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BackgroundVitamin D deficiency has been a critical global health issue within the pediatric population. Closed-off management brought about by the COVID-19 pandemic has drastically impacted outdoor activities and sunlight exposure, however, whether it indirectly further exacerbated the vitamin D deficiency has not been largely investigated, especially among children in China. The purpose of this study was to evaluate 25(OH)D concentrations in children before and during the COVID-19 lockdown and to analyze the factors influencing their vitamin D status.MethodsA cross-sectional survey included children aged 1–6 years from Han Zhong Central Hospital in the southern Shanxi Province of China. This study examined healthy children from a pediatric health care department over two periods: before COVID-19 (March 2019–February 2020), and during COVID-19 (March 2020–February 2021). Total 25(OH)D concentrations were compared between the two observation periods. Vitamin D status was determined by 25(OH)D concentrations: deficient (

The National Institute of Standards and Technology (NIST), in collaboration with the National Institutes of Health, Office of Dietary Supplements (NIH ODS), introduced the first Standard Reference Material® (SRM) for determining vitamin D metabolites in 2009 motivated by significant concerns about the comparability and accuracy of different assays to assess vitamin D status. After 14 years, a suite of five serum matrix SRMs and three calibration solution SRMs are available. Values were also assigned for vitamin D metabolites in five additional SRMs intended primarily to support measurements of other clinical diagnostic markers. Both the SRMs and the certification approach have evolved from significant exogenous serum content to primarily endogenous content and from value assignment by combining the results of multiple analytical methods to the use of measurements exclusively from reference measurement procedures (RMPs). The impact of the availability of these SRMs can be assessed by both the distribution information (sales) and by reports in the scientific literature describing their use for method validation, quality control, and research. In this review, we describe the development of these SRMs, the evolution in design and value assignment, the expansion of information reported, and SRM use in validating analytical methods and providing quality assurance within the vitamin D measurement community. [ABSTRACT FROM AUTHOR]

BackgroundHearing loss (HL) is increasingly recognized as a significant global public health issue, and research on its relationship with vitamin D levels has gained wider attention. However, the association between serum biomarkers 25-hydroxyvitamin D2 (25(OH)D2) and D3 (25(OH)D3) with different types of HL remains unclear. This study aimed to investigate the potential association of serum 25(OH)D2 and 25(OH)D3 with HL in US adults.MethodsA sample of 3,684 individuals aged 20–69 years from the 2015–2016 National Health and Nutrition Examination (NHANES) was analyzed in this study. HL was defined as a pure tone average > 25 dB in either ear at low frequencies (500, 1,000, 2000 Hz), speech frequencies (500, 1,000, 2000, 4,000 Hz), and high frequencies (3,000, 4,000, 6,000, 8,000 Hz). Logistic regression was employed to examine the association between serum 25(OH)D2 and 25(OH)D3 and HL. The study population was then stratified by age, gender, race, and education level to analyze potential differences between adults in different subgroups.ResultsIn the multivariate analysis, it was found that serum 25(OH)D2 was independently associated with low-frequency hearing loss (LFHL) (OR: 1.012 [95% CI, 1.005–1.020]) and speech-frequency hearing loss (SFHL) (OR: 1.011 [95% CI, 1.003–1.018]). Restrictive cubic spline analysis demonstrated a linear dose–response relationship between serum 25(OH)D2 levels and LFHL (p for linearity

Introduction: Bladder neoplasm ranks as the second most prevalent reproductive system malignancy worldwide. On the other hand, the vitamin D as an anti-cancer agent has been a subject of long-standing speculation. Hence, the objective of this study is to explore the correlation between vitamin D and bladder neoplasm risk. Materials and Methods: This systematic review and meta-analysis were designed following the PRISMA checklist. Eligible studies were identified through searches on ProQuest, PubMed, Web of Science, Google Scholar, and Cochrane, with no time restrictions until November 18, 2023. Data analysis was conducted utilizing the STATA 14 software. Results: Serum vitamin D levels less than 50 nmol/L increased the risk of bladder neoplasm (OR: 1.33; 95% CI: 1.08, 1.64), muscle-invasive bladder cancer (MIBC) (OR: 2.73; 95% CI: 1.80, 4.14) and non-muscle invasive bladder cancer (NMIBC) (OR: 1.87 (95% CI: 1.39, 2.52)). However, the risk of bladder neoplasm in people whose serum vitamin D level was less than 50 nmol/L did not increase with age. Vitamin D serum levels greater than or equal to 50 nmol/L in people aged 40 to 49 (OR: 0.51; 95% CI: 0.27, 0.99) prevented bladder neoplasm, but no significant association was seen in people over 50 years old. In addition, there was no significant association between daily vitamin D intake and the risk of bladder neoplasm (OR: 1.13; 95% CI: 0.63, 2.06). Conclusion: The serum vitamin D less than 50 nmol/L was correlated with bladder cancer risk increasing, including MIBC and NMIBC. Registration: This study has been compiled based on the PRISMA checklist, and its protocol was registered on the PROSPERO (CRD42023487519) and Research Registry (UIN: reviewregistry1754) Websites.

Vitamin D is involved in many different functions in the human body. Despite the well-known benefits of vitamin D and increasing trends of testing and supplementation, there is still a high prevalence of vitamin D deficiency worldwide, present in the general but also in the highly-active population such as athletes and dancers. Dancers are at a higher risk of vitamin D deficiency due to their long working hours spent indoors, without exposure to sunlight. The high level of workload and physical demands also put dancers at a high risk of injuries. The aim of this review was to systematically analyze the existing evidence on the association of vitamin D (serum level and supplementation) and injuries in dancers. Medline, Scopus, SportDiscus and Web of Science were searched to identify the available peer-reviewed articles. Five articles met the inclusion criteria (two interventional and three observational studies), three of which additionally investigated the relationship between vitamin D and muscle function. The reported results on positive influence of vitamin D serum level or vitamin D supplementation effect on reduced injury occurrence and enhanced muscular function in adolescent and elite professional ballet dancers are promising but the evidence is limited due to a low number of studies, small samples, and methodological limitations.

