Plasma Vitamin D3 Research Articles

Vitamin D3 reduces the expression of M1 and M2 macrophage markers in breast cancer patients.

Vitamin D3 (VD) is known for its immunomodulatory and anticancer effects. This study aimed to characterize tumor-associated macrophages (TAMs) in breast cancer (BC) and assess the influence of VD and its active metabolite, calcitriol, on their polarization. TAMs were isolated from BC patients and characterized. Monocytes were differentiated into macrophage classes (M0, M1, M2a, M2c) and treated ex vivo with calcitriol. The expression of VD-related proteins in tumor tissue was correlated with TAMs and monocyte-derived macrophages (MDMs) characteristics. TAM expression of CD200R, CD204, CD80, HLA-DR, and CD44 was negatively correlated with CYP27B1 in selected patient groups. Patients with high CYP27B1 tumor expression showed significantly lower CD200R, CD204, and CD44 expression. In patients with normal VD levels and premenopausal, CD80 expression in M2a and M2c MDMs (control, untreated ex vivo with calcitriol) was negatively correlated with plasma VD. Calcitriol reduced HLA-DR during MDM differentiation in all patients; CD80 decrease significantly except in patients with normal VD levels or metastasis. Calcitriol also decreased CD163 expression. The decrease in both M1 and M2 macrophage markers by calcitriol or their negative correlation with CYP27B1 indicate the modulatory, but rather anticancer activity of VD. The intensity of these effects was the strongest in postmenopausal patients and those without metastases.

Corrigendum: The effects of sex, season, and natural sunlight on plasma vitamin D3 levels in two chelonian species (Testudo hermanni, Trachemys scripta) and their interaction with calcium, phosphate, and magnesium as associated plasma compounds

Corrigendum: The effects of sex, season, and natural sunlight on plasma vitamin D3 levels in two chelonian species (Testudo hermanni, Trachemys scripta) and their interaction with calcium, phosphate, and magnesium as associated plasma compounds

Dietary Super-Doses of Cholecalciferol Fed to Aged Laying Hens Illustrates Limitation of 24,25-Dihydroxycholecalciferol Conversion

BackgroundOlder humans taking high concentrations of vitamin D3 supplementation for a prolonged time may be at risk of vitamin D toxicity. It is unclear how dietary super-doses (10,000 times greater than the requirement) can affect vitamin D3 status in aged animals. Aged laying hens could be a model to compare vitamin D3 supplementation effects with women in peri- or postmenopausal stages of life. ObjectivesWe investigated the dietary super-dose impacts of cholecalciferol (vitamin D3) on vitamin D3 status in aged laying hens in production. MethodsForty-eight 68-wk-old Hy-Line Brown laying hens were individually housed in cages with 8 hens per dietary treatment for 11 wk. Hens were randomly assigned to 1 of 6 treatment groups of dietary vitamin D3 supplementation and consumed ad libitum. Supplementation concentrations were 400, 800, 7400, 14,000, 20,000, and 36,000 IU D3/kg of feed. At the end of the study, all hens were sacrificed, and tissue samples and feces were collected. Plasma and egg yolk vitamin D3 metabolites, calcium and phosphorus composition of eggshells, ileal digesta, and feces were measured. Duodenal, ileal, liver, and kidney gene expression levels were also measured. ResultsWe observed that increasing dietary vitamin D3 increased plasma vitamin D3 and egg yolk vitamin D3 (P < 0.0001 for both sites). We also observed an increase in plasma 24,25-dihydroxycholecalciferol as dietary vitamin D3 concentrations increased (P < 0.0001). The plasma 25-hydroxycholecalciferol:24,25-dihydroxycholecalciferol ratio exhibited an asymptotic relationship starting at the 14,000 IU/kg D3 treatment. ConclusionsDietary super-doses of vitamin D3 led to greater plasma and egg yolk vitamin D3 concentrations, which shows that aged laying hens can deposit excess vitamin D3 in egg yolk. We suggest future research should explore how 24-hydroxylation mechanisms are affected by vitamin D3 supplementation. Further understanding of 24-hydroxylation can help ascertain ways to reduce the risk of vitamin D toxicity.

