The objective was to investigate associations of serum vitamin D concentration with depressive symptoms and assess the impact that vitamin D concentration has on the occurrence of depressive symptoms in 20-44-year-old pregnant women, postpartum women, non-pp women (non-pregnant/postpartum women), and men, including a separate subgroup analysis of postpartum breastfeeding and non-breastfeeding women. The study populations were selected from the 2007-2018 NHANES public data. Subjective interview data and objective laboratory data including depressive symptoms, serum vitamin D concentration, nutrient intake, and demographic information were utilized. Two diet patterns were created using principal component analysis, and a Bayesian multinomial model was fit to predict the depression outcomes for each subpopulation. The estimates for the log vitamin D slope parameter were negative for all cohorts; as vitamin D increased, the probability of having no depression increased, while the probability of depression decreased. The pregnant cohort had the steepest vitamin D slope, followed by postpartum women, then non-pp women and men. Higher vitamin D concentration had more impact on decreasing depression risk in pregnant and postpartum women compared to non-pp women and men. Among postpartum women, higher vitamin D concentration had a greater influence on decreasing breastfeeding women\'s depression risk than non-breastfeeding women.

Vitamin D hydroxylation in the liver/kidney results in conversion to its physiologically active form of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]. 1,25(OH)2D3 controls gene expression through the nuclear vitamin D receptor (VDR) mainly expressed in intestinal epithelial cells. Cytochrome P450 (CYP) 24A1 is a catabolic enzyme expressed in the kidneys. Interestingly, a recently identified mutation in another CYP enzyme, CYP3A4 (gain-of-function), caused type III vitamin D-dependent rickets. CYP3A are also expressed in the intestine, but their hydroxylation activities towards vitamin D substrates are unknown. We evaluated CYP3A or CYP24A1 activities on vitamin D action in cultured cells. In addition, we examined the expression level and regulation of CYP enzymes in intestines from mice. The expression of CYP3A or CYP24A1 significantly reduced 1,25(OH)2D3-VDRE activity. Moreover, in mice, Cyp24a1 mRNA was significantly induced by 1,25(OH)2D3 in the intestine, but a mature form (approximately 55 kDa protein) was also expressed in mitochondria and induced by 1,25(OH)2D3, and this mitochondrial enzyme appears to hydroxylate 25OHD3 to 24,25(OH)2D3. Thus, CYP3A or CYP24A1 could locally attenuate 25OHD3 or 1,25(OH)2D3 action, and we suggest the small intestine is both a vitamin D target tissue, as well as a newly recognized vitamin D-metabolizing tissue.

The objective was to explore the patterns of contribution from vitamin D metabolites (D2 and D3) to total vitamin D concentrations in Indian families. This cross-sectional study was carried out in slum-dwelling families residing in Pune city. Data on demography, socio-economic status, sunlight exposure, anthropometry, and biochemical parameters (serum 25OHD2, 25OHD3) via the liquid chromatography-tandem mass spectrometry method were collected. The results are presented for 437 participants (5 to 80 years). One-third were vitamin-D-deficient. Intake of foods containing vitamin D2 or D3 was rarely reported. Irrespective of gender, age, and vitamin D status, the contribution of D3 to total 25OHD concentrations far exceeded that of D2 (p < 0.05). The contribution of D2 ranged from 8% to 33% while that of D3 to 25OHD concentrations ranged from 67% to 92%. 25OHD3 is a major contributor to overall vitamin D concentrations, and the contribution of 25OHD2 was found to be negligible. This implies that sunlight and not diet is currently the major source of vitamin D. Considering that lifestyle and cultural practices may lead to insufficient sunlight exposure for large sections of the society, especially women, dietary contribution to vitamin D concentrations through fortification may play an important role in improving the vitamin D status of Indians.