Simple Summary: Vitamin D biology in equids is unique and poorly understood. Naturally managed (grazing) horses rely on dietary vitamin D2 (ergocalciferol) to provide adequate vitamin D, because endogenous ultraviolet radiation-mediated synthesis of vitamin D3 (cholecalciferol) is ineffective in this species. To test the hypothesis that the management of stabled, non-grazing racehorses is a risk factor for low vitamin D status, the vitamin D status of non-grazing Thoroughbred racehorses in Hong Kong (HK) and grazing Thoroughbred racehorses in the United Kingdom (UK) was compared. The HK horses had lower serum concentrations of 25-hydroxyvitamin D2 (25OHD2) and total 25-hydroxyvitamin D (total 25OHD: the index of vitamin D status), reflecting reduced dietary vitamin D2 intake. These data indicate that HK racehorses required dietary vitamin D3 supplementation to maintain adequate vitamin D status. The inverse relationship between the serum concentrations of 25OHD2 and 25OHD3, previously identified in humans, was observed for the first time in horses, indicating that further study is needed to determine the optimal form of dietary vitamin D supplementation for Thoroughbred racehorses. There is a paucity of data relating to the vitamin D status of racehorses. We hypothesised that the management of racehorses in Hong Kong (HK) predisposes to low vitamin D status unless they receive dietary supplementation. Serum concentrations of 25-hydroxyvitamin D2 (25OHD2), 25-hydroxyvitamin D3 (25OHD3) and total 25-hydroxyvitamin D (total 25OHD) for 79 non-grazing HK racehorses were compared with those for 22 racehorses training in the United Kingdom (UK) that grazed for ≥1 h/d, and for which published data exists. A nested group of 41 HK horses was sampled twice to determine the effect of the duration in HK on vitamin D status. The HK horses had significantly lower serum concentrations of total 25OHD and 25OHD2 than the UK horses; 25OHD2 was undetectable in 15/79 HK sera and serum concentrations of 25OHD2 declined with the duration in HK. The main determinants of vitamin D status were assessed using linear regression; the retained variables were the 25OHD3 concentration and the duration in HK. The inverse relationship between the serum concentrations of 25OHD2 and 25OHD3, previously identified in humans, was observed for the first time in horses. In conclusion, HK racehorses have low serum 25OHD2 and total 25OHD concentrations and rely on D3 supplementation to maintain adequate vitamin D status. Further study is required to determine the optimal form of dietary vitamin D supplementation for Thoroughbred racehorses. [ABSTRACT FROM AUTHOR]

Fresh mushrooms exposed to ultraviolet (UV) radiation prior to drying generate high concentrations of vitamin D2. The aim of this study was to determine the retention of D vitamers in mushrooms that were pulse UV irradiated, then air dried, and stored for up to 12 months. Fresh button mushrooms (A. bisporus) were exposed to pulsed UV radiation (dose 200 mJ/cm2, peak of 17.5 W/cm2), air dried and vacuum sealed before being stored in the dark at room temperature. After storage, samples were freeze dried and quantified for D vitamers using triple quadrupole mass spectrometry. After 3, 6 and 12 months of storage, there was 100% (11.0 ± 0.8 µg/g dry weight (DW), 93% (10.1 ± 0.6 µg/g DW) and 58% (5.5 ± 0.6 µg/g DW) retention of vitamin D2 and 88% (0.14 ± 0.01 µg/g DW), 71% (0.11 ± 0.01 µg/g DW) and 68% (0.1 ± 0.01 µg/g DW) retention of 25-hydroxyvitamin D2 (25(OH)D2), respectively. Compared to the irradiated dried mushrooms that were not stored, the D vitamer concentration was statistically significantly lower (p < 0.05) at 6 and 12 months for 25(OH)D2 and at 12 months for vitamin D2. Sufficient vitamin D2 (99 µg) remained after 12 months storage to provide at least 100% of daily dietary vitamin D requirements in a 20 g serving. [ABSTRACT FROM AUTHOR]

BackgroundEmerging evidence suggests sarcopenia, which is involved in the serum vitamin D deficiency and development of abnormal muscle metabolism, is predominately centered in the general older population. In the present study, we aimed to explore the relationship between the level of serum vitamin D and irisin concentrations in the older adults with sarcopenia.MethodsA cross-sectional study was conducted which included 422 sarcopenia participants (146 males and 276 females). Sarcopenia was assessed according to the recommended diagnostic criteria of the Asia Working Group for Sarcopenia (AWGS). The levels of serum 25-hydroxyvitamin D (25(OH)D), 25-hydroxyvitamin D2 (25(OH)D2) and 25-hydroxyvitamin D3 (25(OH)D3) were determined by LC-MS/MS. Irisin levels were measured by enzyme-linked immunosorbent assay (ELISA). The relationship between serum concentration of vitamin D and irisin were determined using multiple linear regression analysis.ResultsAfter adjustment for potential confounding factors, a significant and positive relationship between changes in irisin across 25(OH)D, and 25(OH)D3 was observed (standard regression coefficients of 0.150 and 0.151, respectively, P < 0.05). However, no significant relationship was observed between serum vitamin D concentrations and irisin levels in males.ConclusionsThis study demonstrated that a higher level of serum vitamin D is independently related to the increment of irisin in sarcopenia females, not in males. These investigations need to be verified in other large-scale prospective studies.

Fresh mushrooms exposed to ultraviolet (UV) radiation prior to drying generate high concentrations of vitamin D2. The aim of this study was to determine the retention of D vitamers in mushrooms that were pulse UV irradiated, then air dried, and stored for up to 12 months. Fresh button mushrooms (A. bisporus) were exposed to pulsed UV radiation (dose 200 mJ/cm2, peak of 17.5 W/cm2), air dried and vacuum sealed before being stored in the dark at room temperature. After storage, samples were freeze dried and quantified for D vitamers using triple quadrupole mass spectrometry. After 3, 6 and 12 months of storage, there was 100% (11.0 ± 0.8 µg/g dry weight (DW), 93% (10.1 ± 0.6 µg/g DW) and 58% (5.5 ± 0.6 µg/g DW) retention of vitamin D2 and 88% (0.14 ± 0.01 µg/g DW), 71% (0.11 ± 0.01 µg/g DW) and 68% (0.1 ± 0.01 µg/g DW) retention of 25-hydroxyvitamin D2 (25(OH)D2), respectively. Compared to the irradiated dried mushrooms that were not stored, the D vitamer concentration was statistically significantly lower (p < 0.05) at 6 and 12 months for 25(OH)D2 and at 12 months for vitamin D2. Sufficient vitamin D2 (99 µg) remained after 12 months storage to provide at least 100% of daily dietary vitamin D requirements in a 20 g serving.

The Vitamin D External Quality Assessment Scheme (DEQAS) distributes human serum samples four times per year to over 1000 participants worldwide for the determination of total serum 25-hydroxyvitamin D [25(OH)D)]. These samples are stored at −40 °C prior to distribution and the participants are instructed to store the samples frozen at −20 °C or lower after receipt; however, the samples are shipped to participants at ambient conditions (i.e., no temperature control). To address the question of whether shipment at ambient conditions is sufficient for reliable performance of various 25(OH)D assays, the equivalence of DEQAS human serum samples shipped under frozen and ambient conditions was assessed. As part of a Vitamin D Standardization Program (VDSP) commutability study, two sets of the same nine DEQAS samples were shipped to participants at ambient temperature and frozen on dry ice. Twenty-eight laboratories participated in this study and provided 34 sets of results for the measurement of 25(OH)D using 20 ligand binding assays and 14 liquid chromatography–tandem mass spectrometry (LC–MS/MS) methods. Equivalence of the assay response for the frozen versus ambient DEQAS samples for each assay was evaluated using multi-level modeling, paired t-tests including a false discovery rate (FDR) approach, and ordinary least squares linear regression analysis of frozen versus ambient results. Using the paired t-test and confirmed by FDR testing, differences in the results for the ambient and frozen samples were found to be statistically significant at p < 0.05 for four assays (DiaSorin, DIAsource, Siemens, and SNIBE prototype). For all 14 LC–MS/MS assays, the differences in the results for the ambient- and frozen-shipped samples were not found to be significant at p < 0.05 indicating that these analytes were stable during shipment at ambient conditions. Even though assay results have been shown to vary considerably among different 25(OH)D assays in other studies, the results of this study also indicate that sample handling/transport conditions may influence 25(OH)D assay response for several assays. [ABSTRACT FROM AUTHOR]

