Maternal and neonatal one-carbon metabolites and the epigenome-wide infant response.

Maternal prenatal status, as encapsulated by that to which a mother is exposed through diet and environment, is a key determinant of offspring health and disease. Alterations in DNA methylation (DNAm) may be a mechanism through which suboptimal prenatal conditions confer disease risk later in life. One-carbon metabolism (OCM) is critical to both fetal development and in supplying methyl donors needed for DNAm. Plasma concentrations of one-carbon metabolites across maternal first trimester (M1), maternal term (M3), and infant cord blood (CB) at birth were tested for association with DNAm patterns in CB from the Michigan Mother and Infant Pairs (MMIP) pregnancy cohort. The Illumina Infinium MethylationEPIC BeadChip was used to quantitatively evaluate DNAm across the epigenome. Global and single-site DNAm and metabolite models were adjusted for infant sex, estimated cell type proportions, and batch as covariates. Change in mean metabolite concentration across pregnancy (M1 to M3) was significantly different for S-adenosylhomocysteine (SAH), S-adenosylmethionine (SAM), betaine, and choline. Both M1 SAH and CB SAH were significantly associated with the global distribution of DNAm in CB, with indications of a shift toward less methylation. M3 SAH and CB SAH also displayed significant associations with locus-specific DNAm in infant CB (FDR<0.05). Our findings underscore the role of maternal one-carbon metabolites in shifting the global DNAm pattern in CB and emphasizes the need to closely evaluate how dietary status influences cellular methylation potential and ultimately offspring health.

One-carbon metabolites, B vitamins and associations with systemic inflammation and angiogenesis biomarkers among colorectal cancer patients: results from the ColoCare Study.

B vitamins involved in one-carbon metabolism have been implicated in the development of inflammation- and angiogenesis-related chronic diseases, such as colorectal cancer (CRC). Yet, the role of one-carbon metabolism in inflammation and angiogenesis among CRC patients remains unclear. The objective of this study was to investigate associations of components of one-carbon metabolism with inflammation and angiogenesis biomarkers among newly diagnosed CRC patients (n 238) in the prospective ColoCare Study, Heidelberg. We cross-sectionally analysed associations between twelve B vitamins and one-carbon metabolites and ten inflammation and angiogenesis biomarkers from pre-surgery serum samples using multivariable linear regression models. We further explored associations among novel biomarkers in these pathways with Spearman partial correlation analyses. We hypothesised that pyridoxal-5'-phosphate (PLP) is inversely associated with inflammatory biomarkers. We observed that PLP was inversely associated with C-reactive protein (CRP) (r -0·33, Plinear < 0·0001), serum amyloid A (SAA) (r -0·23, Plinear = 0·003), IL-6 (r -0·39, Plinear < 0·0001), IL-8 (r -0·20, Plinear = 0·02) and TNFα (r -0·12, Plinear = 0·045). Similar findings were observed for 5-methyl-tetrahydrofolate and CRP (r -0·14), SAA (r -0·14) and TNFα (r -0·15) among CRC patients. Folate catabolite acetyl-para-aminobenzoylglutamic acid (pABG) was positively correlated with IL-6 (r 0·27, Plinear < 0·0001), and pABG was positively correlated with IL-8 (r 0·21, Plinear < 0·0001), indicating higher folate utilisation during inflammation. Our data support the hypothesis of inverse associations between PLP and inflammatory biomarkers among CRC patients. A better understanding of the role and inter-relation of PLP and other one-carbon metabolites with inflammatory processes among colorectal carcinogenesis and prognosis could identify targets for future dietary guidance for CRC patients.

A selective adsorption-based separation of low-mass molecules from biological samples towards high-throughput mass spectrometry analysis in a single drop of human whole blood.

