Plasma Retinol Concentrations Research Articles

Determination of a plasma retinol reference interval in leopard geckos (Eublepharis macularius)

BackgroundLeopard geckos (Eublepharis macularius) are a popular captive bred species of reptile in the exotic pet trade that are commonly seen for clinical presentations consistent with hypovitaminosis A. There has been a lack of antemortem diagnostics available for minimally invasive and appropriate diagnosis of this disease. Reference intervals for retinol have not been published or validated in leopard geckos or any closely related species. MethodsPlasma, with a sample volume of only 50 µl, from 40 healthy captive bred leopard geckos, 20 males and 20 females were analyzed using ultra-high-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS) to determine plasma retinol concentrations. ResultsUsing UPLC/MS analysis data was normally distributed and a reference interval for plasma retinol in the leopard gecko was 0.06 to 0.55µg/mL (0.01–0.08µmol/L). A mean of 0.31µg/mL with a lower limit 95% confidence interval being 0.03 to 0.10 µg/mL and upper limit of 0.52 to 0.59 µg/mL. No significant variations of plasma retinol were found based on gecko color morph, age or weight. Female leopard geckos had a slightly higher mean plasma retinol compared to males, but was not significantly different. Conclusions and clinical relevanceThis study will allow veterinary clinicians to better diagnose and treat hypovitaminosis A and to provide advice on dietary supplementation and dietary improvement for the leopard gecko and possibly other gecko species as well. In addition, these values can be used to design and evaluate other studies for nutritional needs of this species. Limitations of this study include only a single season of collection, no same time comparison to liver values, and the evaluation of only captive bred individuals.

A Correlation Study of Plasma and Breast Milk Retinol Concentrations in Breastfeeding Women in China.

Retinol in breast milk is related to plasma concentration among breastfeeding women, but the linear or curvilinear relationships between the two remains unclear. We conducted a cross-sectional study in 403 Chinese breastfeeding women at 42 ± 7 days postpartum. Plasma and breast milk samples were assayed using high performance liquid chromatography to determine the concentration of retinol. Partial Spearman correlation and multivariable fractional polynomial regression were used to examine the relationships between the two retinol concentrations and between plasma retinol concentration and milk-to-plasma (M/P) retinol. The median (interquartile range, IQR) of the retinol concentration in the plasma was 1.39 (1.21, 1.63) μmol/L and 1.15 (0.83, 1.49) μmol/L in the breast milk, respectively. The partial correlation coefficient between them was 0.17 (p < 0.01). A linear relationship was observed with an adjusted regression coefficient of 0.34 (95% CI: 0.19, 0.49). The relationship between the plasma retinol and M/P ratio was nonlinear and segmented at 1.00 μmol/L of plasma retinol. The regression coefficients, below and above the segmented point, were -1.69 (95% CI: -2.75, -0.62) and -0.29 (95% CI: -0.42, -0.16), respectively. Plasma and breast milk retinol were positively correlated, whereas women with a low concentration of plasma retinol showed a stronger capacity of transferring retinol to breast milk.

290 Vitamin A and Beta-Carotene Supplementation on Growth Performance, Immune System, and Mammary Gland Development in Finishing Pigs

