defense against bacterial infection.

Variability in 25-hydroxyvitamin D (25(OH)D) change following vitamin D supplementation exists. Vitamin D metabolite measurement might assist in predicting 25(OH)D response and also contribute to defining vitamin D adequacy. This study assessed utility of vitamin D metabolite measurements to predict 25(OH)D response and explored the relationship between parathyroid hormone (PTH) and a "vitamin D composite index" comprised of the sum of serum 25(OH) D, cholecalciferol (vitamin DSixty-two postmenopausal women were randomized to daily vitamin DBaseline 25(OH)D was positively correlated (P.05) with vitamin D3, 24,25(OH)This study does not support measurement of vitamin D metabolites in a composite index to assist in prediction of 25(OH)D response to supplementation. Overweight individuals have less robust 25(OH) D response to supplementation, but variability precludes prediction of the result following daily supplementation.BMI = body mass index DXA = dual-energy X-ray absorptiometry LC-MS/MS = liquid chromatography tandem mass spectroscopy 25(OH)D = 25-hydroxyvitamin D 24,25(OH)

Cocoa beans are susceptible to fungal contamination and often contain substantial amounts of ergosterol, the precursor to vitamin D2. We hypothesized that sun-drying the fermented cocoa beans might lead to the conversion of ergosterol to vitamin D2. We quantified vitamin D in cocoa and cocoa-based foods by liquid chromatography-tandem mass spectrometry. Here, we show that cocoa beans from different growing regions contain vitamin D2. Particularly high vitamin D2 content was found in cocoa powder and butter. Among the chocolates, dark chocolate had the highest vitamin D2 content (ranging from 1.90 to 5.48 µg/100 g), white chocolate had the lowest vitamin D2 content (ranging from 0.19 to 1.91 µg/100 g), and chocolate nut spreads had a comparatively low vitamin D2 content, with an average of 0.15 µg/100 g. Thus, cocoa and chocolate are clearly a dietary source of vitamin D, therefore, it is necessary to update food composition databases.

A convergent approach to 25S,26-dihydroxyvitamin D3 (1) has been developed in our laboratories. The A-ring and the CD-fragment are constructed from ergocalciferol and Inhoffen-Lythgoe diol, respectively. The triene system is assembled by a Wittig-Horner coupling. With this convergent synthesis, a novel hydroxylated vitamin D metabolite in our laboratory is available for biological testing.

Vitamin D deficiency has been linked to various diseases, but could be rectified via fortified food stuffs or supplementation. In this study 39 different hydrophobic deep eutectic solvents were evaluated for green extraction of ergosterol from mushroom. Extraction parameters (e.g. time, solvent volume) were optimized using response surface methodology (RSM) and a maximum extraction yield of 6995.00 µg ergosterol/g dry weight mushroom was attained with menthol: pyruvic acid. The extracted ergosterol was purified using a novel methodology and the extraction solvent was reused for six cycles, while retaining extraction efficiency (up to 28%). The ergosterol was exposed to ultra-violet radiation for conversion to ergocalciferol (vitamin D2) resulting in a yield of ergocalciferol that was equivalent to 2142.01 µg/g dry weight mushroom.

An ultra-performance liquid chromatography coupled to atmospheric pressure chemical ionization-quadrupole time-of-flight mass spectrometry method has been optimized and validated for the determination of ergosterol and ergocalciferol in mushroom samples, using cholecalciferol as surrogate standard. The separation was carried out with a Synergi Hydro-RP column (100 mm x 3.00 mm i.d, 2.5 μm particle size), (Phenomenex, CA, USA) column, thermostated at 35 °C. The mobile phase was 0.1 % formic acid aqueous solution and methanol in gradient elution mode and it was achieved in 5 min approximately. Detection was achieved by atmospheric pressure chemical ionization in positive mode and quadrupole time-of-flight mass spectrometry. Desolvation and interface temperatures were set at 500 °C and 150 °C, respectively. The recoveries obtained were within 92-105 % for ergosterol, 77-81 % for ergocalciferol and 83-87 % for cholecalciferol. Method limits of detection were 0.4 and 0.5 μg g-1 for ergosterol and ergocalciferol, respectively, and method limits of quantitation were 1.2 and 1.3 μg g-1 for ergosterol and ergocalciferol, respectively. A rapid and simple extraction procedure using small amount of sample (100 mg) with hexane was optimized and the method was applied to the determination of ergosterol and ergocalciferol in white button mushrooms (Agaricus bisporus var. bisporus) exposed to UV irradiation. Results were compared to the corresponding non-irradiated mushrooms.

The main objective of this work was to determine the stability of vitamin D2 in dried mushrooms Agaricus bisporus, Pleurotus ostreatus and Lentinula edodes during storage, as well as to examine the possibility of inducing vitamin D2 production in dried mushrooms by UVB irradiation. After 1.5 year storage of dried mushrooms, the level of vitamin D2 in button mushrooms was found to be 6.90 μg/g dw, which is a 48.32% of initial level of vitamin D2. In the case of dried oyster and shiitake mushrooms there was a decrease to the level of 66.90% and 68.40%, respectively. It was determined that dried mushrooms can produce ergocalciferol under UVB irradiation. The highest content of vitamin D2 was observed in A. bisporus. Freeze-dried A. bisporus contained from 42.08 to 119.21 μg/g dw and hot-air dried mushrooms contained from 21.51 to 81.17 μg/g dw vitamin D2.