• Vitamin d is known to play a pivotal role in depression. However the two independent metabolites, 25(OH)D 3 and 25(OH)D 2 have totally different associations with depression. • Serum 25(OH)D 3 level has an inverse linear association with depressive symptoms in US females. • The association between 25(OH)D 2 and depression follows an inverted U-shaped curve. The role of vitamin D in depression has been gaining increased research interest. However, little is known about the independent associations of serum 25-hydroxyvitamin D 3 (25(OH)D 3) and D 2 (25(OH)D 2) with depressive symptoms. The goal of this study was to examine the metabolites of vitamin D and their associations with depression. This study was conducted in US females using data from the National Health and Nutrition Examination Survey 2011–2014. Depressive symptoms were assessed using a nine-item Patient Health Questionnaire, and serum 25(OH)D 3 and 25(OH)D 2 levels were measured using liquid chromatography-tandem mass spectrometry. Linear regression and generalized additive models were applied to evaluate the associations between 25(OH)D 3 , 25(OH)D 2 and depression. A negative association between serum 25(OH)D 3 and depressive symptoms was observed in the fully adjusted model. This association was also found among widowed, divorced, separated, and never-married individuals. The association between 25(OH)D 2 and depressive symptoms was not statistically significant, but the dose-response analysis revealed an inverted U-shaped relationship between them with an inflection point at 56.2 nmol/L. To the left of the inflection point, we detected a positive association between 25(OH)D 2 and depressive symptoms, whereas a negative association was observed to the right of the inflection point. The study used a cross-sectional approach, so causation cannot be determined. Our study shows an inverse linear association between serum 25(OH)D 3 and depressive symptoms in US females. The association between 25(OH)D 2 and depression follows an inverted U-shaped curve with the inflection point at 56.2 nmol/L. [ABSTRACT FROM AUTHOR]

ObjectiveVitamin D is critical for calcium and bone metabolism. Vitamin D insufficiency impairs skeletal mineralization and bone growth rate during childhood, thus affecting height and health. Vitamin D status in children with short stature is sparsely reported. The purpose of the current study was to investigate various vitamin D components by high-performance liquid chromatography–tandem mass spectrometry (LC-MS/MS) to better explore vitamin D storage of short-stature children in vivo.MethodsSerum circulating levels of 25-hydroxyvitamin D2 [25(OH)D2], 25-hydroxyvitamin D3 [25(OH)D3], and 3-epi-25-hydroxyvitamin D3 [3-epi-25(OH)D3, C3-epi] were accurately computed using the LC-MS/MS method. Total 25(OH)D [t-25(OH)D] and ratios of 25(OH)D2/25(OH)D3 and C3-epi/25(OH)D3 were then respectively calculated. Free 25(OH)D [f-25(OH)D] was also measured.Results25(OH)D3 and f-25(OH)D levels in short-stature subgroups 2 (school age: 7~12 years old) and 3 (adolescence: 13~18 years old) were significantly lower compared with those of healthy controls. By contrast, C3-epi levels and C3-epi/25(OH)D3 ratios in all the three short-stature subgroups were markedly higher than the corresponding healthy cases. Based on cutoff values developed by Endocrine Society Recommendation (but not suitable for methods 2 and 3), sufficient storage capacities of vitamin D in short-stature subgroups 1, 2, and 3 were 42.8%, 23.8%, and 9.0% as determined by Method 3 [25(OH)D2/3+25(OH)D3], which were lower than those of 57.1%, 28.6%, and 18.2% as determined by Method 1 [25(OH)D2+25(OH)D3+C3-epi] and 45.7%, 28.5%, and 13.6% as determined by Method 2 [25(OH)D2/3+25(OH)D3+C3-epi]. Levels of 25(OH)D2 were found to be weakly negatively correlated with those of 25(OH)D3, and higher 25(OH)D3 levels were positively correlated with higher levels of C3-epi in both short-stature and healthy control cohorts. Furthermore, f-25(OH)D levels were positively associated with 25(OH)D3 and C3-epi levels in children.ConclusionsThe current LC-MS/MS technique can not only separate 25(OH)D2 from 25(OH)D3 but also distinguish C3-epi from 25(OH)D3. Measurement of t-25(OH)D [25(OH)D2+25(OH)D3] alone may overestimate vitamin D storage in children, and short-stature children had lower vitamin D levels compared with healthy subjects. Ratios of C3-epi/25(OH)D3 and 25(OH)D2/25(OH)D3 might be alternative markers for vitamin D catabolism/storage in short-stature children. Further studies are needed to explore the relationships and physiological roles of various vitamin D metabolites.

Vitamin D (VD) deficiency is associated with muscle weakness. A reduction in the incidence of falls in the elderly following VD supplementation and identification of the VD receptor within muscle cells suggests a direct effect of VD on muscle, but little is known about the underlying mechanisms. Here we systematically searched the literature to identify effects of active VD [1,25(OH)2D3] on skeletal muscle myogenesis in vitro, with no restriction on year of publication. Eligibility was assessed by strict inclusion/exclusion criteria and agreed by two independent investigators. Twelve relevant pa-pers were identified using four different cell types (C2C12, primary mouse satellite cells, primary chick myoblasts, and primary human myoblasts) and a range of myogenic markers (myoD, myogenin, creatine kinase, myosin heavy chain, and myotube size). A clear inhibitory effect of 1,25(OH)2D3 on proliferation was reported, while the effects on the different stages of differentiation were less consistent probably due to variation in cell type, time points and doses of 1,25(OH)2D3 used. However, myotube size was consistently increased by 1,25(OH)2D3. Overall, the evidence suggests that 1,25(OH)2D3 inhibits proliferation and promotes differentiation of myoblasts, but future studies should use time courses to gain a clearer understanding.