Abstract A total of 224 finishing pigs were randomly assigned to split-sex pens (n = 4 pigs per pen). Each pen was assigned to one of four diets: control (7,656 IU vitamin A/kg), control diet supplemented with vitamin A (4.36 ppm; Rovimix A 1000, DSM, NJ, US), control diet supplemented with beta-carotene (163.28 ppm; Rovimix β-Carotene 10%, DSM, NJ, US), or control supplemented with oxidized beta-carotene (40 ppm; 10% active ingredient, Avivagen, Ottawa, ON, Canada). Pigs and feeder weights were recorded at the start of the study (d 0) and end of each phase (days 21, 42, and 63). A subset of gilts was randomly selected and a blood sample for vitamin A (retinol) analysis was collected on days 0, 18, 39, 60, and 63. The same gilts were vaccinated against Lawsonia intracellularis and porcine circovirus. Antibodies against each vaccination were measured on days 18, 39, and 60. At the end of the study, the subset of gilts was euthanized to obtain a liver sample for vitamin A (retinyl palmitate is converted to retinol for measurement) analysis and a jejunum sample to measure the mRNA abundance of genes involved in vitamin A metabolism (alcohol dehydrogenase class 1, lecithin retinol acyltransferase phosphatidylcholine-retinol O-acyltransferase, retinol binding protein, and beta-carotene oxygenase 1). Mammary tissue from the second and fourth right anterior mammary glands were collected to assess gland development via immunohistochemical staining for T cells, B cells, and macrophages (CD3, CD20, and IBA1, respectively) and DNA extraction. Vaccine antibodies measures, vitamin A analysis, and mammary immunohistochemical staining was completed at Iowa State University Veterinarian Diagnostic Laboratory (Ames, IA). Data were analyzed in SAS 9.4 (Statistical Analysis System, Cary, NC) via GLIMMIX procedure. Pen was the experimental unit and fixed effects were diet and time. Gilts were nested within pen when appropriate to account for more than one gilt per pen. Oxidized beta-carotene supplementation increased (P = 0.02) average daily gain across phases; however, there were no differences (P = 0.18) in the body weight of pigs. There was no effect (P &amp;gt; 0.05) of diet on plasma or hepatic retinol, IgG or IgM concentrations, immune cell presence, or DNA in mammary tissue. Supplemented vitamin A tended (P = 0.05) to increase the expression of retinol binding protein in the jejunum, but other genes were not affected (P &amp;gt; 0.05) by dietary treatments. A diet by time interaction (P = 0.04) was noted for circovirus sample-to-positive ratios and titer levels, where vitamin A supplementation had better vaccine responses at the end of the study. Thus, pigs supplemented with oxidized beta-carotene had an improved average daily gain over vitamin A supplemented pigs, but pigs supplemented with vitamin A appeared to have a more robust immune system.

Whole cottonseed inclusion in starter feeds improves performance, inflammometabolic profile, and rumination behavior in Holstein dairy calves

The high energy, protein, and fiber contents of whole cottonseed make it a potential candidate for the inclusion in calf starters to promote the rumen development. This study aimed at assessing whether the inclusion of whole cottonseed in the starter would affect performance, metabolic profile, and rumination time in Holstein dairy calves. From 2 to 55 d of age, 12 heifer calves were fed a constant amount of milk replacer twice daily (8 L/d), whereas from 56 to 65 d (weaning) milk replacer was gradually reduced (from 4 to 1 L/d) and fed in a single meal. Calves were blocked by birth body weight and % Brix of colostrum received and randomly assigned to 1 out of 2 dietary treatments: (1) control starter (CTR); (2) starter with 8% inclusion of whole cottonseed (WCS). Treatments were fed for ad libitum intake. From d 56, TMR and hay were offered ad libitum. At 0, 2, 7, 21, 65, and 80 d, BW was measured and blood samples were collected. Feed intake and rumination time were automatically recorded. Compared with CTR, WCS calves were heavier at weaning and after weaning, and consumed more starter from 59 to 72 d. In the immediate postweaning (from 66 to 72 d of age), rumination time increased more in WCS calves. Overall, WCS calves had greater plasma glucose, β-carotene, and retinol concentrations, whereas ceruloplasmin and myeloperoxidase were lower. Calves in the WCS group had greater glucose concentration at 21 and 65 d and lower urea at 65 d (weaning). Plasma β-carotene concentration was greater at 65 and 80 d in calves of the WCS group compared with CTR calves. At 80 d, WCS calves had lower plasma alkaline phosphatase and greater β-hydroxybutyrate, paraoxonase, and tocopherol. These results suggest that inclusion of WCS in the calf starter might be beneficial for rumen development, leading to greater feed intake and BW. Moreover, WCS inclusion was associated with lower oxidative stress and inflammation, improved energy metabolism and liver functionality, and likely quicker rumen development, as might be indicated by the higher plasma β-hydroxybutyrate and rumination time. These changes occurred mainly after weaning, when no differences in starter intake and average daily gain were detected, suggesting a better efficiency of nutrient utilization at this age.

Use of Compartmental Modeling and Retinol Isotope Dilution to Determine Vitamin A Stores in Young People with Sickle Cell Disease Before and After Vitamin A Supplementation