A wide range of nutrients and health-promoting non-nutrient components in mushrooms are a subject of international research, but specific reference materials to facilitate comparison of results among laboratories are lacking. Commercially available food matrix reference materials do not contain components unique to mushrooms (e.g., ergosterol, vitamin D2, chitin, beta-glucans, agaritine, ergothioneine). A mixed mushroom control material (CM) (homogeneous mixture of 15 types of mushrooms) was prepared and characterized for selected components, including proximates (moisture, protein, ash), total folate, folate vitamers, ergosterol, ergosterol metabolites, vitamin D2 (ergocalciferol), amino acids, total dietary fiber, agaritine, elements (sodium, potassium, phosphorous, magnesium, calcium, iron, copper, manganese, zinc), riboflavin, niacin, thiamin, vitamin B6, pantothenic acid. Subsamples of the CM are available to qualified laboratories from the Food Analysis Laboratory Control Center at Virginia Tech (Blacksburg, VA, USA), to be assayed concurrently with mushroom samples for which food composition data will be published along with results for the CM. Implementation of this CM should facilitate comparison of published data on mushroom composition and health benefit among species, and biodiversity within species by serving as common control sample that allows the separation of analytical variability from true differences in sample composition determined at different laboratories.

Our previous studies revealed that the double variants of CYP105A1- R73A/R84A and R73V/R84A-show high levels of activity with respect to conversion of vitamin D3 to its biologically active form, 1α,25-dihydroxyvitamin D3 (1α,25(OH)2D3). In this study, we found that both the double variants were also capable of converting vitamin D2 to its active form, that is, 1α,25-dihydroxyvitamin D2 (1α,25(OH)2D2), via 25(OH)D2, whereas its 1α-hydroxylation activity toward 25(OH)D2 was much lower than that toward 25(OH)D3. Comparison of the wild type and the double variants revealed that the amino acid substitutions remarkably enhanced both 25- and 26-hydroxylation activity toward vitamin D2. After 25-hydroxylation of vitamin D2, further hydroxylation at C26 may occur frequently without the release of 25(OH)D2 from the substrate-binding pocket. Thus, the double variants of CYP105A1 are quite useful to produce 25,26(OH)2D2 that is one of the metabolites of vitamin D2 detected in human serum.

Vitamin D insufficiency is common in children with cystic fibrosis (CF), yet data are sparse regarding the most effective form of vitamin D supplementation. The aim of this study was to compare two different vitamin D replacement regimens.We conducted a randomized controlled trial comparing 50,000 IU of ergocalciferol (vitamin D2) twice weekly for 8 weeks versus 50,000 IU of cholecalciferol (vitamin D3) weekly in patients with CF, pancreatic insufficiency, age 6-21 years and a 25(OH)D30 ng/mL. The primary outcome was change in serum 25(OH)D concentration. For secondary analyses, we examined changes in IgG, IgE and CRP in patients who normalized their vitamin D levels.A total of 47 patients completed the trial. The mean pre-treatment 25(OH)D concentration was 23.1 (SD 4.7) ng/mL. The overall mean increase in 25(OH)D was 11.1 (11.9) ng/mL and 31/47 (66%) achieved a 25(OH)D concentration ≥ 30 ng/mL; of the 26 participants who received D2, 18 (69%) achieved sufficiency while 13/21 (62%) participants treated with D3 achieved sufficiency. There was no difference between groups in change of 25(OH)D (p=0.65). Similarly, there was no difference in the number of patients to achieve vitamin D sufficiency between treatments (p=0.6).Ergocalciferol administered as 50,000 IU twice weekly is as effective as cholecalciferol 50,000 IU weekly for 8 weeks in pediatric patients with CF and vitamin D insufficiency. Only 66% of the patients studied achieved the desired 25(OH)D concentration.

Encapsulation may protect unstable, fat soluble vitamins such as vitamin D2 (ergocalciferol). However, encapsulation by the solvent extraction and/or evaporation techniques can require toxic organic solvents, which greatly increase processing costs. The objective of this study was to evaluate the effect on ergocalciferol encapsulation by a combination of the ionic gelation method with the ultrasonic atomization and microwave drying. Optimization of manufacturing parameters included the addition of pluronic-F127 to the core solution at 1.5% w/w to increase the encapsulation efficiency to nearly 92%, greatly improving performance compared to Tween 80 at 0.5% w/w. Microwave treatment at 230 W promoted the recovery of 100% of the ergocalciferol and reduced drying times to about 30 min, while 690 W degraded 40% of the D2. In contrast, the conventional heating degraded 17% of the ergocalciferol during 12 h of processing. By all the applied methods, microparticles were produced with similar gastoresistance properties of less than 10% release at pH of 1.0, to nearly 100% release at pH of 6.8 and 240 min of dissolution. Analysis showed limited ergocalciferol degradation after 5 months of storage.