Vitamin D3 deficiency and coronavirus-related health problems

Introduction: In addition to boosting calcium absorption in the small intestine and preserving sufficient levels of calcium and phosphate in the serum to support proper bone mineralization, Vitamin D3 fortifies the immune system. Vitamin D3 deficiency can occur when regular intakes are gradually below recommended levels and when people get little sunshine exposure. Materials and Methods: The study was conducted using a case series study approach. A structured checklist was utilized on 107 individuals who had recovered from corona disease to identify the causes behind the unfavorable health effects they continue to experience. In addition, the patients underwent extensive laboratory blood testing in a specialized laboratory and received follow-up care for a period of 6 months. The results were analyzed using the Chi-square test and the Paired t-test using the SPSS software. Results: Despite the finding that 84.1% of the study group had Vitamin D3 levels below the normal range, their physical health conditions had considerably improved as a result of the treatment they received. Conclusion: The study found that patients with coronavirus infection had a high prevalence of Vitamin D3 deficiency. After 6 months, this percentage dropped considerably, raising the possibility that the coronavirus may have contributed to the drop in plasma Vitamin D3 levels. To find out how long-lasting COVID-19 symptoms are related to Vitamin D3 deficiency, more research is required. The following core competencies are addressed in this article: Medical knowledge, Patient care and procedural skills.

Circadian regulation of calcium and phosphorus homeostasis during the oviposition cycle in laying hens

Maintenance of calcium and phosphorus homeostasis in laying hens is crucial for preservation of skeletal integrity and eggshell quality, though physiological regulation of these systems is incompletely defined. To investigate changes in mineral and vitamin D3 homeostasis during the 24-h egg formation cycle, 32-wk-old commercial laying hens were sampled at 1, 3, 4, 6, 7, 8, 12, 15, 18, 21, 23, and 24 h post-oviposition (HPOP; n ≥ 4). Ovum location and egg calcification stage were recorded, and blood chemistry, plasma vitamin D3 metabolites, circulating parathyroid hormone (PTH), and expression of genes mediating uptake and utilization of calcium and phosphorus were evaluated. Elevated levels of renal 25-hydroxylase from 12 to 23 HPOP suggest this tissue might play a role in vitamin D3 25-hydroxylation during eggshell calcification. In shell gland, retinoid-x-receptor gamma upregulation between 6 and 8 HPOP followed by subsequently increased vitamin D receptor indicate that vitamin D3 signaling is important for eggshell calcification. Increased expression of PTH, calcitonin, and fibroblast growth factor 23 (FGF23) receptors in the shell gland between 18 and 24 HPOP suggest elevated sensitivity to these hormones toward the end of eggshell calcification. Shell gland sodium-calcium exchanger 1 was upregulated between 4 and 7 HPOP and plasma membrane calcium ATPase 1 increased throughout eggshell calcification, suggesting the primary calcium transporter may differ according to eggshell calcification stage. Expression in shell gland further indicated that bicarbonate synthesis precedes transport, where genes peaked at 6 to 7 and 12 to 18 HPOP, respectively. Inorganic phosphorus transporter 1 (PiT-1) expression peaked in kidney between 12 and 15 HPOP, likely to excrete excess circulating phosphorus, and in shell gland between 18 and 21 HPOP. Upregulation of FGF23 receptors and PiT-1 during late eggshell calcification suggest shell gland phosphorus uptake is important at this time. Together, these findings identified potentially novel hormonal pathways involved in calcium and phosphorus homeostasis along with associated circadian patterns in gene expression that can be used to devise strategies aimed at improving eggshell and skeletal strength in laying hens.

The effects of sex, season, and natural sunlight on plasma vitamin D3 levels in two chelonian species (Testudo hermanni, Trachemys scripta) and their interaction with calcium, phosphate, and magnesium as associated plasma compounds

Species specific physiology, seasonal changes, sex, and husbandry factors all influence the blood chemistry of chelonians, including vitamin D3, calcium, phosphate and magnesium levels. Problems in the supply of many of these are commonly seen in captive reptiles. The goal of this study was to measure vitamin D3, calcium, phosphate and magnesium in plasma from captive, healthy, adult Hermann’s tortoises (Testudo hermanni) and pond sliders (Trachemys scripta). Samples were categorized and compared based on species, sex, season, and access to sunlight as a central husbandry element. Blood samples of 522 Hermann’s tortoises and 188 pond sliders, taken from March to September 2022, were included in the study. New reference intervals for vitamin D3, calcium, phosphate and magnesium were established, with specific reference intervals calculated for each species, sex, and season in those cases in which significant differences were found based on these factors. For the calculation of reference intervals for vitamin D3 the factors species, season and access to sunlight were considered. Vitamin D3 levels differed by access to sunlight in some seasons depending on the species and were generally higher in Hermann’s tortoises. Plasma vitamin D3 levels did not correlate with calcium, phosphorus or magnesium levels on a larger scale in either species. Calcium, phosphate, and magnesium were higher in females than in males of both species, and magnesium was higher in Hermann’s tortoises than in pond sliders. Our results can be helpful in a more specific interpretation of blood results and support previous findings that direct sunlight is an important factor in chelonian health. More studies are needed to better understand the role of other hormonal influence on the vitamin D3, calcium, phosphate and magnesium metabolism in chelonians.