BackgroundSuboptimal plasma retinol concentrations have been documented in US children with sickle cell disease (SCD) hemoglobin SS type (SCD-HbSS), but little is known about vitamin A kinetics and stores in SCD. ObjectivesThe objectives were to quantify vitamin A total body stores (TBS) and whole-body retinol kinetics in young people with SCD-HbSS and use retinol isotope dilution (RID) to predict TBS in SCD-HbSS and healthy peers as well as after vitamin A supplementation in SCD-HbSS subjects. MethodsComposite plasma [13C10]retinol response data collected from 22 subjects with SCD-HbSS for 28 d after isotope ingestion were analyzed using population-based compartmental modeling (“super-subject” approach); TBS and retinol kinetics were quantified for the group. TBS was also calculated for the same individuals using RID, as well as for healthy peers (n = 20) and for the subjects with SCD-HbSS after 8 wk of daily vitamin A supplements (3.15 or 6.29 μmol retinol/d [900 or 1800 μg retinol activity equivalents/d]). ResultsModel-predicted group mean TBS for subjects with SCD-HbSS was 428 μmol, equivalent to ∼11 mo of stored vitamin A; vitamin A disposal rate was 1.3 μmol/d. Model-predicted TBS was similar to that predicted by RID at 3 d postdosing (mean, 389 μmol; ∼0.3 μmol/g liver); TBS predictions at 3 compared with 28 d were not significantly different. Mean TBS in healthy peers was similar (406 μmol). RID-predicted TBS for subjects with SCD-HbSS was not significantly affected by vitamin A supplementation at either dose. ConclusionsDespite differences in plasma retinol concentrations, TBS was the same in subjects with SCD-HbSS compared with healthy peers. Because 56 d of vitamin A supplementation at levels 1.2 to 2.6 times the Recommended Dietary Allowance did not increase TBS in these subjects with SCD-HbSS, further work will be needed to understand the effects of SCD on retinol metabolism. This trial was registered as NCT03632876 at clinicaltrials.gov.

Association of Plasma Vitamins and Carotenoids, DNA Methylation of LCAT, and Risk of Age-Related Macular Degeneration.

Dysregulation of lipid metabolism has been implicated in age-related macular degeneration (AMD), the leading cause of blindness among the elderly. Lecithin cholesterol acyltransferase (LCAT) is an important enzyme responsible for lipid metabolism, which could be regulated by DNA methylation during the development of various age-related diseases. This study aimed to assess the association between LCAT DNA methylation and the risk of AMD, and to examine whether plasma vitamin and carotenoid concentrations modified this association. A total of 126 cases of AMD and 174 controls were included in the present analysis. LCAT DNA methylation was detected by quantitative real-time methylation-1specific PCR (qMSP). Circulating vitamins and carotenoids were measured using reversed-phase high-performance liquid chromatography (RP-HPLC). DNA methylation of LCAT was significantly higher in patients with AMD than those in the control subjects. After multivariable adjustment, participants in the highest tertile of LCAT DNA methylation had a 5.37-fold higher risk (95% CI: 2.56, 11.28) of AMD compared with those in the lowest tertile. Each standard deviation (SD) increment of LCAT DNA methylation was associated with a 2.23-fold (95% CI: 1.58, 3.13) increased risk of AMD. There was a J-shaped association between LCAT DNA methylation and AMD risk (Pnon-linearity = 0.03). Higher concentrations of plasma retinol and β-cryptoxanthin were significantly associated with decreased levels of LCAT DNA methylation, with the multivariate-adjusted β coefficient being -0.05 (95% CI: -0.08, -0.01) and -0.25 (95% CI: -0.42, -0.08), respectively. In joint analyses of LCAT DNA methylation and plasma vitamin and carotenoid concentrations, the inverse association between increased LCAT DNA methylation and AMD risk was more pronounced among participants who had a lower concentration of plasma retinol and β-cryptoxanthin. These findings highlight the importance of comprehensively assessing LCAT DNA methylation and increasing vitamin and carotenoid status for the prevention of AMD.

Dietary and plasma retinoids are not associated with fatty acid desaturase indices in healthy young adults.

Past research in rodents suggests that fatty acid (FA) desaturase expression and activity may be modified by vitamin A; however, this has not been investigated in humans. The primary objective of this study was to examine associations between dietary retinoid intakes, plasma retinoid concentrations, and FA desaturase indices in young adults. As a secondary objective, biological sex and estrogen-containing contraceptive (EC) use were investigated due to prior evidence demonstrating that both can influence plasma retinol concentration and FA desaturase indices. Dietary retinoid intake (food frequency questionnaire), plasma retinoid concentrations (high-performance liquid chromatography-tandem mass spectrometry), plasma FA (gas chromatography), and FA desaturase indices (product-to-precursor ratios) from 945 adults recruited for the cross-sectional Toronto Nutrigenomics and Health study were analyzed. Participants were stratified into quartiles based on plasma retinol concentration and data analyzed by one-way analysis of covariance. Dietary retinoid intakes were not associated with the overall n-3 pathway, overall n-6 pathway, delta-5 desaturase, delta-6 desaturase, or delta-9 desaturase indices (all r < 0.10, p > 0.05). The overall n-6 pathway index was significantly higher (p = 0.0004) and the delta-5 desaturase index was significantly lower (p = 0.0003) in individuals with higher plasma retinol levels; however, these differences were lost when participants were grouped by biological sex and EC use. Although weak relationships were observed between plasma retinol and some FA desaturase indices in the total population, these associations appear to be driven by biological sex and EC usage rather than retinoids. We therefore find little evidence of a relationship between retinoids and FA desaturase indices in young, healthy adults.