Studies on the relationship between vitamin D (VitD) and glucose homeostasis usually consider either total VitD or 25OHD3 but not 25OHD2 and epimers. We aimed to evaluate the cross-sectional association of VitD compounds with glucose homeostasis measurements in pregnant women with overweight/obesity participating in the Vitamin D And Lifestyle Intervention for Gestational Diabetes Mellitus Prevention study. Methods: The analysis included 912 women. Inclusion criteria: <20 weeks gestation, body mass index ≥29 kg/m2 and information on exposure and outcome variables at baseline. Measurements: A 75 g OGTT at <20, 24−28 and 35−37 weeks gestation (except if previous diabetes diagnosis). Exposure variables: 25OHD2, 25OHD3 and C3-epimer. Outcome variables: fasting and post-challenge insulin sensitivity and secretion indices, corresponding disposition indices (DI), plasma glucose at fasting and 1 and 2 h, hyperglycemia in pregnancy (HiP). Statistics: Multivariate regression analyses with adjustment. Results: Baseline VitD sufficiency was 66.3%. Overall, VitD compounds did not show strong associations with any glucose homeostasis measures. 25OHD3 showed direct significant associations with: FPG at <20 and 24−28 weeks (standardized β coefficient (β) 0.124, p = 0.030 and 0.111, p = 0.026 respectively), 2 h plasma glucose at 24−28 weeks (β 0.120, p = 0.018), and insulin sensitivity (1/HOMA-IR, β 0.127, p = 0.027) at 35−37 weeks; it showed an inverse association with fasting DI (QUCKI*HOMA-β) at <20 and 24−28 weeks (β −0.124, p = 0.045 and β −0.148, p = 0.004 respectively). 25OHD2 showed direct associations with post-challenge insulin sensitivity (Matsuda, β 0.149, p = 0.048) at 24−28 weeks) and post-challenge DI (Matsuda*Stumvoll phase 1) at 24−28 and 35−37 weeks (β 0.168, p = 0.030, β 0.239, p = 0.006). No significant association with C3-epimer was observed at any time period. Conclusions: In these women with average baseline VitD in sufficiency range, VitD compounds did not show clear beneficial associations with glucose homeostasis measures.

The effect of metabolically active bariatric surgery treatment on lipid metabolism is inconclusive. The authors of this study presume that initial vitamin D status may play a regulating role in influencing the beneficial post-effects of bariatric surgery, especially the lipid profile. The biochemical data obtained from 24 patients who had undergone laparoscopic one-anastomosis gastric bypass (OAGB) at baseline, 3 months before the surgery, at the time of surgery, and 6 months later, demonstrate that vitamin D status influenced the postoperative lipid profile. The baseline established the partition line which divided patients into two groups according to the stated calcidiol initial concentration level of 32 ng/mL. The data shows that OAGB induces a decrease in TG and hsCRP while increasing HDL. Conversely, in patients whose 25(OH)D3 was below 32 ng/mL TC significantly increased while those above this concentration remained in the normal physiological range. The changes induced by OAGB in TG, glucose, and hsCRP were similar in both groups. Unexpectedly, the surgery did not affect vitamin D metabolites. In conclusion, the results of the study suggest that a higher concentration of serum 25(OH)D3 may enhance the protective effects of OAGB.

This dataset demonstrates the content of vitamin D3 and 25-hydroxyvitamin D3 (25OHD3) in boiled, grilled, and raw pork for both lean and whole cuts including ribs, tenderloin, shoulder, neck, belly, and center chops. Total fat content in raw pork cuts was determined by the gravimetric method. Quantification analysis using reverse phase high performance liquid chromatography with UV-DAD connected. Purification of fatty acids from proteins was performed using a c18 cartridge, resolving vitamin D3 from its metabolites by subjecting purified samples to polar silica cartridge, purification from sugars by a series of two amino columns, and quantification of vitamin D3 and 25-hydroxyvitamin D3 by injecting purified extracts on C18 SPE column. Grilling samples was conducted in an electric combi oven.

Neonatal dried blood spots (DBS) provide a remarkable resource for biobanks. These microsamples can provide information related to the genetic correlates of disease and can be used to quantify a range of analytes, such as proteins and small molecules. However, after routine neonatal screening, the amount of DBS sample available is limited. To optimize the use of these samples, there is a need for sensitive assays which are integrated across different analytic platforms. For example, after DNA extraction, protein extracts are available for additional analyses. We describe a sensitive and robust LC-MS/MS method for 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 optimized for leftover protein extracts from DBS, which has excellent recovery, precision, and accuracy.

OBJECTIVE: Lipemia is one of the causes of interference in immunoassay and LC-MS/MS methods. Increased prevalence of vitamin D deficiency in the US, where obesity is gradually increasing, raises the suspicion that high levels of fat diet and blood lipid levels interfere with vitamin D measurement results. The focus of this study was to investigate the effect of blood lipid profiles on vitamin D results and prevent the matrix effect.
METHODS: In this study, 25OH vitamin D3 (25OHD3) levels of 100 samples consecutively accepted to biochemistry laboratory regardless of age and sex were measured by the LC-MS/MS method, and each sample was restudied after 1/10 dilution. After dilution restudy, two groups were obtained-group 1 (results deviating below 20%) and group 2 (results deviating above 20%)-and the difference between the groups was investigated. There were 79 patients in group 1 and 21 patients in group 2. In our study, lipid profiles (triglyceride, total cholesterol, HDL, LDL) from the same samples of consecutive vitamin D patients were studied.
RESULTS: It was observed that the triglyceride, total cholesterol HDL, LDL, and 25OHD3 measurements of group 1 and group 2 were similar (p > 0.05). While the mean vitamin D value in the second group was 9.94 ± 7.85, the mean vitamin D value after dilution was measured as 39.23 ± 18.13 and was statistically significant. 25OHD3 concentrations of 21 patients out of 100 were found to be falsely low. Measurements were repeated to confirm the results.
CONCLUSIONS: The matrix effect caused by exogenous and endogenous interferences in the blood could be a hidden factor increasing the prevalence of vitamin D deficiency by causing falsely low 25OHD3 values. Suspicious results should be remeasured by a dilution study.