A manganese dioxide nanoparticle-bimetallic metal organic framework composite for selective and sensitive detection of vitamin D3 in human plasma.

For the first time a metal organic framework nanomaterial has been developedcomprising manganese dioxide nanoparticle and iron and zinc metal ions interlinked with each other via terephthalic acid. The framework shape was identified as an elongated hexagonal nanorod (TEM) with varying functional groups (FT-IR) and diffraction patterns (XRD). The framework nanocomposite as such in aqueous acidic electrolyte solution has displayed an excellent conductivity (redox behavior) and surface excess (3.08 × 10-8 cm-2). Under the optimized conditions (0.1 M H2SO4 as electrolyte, 50 mV/s scan rate, +1.26 V (vs Ag/AgCl)), the metal organic framework coated electrode has selectively identified vitamin D3 (VD3) in the presence of various other interfering molecules and displayed excellent limit of detection (1.9 ng mL-1). The developed sensor has been applied to the determination of VD3 in extracted human plasma samples (RSD of 0.3-2.6 % and recovery of 96-102 %), and the obtained VD3 values are similar to HPLC-UV method.

ÇOCUKLARDA HELİKOBAKTER PYLORİ ENFEKSİYONU İLE PLAZMA 25 HİDROKSİ VİTAMİN D3 DÜZEYİ ARASINDAKİ İLİŞKİ

OBJECTIVE: The severity of the Helicobacter pylori (H.pylori) infection and the presence H.pylori related diseases are affected by host, bacterial and environmental factors. In this study it is aimed to show relationship between H.pylori infection and plasma Vitamin D3 (vitD3) has significant role in regulation inflamatory response in children with H.pylori infection. MATERIAL AND METHODS: Two hundred one patients aged between 3-18 years, referred to pediatric gastroentrology department of Celal Bayar University Medical Faculty between March 2010 to March 2011 and performed upper gastrointestinal endoscopy because of gastrointestinal symptoms and iron deficiency anemia refractory to medical therapy were enrolled in this study. Histopathologic diagnosis of biopsy speciments of the patients were evaluated. There were 98 patients in H.pylori positive group and 103 patients in H.pylori negative group. Plasma level of vitD3 of H.pylori positive and H.pylori negative cases were compared. Relationship between gastric mucosal activity the degree of inflammation severity and vitD3 level were evaluated in the H.pylori positive group. RESULTS: Vitamin D3 levels were low in 80 (81.6%) H.pylori positive patients and in 76 (73%) H.pylori negative patients. This difference was not statistically significant (p&amp;gt;0.05). The mean value of plasma level of vitD3 H.pylori positive and negative groups were 15.64±8.9 ng/mL and 16.36±11.35 ng/mL respectively. The difference between the groups was not statistically significant (p&amp;gt;0.05). When patients in H.pylori positive and negative groups were classified according to plasma vitD3 level in four different groups as severe deficiency, deficiency, insuficiency and sufficiency. Vitamin D3 severe deficiency in H.pylori positive group was statistically more frequent than H.pylori negative group (p&amp;lt;0.05). Severity of chronic inflamation and tissue H.pylori activity increased inversely with the mean vitD3 level in both groups (p&amp;lt;0.05). CONCLUSIONS: Results of this study suggests that vitD3 deficiency is common in children in our region. Vitamin D3 deficiency is risk factor for H.pylori infection. The findings of this study shows vitD3 increasing antibacterial effect and reducing the severity of inflammation in H.pylori infection.

Effects of Dietary Vitamin E Supplementation on Reproductive Performance, Egg Characteristics, Antioxidant Capacity, and Immune Status in Breeding Geese during the Late Laying Period.