Plasma Retinol Concentrations and Dietary Intakes of Mother–Infant Sets in Singleton versus Twin Pregnancy

Vitamin A (retinol) is essential for normal fetal development, but the recommendation for maternal dietary intake (Retinol Activity Equivalent, RAE) does not differ for singleton vs. twin pregnancy, despite the limited evaluation of retinol status. Therefore, this study aimed to evaluate plasma retinol concentrations and deficiency status in mother–infant sets from singleton vs. twin pregnancies as well as maternal RAE intake. A total of 21 mother–infant sets were included (14 singleton, 7 twin). The HPLC and LC-MS/HS evaluated the plasma retinol concentration, and data were analyzed using the Mann–Whitney U test. Plasma retinol was significantly lower in twin vs. singleton pregnancies in both maternal (192.2 vs. 312.1 vs. mcg/L, p = 0.002) and umbilical cord (UC) samples (102.5 vs. 154.4 vs. mcg/L, p = 0.002). The prevalence of serum-defined vitamin A deficiency (VAD) &lt;200.6 mcg/L was higher in twins vs. singletons for both maternal (57% vs. 7%, p = 0.031) and UC samples (100% vs. 0%, p &lt; 0.001), despite a similar RAE intake (2178 vs. 1862 mcg/day, p = 0.603). Twin pregnancies demonstrated a higher likelihood of vitamin A deficiency in mothers, with an odds ratio of 17.3 (95% CI: 1.4 to 216.6). This study suggests twin pregnancy may be associated with VAD deficiency. Further research is needed to determine optimal maternal dietary recommendations during twin gestation.

Ovariectomy Increases Circulating Retinol-Binding Protein Concentrations Independently of Sex-Dependent Differences in Retinol Concentrations in Rats

BackgroundThere is a sex-dependent difference in blood retinol and RBP concentrations, and plasma RBP is associated with insulin resistance. ObjectivesWe aimed to clarify sex-dependent variations in body concentrations of retinol and RBPs and their association with sex hormones in rats. MethodsPlasma and liver retinol concentrations and hepatic mRNA and plasma concentrations of RBP4 were analyzed in 3- and 8-wk-old male and female Wistar rats before and after sexual maturity (experiment 1) and in orchiectomized male Wistar rats (experiment 2) and ovariectomized female Wistar rats (experiment 3). Furthermore, the mRNA and protein concentrations of RBP4 in adipose tissue were measured in ovariectomized female rats (experiment 3). ResultsThere were no sex-dependent differences in liver retinyl palmitate and retinol concentrations; however, the plasma retinol concentration was significantly higher in male rats than that in female rats after sexual maturity. Furthermore, the plasma retinol concentrations did not differ between the ovariectomized or orchiectomized rats and the control rats. Plasma Rbp4 mRNA concentrations were higher in male rats than those in female rats but not in castrated and control rats, a change consistent with plasma retinol concentration. Plasma RBP4 concentrations were also higher in male rats than those in female rats; however, unlike liver Rbp4 gene expression, plasma RBP4 concentrations were 7-fold higher in the ovariectomized rats than those in the control rats. Moreover, the Rbp4 mRNA concentrations in inguinal white adipose tissue was significantly higher in the ovariectomized rats than those in the control rats and correlated with plasma RBP4 concentrations. ConclusionsHepatic Rbp4 mRNA is higher in male rats through a sex hormone–independent mechanism, which may contribute to sex differences in blood retinol concentrations. Furthermore, ovariectomy leads to an increase in adipose tissue Rbp4 mRNA and blood RBP4 concentrations, which may contribute to insulin resistance in ovariectomized rats and postmenopausal women.