1 H-NMR Chemical compounds studied in this article: Beta-carotene (PubChem CID: 5280489) Curcumin (PubChem CID: 969516) Ergocalciferol (PubChem CID: 5280793) Folic acid (PubChem CID: 6037) abstract The main purpose of the current work was exploring the potential application of the proteinepoly- saccharide soluble nanocomplexes as delivery systems for nutraceuticals in liquid foods. In this study, the intrinsic transporting property of b-lactoglobulin (BLG) was utilized to develop nanoscale green delivery systems. The binding analysis using fluorescence spectroscopy suggested that the complexation between BLG and four nutraceutical models including b-carotene, folic acid, curcumin and ergocalciferol occurred under all conditions but varied as a function of pH and nutraceutical type. The 1 H-NMR study of hy- drophilic ligands binding to BLG provided complementary information on the interactions between protein and water soluble ligands. These findings resulted in designing nanoscopic delivery systems for encapsulation of both hydrophobic and hydrophilic bioactives in clear liquid food products of acidic pH. The stability experiments demonstrated that the nutraceuticals of low solubility in water were suc- cessfully entrapped within electrostatically stable nanocomplexes arising from BLG-sodium alginate interactions. The electrophoretic mobility analysis showed that soluble nanocomplexes had good sta- bility against aggregation.

The objective of the present investigation was to determine bioavailability of calcium and vitamin D₂ from milk fortified with either calcium or vitamin D₂ alone or when both were used for preparation of multiple micronutrient fortified milk and also to study its interaction with iron and zinc bioavailability. 32 weanling male rats (aged 21-28 days) were assigned into four groups and were fed milk and milk fortified with calcium, vitamin D₂ and calcium+vitamin D₂. Vitamin D₂ increased calcium bioavailability. In multiple micronutrient fortified milk, the bioavailability of both calcium+vitamin D₂ increased in comparison to single fortification. Calcium fortification decreased, whereas vitamin D₂ increased the absorption of iron and zinc. However, calcium and vitamin D₂ when fortified in combination, the iron and zinc bioavailability was similar to control group. There was positive association between bioavailability of calcium and vitamin D₂.

Objective To report an uncommon cause of 1,25-dihydroxyvitamin D (1,25[OH]2D)-mediated hypercalcemia associated with splenic sarcoidosis and illustrate the evaluation and potential role of fluorodeoxyglucose (FDG) positron emission tomography (PET)/computed tomography (CT) in such patients. Methods We present detailed clinical features, laboratory results, imaging results, and pathology results for this rare entity, discuss evaluation and management options, and review previous literature. Results A 65-year-old male presented with symptomatic hypercalcemia, with a serum calcium level of 14.1 mg/dL 3 months after being initiated on ergocalciferol for vitamin D deficiency. He was found to have a suppressed parathyroid hormone level, normal 25-hydroxyvitamin D (25[OH]D) level, and elevated 1,25(OH)2D level. Extensive evaluation did not yield a definitive diagnosis. Hiscalcium levels normalized and symptoms resolved on prednisone then recurred when prednisone was discontinued. FDG PET/CT showed intense uptake in the spleen. Splenectomy was performed, which resulted in resolution of hypercalcemia and yielded a diagnosis of splenic sarcoidosis. Conclusion: Splenic sarcoidosis causing hypercalcemia has been rarely reported. Our case is unique in that the spleen lacked typical focal nodularity on cross-sectional CT imaging, which is expected in sarcoid involvement of the spleen. Our case adds to an emerging literature documenting the potential value of FDG PET/CT in localizing otherwise occult 1,25(OH)2D-mediated hypercalcemia. (Endocr Pract. 2014;20:e28-e33)

The optimal treatment regimen for correcting vitamin D insufficiency in diabetic patients has not been established.Two hundred and forty four adult diabetic patients with vitamin D insufficiency were enrolled to receive: Ergocalciferol (D2) 50,000 IU daily over 10 days (500,000 IU) followed by Calcichew D3 (calcium carbonate/Cholecalciferol) BID (~24,000 IU cholecalciferol/month) (ECC) (n=53); Cholecalciferol (D3) 40,000 IU daily over 10 days (400,000 IU) followed by Calcichew D3 BID (~24,000 IU cholecalciferol/month) (CCC) (n=94) or Cholecalciferol 40,000 IU daily over 10 days (400,000 IU) followed by Cholecalciferol 40,000 IU monthly (CC) (n=97). The 25(OH)D, HbA1c, lipids, blood pressure and eGFR were assessed at baseline and after a mean follow up of 8.0±4.0 months.Treatment increased 25(OH)D concentrations significantly in ECC (17.4±13.8 vs 29.9±9.6 ng/ml, P0.0001), CCC (14.2±6.6 vs 30.9±13.1 ng/ml, p0.0001) and CC (13.5±8.4 vs 33.9±14.4 ng/ml, P0.0001). The relative increase in 25(OH)D was significantly lower with ECC compared to CC (+14.6±12.2 vs +20.6±15.0, P=0.01) and the majority of subjects in the ECC group (63%) remained vitamin D deficient (25(OH)D30 ng/ml) compared to CCC (46%) and CC (36%) (P=0.0005).This study demonstrates that relatively aggressive treatment regimens of both vitamin D2 and D3 increase 25(OH)D concentrations in diabetic patients, but the ability to raise 25(OH)D status to 'sufficient' levels is inadequate in a large proportion of individuals.