This study aimed to tentatively evaluate the effects of dietary vitamin E (VE) on goose reproductive physiology through the investigation of reproductive performance, egg characteristics, antioxidant capacity, and immune status in breeding geese. A total of 480 female and 96 male Jiangnan White breeding geese were randomly assigned to four treatments with four replicates, and each replicate had 30 females and six males. Four levels of VE were successively added to four treatment diets from 48 to 54 weeks of age, representing the effects of VE deficiency (0 IU/kg), basic-dose VE (40 IU/kg), middle-dose VE (200 IU/kg), and high-dose VE (2000 IU/kg). Neither the egg-laying rate nor the healthy-gosling rate were affected by any of the VE supplementations (p > 0.05). The qualified egg rate, hatchability of fertilized eggs, and spleen index were increased by each VE supplementation (p < 0.05). Egg fertility, the concentration of plasma reproductive hormones (i.e., the follicle-stimulating hormone, estradiol, and progesterone), follicular development, and antioxidant enzyme activities—i.e., the concentration of malondialdehyde (MDA), total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px)—in the liver and ovary were improved by 200 IU/kg of dietary VE (p < 0.05). Plasma VE concentration, immunoglobulin A, and immunoglobulin G content were increased, whereas plasma vitamin D3 concentration was reduced by increasing dietary VE levels to 2000 IU/kg (p < 0.05). The VE deposition of yolk, the yolk color depth, and the albumen rate were increased by each VE supplementation (p < 0.05). Antioxidant enzyme activities (i.e., MDA concentration, T-AOC, SOD, and GSH) in yolk were improved by 200 IU/kg and 2000 IU/kg of dietary VE (p < 0.05), compared with 0 IU/kg. The VE deposition was significantly correlated with GSH activity and the MDA concentration in egg yolk (p < 0.05). However, the high intake of dietary VE (2000 IU/kg vs. 200 IU/kg) decreased egg fertility (p < 0.05) and reduced the antioxidant capacity in the liver and ovary (p < 0.05). The qualified egg rate was positively correlated to immunoglobulin production (p < 0.05). Egg fertility and hatchability were correlatively improved by increased antioxidant enzyme activity; decreased MDA in the liver and ovary; hatchability; and enhanced immune status (p < 0.05). To sum up, both VE deficiency and high-dose VE (2000 IU/kg) reduced reproductive performance, whereas a dose of 200 IU/kg VE achieved optimal fertility, possibly through enhancing antioxidant capacity and immune status.

Microalgae-derived sterols do not reduce the bioavailability of oral vitamin D3 in mice.

Microalgae have drawn increasing attention as sustainable food sources, also because of their lipid-lowering phytosterols. As phytosterols are also discussed critically regarding their effect on the availability of fat-soluble vitamins, this study aimed to investigate microalgae-derived phytosterols and their effect on vitamin D status. GC-MS analysis showed large variations in the phytosterol profiles of microalgal species. The most frequent sterols were β-sitosterol and stigmasterol. To investigate their effects on vitamin D status, 40 mice were randomized to four groups and fed a vitamin D3-adequate (25 μg/kg) Western-style diet with 0% phytosterols (control) or 1% ergosterol (a fungal sterol not typical for microalgae), β-sitosterol or stigmasterol for four weeks. Contrary to the hypothesis that phytosterols adversely affect vitamin D uptake, mice fed β-sitosterol had significantly higher concentrations of vitamin D3 in plasma (3.15-fold, p<0.01), liver (3.15-fold, p<0.05), and skin (4.12-fold, p<0.005) than the control group. Small increases in vitamin D3 in plasma and skin were also observed in mice fed stigmasterol. In contrast, vitamin D3 levels in the ergosterol and control groups did not differ. The increased tissue levels of vitamin D3 in mice fed β-sitosterol and stigmasterol were not attributable to the observed reduction in liver triglycerides in these groups. The data rather suggest that changes in bile acid profiles were responsible for the beneficial effect of microalgae sterols on the bioavailability of vitamin D3. In conclusion, consumption of microalgae might not adversely affect vitamin D status.