Effects of different vitamin A supplies on performance and the risk of ketosis in transition cows

This experiment investigated the effects of feeding low and high supplies of vitamin A (VA) during the transition period on plasma metabolites, prevalence of ketosis, and early milk production. In a randomized complete block design, 42 prefresh Holstein cows and 21 heifers were blocked by parity and calving date and assigned to 1 of 3 dietary treatments (n = 21 per treatment unless noted): CON, a transition diet with supplemental VA (75,000 IU/d) to meet the requirement; LVA, a transition diet with no supplemental VA; or HVA, a transition diet receiving supplemental VA (187,500 IU/d) 2.5 times greater than the requirement. Experimental periods were prepartum (-14 d prepartum), postpartum (1 to 30 d in milk), and carryover period (31 to 58 d in milk; common lactating diet with adequate VA was fed). Differences in dry matter intake in the pre- and postpartum periods and milk yield were not detected among treatment. Milk fat, protein, and lactose yields were similar among treatments and not affected by VA. Somatic cell count increased linearly with increasing VA. Body weight and body condition score decreased postpartum, but no VA effect was observed. Plasma retinol concentrations (n = 10 per treatment) decreased at d 2 postpartum and increased as lactation progressed, but the concentrations were unaffected by treatment. Plasma β-carotene (n = 10 per treatment) had a treatment by time interaction and its concentration decreased after parturition and remained low for 2 wk. Plasma fatty acids and β-hydroxybutyrate did not differ among treatments. Milk retinol concentration and yield (n = 10 per treatment) increased as VA supply increased. Segmented neutrophils (%) decreased, and lymphocytes (%) increased in blood with increasing VA supply. In conclusion, providing different supplies of VA did not affect production, mobilization of body fat, and risk of ketosis; however, excessive VA supply may have negatively affected the immune response, in part contributing to increased milk somatic cell counts during early lactation.

Vitamin A Intake and Status Influence Retinol Balance, Utilization and Dynamics in Rats ,

To study effects of vitamin A status on retinol dynamics, male rats were fed purified diets varying in vitamin A concentration. Group 1 rats had marginal liver vitamin A levels (approximately 500 nmol) and were in a slight positive vitamin A balance; Group 2 had similar liver levels but were in a slight negative balance; Group 3 had lower liver levels (approximately 370 nmol) and were in a slight negative balance; Group 4 had depleted liver reserves (<10 nmol) and were in vitamin A balance. [3H]Retinol-labeled plasma was injected intravenously, and serial plasma samples were collected for 41 d while rats (six per group) consumed approximately 50 nmol retinol/d (Group 1) or -25 nmol/d (Groups 2-4). Plasma retinol was normal in Groups 1-3 (1.9-2.0 micromol/L) and lower in Group 4 (0.96 micromol/L). Plasma tracer data were fit to a three-compartment model. The central plasma retinol compartment (transit time, 1.5-1.7 h) exchanged with a fast turning-over extravascular vitamin A pool (transit time, 3-4.5 h; -40 nmol) and with a larger, slow turning-over extravascular pool (transit time, 5.5-10 d) that was the site of irreversible utilization of vitamin A. Irreversible utilization was 36 nmol/d (Group 1), 29 nmol/d (Groups 2 and 3) and 20 nmol/d (Group 4). The data indicate that in rats with low or marginal vitamin A status, vitamin A intake, vitamin A reserves and plasma retinol concentration all influence vitamin A utilization and other aspects of retinol dynamics.

Dietary Vitamin A Restriction Produces Marginal Vitamin A Status in Young Rats

A model of marginal vitamin A deficiency was developed in young rats by limiting the vitamin A intake of dams and of offspring postweaning. Prior to and throughout pregnancy and lactation, female rats were fed diets in which the vitamin A concentration was either low marginal [0.18 retinol equivalent (RE)/g diet], high marginal (0.4 RE/g) or sufficient (4 RE/g). Vitamin A restriction had no effect on dams' reproduction or litter sizes, but total retinol in liver was depleted by the end of lactation. Pups fed all diets grew steadily from birth through 35 d. The milk curd total retinol concentration of 9-d-old pups' stomachs was significantly different among the low marginal, high marginal and vitamin A-sufficient groups. By 35 d, plasma retinol concentrations of pups in the low marginal group were less than half of those in the high marginal and vitamin A-sufficient groups. The liver total retinol concentrations and lecithin retinol:acyltransferase activity of 35-d-old pups in the low and high marginal groups were much lower than those of vitamin A-sufficient pups. When pups from dams fed low marginal diet were weaned onto vitamin A-free diet, frank vitamin A deficiency was evident by 33 d of age as judged by physical signs and biochemical alterations in vitamin A status. Thus, these dietary protocols are useful in inducing early onset of either marginal or frank vitamin A deficiency.