In this work, a newly designed multifunctional adsorbent material, octadecyl-modified ordered mesoporous carbon (C18-CMK-8), was synthesized and employed for simultaneous analysis of multiple small molecules by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The C18-CMK-8 was facilely prepared by one-step chemical binding of octadecyltrimethoxysilane to an ordered mesoporous carbon CMK-8. The nanosized pore structure of C18-CMK-8 can selectively enrich low-mass molecules while excluding interferences from large molecules in whole blood; meanwhile, the C18 chain can greatly enhance the affinity for a wide range of small molecules. Then, the C18-CMK-8-based solid-phase extraction (SPE) combined with MALDI-TOF MS measurement was applied in rapid and high-throughput screening of six doping agents (triamterene, trenbolone, testosterone, methyltestosterone, clenbuterol, and strychnine)) in single-drop human whole blood samples. The method showed high analytical sensitivity (detection limit 0.05-0.1 ng mL-1), good recoveries (67.2-114.3%), minimal sample requirement (∼20 µL), and robust anti-interference ability. With distinct advantages such as high throughput, rapidness, minimal sample requirement, and high sensitivity, this study not only offers a versatile enrichment material for complex sample preparation, but also demonstrates a promising tool for minimal whole blood analysis.

Glucose feeds the TCA cycle via circulating lactate.

Mammalian tissues are fuelled by circulating nutrients, including glucose, amino acids, and various intermediary metabolites. Under aerobic conditions, glucose is generally assumed to be burned fully by tissues via the tricarboxylic acid cycle (TCA cycle) to carbon dioxide. Alternatively, glucose can be catabolized anaerobically via glycolysis to lactate, which is itself also a potential nutrient for tissues and tumours. The quantitative relevance of circulating lactate or other metabolic intermediates as fuels remains unclear. Here we systematically examine the fluxes of circulating metabolites in mice, and find that lactate can be a primary source of carbon for the TCA cycle and thus of energy. Intravenous infusions of 13C-labelled nutrients reveal that, on a molar basis, the circulatory turnover flux of lactate is the highest of all metabolites and exceeds that of glucose by 1.1-fold in fed mice and 2.5-fold in fasting mice; lactate is made primarily from glucose but also from other sources. In both fed and fasted mice, 13C-lactate extensively labels TCA cycle intermediates in all tissues. Quantitative analysis reveals that during the fasted state, the contribution of glucose to tissue TCA metabolism is primarily indirect (via circulating lactate) in all tissues except the brain. In genetically engineered lung and pancreatic cancer tumours in fasted mice, the contribution of circulating lactate to TCA cycle intermediates exceeds that of glucose, with glutamine making a larger contribution than lactate in pancreatic cancer. Thus, glycolysis and the TCA cycle are uncoupled at the level of lactate, which is a primary circulating TCA substrate in most tissues and tumours.

Association of TCN2 rs1801198 c.776G>C polymorphism with markers of one-carbon metabolism and related diseases: a systematic review and meta-analysis of genetic association studies.

Background: Vitamin B-12 (cobalamin) deficiency may produce severe neurologic and hematologic manifestations. Approximately 20-25% of circulating cobalamin binds to transcobalamin 2 (TCN2), which is referred to as active vitamin B-12. The G allele of the TCN2 c.776G>C (rs1801198) polymorphism has been associated with a lower plasma concentration of holotranscobalamin. However, genotype association studies on rs1801198 have led to conflicting results regarding its influence on one-carbon metabolism (OCM) markers or its association with pathologic conditions.Objective: We assessed the association of rs1801198 genotypes with OCM marker concentrations and primary risks of congenital abnormalities, cancer, and Alzheimer disease.Design: We conducted a systematic review of the literature that was published from January 1966 to February 2017 and included all studies that assessed the association between rs1801198 and OCM markers or a pathologic condition.Results: Thirty-four studies met the inclusion criteria. Subjects with the rs1801198 GG genotype had significantly lower concentrations of holotranscobalamin [standardized mean difference (SMD): -0.445 (95% CI: -0.673, -0.217; P < 0.001); I2 = 48.16% (95% CI: 0.00%, 78.10%; P = 0.07)] and higher concentrations of homocysteine (European descent only) [SMD: 0.070 (95% CI: 0.020, 0.120; P = 0.01); I2 = 0.00% (95% CI: 0.00%, 49.59%; P = 0.73)] than did subjects with the rs1801198 CC genotype. The meta-analysis on the association between rs1801198 and methylmalonic acid (MMA) lacked statistical power. No significant difference was observed regarding cobalamin, folate, and red blood cell folate. No significant association was observed between rs1801198 and primary risks of congenital abnormalities, cancer, or Alzheimer disease.Conclusions: Meta-analysis results indicate an influence of rs1801198 on holotranscobalamin and homocysteine concentrations in European-descent subjects. In addition, well-designed and -powered studies should be conducted for assessing the association between rs1801198 and MMA and clinical manifestations that are linked to a decreased availability of cobalamin. This review was registered at www.crd.york.ac.uk/prospero as CRD42017058504.