Objective: Vitamin D is critical for calcium and bone metabolism. Vitamin D insufficiency impairs skeletal mineralization and bone growth rate during childhood, thus affecting height and health. Vitamin D status in children with short stature is sparsely reported. The purpose of the current study was to investigate various vitamin D components by high-performance liquid chromatography–tandem mass spectrometry (LC-MS/MS) to better explore vitamin D storage of short-stature children in vivo. Methods: Serum circulating levels of 25-hydroxyvitamin D2 [25(OH)D2], 25-hydroxyvitamin D3 [25(OH)D3], and 3-epi-25-hydroxyvitamin D3 [3-epi-25(OH)D3, C3-epi] were accurately computed using the LC-MS/MS method. Total 25(OH)D [t-25(OH)D] and ratios of 25(OH)D2/25(OH)D3 and C3-epi/25(OH)D3 were then respectively calculated. Free 25(OH)D [f-25(OH)D] was also measured. Results: 25(OH)D3 and f-25(OH)D levels in short-stature subgroups 2 (school age: 7~12 years old) and 3 (adolescence: 13~18 years old) were significantly lower compared with those of healthy controls. By contrast, C3-epi levels and C3-epi/25(OH)D3 ratios in all the three short-stature subgroups were markedly higher than the corresponding healthy cases. Based on cutoff values developed by Endocrine Society Recommendation (but not suitable for methods 2 and 3), sufficient storage capacities of vitamin D in short-stature subgroups 1, 2, and 3 were 42.8%, 23.8%, and 9.0% as determined by Method 3 [25(OH)D2/3+25(OH)D3], which were lower than those of 57.1%, 28.6%, and 18.2% as determined by Method 1 [25(OH)D2+25(OH)D3+C3-epi] and 45.7%, 28.5%, and 13.6% as determined by Method 2 [25(OH)D2/3+25(OH)D3+C3-epi]. Levels of 25(OH)D2 were found to be weakly negatively correlated with those of 25(OH)D3, and higher 25(OH)D3 levels were positively correlated with higher levels of C3-epi in both short-stature and healthy control cohorts. Furthermore, f-25(OH)D levels were positively associated with 25(OH)D3 and C3-epi levels in children. Conclusions: The current LC-MS/MS technique can not only separate 25(OH)D2 from 25(OH)D3 but also distinguish C3-epi from 25(OH)D3. Measurement of t-25(OH)D [25(OH)D2+25(OH)D3] alone may overestimate vitamin D storage in children, and short-stature children had lower vitamin D levels compared with healthy subjects. Ratios of C3-epi/25(OH)D3 and 25(OH)D2/25(OH)D3 might be alternative markers for vitamin D catabolism/storage in short-stature children. Further studies are needed to explore the relationships and physiological roles of various vitamin D metabolites. [ABSTRACT FROM AUTHOR]

Vitamin D (VD) deficiency is associated with muscle weakness. A reduction in the incidence of falls in the elderly following VD supplementation and identification of the VD receptor within muscle cells suggests a direct effect of VD on muscle, but little is known about the underlying mechanisms. Here we systematically searched the literature to identify effects of active VD [1,25(OH)2D3] on skeletal muscle myogenesis in vitro , with no restriction on year of publication. Eligibility was assessed by strict inclusion/exclusion criteria and agreed by two independent investigators. Twelve relevant pa-pers were identified using four different cell types (C2C12, primary mouse satellite cells, primary chick myoblasts, and primary human myoblasts) and a range of myogenic markers (myoD, myogenin, creatine kinase, myosin heavy chain, and myotube size). A clear inhibitory effect of 1,25(OH)2D3 on proliferation was reported, while the effects on the different stages of differentiation were less consistent probably due to variation in cell type, time points and doses of 1,25(OH)2D3 used. However, myotube size was consistently increased by 1,25(OH)2D3. Overall, the evidence suggests that 1,25(OH)2D3 inhibits proliferation and promotes differentiation of myoblasts, but future studies should use time courses to gain a clearer understanding. [ABSTRACT FROM AUTHOR]

An interlaboratory study was conducted through the Vitamin D Standardization Program (VDSP) to assess commutability of Standard Reference Materials® (SRMs) and proficiency testing/external quality assessment (PT/EQA) samples for determination of serum total 25-hydroxyvitamin D [25(OH)D] using ligand binding assays and liquid chromatography-tandem mass spectrometry (LC-MS/MS). A set of 50 single-donor serum samples were assigned target values for 25-hydroxyvitamin D2 [25(OH)D2] and 25-hydroxyvitamin D3 [25(OH)D3] using reference measurement procedures (RMPs). SRM and PT/EQA samples evaluated included SRM 972a (four levels), SRM 2973, six College of American Pathologists (CAP) Accuracy-Based Vitamin D (ABVD) samples, and nine Vitamin D External Quality Assessment Scheme (DEQAS) samples. Results were received from 28 different laboratories using 20 ligand binding assays and 14 LC-MS/MS methods. Using the test assay results for total serum 25(OH)D (i.e., the sum of 25(OH)D2 and 25(OH)D3) determined for the single-donor samples and the RMP target values, the linear regression and 95% prediction intervals (PIs) were calculated. Using a subset of 42 samples that had concentrations of 25(OH)D2 below 30 nmol/L, one or more of the SRM and PT/EQA samples with high concentrations of 25(OH)D2 were deemed non-commutable using 5 of 11 unique ligand binding assays. SRM 972a (level 4), which has high exogenous concentration of 3-epi-25(OH)D3, was deemed non-commutable for 50% of the LC-MS/MS assays. [ABSTRACT FROM AUTHOR]

Purpose: Paclitaxel (PTX) has transpired as a significant agent in the treatment of breast cancer. Meanwhile, polylactic glycolic acid (PLGA) nanoparticles (NPs) are able to increase the anticancer effect of the PTX in the blood. Methods: Nano-precipitation was used to prepare the PLGA-PTX-VitD3 co-delivery NPs. Drug loading, encapsulation efficiency, in vitro release profile, cell viability, migration, apoptosis, and bcl2 expression of NPs were evaluated. Results: The average size of co-delivery NPs was 231 ± 46 nm. Observed was a controlled release of the PTX and vitamin D3 from co-delivery NPs between 0.5 and 240 hours. MTT showed the ability of 8 μg.mL-1 of co-delivery NPs to kill 50 % of the MCF-7; likewise, the co-delivery NPs prevented MCF-7 migration. The co-delivery NPs led 46.35 % MCF-7 to enter primary apoptosis. 60.8% of MCF-7 in the control group were able to enter the G (1) phase of the cell cycle. The co-delivery NPs increased expression of bax. In addition to its higher toxicity against MCF-7 than that of PTX, co-delivery NPs were able to release drugs continuously for a long period, which indeed increased the efficiency of the drugs. Conclusion: The effect of co-delivery NPs on MCF-7 cell viability was different from that in other drugs. In fact, the co-deliver NPs were able to release drugs continuously for a long time, this could induce primary apoptosis in the MCF-7 and decrease the metastasis and toxicity of drugs.

Aim of the study: Vitamin D deficiency is known to be associated with disease severity, unresponsiveness to treatment, and morbidity among patients with chronic viral hepatitis B and C, autoimmune hepatitis, and alcoholic hepatitis. This study aims to research vitamin D levels in patients suffering from cirrhotic and non-cirrhotic phases of hepatitis D. Material and methods: 170 individuals in total were included in the study in the form of two groups: the first group of 100 patients with chronic hepatitis D (CHD), 30 of whom had cirrhosis, and the second control group of 70 individuals with similar characteristics to those of the first group in terms of age, type, and seasonal sampling. Levels of 25-hydroxy vitamin D [25(OH)D] were measured in the serum collected from patients and the control group. Results: The lowest 25(OH)D levels were identified in patients with cirrhotic CHD. When these levels were compared with those of the control group, they were found to be significant (15.30 ±6.92 and 18.90 ±8.30 ng/ml, respectively, p = 0.04). 25(OH)D deficiency (< 10 ng/ml) was detected at significantly higher rates in patients with both cirrhotic and non-cirrhotic CHD compared to the healthy controls (30%, 25%, and 8.5%, respectively, p = 0.01). A significant correlation was established between 25(OH)D levels and bilirubin in patients with CHD (r = 0.252, p = 0.012). Multivariate analysis showed that chronic hepatitis D (odds ratio [OR] = 3.608, 95% confidence interval [CI]: 1.31-9.89, p = 0.013) and age (OR = 1.04, 95% CI: 1.00-1.08, p = 0.033) were associated with vitamin D deficiency. Conclusions: Frequency of 25(OH)D vitamin deficiency is higher in patients with CHD. The identification of vitamin D levels and the replacement of any deficiency may create a positive effect on disease progression, morbidity, and mortality levels. [ABSTRACT FROM AUTHOR]