Temporal changes in the systemic concentrations of retinoids in pregnant and postpartum women.

Retinoids and vitamin A are essential for multiple biological functions, including vision and immune responses, as well as the development of an embryo during pregnancy. Despite its importance, alterations in retinoid homeostasis during normal human pregnancy are incompletely understood. We aimed to characterize the temporal changes in the systemic retinoid concentrations across pregnancy and postpartum period. Monthly blood samples were collected from twenty healthy pregnant women, and plasma concentrations of retinol, all-trans-retinoic acid (atRA), 13-cis-retinoic acid (13cisRA), and 4-oxo-retinoic acids were measured using liquid chromatography-tandem mass spectrometry. Significant decreases in 13cisRA concentrations over the pregnancy were observed, with rebound increases in retinol and 13cisRA levels after delivery. Of note, atRA concentrations exhibited a unique temporal pattern with levels peaking at mid-pregnancy. While the 4-oxo-atRA concentration was below the limit of quantification, 4-oxo-13cisRA was readily detectable, and its temporal change mimicked that of 13cisRA. The time profiles of atRA and 13cisRA remained similar after correction by albumin levels for plasma volume expansion adjustment. Together, the comprehensive profiling of systemic retinoid concentrations over the course of pregnancy provides insights into pregnancy-mediated changes in retinoid disposition to maintain its homeostasis.

Retinyl Ester Storage Is Normal in Transgenic Mice with Enhanced Expression of Cellular Retinol-Binding Protein Type I

This report describes the production and characterization of transgenic mice with high expression of human cellular retinol-binding protein type I [hCRBP(I)]. In initial experiments, overexpression of hCRBP(I) was driven by the strong promoter SRα, but no transgenic offspring were produced. When we used the less efficient mouse metallothionein I promoter fused to the hCRBP(I) cDNA for microinjection, we obtained 12% transgenic offspring. Two of these transgenic mice (409/1 and 401/2) expressed mRNA and immunoreactive hCRBP(I) in several organs. Both lines had relatively high contents of hCRBP(I) in intestine, testis and epididymis. On the other hand, only 401/2 transgenic mice had high contents of hCRBP(I) in kidney. Effects on storage of vitamin A were studied by measuring the concentration of retinyl esters in different organs. The concentrations of retinyl esters in liver, lung and kidney did not significantly differ between transgenic and control mice, and the concentration of total retinol in plasma was within the normal range in transgenic mice. Furthermore, feeding mice a diet with high or low concentrations of vitamin A for 2 wks resulted in no marked differences in the concentrations of retinyl esters in liver, kidney, lung, intestine and testis in transgenic mice compared with control mice. Therefore, in spite of high expression of hCRBP(I) in several organs, the transgenic mice had normal storage of retinyl esters in all organs studied. The present in vivo study indicates that the CRBP(I) content alone does not control retinyl ester storage.

Retinol and Pro-Vitamin A Carotenoid Nutritional Status during Pregnancy Is Associated with Newborn Hearing Screen Results

The prenatal period is critical for auditory development; thus, prenatal influences on auditory development may significantly impact long-term hearing ability. While previous studies identified a protective effect of carotenoids on adult hearing, the impact of these nutrients on hearing outcomes in neonates is not well understood. The purpose of this study is to investigate the relationship between maternal and umbilical cord plasma retinol and carotenoid concentrations and abnormal newborn hearing screen (NHS) results. Mother-infant dyads (n = 546) were enrolled at delivery. Plasma samples were analyzed using HPLC and LC-MS/MS. NHS results were obtained from medical records. Statistical analysis utilized Mann-Whitney U tests and logistic regression models, with p ≤ 0.05 considered statistically significant. Abnormal NHS results were observed in 8.5% of infants. Higher median cord retinol (187.4 vs. 162.2 μg/L, p = 0.01), maternal trans-β-carotene (206.1 vs. 149.4 μg/L, p = 0.02), maternal cis-β-carotene (15.9 vs. 11.2 μg/L, p = 0.02), and cord trans-β-carotene (15.5 vs. 8.0 μg/L, p = 0.04) were associated with abnormal NHS. Significant associations between natural log-transformed retinol and β-carotene concentrations and abnormal NHS results remained after adjustment for smoking status, maternal age, and corrected gestational age. Further studies should investigate if congenital metabolic deficiencies, pesticide contamination of carotenoid-rich foods, maternal hypothyroidism, or other variables mediate this relationship.