One-carbon metabolism, cognitive impairment and CSF measures of Alzheimer pathology: homocysteine and beyond.

BACKGROUND: Hyperhomocysteinemia is a risk factor for cognitive decline and dementia, including Alzheimer disease (AD). Homocysteine (Hcy) is a sulfur-containing amino acid and metabolite of the methionine pathway. The interrelated methionine, purine, and thymidylate cycles constitute the one-carbon metabolism that plays a critical role in the synthesis of DNA, neurotransmitters, phospholipids, and myelin. In this study, we tested the hypothesis that one-carbon metabolites beyond Hcy are relevant to cognitive function and cerebrospinal fluid (CSF) measures of AD pathology in older adults. METHODS: Cross-sectional analysis was performed on matched CSF and plasma collected from 120 older community-dwelling adults with (n = 72) or without (n = 48) cognitive impairment. Liquid chromatography-mass spectrometry was performed to quantify one-carbon metabolites and their cofactors. Least absolute shrinkage and selection operator (LASSO) regression was initially applied to clinical and biomarker measures that generate the highest diagnostic accuracy of a priori-defined cognitive impairment (Clinical Dementia Rating-based) and AD pathology (i.e., CSF tau phosphorylated at threonine 181 [p-tau181]/ß-Amyloid 1-42 peptide chain [Aß1-42] >0.0779) to establish a reference benchmark. Two other LASSO-determined models were generated that included the one-carbon metabolites in CSF and then plasma. Correlations of CSF and plasma one-carbon metabolites with CSF amyloid and tau were explored. LASSO-determined models were stratified by apolipoprotein E (APOE) ε4 carrier status. RESULTS: The diagnostic accuracy of cognitive impairment for the reference model was 80.8% and included age, years of education, Aß1-42, tau, and p-tau181. A model including CSF cystathionine, methionine, S-adenosyl-L-homocysteine (SAH), S-adenosylmethionine (SAM), serine, cysteine, and 5-methyltetrahydrofolate (5-MTHF) improved the diagnostic accuracy to 87.4%. A second model derived from plasma included cystathionine, glycine, methionine, SAH, SAM, serine, cysteine, and Hcy and reached a diagnostic accuracy of 87.5%. CSF SAH and 5-MTHF were associated with CSF tau and p-tau181. Plasma one-carbon metabolites were able to diagnose subjects with a positive CSF profile of AD pathology in APOE ε4 carriers. CONCLUSIONS: We observed significant improvements in the prediction of cognitive impairment by adding one-carbon metabolites. This is partially explained by associations with CSF tau and p-tau181, suggesting a role for one-carbon metabolism in the aggregation of tau and neuronal injury. These metabolites may be particularly critical in APOE ε4 carriers.

Circulating concentrations of biomarkers and metabolites related to vitamin status, one-carbon and the kynurenine pathways in US, Nordic, Asian, and Australian populations.

Background: Circulating concentrations of biomarkers that are related to vitamin status vary by factors such as diet, fortification, and supplement use. Published biomarker concentrations have also been influenced by the variation across laboratories, which complicates a comparison of results from different studies.Objective: We robustly and comprehensively assessed differences in biomarkers that are related to vitamin status across geographic regions.Design: The trial was a cross-sectional study in which we investigated 38 biomarkers that are related to vitamin status and one-carbon and tryptophan metabolism in serum and plasma from 5314 healthy control subjects representing 20 cohorts recruited from the United States, Nordic countries, Asia, and Australia, participating in the Lung Cancer Cohort Consortium. All samples were analyzed in a centralized laboratory.Results: Circulating concentrations of riboflavin, pyridoxal 5'-phosphate, folate, vitamin B-12, all-trans retinol, 25-hydroxyvitamin D, and α-tocopherol as well as combined vitamin scores that were based on these nutrients showed that the general B-vitamin concentration was highest in the United States and that the B vitamins and lipid soluble vitamins were low in Asians. Conversely, circulating concentrations of metabolites that are inversely related to B vitamins involved in the one-carbon and kynurenine pathways were high in Asians. The high B-vitamin concentration in the United States appears to be driven mainly by multivitamin-supplement users.Conclusions: The observed differences likely reflect the variation in intake of vitamins and, in particular, the widespread multivitamin-supplement use in the United States. The results provide valuable information about the differences in biomarker concentrations in populations across continents.