Background Aortic valve (AV) calcification is the major characteristic of AV sclerosis. Several forms of calcium salts accumulate including hydroxyapatite in late phases. Purpose We sought to examine whether a modified cholesterol – Vitamin D2 diet can produce AV calcification and stenosis in a rabbit model. Methods AV calcification was introduced in New Zealand male rabbits using the modified diet: normal chaw+ 0.5%w/w cholesterol+ 3.500IU Vitamin D2/kg/d (ergocalciferol). Thirty six rabbits were equally randomized into control (normal chaw) and experimental group (modified diet) for 12 weeks. Echocardiography was performed at baseline and the day before the sacrifice. AV Vmax, Transaortic pressure gradient and AV area (AVA) were measured (Vivid 7, GE). AV cusps (collected every 2 weeks, 3 per group) were processed for Fourier-Transformed Infrared Spectroscopy (FT-IR) to detect and characterize calcium salt deposition longitudinally (Nicolet 6700 spectrometer, OMNIC 7.3 software). AVs from surgical patients with severe AV stenosis were used for qualitative comparisons. Results None of the echocardiographic parameters was significantly altered with modified diet; baseline AVA: 0.32 ± 0.06 cm2 vs. 12 weeks: 0.37 ± 0.07, P = 0.18. FT-IR vibrations in the region of 1800-800 cm-1 demonstrated the peroxidation of lipids, advanced glycation end products (AGEs), deposition of CaCO3 and non-hydroxyapatite Ca3 (PO4)2. Further, the 840 cm-1 (HCO3-) band is in accordance to those obtained from human AVs. Conclusion Overall, our rabbit Vitamin D2 model did not produce any AV hemodynamic changes. Interestingly, FT-IR spectroscopy showed that Vitamin D2 stimulated formation of AGEs, deposition of carbonic and phosphate calcium salts in the rabbit AV, both present in human AVs. This study demonstrates that acidosis is an important pathway of AV calcification. Further analysis is needed to elucidate molecular mechanisms in this particular animal model.

Objective Pruritus is a common problem among hemodialyzed patients. Its causes are poorly understood, and, as a result, itching is often attributed to elevated serum phosphorus and other disorders of bone and minerals. The primary purpose of this study was to analyze the relationship between pruritus and common tests of bone and mineral disease. Methods This study was a post hoc analysis of data from a randomized controlled trial of 3 months of ergocalciferol versus placebo treatment in 50 hemodialysis patients with uremic pruritus. A pruritus survey was administered at baseline and then every 2 weeks for 12 weeks. Concurrent serum phosphorus, intact parathyroid hormone (PTH), serum calcium, and calcium-phosphate product were measured. Results Pruritus score was not found to be associated or correlated with serum phosphate, intact PTH, serum calcium, or calcium-phosphate product at each time interval or over time. Likewise, when analyzed by original study group (placebo or ergocalciferol), no association or correlation between the mineral and bone indicators and itching were found. Conclusion Neither serum phosphate nor other tests of bone and mineral status were found to be significant predictors of pruritus at any point in time or over time.

Calcitriol and analogs inhibit renin-angiotensin system, which has a pivotal role in glomerular and tubulo-interstitial damages and proteinuria, and inhibit NF-κB activation which is known to play an important role in renal diseases by promoting inflammation and fibrogenesis. Vitamin D presents interesting pleiotropic effects for the CKD patient (reduction of mortality, antiproteinuric effect and anti-inflammatory properties). "Native" vitamin D (cholecalciferol or ergocalciferol) administration in these patients also decrease parathyroid hormone levels. Native vitamin D administration in CKD patients is safe and does not lead to increased risk of vascular calcification, despite the known hypercalcemic and hyperphosphoremic properties of the molecule in its active form. Native vitamin D administration is not associated with an increased risk of renal stones, at pharmacological doses and without important concomitant administration of calcium salts. In the field of renal transplantation, experimental studies show that vitamin D analogs have a protective role against acute rejection but clinical studies remain mainly observational.

A simple and rapid method was developed using reversed-phase liquid chromatography (LC) with both diode array (DAD) and atmospheric pressure chemical ionization mass spectrometric (APCI-MS) detection, for the simultaneous analysis of the vitamins ergocalciferol (D2), cholecalciferol (D3), phylloquinone (K1), menaquinone-4 (K2) and a synthetic form of vitamin K, menadione (K3). The Taguchi experimental method, an orthogonal array design (OAD), was used to optimize an efficient and clean preconcentration step based on dispersive liquid-liquid microextraction (DLLME). A factorial design was applied with six factors and three levels for each factor, namely, carbon tetrachloride volume, methanol volume, aqueous sample volume, pH of sample, sodium chloride concentration and time of the centrifugation step. The DLLME optimized procedure consisted of rapidly injecting 3 mL of acetonitrile (disperser solvent) containing 150 µL carbon tetrachloride (extraction solvent) into the aqueous sample, thereby forming a cloudy solution. Phase separation was performed by centrifugation, and the sedimented phase was evaporated with nitrogen, reconstituted with 50 µL of acetonitrile, and injected. The LC analyses were carried out using a mobile phase composed of acetonitrile, 2-propanol and water, under gradient elution. Quantification was carried out by the standard additions method. The APCI-MS spectra, in combination with UV spectra, permitted the correct identification of compounds in the food samples. The method was validated according to international guidelines and using a certified reference material. The validated method was applied for the analysis of vitamins D and K in infant foods and several green vegetables. There was little variability in the forms of vitamin K present in vegetables, with the most abundant vitamer in all the samples being phylloquinone, while menadione could not be detected. Conversely, cholecalciferol, which is present in food of animal origin, was the main form in infant foods, while ergocalciferol was not detected.