EFFECT OF ADDING VITAMIN D3 AND VARIOUS PARTICLE SIZE OF OYSTER SHELL TO DIETS IN CALCIUM AND VITAMIN D3 CONCENTRATION, EGG SHELL AND BONE BREAKING FORCE IN LAYING HENS

This study was conducted in the poultry farm belong to Department of Animal Production at College of Agriculture, University of Anbar, from 20/7/2021 to 15/10/2021, to show the role of vitamin D3 and multi particle size of oyster shells to providing the needed calcium for laying hens in order to manufacture the eggshell throughout the hours of the day and its impact on shell quality and bone health. one hundred-eight Lohman Brown laying hens, 43 weeks of age, were distributed randomly to nine treatments, four replicates for each treatment and three hens for each replicate. The experimental treatments were as follows: T1/ control diet without any additives, T2/ calcium source 50% limestone + 50% oyster shell size less than 1 mm + vitamin D3 4000 IU kg-1, T3/ calcium source 50% limestone +50% oyster shell size 1-2 mm + vitamin D3 4000 IU kg-1, T4/ calcium source 50% limestone + 50% oyster shell size 2-3 mm + vitamin D3 4000 IU kg-1, T5/ calcium source 50% Limestone + 50% oyster shell size 3–5 mm + Vitamin D3 4000 IU kg-1, T6/ Calcium source 50% limestone + 50% oyster shell size less than 1 mm + Vitamin D3 8000 IU kg-1, T7 / Calcium source 50% limestone + 50% oyster shell 1-2 mm +Vitamin D3 8000 IU kg-1, T8 Calcium source 50% limestone + 50% oyster shell 2-3 mm + Vitamin D3 8000 IU kg-1, T9/ Calcium source 50% limestone +50% oyster shell size 3-5mm + Vitamin D3 8000 IU kg-1. The results showed a significant increase (P≤0.05) in levels of calcium in blood plasma for fifth, sixth, seventh, eighth and ninth treatments. The results indicated a Significant increase (P≤0.05) in level of vitamin D3 in blood plasma for the sixth, seventh, eighth and ninth treatments. The results showed a significant increase (P≤0.05) in tibia bone breaking strength and Calcium content in tibia bone for the sixth, seventh, eighth and ninth treatments. The results of eggshell breaking force trait indicated significant increase (P≤0.05) for fourth, fifth, sixth, seventh, eighth and ninth treatments.

Dietary branched-chain amino acids modulate the dynamics of calcium absorption and reabsorption in protein-restricted pigs

BackgroundVery low-protein (VLP) diets negatively impact calcium (Ca) metabolism and absorption. The objective of this study was to investigate the effect of supplemental branched-chain amino acids (BCAA) and limiting amino acids (LAA) on Ca digestibility, absorption and reabsorption in pigs fed with VLP diets. Forty-eight piglets were assigned to six treatments: positive control (PC), negative control (NC), and NC containing LAA 25%, LAA 50%, LAA + BCAA 25% (LB25) and LAA + BCAA 50% (LB50) more than recommendations.ResultsRelative to PC or NC, LB25 and LB50 had higher digestibility of Ca and plasma Ca and phosphorus (P), but lower plasma vitamin D3. LB50 tended to increase vitamin D receptor transcript and protein in the gut, but decreased mRNA or protein abundance of parathyroid hormone 1 receptor (PTH1R), calbindin 1 (CALB1), cytochrome P450 family 27 subfamily B member 1 and occludin in small intestine. LB50 increased the transcript of cytochrome P450 family 24 subfamily A member 1 and PTH1R but decreased the transcript of transient receptor potential cation channel subfamily V member 5, CALB1 and solute carrier family 17 member 4 in kidney.ConclusionOverall, BCAA increased Ca digestibility through regulating the transcellular and paracellular Ca absorption in the gut and reabsorption in kidney during protein restriction.

Correlation between Levels of Vitamins D3 and E in Type 2 Diabetes Mellitus: A Case-Control Study in Serdang, Selangor, Malaysia.