Preparation of a novel starch-derived three-dimensional ordered macroporous carbon for improving the dissolution rate and oral bioavailability of water-insoluble drugs.

In our study, soluble starch was applied as a novel carbon source for preparing three-dimensional ordered macroporous carbon (3DOMC) using monodisperse silica nanospheres as the hard template. The 3DOMC was used as an insoluble drug carrier when it was found that it could markedly improve the water solubility of felodipine (FDP). The structural features of 3DOMC were characterized by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The 3DOMC structure was found to have a higher drug loading than microporous and mesoporous structures, and the interconnected nanostructure effectively inhibited the formation of drug crystals. FDP, belonging to the Biopharmaceutics Classification System II (BCSII), was chosen as the model drug and was loaded into the 3DOMC structure by solvent evaporation. The state of FDP in the 3DOMC structure was characterized by powder X-ray diffractometry (PXRD), differential scanning calorimetry (DSC) and Fourier-transform infrared spectroscopy (FTIR). The results obtained showed that FDP was present in the pores in an amorphous or microcrystalline state. In vivo and in vitro experiments indicated that 3DOMC could significantly improve the drug dissolution rate, but the FDP-3DOMC self-made common tablets had the disadvantage of a burst effect. For this reason, osmotic pump technology was used to control the drug release rate. We developed a potentially useful insoluble drug carrier for pharmaceutical applications.

Analysis of metabolic pools in broilers chicks.

This paper shows the possibility of obtaining new parameters for the mathematical modelling of data on stable isotopes in biological systems and its application in obtaining data on metabolic pools of blood plasma, blood serum, liver and muscle of broilers. This theory states that the modelling of turnover used for studies of isotopic incorporation when the metabolism has a single metabolic pool is feasible by the technique of setting an exponential. However, when the metabolism has more than one metabolic pool, it is necessary to apply the linearization technique, linear regression adjustment and evaluation of the assumptions of regression to obtain the kinetic parameters such as half-life (T1/2) and isotope exchange rate (k). The application of this technique on carbon-13 data from 100 one-day-old chicks, with the change of diet composed of grains of the photosynthetic cycle of plants from C4 to C3, in broilers has enabled the discovery that the liver, blood plasma and blood serum have a single metabolic pool; however, the pectoral muscle has two metabolic pools. For the liver, blood plasma and blood serum, the half-life values were found by the exponential fit being T1/2 = 1.4 days with the rate of exchange of k = 0.502, T1/2 = 2.4 days with k = 0.293 and T1/2 = 2.0 days with k = 0.348, respectively. For the pectoral muscle, after linearization, the half-life values were found for T1/2(1) = 1.7 and T1/2(2) = 3 days, with exchange rates of k1 = 0.405 and k2 = 0.235, representing approximately 66 and 34%, respectively.

Serum pneumoproteins in tunnel construction workers.

PURPOSE: The aim was to study inflammatory biomarkers in tunnel construction workers (TCW). METHODS: Surfactant protein D (SP-D), Clara cell protein 16 (CC-16) and C-reactive protein (CRP) were studied in serum of 90 TCW and 50 referents before and at the end of an 11-day work period. Personal air sampling was carried out on the two consecutive days before follow-up. RESULTS: The TCW's geometric mean exposure to particulate matter and α-quartz were 604 and 74 µg/m(3), respectively. The arithmetic mean concentration of elemental carbon was 51 µg/m(3). The arithmetic mean concentration of SP-D was reduced by 7.6 µg/L in the TCWs and 0.6 µg/L in the referents (p = 0.04) at the end as compared to before the work period. Subjects who had ever been TCW had lower arithmetic mean CC-16 concentrations at baseline (5.4 µg/L) than subjects who had never worked as TCW (6.4 µg/L). Years worked as TCW was significantly associated with an annual mean decline of the CC-16 concentration of 0.04 µg/L. The concentrations of the biomarker of systemic inflammation, CRP, were not affected by exposure in the TCWs. Current smoking and body mass index have a large impact on the measured biomarker concentrations. CONCLUSIONS: The results suggest that former and current TCWs have lower serum CC-16 concentrations than referents, while the concentrations of SP-D decreased during exposure. The serum biomarker of systemic inflammation, CRP, was not altered during exposure. Current smoking and BMI were related to the concentrations of all measured biomarkers.