Inadequate data on tissue distribution of vitamin D and its metabolites remains a barrier to defining health outcomes of vitamin D intake and 25-hydroxyvitamin D (25(OH)D) status. The purpose of this study was to develop a method for the analysis of vitamin D2 (ergocalciferol), vitamin D3 (cholecalciferol), 25(OH)D2, and 25(OH)D3 in soft tissues, and determine distribution in select tissues from a dose-response study of vitamin D2 and vitamin D3 in rats. Liver, gastrocnemius muscle, and epididymal fat homogenates were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with electrospray ionization following liquid-liquid extraction, solid-phase extraction, and derivatization with 4-phenyl-1,2,4-triazoline-3,5-dione (PTAD). A dose-response was observed in most tissues for vitamin D and 25(OH)D from both vitamers. Vitamin D concentration was greater in epididymal fat than gastrocnemius muscle and liver, but 25(OH)D concentration was not significantly different between tissues. Soft tissues of rats fed crystalline vitamin D3 had higher concentrations of total vitamin D than those of rats fed yeast-derived vitamin D2, while total 25(OH)D concentrations were similar between vitamin D sources. This method is well suited to more complete studies of vitamin D bioavailability and metabolite tissue distribution.

Vitamin D3 (cholecalciferol) levels were determined in finfish and shellfish using UV detection at 265 nm (combined with auxiliary full scan UV detection) and selected ion monitoring (SIM) mass spectrometry (MS), using vitamin D2 (ergocalciferol) as an internal standard. Analysis of standard reference material (SRM) NIST 1849 (Infant/Adult Nutritional Formula) was included to validate the method. Three-point calibration curves were employed, allowing values to be determined over a range of species, from those having little or no detectable vitamin D3 (e.g., pollock, shrimp) to those with high levels (e.g., salmon with up to 33.23 μg/100 g). The limit of detection (LOD) and limit of quantitation (LOQ) calculated from the uncertainty and intercept of the calibration curves were 1.22 μg/100 g and 5.30 μg/100 g, respectively, based on all analyses (n = 27 sequences). Use of response factors (RF) allowed quantitation at lower levels of vitamin D3, with an LOQ of

25-Hydroxyvitamin D [25(OH)D] is a marker of nutritional status; however, chronic kidney disease (CKD) results in alterations in vitamin D metabolism, including the loss of vitamin D-binding proteins and alterations in CYP27B1 and CYP24 enzymes that metabolize 25(OH)D. This study was designed to determine the predictors of responsiveness to correction of vitamin D deficiency with oral vitamin D2 (ergocalciferol) in adults.A retrospective study of 183 veterans with 25(OH)D level30 ng/mL, who were treated with 50,000 IU per week of vitamin D2, was performed. Logistic regression models were developed to determine the factors predicting the response to treatment, defined as either the change in serum 25(OH)D level/1000 IU of vitamin D2 or the number of vitamin D2 doses (50,000 IU per dose) administered.The mean age of the patients was 63 ± 12 years. About 87% were men and 51% diabetic, and 29% had an estimated glomerular filtration rate of60 mL/min/1.73 m. The average number of vitamin D2 doses was 10.91 ± 5.95; the average increase in 25(OH)D level was 18 ± 10.80 ng/mL. 25(OH)D levels remained30 ng/mL in 61 patients after treatment. A low estimated glomerular filtration rate and the presence of diabetes mellitus were significant independent predictors for inadequate response to vitamin D2 treatment in logistic regression models. Patients with CKD required greater amounts of vitamin D2 to achieve similar increases in 25(OH)D levels, versus non-CKD patients.The presence of CKD and diabetes mellitus is associated with resistance to correction of 25(OH)D deficiency with vitamin D2 therapy. The underlying mechanism needs to be evaluated in prospective studies.

The biological equivalency of ergocalciferol (D2) and cholecalciferol (D3) has been debated; several comparisons have appeared in the adult literature but are scarce in pediatrics. The objective of this study was to compare increases in plasma 25-hydroxyvitamin D [25(OH)D] concentrations and attainment of 50 and 75 mol/L status cutoffs following 3 mo of daily supplementation with D2 compared with D3. Healthy, breast-fed, 1-mo-old infants (n = 52) received 10 μg (400 ic) of either D2 or D3 daily. At 1 and 4 mo of age, plasma 25-hydroxyergocalciferol and 25-hydroxycholecalciferol concentrations were determined by liquid chromatography tandem MS (LC-MS/MS) and total 25(OH)D by chemiluminescent immunoassay (DiaSorin Liaison). Data were analyzed using t tests and χ2 by intent to treat. A total of 23% of infants were deficient (≤24.9 nmol/L) at baseline and 2% at follow-up on the basis of LC-MS/MS. At 4 mo, 96% were breastfed and there were no differences in compliance, breastfeeding rates, or sun exposure among groups. The change in total 25(OH)D measured by LC-MS/MS did not differ between the D2 (17.6 ± 26.7 nmol/L) and D3 (22.2 ± 20.2 nmol/L) groups. In the combined groups, the baseline plasma 25(OH)D concentration was inversely related to the change in total 25(OH)D (r = −0.52; P < 0.001). Overall, 86% of infants met the 50 nmol/L cutoff at follow-up; however, fewer infants in the D2 group (75%) met this level compared with the D3 group (96%) (P < 0.05). Similar results were obtained by immunoassay. In conclusion, the increase in the 25(OH)D concentration among the D2 and D3 groups did not differ, suggesting daily intake of either isoform is acceptable for infants