An overview of vitamins D3 and E suggests micronutrient deficiency contributes to type 2 diabetes mellitus (T2DM). A case-control study was conducted to determine the status of plasma vitamins D3 and E isomers amongst diabetic Malaysians. Two groups were recruited for participation, one comprising fifty diabetic subjects (DM) and one comprising fifty non-diabetic (non-DM) subjects, in order to assess their plasma vitamin D3, calcium and vitamin E status. Glycaemic status (haemoglobin A1c, HbA1c; fasting blood glucose, FBG; C-Peptide) and lipid profiles (total cholesterol, TC; triglycerides, TG; low-density lipoprotein-cholesterol, LDL-C; high-density lipoprotein-cholesterol, HDL-C) were assessed, followed by anthropometric measurements. The Mann–Whitney U-test, Kruskal–Wallis and Spearman’s correlation coefficient were used to elucidate the association between levels of plasma vitamins D3 and E and T2DM. The vitamin D3 deficiency group (<20 ng/mL) showed a significant correlation (p < 0.05) with glycaemic status (HbA1c and FBG) and lipid profiles (HDL-C, LDL and TC). Spearman’s correlation demonstrated that vitamin D3 status is strongly correlated with HDL levels (p < 0.05). Similarly, plasma total vitamin E levels >4.9 μg/mL revealed significantly different FBG, HbA1c, C-Peptide, LDL, HDL and TC levels across both groups. Moreover, family history, smoking, waist circumference and HbA1c levels demonstrated a significant association (p < 0.05) with levels of vitamins D and E but not FBG and lipid profiles. This could be because the pre-diabetic status among the non-DM group influenced the outcomes of this study.

Long-term effects of laparoscopic sleeve gastrectomy and Roux-Y-gastric bypass on body composition and bone mass density

Abstract Objective Currently, the two most common bariatric procedures are laparoscopic sleeve gastrectomy (LSG) and laparoscopic gastric bypass (LRYGB). Long-term data comparing the two interventions in terms of their effect on body composition and bone mass density (BMD) is scarce. The aim of this study was to assess body composition and BMD at least five years after sleeve gastrectomy (LSG) and gastric bypass (LRYGB). Methods Bariatric patients at least five years post-surgery (LSG or LRYGB) were recruited and body composition and BMD was measured by means of DEXA. Data from body composition before surgery was included in the analysis. Blood samples were taken for determination of plasma calcium, parathyroid hormone (PTH), Vitamin D3, alkaline phosphatase and C-terminal telopeptide (CTX), and individual risk for osteoporotic fracture assessed by The Fracture Risk Assessment Tool (FRAX) score was calculated. After surgery, all patients received multivitamins, vitamin D3, and zinc. In addition, LRYGB-patients were prescribed calcium. Results A total of 142 patients were included, 72 LSG and 70 LRYGB, before surgery: median BMI 43.1 kg/m2, median age 45.5 years, 62.7% females. Follow-up after a median of 6.7 years. For LRYGB, percentage total weight loss (%TWL) at follow up was 26.3%, and for LSG 24.1%, (p = 0.243). LRYGB lead to a slightly lower fat percentage in body composition. At follow-up, 45% of both groups had a T-score at the femoral neck below -1, indicating osteopenia. No clinically relevant difference between the groups in BMD was found. Conclusion At 6.7 years post-surgery, no difference in body composition and BMD between LRYGB and LSG was found. Deficiencies and bone loss remain an issue after both interventions and should be monitored.

The Positive Impact of Vitamin D on Glucocorticoid-Dependent Skeletal Muscle Atrophy.

(1) The study aimed to investigate whether vitamin D3 supplementation would positively affect rats with glucocorticoids-induced muscle atrophy as measured by skeletal muscle mass in two experimental conditions: chronic dexamethasone (DEX) administration and a model of the chronic stress response. (2) The study lasted 28 consecutive days and was performed on 45 male Wistar rats randomly divided into six groups. These included two groups treated by abdominal injection of DEX at a dose of 2 mg/kg/day supplemented with vegetable oil (DEX PL; n = 7) or with vitamin D3 600 IU/kg/day (DEX SUP; n = 8), respectively, and a control group treated with an abdominal injection of saline (CON; n = 6). In addition, there were two groups of rats chronically stressed by cold water immersion (1 hour/day in a glass box with 1-cm-deep ice/water mixture; temperature ~4 °C), which were supplemented with vegetable oil as a placebo (STR PL; n = 9) or vitamin D3 at 600 IU/kg/day (STR SUP; n = 9). The last group was of sham-stressed rats (SHM; n = 6). Blood, soleus, extensor digitorum longus, gastrocnemius, tibialis anterior, and quadriceps femoris muscles were collected and weighed. The heart, liver, spleen, and thymus were removed and weighed immediately after sacrifice. The plasma corticosterone (CORT) and vitamin D3 metabolites were measured. (3) We found elevated CORT levels in both cold water-immersed groups; however, they did not alter body and muscle weight. Body weight and muscle loss occurred in groups with exogenously administered DEX, with the exception of the soleus muscle in rats supplemented with vitamin D3. Decreased serum 25(OH)D3 concentrations in DEX-treated rats were observed, and the cold water immersion did not affect vitamin D3 levels. (4) Our results indicate that DEX-induced muscle loss was abolished in rats supplemented with vitamin D3, especially in the soleus muscle.