Adsorption properties of an activated carbon for 18 cytokines and HMGB1 from inflammatory model plasma.

The ability of an activated carbon (AC) to adsorb 18 different cytokines with molecular weights ranging from 8 kDa to 70 kDa and high mobility group box-1 (HMGB1) from inflammatory model plasma at 310 K and the mechanisms of adsorption were examined. Porosity analysis using N2 gas adsorption at 77K showed that the AC had micropores with diameters of 1-2 nm and mesopores with diameters of 5-20 nm. All 18 cytokines and HMGB1 were adsorbed on the AC; however, the shapes of the adsorption isotherms changed depending on the molecular weight. The adsorption isotherms for molecules of 8-10 kDa, 10-20 kDa, 20-30 kDa, and higher molecular weights were classified as H-2, L-3, S-3, and S-1 types, respectively. These results suggested that the adsorption mechanism for the cytokines and HMGB1 in the mesopores and on the surface of the AC differed as a function of the molecular weight. On the basis of these results, it can be concluded that AC should be efficient for cytokine adsorption.

Turnover do carbono em sangue e plasma, nas fases crescimento e postura, de codornas japonesas (Coturnix coturnix japonica) / Turnover of carbon in blood and plasma, growth stages and posture, japanese quails (Coturnix coturnix japonica)

O estudo objetivou avaliar o turnover do 13C no sangue e plasma de codornas japonesas utilizando a técnica de isótopos estáveis, para a obtenção do patamar de equilíbrio isotópico que servirá de fundamento para estudos de rastreabilidade. Foram utilizadas 300 aves durante o período experimental de 1-42 e 49-97 dias de idade. Os tratamentos da primeira fase foram constituídos de dietas à base de arroz (C3), contendo ou não farinha de carne e ossos bovinos e um com dieta à base de milho (C4). Nessa primeira fase foi analisado o turnover do sinal isotópico do matrizeiro à base de dietas C4 para dietas à base de C3, como também as diferenças isotópicas das dietas contendo ou não farinha de origem animal. Na segunda fase houve uma substituição de dietas, ou seja, as aves no tratamento C4 da primeira fase passaram a consumir dieta C3, e o tratamento que antes consumia dieta C3 passou para dieta C4. Para determinar a taxa de turnover e o percentual estimado de participação da farinha na composição do material coletado, foi empregado o modelo de diluição isotópica utilizando valores do δ13C. A comparação entre as meias-vidas do sangue e plasma da primeira fase revelou o enriquecimento do δ13C na dieta; já na segunda fase foi possível observar as velocidades de incorporação após a troca das dietas...

The study aimed to evaluate the turnover of 13C in the blood and plasma of Japanese quail using the technique of stable isotopes to obtain the level of isotopic equilibrium that will be the foundation for studies of traceability. A total of 300 birds during the experimental period of 1-42 and 49-97 days of age. The first phase of the treatments consisted of diets based on rice (C3), with or without meat and beef bones and a diet based on corn (C4). This was first examined the turnover signal farm matrix isotope-based diets based diets C4 to C3, as well as isotopic differences of diets with or without animal meal In the second stage there was a substitution of diets, or C4 treatment of the first stage passes to consume C3-based diet than before treatment and diet consumed C3 to C4 diet. To determine the turnover rate and the estimated percentage of participation of flour in the composition of the collected material was used isotope dilution model using δ13C values. A comparison of the half-lives of blood and plasma from the first phase discloses the enrichment of dietary δ 13C, in the second phase was observed after incorporation speeds exchange diets...

Alcohol consumption, one-carbon metabolites, liver cancer and liver disease mortality.

BACKGROUND: Excess alcohol consumption adversely affects one-carbon metabolism and increases the risk of liver disease and liver cancer. Conversely, higher folate levels have been inversely associated with liver damage. The current study investigated the effects of alcohol and one-carbon metabolite intake on liver cancer incidence and liver disease mortality within the Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study. METHODS: Cox proportional hazards modeling was used to calculate hazard ratios and 95% confidence intervals (CIs) in a population of 27,086 Finnish males with 194 incident liver cancers and 213 liver disease deaths. In a nested case-control subset (95 liver cancers, 103 controls), logistic regression was used to calculate odds ratios and 95% CIs for serum one-carbon metabolites in relation to liver cancer risk. RESULTS: Daily alcohol consumption of more than 20.44 g was associated with an increased risk of both liver cancer incidence (Hazard Ratio (HR) 1.52, 95%CI 1.06-2.18) and liver disease mortality (HR 6.68, 95%CI 4.16-10.71). These risks were unaffected by one-carbon metabolite intake. Similarly, in the case-control study, none of the serum one-carbon metabolites were associated with liver cancer. CONCLUSIONS: The current study provided no convincing evidence for a protective association of one-carbon metabolite intake or serum level on the risk of liver cancer or liver disease mortality.