Objectives Vitamin D deficiency is more common in African Americans than in the general population or other ethnicities. Vitamin D deficiency also occurs more frequently in patients with primary hyperparathyroidism (PHPT) than in the general population. Currently, the limited data on vitamin D deficiency in African Americans with primary hyperparathyroidism (PHPT) is inconsistent as to whether the vitamin D deficiency observed in PHPT is yet even more pronounced in Africans with PHPT relative to non-African Americans with PHPT. Methods On the basis of biochemical, radiological, and surgical (adenoma weight) parameters, African Americans have been reported to have a more severe form of PHPT than non-African Americans. However, comparative clinical manifestations of PHPT in African Americans have not been well described. Results Current guidelines recommend vitamin D repletion in mild, asymptomatic PHPT when levels of 25-hydroxyvitamin D are less than 20 ng/mL. Studies that reported vitamin D repletion with ergocalciferol or cholecalciferol in PHPT have not stratified data according to ethnicity. Discrepancies therefore exist between repleting vitamin D in African Americans who may potentially have a more severe PHPT profile, but simultaneously a more pronounced vitamin D deficiency. Conclusion Effectively designed clinical trials are necessary to evaluate the indications, efficacy, and safety of vitamin D in African Americans with PHPT. (Endocr Pract. 2012;18:947-953)

Few foods contain ergocalciferol or cholecalciferol. Treatment of mushrooms with UV light increases ergocalciferol content and could provide a dietary source of vitamin D. We evaluated the impact of consuming UV-treated white button mushrooms (Agaricus bisporus) on the vitamin D status of healthy adults. Thirty-eight volunteers were randomized to 4 treatments consumed with a standard meal for 6 wk: the control (C) group received untreated mushrooms providing 0.85 mg/d ergocalciferol (n = 10); groups M1 and M2 received UV-treated mushrooms providing 8.8 (n = 10) and 17.1 mg/d (n =9 ), respectively; and the supplement (S) group received purified ergocalciferol plus untreated mushrooms, providing a total of 28.2 mg/d (n = 9). Serum total 25-hydroxyvitamin D [25(OH)D] and 25-hydroxyergocalciferol [25(OH)D2] were 83 6 38 and 2.4 6 2.0 nmol/L, respectively, at baseline (mean 6 SD). At wk 6, 25(OH)D2 had increased and was higher in all treatment groups than in the C group, whereas 25-hydroxycholecalciferol [25(OH)D3] had decreased and was lower in the M2 and S groups than in the C group. Increases in 25(OH)D2 for groups C, M1, M2, and S were 1.2 6 5.2, 13.8 6 7.3, 12.7 6 3.7, and 32.8 6 3.3 nmol/L and decreases in 25(OH)D3 were 23.9 6 16.3, 210.4 6 6.4, 220.6 6 14.6, and 229.5 6 15.9 nmol/L, respectively. Concentrations did not change in group C. In summary, ergocalciferol was absorbed and metabolized to 25(OH) D2 but did not affect vitamin D status, because 25(OH)D3 decreased proportionally. J. Nutr. 142: 1246‐1252, 2012.

Uncertainty remains regarding the efficacy of low intakes of ergocalciferol (vitamin D2 or D2) and cholecalciferol (vitamin D3 or D3) provided in food to increase serum 25-hydroxy-vitamin D (25-OH-D) metabolite concentrations when UV-B exposure is low. We recruited 40 healthy men and women into a double-blind, parallel design, randomized controlled trial. Participants received placebo or 1 of 4 experimental treatments (D2 or D3 at 5 or 10 mg/d) supplied as a malted milk drink for 4 wk during a period of minimal UV-B exposure in the UK. The primary outcome was a change in serum 25-OH-D2 and 25-OH-D3 concentrations measured by ultra-performance liquid chromatography tandem MS. The secondary outcomes were changes in concentrations of plasma parathyroid hormone and serum calcium (Ca 2+ ). Baseline concentrations (geometric mean 6 SD) of 25-OH-D2, 25-OH-D3, and total 25-OH-D were 3 6 4, 32 6 22, and 37 6 22 nmol/L, respectively. Both D2- and D3-fortified drinks resulted in dose-dependent increases (P , 0.001) in their respective 25-OH metabolites that did not significantly differ in size. Increments from baseline compared with the placebo group following 5 and 10 m g/d of D2 were (mean 6 SEM) 9.4 6 2.5 and 17.8 6 2.4 nmol/L for 25-OH-D2 and following 5 and 10 m g/d of D3 were 15.16 4.7 and 22.9 6 4.6 nmol/L for 25-OH-D3, respectively. There was no difference between D2 and D3 groups in the incremental AUC of their respective metabolites. These findings suggest that D2 and D3 are equipotent in increasing 25-OH-D in healthy men and women with negligible UV-B exposure. J. Nutr. 142: 1286‐1290, 2012.