Lyophilization as pre-processing for sample storage in the determination of vitamin D3 and metabolites in serum and plasma

Determination of vitamin D levels in human biological specimens has gained a high relevance over the last decades, essentially because low levels have been associated with several biological disorders. In fact, vitamin D deficiency has become a worldwide health concern covering all ages and genders. The storage of biofluids has to be considered for determination of vitamin D and metabolites in order to fully preserve matrices status. This study attempts to evaluate lyophilization of serum and plasma as a pre-processing step for sample storage prior to quantitative analysis of vitamin D3 and its main hydroxylated metabolites −25(OH)D3, 24,25(OH)2D3 and 1,25(OH)2D3. The protocol including sample lyophilization was characterized in terms of analytical features and compared to the same method, based on SPE–LC–MS/MS, without lyophilization. Sensitivity, precision and accuracy were not affected when we operated with lyophilized serum and plasma and results provided by a set of twenty-four serum samples from DEQAS (Vitamin D External Quality Assessment Scheme) were in agreement with reported concentrations for 25(OH)D3 and 1,25(OH)2D3. A stability study programmed for 9 months allowed ensuring that the concentration of vitamin D3 and metabolites in lyophilized serum and plasma stored at room temperature was not affected during this period. This research has demonstrated that the quantitation of target metabolites is not under the influence of lyophilization. Therefore, including lyophilization prior to analysis could reduce shipment and storage costs, avoid delays of sample processing, and increase the stability of the target analytes due to an effective quenching process.

Markers Indicating Body Vitamin D Stores and Responses of Liver and Adipose Tissues to Changes in Vitamin D Intake in Male Mice.

Circulating 25-hydroxyvitamin D (25(OH)D) is regarded as the most reliable biomarker of vitamin D status. However, limited data exist concerning the suitability of 25(OH)D as an indicator of body vitamin D stores and the ability of adipose tissue to mobilize vitamin D. In the first study, in which male mice received different vitamin D3 doses for three weeks, we found strong linear response relationships between vitamin D3 intake and levels of vitamin D3 in the plasma (p < 0.001), liver (p < 0.001) and adipose tissues (p < 0.001), and strong positive correlations between plasma and tissue stores of vitamin D3 (p < 0.001). Plasma levels of 25(OH)D3 and 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) showed weak or no correlations with tissue vitamin D3 stores. Data from a second study demonstrate a strong and rapid response of plasma 25(OH)D3 in vitamin D3-treated mice with a low vitamin D status. Additionally, mice fed a vitamin D-free diet showed a strong and rapid decline in vitamin D3 in the liver, whereas the decline in different adipose tissues was distinctly lower than that in the liver. To conclude, tissue stores of vitamin D3 were best reflected by plasma vitamin D3. In contrast to the liver, adipose tissues responded less sensitively to an absence of vitamin D intake.

MON-379 Diagnostically Significant Relationship Between the Results of Determination of Vitamin D3 and Its Metabolites by the EIA Method and Pure Vitamin D3 by the LC-MS Method