Monoclonal immunoglobulins can cause interference in many laboratory analyses. During a 4 month period we observed seven patients with monoclonal disease and falsely extremely elevated 25-hydroxyvitamin D (25(OH)D) results above 160 ng/mL (>400 nmol/L) measured using an immunoassay from Abbott Diagnostics. Based on these findings, we studied the occurrence of falsely elevated 25(OH)D in samples with paraproteins and investigated possible mechanisms of the observed interference. 25(OH)D was analyzed using the Architect i2000 platform from Abbott Diagnostics and a higher order method, liquid chromatography-mass spectrometry (LC-MS/MS), in serum samples from 50 patients with known monoclonal disease. Patients with falsely elevated 25(OH)D were included in further studies to elucidate the cause of interference. Spuriously elevated results were in addition analyzed on two alternative platforms (Siemens and Roche). Falsely elevated 25(OH)D levels were present in eight patients on the Abbott analyzer and one on the Siemens platform. Results from Roche were comparable with LC-MS/MS. Additional investigations excluded elevated concentrations of rheumatoid factor and heterophilic antibodies as the cause of interference in the Abbott assay. Laboratories should be aware of the risk of falsely elevated 25(OH)D in samples run on the Architect analyzer from patients with monoclonal disease. Highly elevated vitamin D results should be diluted and if the dilution is non-linear, rerun by a different method, preferably LC-MS/MS. In patients with spuriously elevated 25(OH)D without known monoclonal disease, the laboratory should consider requesting protein electrophoresis to exclude paraprotein interference. [ABSTRACT FROM AUTHOR]

Background: Despite recommendations to limit vitamin D testing to specific clinical scenarios, test volume remains high in many clinical laboratories. Automated total vitamin D immunoassays frequently under- or over-recover 25-hydroxyvitamin D2 [25(OH)D2], making accurate assessment of vitamin D status difficult in patients taking high-dose 25(OH)D2 supplements. Mass spectrometry-based methods offer excellent recovery of 25(OH)D2 but are not practical for use in all laboratories. In this study, we evaluated 2 automated immunoassays against an LC–MS/MS method performed at a national reference laboratory. Methods: A method comparison against LC–MS/MS was performed for the Roche Elecsys Vitamin D total II assay and the IDS-iSYS 25 VitDS immunoassays using 49 patient specimens submitted for clinical 25(OH)D measurement. Mean bias was calculated, and vitamin D status was determined for each specimen according to the 2011 Endocrine Society clinical practice guidelines. Results: Theil–Sen regression lines relative to LC–MS/MS were y = 0.88x + 2.94 for Roche and y = 1.03x + 2.48 for IDS. Mean bias (±SD) in samples with 25(OH)D2 concentrations less than 5 ng/mL was −0.25 ng/mL (±6.30) for Roche and −1.45 ng/mL (±6.82) for the IDS. Mean bias (±SD) in samples with 25(OH)D2 concentrations greater than 5 ng/mL was −3.19 ng/mL (±6.61) for Roche and 5.52 ng/mL (±6.36) for IDS. Median percentage recovery of 25(OH)D2 was 87.1% (interquartile range 76.0–111.3) for Roche and 120.6% (interquartile range: 105.3–133.4) for IDS. Vitamin D status was misclassified in 7 samples by the Roche assay and 3 by the IDS assay. For all but one of the discordant pairs, the immunoassay result was within 1.7 ng/mL of the diagnostic cutoff. Conclusions: The automated immunoassays evaluated here demonstrate improved recovery of 25(OH)D2 relative to previous generations. Both are acceptable for use in the determination of vitamin D status. [ABSTRACT FROM AUTHOR]

Objectives Dry eye disease (DED) is an increasingly important public health problem in Korea. Previous studies conducted in Korea have reported inconsistent results regarding the protective effects of vitamin D on DED, and these discrepancies may be related to the relatively simple questionnaire that has been used. Thus, we evaluated the association of serum vitamin D levels with DED using the ocular surface disease index (OSDI). Methods The present study evaluated data from participants in the Study Group for Environmental Eye Disease (2014-2015). This group included data from 752 participants, and data from 740 participants (253 men and 487 women) were analyzed in the present study. DED severity was evaluated using the OSDI. Results Higher serum vitamin D levels were associated with a non-significantly reduced risk of DED in the crude analysis (odds ratio [OR], 0.991; 95% confidence interval [CI], 0.971 to 1.011) and in the adjusted analysis (OR, 0.988; 95% CI, 0.966 to 1.010). In the crude analysis of no/mild DED vs. moderate/severe DED, men exhibited a decreased risk with increasing serum vitamin D levels (OR, 0.999; 95% CI, 0.950 to 1.051), while women exhibited an increased risk (OR, 1.003; 95% CI, 0.979 to 1.027). In these analyses, we found no significant associations. Conclusions The findings of the present study support previous reports that serum vitamin D levels are not associated with DED.

Purpose: Paclitaxel (PTX) has transpired as a significant agent in the treatment of breast cancer. Meanwhile, polylactic glycolic acid (PLGA) nanoparticles (NPs) are able to increase the anticancer effect of the PTX in the blood. Methods: Nano-precipitation was used to prepare the PLGA-PTX-VitD3 co-delivery NPs. Drug loading, encapsulation efficiency, in vitro release profile, cell viability, migration, apoptosis, and bcl2 expression of NPs were evaluated. Results: The average size of co-delivery NPs was 231 ± 46 nm. Observed was a controlled release of the PTX and vitamin D3 from co-delivery NPs between 0.5 and 240 hours. MTT showed the ability of 8 μg.mL-1 of co-delivery NPs to kill 50 % of the MCF-7; likewise, the co-delivery NPs prevented MCF-7 migration. The co-delivery NPs led 46.35 % MCF-7 to enter primary apoptosis. 60.8% of MCF-7 in the control group were able to enter the G (1) phase of the cell cycle. The co-delivery NPs increased expression of bax. In addition to its higher toxicity against MCF-7 than that of PTX, co-delivery NPs were able to release drugs continuously for a long period, which indeed increased the efficiency of the drugs. Conclusion: The effect of co-delivery NPs on MCF-7 cell viability was different from that in other drugs. In fact, the co-deliver NPs were able to release drugs continuously for a long time, this could induce primary apoptosis in the MCF-7 and decrease the metastasis and toxicity of drugs. [ABSTRACT FROM AUTHOR]