[Hygienic evaluation of immune and endocrine systems and modifications of their relationship in reproductive-age women working under exposure to chemical factors in activated carbon emissions].

The article presents results of the evaluation the changes in the relationships between immune and endocrine systems in reproductive-age women, working under exposure to chemical factors from activated carbon production. A significant increase of some chemical elements and compounds was found in blood that was associated with changes in the endocrine and immune status, as well as the presence of features in correlation parameters of these systems in reproductive-age women, working under exposure to chemical factors.

Slow isotope turnover rates and low discrimination values in the American alligator: implications for interpretation of ectotherm stable isotope data.

Stable isotope analysis has become a standard ecological tool for elucidating feeding relationships of organisms and determining food web structure and connectivity. There remain important questions concerning rates at which stable isotope values are incorporated into tissues (turnover rates) and the change in isotope value between a tissue and a food source (discrimination values). These gaps in our understanding necessitate experimental studies to adequately interpret field data. Tissue turnover rates and discrimination values vary among species and have been investigated in a broad array of taxa. However, little attention has been paid to ectothermic top predators in this regard. We quantified the turnover rates and discrimination values for three tissues (scutes, red blood cells, and plasma) in American alligators (Alligator mississippiensis). Plasma turned over faster than scutes or red blood cells, but turnover rates of all three tissues were very slow in comparison to those in endothermic species. Alligator δ(15)N discrimination values were surprisingly low in comparison to those of other top predators and varied between experimental and control alligators. The variability of δ(15)N discrimination values highlights the difficulties in using δ(15)N to assign absolute and possibly even relative trophic levels in field studies. Our results suggest that interpreting stable isotope data based on parameter estimates from other species can be problematic and that large ectothermic tetrapod tissues may be characterized by unique stable isotope dynamics relative to species occupying lower trophic levels and endothermic tetrapods.

Equivalence of arterial and venous blood for [11C]CO2-metabolite analysis following intravenous administration of 1-[11C]acetate and 1-[11C]palmitate.

PURPOSE: Sampling of arterial blood for metabolite correction is often required to define a true radiotracer input function in quantitative modeling of PET data. However, arterial puncture for blood sampling is often undesirable. To establish whether venous blood could substitute for arterial blood in metabolite analysis for quantitative PET studies with 1-[(11)C]acetate and 1-[(11)C]palmitate, we compared the results of [(11)C]CO2-metabolite analyses performed on simultaneously collected arterial and venous blood samples. METHODS: Paired arterial and venous blood samples were drawn from anesthetized pigs at 1, 3, 6, 8, 10, 15, 20, 25 and 30min after i.v. administration of 1-[(11)C]acetate and 1-[(11)C]palmitate. Blood radioactivity present as [(11)C]CO2 was determined employing a validated 10-min gas-purge method. Briefly, total blood (11)C radioactivity was counted in base-treated [(11)C]-blood samples, and non-[(11)C]CO2 radioactivity was counted after the [(11)C]-blood was acidified using 6N HCl and bubbled with air for 10min to quantitatively remove [(11)C]CO2. RESULTS: An excellent correlation was found between concurrent arterial and venous [(11)C]CO2 levels. For the [(11)C]acetate study, the regression equation derived to estimate the venous [(11)C]CO2 from the arterial values was: y=0.994x+0.004 (r(2)=0.97), and for the [(11)C]palmitate: y=0.964x-0.001 (r(2)=0.9). Over the 1-30min period, the fraction of total blood (11)C present as [(11)C]CO2 rose from 4% to 64% for acetate, and 0% to 24% for palmitate. The rate of [(11)C]CO2 appearance in venous blood appears similar for the pig model and humans following i.v. [(11)C]-acetate administration. CONCLUSION: Venous blood [(11)C]CO2 values appear suitable as substitutes for arterial blood samples in [(11)C]CO2 metabolite analysis after administration of [(11)C]acetate or [(11)C]palmitate ADVANCES IN KNOWLEDGE AND IMPLICATIONS FOR PATIENT CARE: Quantitative PET studies employing 1-[(11)C]acetate and 1-[(11)C]palmitate can employ venous blood samples for metabolite correction of an image-derived tracer arterial input function, thereby avoiding the risks of direct arterial blood sampling.