Glycol chitosan was linked to vitamin D2 hemisuccinate (ergocalciferol hemisuccinate) for controlled release through water-soluble carbodiimide activation. The resulting conjugate formed self-assembled nanoparticles in aqueous solution with particle size of 279 nm and ergocalciferol hemisuccinate content of 8.4% (w/w). Almost spherical 50–90 nm nanoparticles were observed by scanning and transmission electron microscopy upon drying. Drug linking to glycol chitosan was confirmed by FTIR spectroscopy and proton NMR. Particles were also characterized by differential scanning calorimetry and wide-angle X-ray diffraction studies. In vitro vitamin D2 release studies performed in water at acid pH indicated a drug release dependence on the solution acidity. Almost constant release rate was observed during the first 8 h. These results indicate that the obtained nanoparticles could be good candidates for vitamin D2 release to animals and humans. Glycol chitosan was linked to vitamin D2 hemisuccinate (ergocalciferol hemisuccinate) for controlled release through water-soluble carbodiimide activation. The resulting conjugate formed self-assembled nanoparticles in aqueous solution with particle size of 279 nm and ergocalciferol hemisuccinate content of 8.4% (w/w). Almost spherical 50–90 nm nanoparticles were observed by scanning and transmission electron microscopy upon drying. Drug linking to glycol chitosan was confirmed by FTIR spectroscopy and proton NMR. Particles were also characterized by differential scanning calorimetry and wide-angle X-ray diffraction studies. In vitro vitamin D2 release studies performed in water at acid pH indicated a drug release dependence on the solution acidity. Almost constant release rate was observed during the first 8 h. These results indicate that the obtained nanoparticles could be good candidates for vitamin D2 release to animals and humans.

In present communication, effect of in vitro and ex vitro culture conditions was investigated on the yield of fat soluble vitamins in chickpea (Cicer arietinum L.) seedlings analyzed by high performance liquid chromatography (HPLC). Amongst the tested culture conditions (in vitro and ex vitro); in vitro condition proved to be highly effective compared to ex vitro. We noticed a significant difference in vitamins D (ergocalciferol), E (α-tocopherol) and K (phylloquinone) yields in chickpea seedlings grown in two different conditions. Maximum yield with a linear increase in vitamins D and E was noticed upto 9 days old in vitro grown seedlings. Vitamin K yield was also high in in vitro grown seedlings with a linear increase upto 11 days. Although, vitamin A was not detected, the vitamin production in germinating seeds was, therefore, age and culture condition dependent. The study revealed that, in in vitro condition, the level of fat soluble vitamins enhanced in seedlings, which could be used for human consumption with value addition in the diet of vegetarians.

The synthesis of two potent osteoclast-forming suppressing agents isolated from the Chinese mushroom Agrocybe chaxingu , demethylincisterol A 3 and chaxine A, was accomplished using ergocalciferol as the starting material. Our methodology for the synthesis of demethylincisterol A 3 and chaxine A featured the construction of a butenolide moiety by the intramolecular Horner–Wadsworth–Emmons reaction under Masamune–Roush conditions. This is the first reported synthesis of chaxine A.

Introduction There is emerging evidence on the widespread tissue effects of vitamin D. Aims To formulate a position statement on the role of vitamin D in postmenopausal women. Materials and methods Literature review and consensus of expert opinion. Results and conclusions Epidemiological and prospective studies have related vitamin D deficiency with not only osteoporosis but also cardiovascular disease, diabetes, cancer, infections and neurodegenerative disease. However the evidence is robust for skeletal but not nonskeletal outcomes where data from large prospective studies are lacking. The major natural source of vitamin D is cutaneous synthesis through exposure to sunlight with a small amount from the diet in animal-based foods such as fatty fish, eggs and milk. Vitamin D status is determined by measuring serum 25-hydroxyvitamin D [25(OH)D] levels. Optimal serum 25(OH)D levels are in the region of 30–90 ng/mL (75–225 nmol/L) though there is no international consensus. Levels vary according to time of the year (lower in the winter), latitude, altitude, air pollution, skin pigmentation, use of sunscreens and clothing coverage. Risk factors for low serum 25(OH)D levels include: obesity, malabsorption syndromes, medication use (e.g. anticonvulsants, antiretrovirals), skin aging, low sun exposure and those in residential care. Fortified foods do not necessarily provide sufficient amounts of vitamin D. Regular sunlight exposure (without sunscreens) for 15 min, 3–4 times a week, in the middle of the day in summer generate healthy levels. The recommended daily allowance is 600 IU/day increasing to 800 IU/day in those aged 71 years and older. Supplementation can be undertaken with either vitamin D2 (ergocalciferol) or vitamin D3 (cholecalciferol) with monitoring depending on the dose used and the presence of concomitant medical conditions such as renal disease.