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The detection of 25-hydroxyvitamin D (25OHD) from dried blood spots (DBS) has been widely studied. However, the existing pretreatment methods suffer from limitations in terms of throughput (usually exceeding 2 h), complexity (involving liquid-liquid extraction or solid-phase extraction), and contamination (including multiple steps of organic solvent evaporation). We first released 25OHD from DBS samples by 50% acetonitrile solution through ultrasonication. Subsequently, the cold-induced phase separation technique was introduced for in-situ concentration and purification. Afterward, the PTAD derivatization of 25OHD was performed directly, profiting from the high acetonitrile content in the concentrated solution. In the end, the resulting solution was submitted to LC-MS/MS for quantification. The established LC-MS/MS methodology possessed favorable analytical performance, possessing lower limit of quantification of 1 ng/mL pointing to plasma, accuracy of 86.8–110.1% and imprecision of 5.4–16.8%. Method comparison with plasma samples demonstrated that over 93% of the detections met the acceptance limit for cross-validation of ±20%. The novel sample preparation can be finished within 15 min and eliminated the traditional steps of extraction and organic solvent evaporation. Based on this high-throughput, reliable and applicable LC-MS/MS method, the detection of 25OHD in DBS samples can be better achieved for clinical patients and researchers with relevant demands. [Display omitted] • A highly applicable LC-MS/MS method was developed for 25OHD detection in DBS. • Extraction and PTAD derivatization of 25OHD can be completed within 15 min. • The new method can provide consistent results as the plasma-based method. [ABSTRACT FROM AUTHOR]

Objectives: Epidemiological studies have reported that vitamin D deficiency is associated with inflammatory disease. Smoking is a well-known risk factor for inflammation. However, few studies have investigated the interactive effect of vitamin D deficiency and smoking on inflammation. This study aims to investigate the interaction of vitamin D and smoking with inflammatory markers in the urban elderly. Methods: We used data from the Korean Elderly Environmental Panel Study, which began in August 2008 and ended in August 2010, and included 560 Koreans ≥60 years old living in Seoul. Data was collected via questionnaires that included items about smoking status at the first visit. Vitamin D levels, high-sensitivity C-reactive protein (hs-CRP), and white blood cell (WBC) counts were repeatedly measured up to three times. Results: The association of vitamin D and hs-CRP was significant after adjusting for known confounders (β=-0.080, p=0.041). After separate analysis by smoking status, the association of vitamin D deficiency and hs-CRP in smokers was stronger than that in nonsmokers (smokers: β=-0.375, p=0.013; non-smokers: β=-0.060, p=0.150). Smoking status was an effect modifier that changed the association between vitamin D deficiency and hs-CRP (interaction estimate: β=-0.254, p=0.032). Vitamin D was not significantly associated with WBC count (β=0.003, p=0.805). Conclusions: Vitamin D deficiency was associated with hs-CRP in the urban elderly. Smoking status was an effect modifier of this association. Vitamin D deficiency was not significantly associated with WBC count.

Introduction: Vitamin D deficiency is a common problem in Parkinson's disease (PD). We investigated 25-hydroxyvitamin D [25(OH)D] values and related factors in ambulatory patients with PD. Materials and Method: This descriptive study included 48 (25 women, 23 men) patients with idiopathic PD. Serum 25(OH)D and parathormone levels as well as falls within the previous six months were recorded. Disease severity was evaluated by Hoehn and Yahr Scale. Bone mineral density (BMD) was measured using dual-energy X-ray absorptiometry at the L1-L4 spine and femoral neck. Results: Mean age was 64.4±10.2 years. Mean disease duration was 5.5±3.0 years. Median Hoehn and Yahr stage was 2 (Min-max: 1-3). Mean serum 25(OH)D level was 27.35±9.83 ng/mL. 54.1% of the patients with PD had vitamin D deficiency or insufficiency. The falling rate in the last six months was 41.7%. Median number of falls was 1.5 (Min-max: 1-5). There was a statistically significantly negative correlation between serum 25(OH)D level and disease duration, disease severity, number of falls, and serum parathormone level, whereas a significant positive correlation was found between serum 25(OH)D level and femoral neck BMD. Multiple linear regression analysis revealed that disease duration and number of falls were the predictors of the serum 25(OH)D level (Adjusted R²=0.54, F=28.6, p<0.0001). Conclusion: This study suggests that disease duration and number of falls are main predictors for low serum vitamin D level, and that serum parathormone level and femoral neck bone density may be affected by low serum vitamin D level in patients with PD. [ABSTRACT FROM AUTHOR]

Background We evaluated the performance of ultra-performance liquid chromatography-tandem mass spectrometry to measure serum 3-epi-25-hydroxyvitamin D3, 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 concentrations in 519 infant, paediatric and adolescent serum samples in Korea. Methods We used a Kinetex XB-C18 column and isocratic methanol/water (77.5/22.5, v/v) with 0.025% (v/v) high-performance liquid chromatography solvent additive flowing at 0.25 mL/min, yielding an 11 min/sample run time. A TQD triple quadrupole mass spectrometer in electrospray ionization positive ion mode with multiple reaction monitoring transition via an MSMS vitamin D kit was used to evaluate precision, carryover, ion suppression and linearity. Samples were prepared using the 4-phenyl-1,2,4-triazoline-3,5-dione derivatization method. Results Intra- and inter-run precisions were 1.23-13.28% and 1.02-10.08%, respectively. Group carryovers were -0.27% and 0.10%, respectively. There was no ion suppression. The calibration curve showed good linearity from calibrator Level 1 (11.75 nmol/L) to 6 (375 nmol/L) with R2 > 0.9999. The 3-epi-25-hydroxyvitamin D3 and 25-hydroxyvitamin D3 peaks were clearly separated in the extracted ion chromatogram. Infant serum samples 3-epi-25-hydroxyvitamin D3 concentrations were significantly higher than paediatric and adolescent concentrations. Conclusions The ultra-performance liquid chromatography-tandem mass spectrometry assay performed acceptably, clearly separating 3-epi-25-hydroxyvitamin D3 from 25-hydroxyvitamin D3. High 3-epi-25-hydroxyvitamin D3 concentrations were observed in infant but not in paediatric and adolescent serum samples. [ABSTRACT FROM AUTHOR]

Background Maternal vitamin D deficiency is associated with maternal and perinatal adverse effects. This study was conducted to assess the vitamin D status among pregnant Sudanese women. Methods A total of 180 pregnant women were enrolled in a cross-sectional study in Saad Abualila Hospital, Khartoum, Sudan. The medical history of each woman was collected and 25-hydroxyvitamin D [25(OH)D] was measured using an electrochemiluminescence immunoassay. Results The median age, gravidity and gestational age was 27.7 y, 1.0 and 10.7 weeks, respectively. Of the 180 woman, 169 (93.9%) had vitamin D deficiency (≤20 ng/ml). There was no correlation between the 25(OH)D level and body mass index (r=−0.135, p=0.071) or haemoglobin level (r= 0.001, p=0.999). Conclusions The current study showed a high prevalence of vitamin D deficiency. Further studies investigating the risk factors for vitamin D deficiency and the outcome of pregnancy are needed. [ABSTRACT FROM AUTHOR]