In this review, we discuss the sources, formation, metabolism, function, biological activity, and potency of C3-epimers (epimers of vitamin D). We also determine the role of epimerase in vitamin D-binding protein (DBP) and vitamin D receptors (VDR) according to different subcellular localizations. The importance of C3 epimerization and the metabolic pathway of vitamin D at the hydroxyl group have recently been recognized. Here, the hydroxyl group at the C3 position is orientated differently from the alpha to beta orientation in space. However, the details of this epimerization pathway are not yet clearly understood. Even the gene encoding for the enzyme involved in epimerization has not yet been identified. Many published research articles have illustrated the biological activity of C3 epimeric metabolites using an in vitro model, but the studies on in vivo models are substantially inadequate. The metabolic stability of 3-epi-1α,25(OH)2D3 has been demonstrated to be higher than its primary metabolites. 3-epi-1 alpha, 25 dihydroxyvitamin D3 (3-epi-1α,25(OH)2D3) is thought to have fewer calcemic effects than non-epimeric forms of vitamin D. Some researchers have observed a larger proportion of total vitamin D as C3-epimers in infants than in adults. Insufficient levels of vitamin D were found in mothers and their newborns when the epimers were not included in the measurement of vitamin D. Oral supplementation of vitamin D has also been found to potentially cause increased production of epimers in mice but not humans. Moreover, routine vitamin D blood tests for healthy adults will not be significantly affected by epimeric interference using LC-MS/MS assays. Recent genetic models also show that the genetic determinants and the potential factors of C3-epimers differ from those of non-C3-epimers.Most commercial immunoassays techniques can lead to inaccurate vitamin D results due to epimeric interference, especially in infants and pregnant women. It is also known that the LC-MS/MS technique can chromatographically separate epimeric and isobaric interference and detect vitamin D metabolites sensitively and accurately. Unfortunately, many labs around the world do not take into account the interference caused by epimers. In this review, various methods and techniques for the analysis of C3-epimers are also discussed. The authors believe that C3-epimers may have an important role to play in clinical research, and further research is warranted.

The demand for vitamin D testing is increasing in China. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) immunoassay is specific and accurate but requires expensive equipment, experienced operators, and complicated pretreatment of serum. Automated immunoassays are simple and convenient but only determine total 25-hydroxyvitamin D (25OHD). The objective of this study was to quantify 25OHD2 and 25OHD3 in patients to assist clinical physicians and laboratory directors in choosing the most appropriate method to determine 25OHD.
Vitamin D testing was conducted for 23,695 patients in Peking Union Medical College Hospital from May 2015 to January 2017. Using this large data set, the prevalence and levels of 25OHD2 were analyzed. LC-MS/MS was used to separately determine 25OHD2 and 25OHD3.
25OHD2 (≥2.5 ng/mL) was detected in 16.4% (3877/23,695) of patients. Males had a significantly lower incidence of detectable 25OHD2 (p<0.01); 1077 (13.9%) samples contained detectable 25OHD2 (median: 3.7 ng/mL; 2.5%-97.5%: 2.5-17.2 ng/mL). For females, 2800 (17.5%) samples contained detectable 25OHD2 (median: 4.0 ng/mL; range: 2.5-20.6 ng/mL). Of the 3877 patients with detectable 25OHD2, males had a significantly higher level of 25OHD3 (p<0.01). There was no significant difference in total 25OHD. The proportion of 25OHD2 in total 25OHD was 1.3%-100%; 87.5% (3391/3877) of the samples contained <10 ng/mL 25OHD2. 25OHD2 negatively correlated with 25OHD3 (r=-0.197, p<0.01) and positively correlated with total 25OHD (r=0.217, p<0.01).
Prevalence of 25OHD2 in patients tested for vitamin D is relatively high in China. 25OHD2 is significantly negatively correlated with 25OHD3.