Vitamin A Nutritional Status Is a Key Determinant of Bone Mass in Children.

The bone mass increases that occur during the period of childhood are of great significance for maximizing the peak bone mass in adults and preventing for osteoporosis. Studies have reported that VA can improve the bone health in adults. Moreover, limited studies have assessed such associations in children. In this cross-sectional study including 426 children, we assessed the children's plasma retinol concentration by liquid chromatography-mass spectrometry and the dietary intake of VA and carotenoids using a structured Food Frequency Questionnaire. Their bone mineral content and bone mineral density (BMD) were measured using dual-energy X-ray absorptiometry. After adjusting for potential confounders, the restricted cubic spline revealed an inverted U-shaped association between plasma retinol concentration and BMD; the estimated effects on the TBLH BMD per μmol/L increase in the plasma retinol concentration were 1.79 × 10-2 g/cm2 below 1.24 μmol/L and -5.78 × 10-3 g/cm2 above this point (p for non-linearity = 0.046). A multiple linear regression analysis revealed a positive association between the plasma retinol concentration and the TBLH BMC (β = 1.89, 95% CI: 1.64 × 10-1-3.62, p = 0.032). In conclusion, an appropriate plasma retinol concentration and greater intakes of dietary VA and β-carotene may enhance the bone mineral status of children who are aged 6-9 years.

11 Evaluating Relationships Between Plasma and Liver Retinol Concentrations in the Beef Cow and Calf

Abstract In the young calf, vitamin A is particularly important for immune system maturation, and calves rely on colostrum at birth to supply vitamin A. Cow vitamin A status may influence colostral vitamin A concentrations, which can impact vitamin A status of their calves. Very little is understood about this relationship between beef cow and calf vitamin A status. Furthermore, limited data is available on how plasma and liver retinol concentrations in cattle are related to one another. The objective of this study was to examine relationships between liver and plasma retinol concentrations in the cow, calf, and within cow-calf pairs. Multiparous MARC II beef cows (n = 120; 6.4 ± 1.2 SD years of age; 592 ± 58 SD kg BW) in mid-gestation were provided supplemental vitamin A as retinyl acetate in a supplemental pellet at a rate of 21,125 ± 8,560 SD IU/d (current NASEM recommendation = 33,000 IU/d). Cows were individually fed using Calan gates a diet consisting of alfalfa hay, corn silage, and supplemental pellet for 144 d (111 d pre-calving and 32 d post-calving). Basal diet vitamin A concentration was 490 IU/kg of DM. Mean initial liver concentration of cows was 830 µg retinol/g of DM. To assess vitamin A status, liver biopsies and plasma samples were collected on cows and calves at the end of the supplemental period when calves averaged 32 ± 7 days of age (DOA). Pearson correlations were used to test for linear relationships between cow liver and plasma retinol concentrations, calf liver and plasma retinol concentrations, and liver and plasma retinol concentrations between the cow and her calf. No linear relationship (P = 0.10; r=0.16) was observed between liver and plasma retinol in cows. In the present study, mean cow liver retinol concentration (482 ± 182 SD µg/g of DM) fell within the current adequacy reference range of 300–700 µg/g of DM. Cow plasma retinol (272 ± 40 SD ng/mL) was slightly below the reference range of 300–800 ng/mL. A positive correlation (P &amp;lt; 0.01; r=0.37) was detected between calf liver (51 ± 27 SD µg/g of DM) and plasma (190 ± 47 ng/mL) retinol concentrations. Both were below what would be considered adequate (100–350 µg/g of DM in liver; 225–325 ng/mL in plasma) for calves at 32 DOA. There was a positive correlation (P &amp;lt; 0.01; r=0.31) between cow and calf liver retinol, suggesting that as cow retinol liver concentrations increased, calf liver retinol concentrations increased. It appears that despite cows having adequate liver retinol concentrations when 21,125 IU vitamin A/d was fed, it did not result in calf liver retinol stores that would be considered adequate given current reference ranges. USDA is an equal opportunity employer and provider.