Background: The 2007-2010 NHANES provides the first US nationally representative serum 25-hydroxyvitamin D [25(OH)D] concentrations measured by standardized liquid chromatography-tandem mass spectrometry.Objective: We describe patterns for total 25(OH)D and individual metabolites in persons aged ≥1 y stratified by race-ethnicity and grouped by demographic, intake, physiologic, and lifestyle variables.Methods: We measured 25-hydroxycholecalciferol [25(OH)D3], 25-hydroxyergocalciferol [25(OH)D2], and C3-epimer of 25(OH)D3 [C3-epi-25(OH)D3] in serum samples (n = 15,652) from the 2007-2010 cross-sectional NHANES [total 25(OH)D = 25(OH)D3 + 25(OH)D2].Results: Concentrations (median, detection rate) of 25(OH)D3 (63.6 nmol/L, 100%) and C3-epi-25(OH)D3 (3.40 nmol/L, 86%) were generally detectable; 25(OH)D2 was detectable in 19% of the population. Total 25(OH)D, 25(OH)D3, and C3-epi-25(OH)D3 displayed similar demographic patterns and were strongly correlated (Spearman's r > 0.70). Concentrations of 25(OH)D2 (90th percentile) were much higher in persons aged ≥60 y (17.3 nmol/L) than in younger age groups (≤4.88 nmol/L). We noted significant race-ethnicity differences in mean total 25(OH)D [non-Hispanic blacks (NHBs), Hispanics, and non-Hispanic whites (NHWs): 46.6, 57.2, and 75.2 nmol/L, respectively] and in the prevalence of total 25(OH)D <30 nmol/L overall (24% of NHBs, 6.4% of Hispanics, and 2.3% of NHWs) as well as stratified by season (winter months: 30% of NHBs, 7.5% of Hispanics, and 3.8% of NHWs; summer months: 17% of NHBs, 4.4% of Hispanics, and 1.6% of NHWs). Persons with higher vitamin D intakes (diet, supplements, or both) and those examined during May-October had significantly higher total 25(OH)D. Significant race-ethnicity interactions in a multiple linear regression model confirmed the necessity of providing race-ethnicity-specific estimates of total 25(OH)D.Conclusions: Race-ethnicity differences in the prevalence of low total 25(OH)D remained strong even after adjustment for season to account for the NHANES design imbalance between season, latitude, and race-ethnicity. The strong correlation between C3-epi-25(OH)D3 and 25(OH)D3 may be because the epimer is a metabolite of 25(OH)D3. The presence of 25(OH)D2 mainly in older persons is likely a result of high-dose prescription vitamin D2. [ABSTRACT FROM AUTHOR]

There is a need for food-based solutions for preventing vitamin D deficiency. Vitamin D3 (D3) is mainly used in fortified food products, although the production of vitamin D2 (D2) is more cost-effective, and thus may hold opportunities. We investigated the bioavailability of D2 from UV-irradiated yeast present in bread in an 8-week randomised-controlled trial in healthy 20–37-year-old women (n 33) in Helsinki (60°N) during winter (February–April) 2014. Four study groups were given different study products (placebo pill and regular bread=0 µg D2 or D3/d; D2 supplement and regular bread=25 µg D2/d; D3 supplement and regular bread=25 µg D3/d; and placebo pill and D2-biofortified bread=25 µg D2/d). Serum 25-hydroxyvitamin D2 (S-25(OH)D2) and serum 25-hydroxyvitamin D3 (S-25(OH)D3) concentrations were measured at baseline, midpoint and end point. The mean baseline total serum 25-hydroxyvitamin D (S-25(OH)D=S-25(OH)D2+S-25(OH)D3) concentration was 65·1 nmol/l. In repeated-measures ANCOVA (adjusted for baseline S-25(OH)D as total/D2/D3), D2-bread did not affect total S-25(OH)D (P=0·707) or S-25(OH)D3 (P=0·490), but increased S-25(OH)D2 compared with placebo (P<0·001). However, the D2 supplement was more effective than bread in increasing S-25(OH)D2 (P<0·001). Both D2 and D3 supplementation increased total S-25(OH)D compared with placebo (P=0·030 and P=0·001, respectively), but D2 supplementation resulted in lower S-25(OH)D3 (P<0·001). Thus, D2 from UV-irradiated yeast in bread was not bioavailable in humans. Our results support the evidence that D2 is less potent in increasing total S-25(OH)D concentrations than D3, also indicating a decrease in the percentage contribution of S-25(OH)D3 to the total vitamin D pool. [ABSTRACT FROM PUBLISHER]

Objective: Actualization of statement in 25-hydroxyvitamin D measurement Method: Information from websites and other documents of international vitamin D standardization program VDSP and from recent publications deals with 25-hydroxyvitamin D measurements in human serum Results: After 1 year we can see that effect of international standardization program VDSP is negligible both in precision and bias values. Significantly higher problems with analytical quality show immunochemical methods than separate measurements. Standardization process VDSP continues in 2015-2016 by study of the commutability experiments.Their target is to create highly commutable control materials for CAP and DEQAS programs, assured reliable verification of standardization level in routine and research clinical laboratories. Many recent publications are devoted to assessment of components, negatively influenced the results of 25-hydroxyvitamin D measurements in serum samples. Objects of interest are namely 25-hydroxyvitamin D2 and its level of cross immunoreaction, influence on bias level by 24, 25-dihydroxyvitamin D content, problems with 3-epi-25-hydroxyvitamin D3, and also with vitamin D bound protein (VDSP). These influences, resulted to elevated bias values are much more frequented in immunochemical methods than in separate methods and are in different immunochemical methods very different. Discussion: Risk of increased bias values namely in immunochemical methods can decreased reliability of classification of deficiency by vitamin D in individual patients. It also can be the source of problems in analysis of children due to high concentration of 3-epi-25-hydroxyvitamin D3 and in analysis of dialyzed patients due to problems with VDBP. [ABSTRACT FROM AUTHOR]

Fresh and dried mushrooms naturally generate vitamin D 2 when exposed to ultraviolet (UV) radiation. Vitamin D 2 and 25-hydroxyvitamin D 2 (25(OH)D 2) concentrations were compared in dried button mushrooms (Agaricus bisporus) exposed to pulsed UV radiation either before or after air-drying. A further aim was to assess the effect of air-drying on the generation of D vitamers. Fresh button mushrooms were irradiated (Irr) with a total of 200 mJ/cm2 pulsed UV radiation before (Irr/AD) or after (AD/Irr) being air-dried (AD). A third group of fresh button mushrooms was irradiated but not air-dried (Irr). Control mushrooms were fresh and untreated. The D vitamers were quantified in freeze-dried samples using triple quadrupole mass spectrometry. Irr/AD mushrooms had more than double the concentration of vitamin D 2 than AD/Irr mushrooms (9.5 µg/g dry weight (dw) vs 4.6 µg/g dw). However, Irr mushrooms contained 6.3 µg/g dw. The concentration of 25(OH)D 2 in Irr mushrooms was 0.05 μg/g dw, while 0.14 μg/g dw was detected in Irr/AD mushrooms. There was no detectable 25(OH)D 2 in control mushrooms, nor in AD/Irr mushrooms. The sequence of irradiating and drying mushrooms was a key factor in generating vitamin D 2. • Pulsed UV radiation of dried mushrooms generates high amounts of vitamin D 2. • Vitamin D 2 higher when mushrooms are exposed to UV before drying vs after drying. • UV radiation generated 25-hydroxyvitamin D 2 in fresh, but not dried, mushrooms. [ABSTRACT FROM AUTHOR]