Enrichment of vitamin D2 in Agaricus bisporus white button mushroom (WBM) using continuous UV light needs a longer exposure time, which can lead to discoloration. Using a Xenon pulsed UV light source, the yield of vitamin D2 was evaluated in freshly harvested button mushrooms and mushroom slices after exposure to 2.5, 3, 6 and 9 pulses of UV light at an average dose of 1.150 J/cm2 energy per pulse. Increase in vitamin D2 content was proportionate to the number of pulses of UV light. There was no difference in the vitamin D2 content of mushrooms between 200 g and 500 g punnets for the corresponding number of pulses (737 ± 81, 847 ± 38, 1148 ± 182, 1611 ± 444% versus 747 ± 48, 911 ± 35, 1307 ± 109, 2028 ± 181% Daily Value/serving, respectively; P > 0.05). Mushrooms in the top layer showed significantly higher amounts of vitamin D2 content than those in the bottom layer of a 500 g punnet (657 ± 22, 796 ± 76, 1433 ± 138, 1878 ± 178% versus 129 ± 60, 237 ± 117, 403 ± 35, 830 ± 257% DV/serving, respectively; P

Vitamin D deficiency is present in the vast majority of patients with chronic kidney disease (CKD), and correcting a poor vitamin D status is recommended as a treatment of CKD–mineral and bone disorders. In this review, we summarize the molecular and clinical data on the role of vitamin D status for heart diseases and its risk factors, with particular attention to patients with CKD. Experimental studies strongly suggest that vitamin D metabolism has a crucial role in myocardial and vascular pathophysiology. This is supported by observations of vitamin D receptor knockout mice, which suffer from myocardial hypertrophy and arterial hypertension with increased activity of the renin–angiotensin system. In the general population and in particular in CKD patients, a poor vitamin D status is associated with cardiovascular (CV) risk factors and preclinical manifestations of CV disease including coronary calcification. Poor vitamin D status is also associated with prevalent and incident CV diseases, such as heart failure and sudden cardiac death. Native as well as active vitamin D treatments improve CV risk profiles and exert beneficial effects on parameters of myocardial structure and function. Whether vitamin D therapy is effective for the prevention or treatment of CV disease remains to be proven in large-scale randomized controlled trials. Native vitamin D should, however, be supplemented in virtually all CKD patients with reduced 25-hydroxyvitamin D concentrations, and the promising data on antiproteinuric and cardioprotective effects of paricalcitol may extend the future indication spectrum for active vitamin D treatment.

We previously demonstrated that vitamin D₂ (ergocalciferol) triggers axon regeneration in a rat model of peripheral nerve transection. In order to confirm the regenerative potential of this neuroactive steroid, we performed a study in which vitamin D₃ (cholecalciferol) was delivered at various doses to paralytic rats. After spinal cord compression at the T10 level, rats were given orally either vehicle or vitamin D₃ at the dose of 50 IU/kg/day or 200 IU/kg/day. Three months later, M and H-waves were recorded from rat Tibialis anterior muscle in order to quantify the maximal H-reflex (H(max)) amplitude. We also monitored the ventilatory frequency during an electrically induced muscle fatigue known to elicit the muscle metaboreflex and an increase in respiratory rate. Spinal cords were then collected, fixed and immunostained with an anti-neurofilament antibody. We show here that vitamin D-treated animals display an increased number of axons within the lesion site. In addition, rats supplemented with vitamin D₃ at the dose of 200 IU/kg/day exhibit (i) an improved breathing when hindlimb was electrically stimulated; (ii) an H-reflex depression similar to control animals and (iii) an increased number of axons within the lesion and in the distal area. Our data confirm that vitamin D is a potent molecule that can be used for improving neuromuscular adaptive mechanisms and H-reflex responses.

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 (≤60nmol/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.

Vitamin D was discovered and had its chemical structure described in the early years of the last century. Although classified as a nutrient because it was found in small quantities in butter, it soon became clear that exposure of skin to sunlight, supplies most of the vitamin D necessary for good health in human beings. Vitamin D (D3 or cholecalciferol) synthesis in the skin is extremely rapid and remarkably robust despite the complexity of the mechanisms involved. However, a number of factors related to latitude location, season, and skin characteristics can interfere with the photoproduction of vitamin D. The 2 forms of vitamin D (D3 or D2-ergocalciferol) are biologically inactive and require activation in the liver and kidney. The product of the first hydroxylation of vitamin D in the liver, 25-hydroxyvitamin D (25(OH)D), is the marker of vitamin D status. Hypovitaminosis D (serum 25(OH)D

Objective To investigate seasonal variation of vitamin D levels in 148,821 serum samples during a 2 year time period in a northern-latitude city in the United States. Methods Total vitamin D assay testing by chemiluminescence was performed on the DiaSorin Liaison. Vitamin D results were extracted from the laboratory information system without patient identification during 2011 and 2012 and separated by season and vitamin D results: less than 10 ng/mL (deficient), 10–20.0 ng/mL (insufficient), 20.1–30 ng/mL (borderline), 30.1–40 ng/mL (sufficient), 40.1–100 ng/mL, and greater than 100 ng/mL. Results The seasonal winter period constituted the months of January through March; spring, April through June; summer, July through September; and fall, October through December. The data set analyzed included 36,643 samples during the winter, 38,299 in spring, 36,141 in summer, and 37,738 in fall and demonstrated an expected rise and fall in vitamin D levels. Conclusion This retrospective epidemiological study demonstrates seasonal variation of vitamin D levels at clinical decision points. Although not unexpected, this variation has an impact on studies relating low vitamin D levels to higher rates of cancer, cardiovascular disease, multiple sclerosis, diabetes, autoimmune disease, and a host of other health risk assessments.