Multiassay nutritional metabolomics profiling of low vitamin A status versus adequacy is characterized by reduced plasma lipid mediators among lactating women in the Philippines: A pilot study

Low vitamin A (VA) status is common among lactating women in low-income countries. Lactation has substantial effects on mother's metabolism and VA is required in multiple biological processes, including growth, vision, immunity, and reproduction. The objective of this pilot study was to use metabolomics profiling to conduct a broad, exploratory assessment of differences in plasma metabolites associated with low VA status versus VA adequacy in lactating women. Plasma samples from lactating women who participated in a survey in Samar, Philippines, were selected from a cross-sectional study based on plasma retinol concentrations indicating low (VA–; n = 5) or adequate (VA+; n = 5) VA status (plasma retinol <0.8 or >1.05 µmol/L). The plasma results collected from 6 metabolomics assays (oxylipins, endocannabinoids, bile acids, primary metabolomics, biogenic amines, and lipidomics) were compared by group using liquid chromatography mass spectrometry. Twenty-eight metabolites were altered in the VA– versus VA+ status groups, with 24 being lipid mediators (P < .05). These lipid mediators included lower concentrations of arachidonic acid- and eicosapentaenoic acid-derived oxylipins, as well as lysophospholipids and sphingolipids, in the VA– group (P < .05). Chemical similarity enrichment analysis identified hydroxy-eicosatetraenoic acids, hydroxy-eicosapentaenoic acids, and dihydroxy-eicosatetraenoic acids as significantly altered oxylipin clusters (P < .0001, false discovery rate [FDR] P < .0001), as well as sphingomyelins, saturated lysophosphatidylcholines, phosphatidylcholines, and phosphatidylethanolamines (P < .001, FDR P < .01). The multiassay nutritional metabolomics profiling of low VA status compared with adequacy in lactating women was characterized by reduced lipid mediator concentrations. Future studies with stronger study designs and larger sample size are needed to confirm and validate these preliminary results.

Plasma Retinoid Concentrations Are Altered in Pregnant Women.

Vitamin A is vital to maternal–fetal health and pregnancy outcomes. However, little is known about pregnancy associated changes in maternal vitamin A homeostasis and concentrations of circulating retinol metabolites. The goal of this study was to characterize retinoid concentrations in healthy women (n = 23) during two stages of pregnancy (25–28 weeks gestation and 28–32 weeks gestation) as compared to ≥3 months postpartum. It was hypothesized that plasma retinol, retinol binding protein 4 (RBP4), transthyretin and albumin concentrations would decline during pregnancy and return to baseline by 3 months postpartum. At 25–28 weeks gestation, plasma retinol (−27%), 4-oxo-13-cis-retinoic acid (−34%), and albumin (−22%) concentrations were significantly lower, and all-trans-retinoic acid (+48%) concentrations were significantly higher compared to ≥3 months postpartum in healthy women. In addition, at 28–32 weeks gestation, plasma retinol (−41%), retinol binding protein 4 (RBP4; −17%), transthyretin (TTR; −21%), albumin (−26%), 13-cis-retinoic acid (−23%) and 4-oxo-13-cis-retinoic acid (−48%) concentrations were significantly lower, whereas plasma all-trans-retinoic acid concentrations (+30%) were significantly higher than ≥3 months postpartum. Collectively, the data demonstrates that in healthy pregnancies, retinol plasma concentrations are lower, but all-trans-retinoic acid concentrations are higher than postpartum.

The Use of Datasets for Theoretical Subjects to Validate Vitamin A–Related Methods and Experimental Designs

We review recent work in which model-based compartmental analysis has been applied to data for theoretical human subjects in order to study questions related to vitamin A kinetics and metabolism. Using model simulations in this way, one can validate experimental designs, evaluate or improve vitamin A assessment methods, study the influence of perturbations on assessment methods, and/or advance information related to retinol kinetics. We also provide some information on the rationale for assigning physiologically appropriate values for specified characteristics [e.g., plasma retinol concentration, vitamin A total body stores (TBS), vitamin A intake] to hypothetical individuals, and in addition, we outline how one might first select an appropriate compartmental model for whole-body vitamin A metabolism and then specify physiologically reasonable values for the associated retinol kinetic parameters. In the studies discussed here, the Simulation, Analysis, and Modeling software was used to simulate responses in key model compartments for hypothetical subjects so that model predictions could be compared to assigned values or projected outcomes. For example, in the case of the retinol isotope dilution (RID) method that is used to assess vitamin A status, application of this approach has provided a way to evaluate the accuracy of TBS predictions under different steady state and non-steady state conditions, thus increasing confidence about the validity of RID results obtained in the field. Although datasets for theoretical subjects have been used to evaluate protocols in pharmacokinetics, to our knowledge, other nutrition researchers have not previously used approaches such as those described here. Our results to date indicate that this strategy has the potential to provide useful information related not only to vitamin A but perhaps to other nutrients as well.