Background: Low circulating levels of total vitamin D [25(OH)D] and 25(OH)D3 have been associated with vascular complications in few studies on individuals with type 1 diabetes. However, these measures are affected by UV light exposure. Circulating 25(OH)D2, however, solely represents dietary intake of vitamin D2, but its association with complications of diabetes is currently unknown. We investigated the associations between 25(OH)D2 and 25(OH)D3 and the prevalence of albuminuria, retinopathy and cardiovascular disease (CVD) in individuals with type 1 diabetes. Methods: We measured circulating 25(OH)D2 and 25(OH)D3 in 532 individuals (40 ± 10 years old, 51 % men) with type 1 diabetes who participated in the EURODIAB Prospective Complications Study. Cross-sectional associations of 25(OH)D2 and 25(OH)D3 with albuminuria, retinopathy and CVD were assessed with multiple logistic regression analyses adjusted for age, sex, season, BMI, smoking, HbA1c, total-HDL-cholesterol-ratio, systolic blood pressure, antihypertensive medication, eGFR, physical activity, alcohol intake, albuminuria, retinopathy and CVD, as appropriate. Results: Fully adjusted models revealed that 1 nmol/L higher 25(OH)D2 and 10 nmol/L higher 25(OH)D3 were associated with lower prevalence of macroalbuminuria with ORs (95 % CI) of 0.56 (0.43;0.74) and 0.82 (0.72;0.94), respectively. These vitamin D species were not independently associated with microalbuminuria, non-proliferative and proliferative retinopathy or CVD. Conclusions: In individuals with type 1 diabetes, both higher 25(OH)D2 and 25(OH)D3 are associated with a lower prevalence of macroalbuminuria, but not of retinopathy and CVD. Prospective studies are needed to further examine the associations between 25(OH)D2 and 25(OH)D3 and the development of microvascular complications and CVD in type 1 diabetes. [ABSTRACT FROM AUTHOR]

Background Higher sunlight exposure is correlated with lower incidence of breast cancer in ecological studies, but findings from prospective studies regarding the association of circulating levels of vitamin D with the risk of breast cancer have been null. The objective of this study was to examine the relation between plasma levels of vitamin D and the risk of postmenopausal breast cancer. Methods We conducted a nested case-control study within the Multiethnic Cohort Study of five race/ethnic groups (white, African-American, Native Hawaiian, Japanese, and Latino) from Hawaii and Los Angeles between 2001 and 2006. Pre-diagnostic plasma levels of 25- hydroxyvitamin D2 [25(OH)D2], 25-hydroxyvitamin D3 [25(OH)D3] and 25(OH)D (sum of 25(OH)D2 and 25(OH)D3) were examined among 707 postmenopausal breast cancer cases and matched controls. Results Using conditional logistic regression models, 20 ng/mL increases of plasma 25(OH)D3 (odds ratio (OR) 0.28; 95% confidence interval (CI) 0.14-0.56) and 25(OH)D (OR 0.43; 95% CI 0.23-0.80) were inversely associated with breast cancer risk among white women, but not among women in other race/ethnic groups. Using two-segmented, piecewise-linear logistic regression models, the change-points of the ORs, either for 25(OH)D3 or for 25(OH)D, were detected as 20 ng/mL among whites. Conclusions Circulating 25(OH)D3 and 25(OH)D were associated with a reduced risk of postmenopausal breast cancer among whites, but not in other ethnic groups, who reside in low latitude regions. [ABSTRACT FROM AUTHOR]

Abstract: Objectives: For quantification of 25-hydroxyvitamin D3 (25OH-D3), 25-hydroxyvitamin D2 (25OH-D2), 3-epi-25-hydroxyvitamin D3 (3-epi-25OH-D3) and 24R,25-dihydroxyvitamin D3 (24R,25(OH)2-D3) in human serum a high performance liquid chromatography tandem mass spectrometry (HPLC–MS/MS) method was developed and validated. Design and methods: After protein precipitation further purification is achieved with on-line sample preparation using a reversed phase (RP) C-4 column. Chromatographic separation is realized by a RP-column with core shell material and pentafluorophenyl (PFP) selectivity. Atmospheric pressure chemical ionization in the positive ion mode with multi‐reaction monitoring is used for analyte detection. Results: Baseline separation of the analytes is achieved below 10min. The method is linear over the range 4.0–265.3nmol/L for 25OH-D3, 3.9–183.6nmol/L for 25OH-D2, 2.0–133.8nmol/L for 3-epi-25OH-D3 and 2.8–129.9nmol/L for 24R,25(OH)2-D3 (r2 >0.998). The limit of quantification is 4.0nmol/L for 25OH-D3, 3.9nmol/L for 25OH-D2, 2.0nmol/L for 3-epi-25OH-D3 and 2.8nmol/L for 24R,25(OH)2-D3. The CVs for the intra-day and inter-day precision are <5% and <4%, respectively. Metabolite levels for a set of 50 human serum samples have been determined and resulted in the detection of considerable amounts of 3-epi-25OH-D3 and 24R,25(OH)2-D3. Conclusions: This highly specific HPLC–MS/MS method is suitable for vitamin D profiling. There is a correlation between 25OH-D3 and 24R,25(OH)2-D3. Serum concentration of 24R,25(OH)2-D3 increases disproportionally with increasing concentration of 25OH-D3. [Copyright &y& Elsevier]

Abstract: Liquid chromatographic methods with atmospheric pressure chemical ionization mass spectrometry were developed for the determination of the vitamin D metabolites 25-hydroxyvitamin D2 (25(OH)D2), 25-hydroxyvitamin D3 (25(OH)D3), and 3-epi-25-hydroxyvitamin-D3 (3-epi-25(OH)D3) in the four Levels of SRM 972, Vitamin D in Human Serum. One method utilized a C18 column, which separates 25(OH)D2 and 25(OH)D3, and one method utilized a CN column that also resolves the diastereomers 25(OH)D3 and 3-epi-25(OH)D3. Both methods utilized stable isotope labeled internal standards for quantitation of 25(OH)D2 and 25(OH)D3. These methods were subsequently used to evaluate SRM 909c Human Serum, and 25(OH)D3 was the only vitamin D metabolite detected in this material. However, SRM 909c samples contained matrix peaks that interfered with the determination of the [2H6]-25(OH)D3 peak area. The chromatographic conditions for the C18 column were modified to remove this interference, but conditions that separated the matrix peaks from [2H6]-25(OH)D3 on the CN column could not be identified. The alternate internal standard [2H3]-25(OH)D3 did not suffer from matrix interferences and was used for quantitation of 25(OH)D3 in SRM 909c. During the evaluation of SRM 909c samples, a third method was developed using a pentafluorophenylpropyl column that also separates the diastereomers 25(OH)D3 and 3-epi-25(OH)D3. The 25(OH)D3 was measured in SRM 909c using all three methods, and the results were compared. [Copyright &y& Elsevier]