Diagnostically significant relationship between the results of determination of vitamin D3 and its metabolites by the EIA method and pure vitamin D3 by the LC-MS method.Introduction: Vitamin D is an important hormone in the human body. He is involved in many physiological body processes. Measuring the level of vitamin D in the patient’s blood is important, diagnostic criteria for identifying and confirming a number of diseases: obesity, hypogonadism, sarcopenia, autoimmune pathologies.Two methods currently prevail in the laboratory diagnostic market measurement of the level of vitamin D3 in blood plasma: EIA and LC-MS (with its variety - LC-MS / MS).Fundamental differences in the physicochemical nature of these methods are the basis for differing results determined in the same sample.Objective: In this paper, we set the goal of determining how the results correlate for determining vitamin D3 and its metabolites using the EIA method with the results of determination Pure Vitamin D3 through LC-MS / MS.Materials and methods: The study was conducted at the clinic of Professor Kalinchenko. Have been selected patients with a clinical picture of vitamin D deficiency. These patients were referred to determination of the level of vitamin D3 using the above methods, subject to preparation rules before analysis. EIA was performed using LC-MS / MS was done using AB SCIEX QTRAP 4500 apparatus connected to Waters Acquity UPLC system. The results were combined for subsequent statistical processing. For a pair of EIA / LC-MS methods, we determined the reliability value of the approximation r ^ 2 and the linear regression equation.Results: For the presented data, the level of vitamin D3 and its metabolites determined by EIA, and pure vitamin D3, determined by LC-MS, r ^ 2 was 0.9638 (very strong dependence), and the linear regression equation was as follows: LC-MS [nm / ml] = 1.2808 * (EIA) [nmol / ml] + 6.9731.Discussion: Despite studies by foreign colleagues showing a high level bias and low correlation of results between the EIA and LC-MS methods, our data show a very strong relationship between the two values. By our assumptions, the cause of the various findings is monoclonal antibodies, differing in the global market for reagents for determining the level of hormones derived our linear regression equation allows a practicing endocrinologist to quickly and accurately determine the level of true vitamin D3 in the patient.

Maternal circulating Vitamin D3 levels during pregnancy and behaviour across childhood

Vitamin D deficiency during critical periods of development could lead to persistent brain alterations. We aimed to assess the association between maternal vitamin D3, the major circulatory form of vitamin D, at pregnancy and neurodevelopmental outcomes during childhood, namely: behavioural problems, Attention Deficit and Hyperactivity Disorder (ADHD) and Autism Spectrum Disorder (ASD) symptoms, and social competence. This study included 2,107 mother-child pairs of a Spanish population-based birth cohort. Maternal plasma vitamin D3 was measured in pregnancy. The outcomes were measured through questionnaires at 5, 8, 14, and 18 years old. We ran multivariate regression models adjusted for potential confounding variables. We found that per each 10 ng/mL increment of maternal vitamin D3, children obtained higher social competence scores (coefficient = 0.77; 95% CI = 0.19, 1.35) at 5 years old. However, we observed null associations between maternal vitamin D3 and total behavioural problems and ADHD and ASD symptoms in children from 5 to 18 years old. Further studies carried out in countries where the population is exposed to lower vitamin D levels are needed.

The Impact of Thyme and Rosemary on Prevention of Osteoporosis in Rats.

Osteoporosis poses an important public health problem which affects millions of people worldwide. There is a direct link between calcium deficiency in diet and induction of osteoporosis and bone loss. The current study was conducted to evaluate the protective effect of thyme (Thymus vulgaris L.) and rosemary (Rosmarinus officinalis L.) against osteoporosis in rats with low calcium intake. Essential oils of rosemary and thyme were analyzed. The experiment was carried out on growing male Sprague–Dawley rats; the experimental animals were divided into 5 groups: 1, control negative was fed standard balanced diet; 2, control positive was fed balanced diet with low calcium level (L Ca) (Ca 0.1% w/w); 3, (L Ca) + thyme powder (5% w/w); 4, (L Ca) + rosemary powder (5% w/w); 5, (L Ca) + orally administration with CaCO3 (27 mg/kg body weight). Blood samples were collected for different biochemical analyses in plasma (calcium (Ca), phosphorus (P), magnesium (Mg), tumor necrosis factor-alpha (TNF-α), C-reactive protein (CRP), malondialdehyde (MDA), parathyroid hormone (PTH), C-terminal telopeptide (CTX), and 1,25-(OH)2-vitamin D3). Femur mass, length, and bone mineral density (BMD) were recorded, and histopathological studies for femurs were examined. Low-calcium diet induced osteoporotic changes in positive control rats (decrease in Ca, vitamin D3, and BMD and increase in CTX, PTH, TNF-α, CRP, and MDA). Supplementation with thyme and rosemary inhibited significantly the development of bone loss, increased Ca and vitamin D3 in plasma, improved BMD, and also prevented the inflammation and oxidative stress (improved TNF-α, CRP and MDA) compared to the positive control. The histopathological examination of treated groups showed an improvement in bone histology and protection against bone loss. However, thyme powder showed more effective impact than rosemary. Our study demonstrates that thyme and rosemary effectively mitigated calcium deficiency-induced bone loss and maybe considered as promising candidates for preventing bone resorption and osteoporosis.