Inherent variation in stable isotope values and discrimination factors in two life stages of green turtles.

We examine inherent variation in carbon and nitrogen stable isotope values of multiple soft tissues from a population of captive green turtles Chelonia mydas to determine the extent of isotopic variation due to individual differences in physiology. We compare the measured inherent variation in the captive population with the isotopic variation observed in a wild population of juvenile green turtles. Additionally, we measure diet-tissue discrimination factors to determine the offset that occurs between isotope values of the food source and four green turtle tissues. Tissue samples (epidermis, dermis, serum, and red blood cells) were collected from captive green turtles in two life stages (40 large juveniles and 30 adults) at the Cayman Turtle Farm, Grand Cayman, and analyzed for carbon and nitrogen stable isotopes. Multivariate normal models were fit to the isotope data, and the Bayesian Information Criterion was used for model selection. Inherent variation and discrimination factors differed among tissues and life stages. Inherent variation was found to make up a small portion of the isotopic variation measured in a wild population. Discrimination factors not only are tissue and life stage dependent but also appear to vary with diet and sea turtle species, thus highlighting the need for appropriate discrimination factors in dietary reconstructions and trophic-level estimations. Our measures of inherent variation will also be informative in field studies employing stable isotope analysis so that differences in diet or habitat are more accurately identified.

Use of relative 12C/14C isotope ratios to estimate metabolite concentrations in the absence of authentic standards.

BACKGROUND: There is considerable interest in the determination of relative abundances of human metabolites in plasma (and potentially excreta) with reasonable accuracy early on in the drug development process in order to make scientifically sound decisions with regard to the presence of potentially active or toxic disproportionate metabolites. At this point, authentic metabolite standards are generally not available. RESULTS: A new methodology is proposed for the estimation of metabolite concentrations in the absence of authentic standards. A reference sample containing radiolabeled metabolites of interest is produced by incubating the (14)C-labeled drug in vitro, and mixed with a sample to be quantitated containing the unlabeled metabolites. The (12)C/(14)C isotope ratio is measured with high-resolution ESI-MS for each metabolite, and used as a basis for quantitation of the cold metabolite based on the concentration of radioactive metabolite, determined from independent analysis of the radioactive sample with LC-radiochemical detection. The (14)C-labeled metabolite serves as an isotopically labeled internal standard, which corrects for any variations in injection volume, sample preparation, MS intensity drift, matrix effects and/or saturation of electrospray ionization. The approach was validated by the analysis of solutions containing variable amounts of the analyte with a fixed amount of radioactive standard on a QToF Synapt(®) G2 MS system. The same methodology was also successfully applied to first-in-human plasma samples analyzed on a LTQ-Orbitrap(®). CONCLUSION: The metabolite abundances obtained by (12)C/(14)C isotope ratio measurements showed suitable accuracy and precision and were very close to those obtained with matrix mixing. The parent drug concentrations also corresponded well with the bioanalytical results obtained with a validated LC-MS/MS method.

Isotopic segregation between sympatric seabird species increases with nutritional stress.

Dietary segregation is essential for the coexistence of closely related species of animals. However, little is known about how changes in availability of food resources might affect trophic interactions of wild animals breeding in sympatry. Here, we examined how interannual variations in relative food availability (as reflected in blood levels of stress hormone corticosterone, CORT) affect food partitioning (assessed via a comparison of stable isotope δ(15)N and δ(13)C ratios of blood) between the common murre (Uria aalge) and thick-billed murre (Uria lomvia), breeding on a single colony in the Bering Sea. During a 6-year study, CORT varied among years but not between species, whereas stable isotope ratios varied among years and between species. Isotopic distance between species increased with increasing CORT. These results indicate that, when food was not limiting, both species relied on similar food resources. As foraging conditions deteriorated, murres diverged in their diets. We conclude that the degree of dietary segregation between Uria spp. varies with changes in the availability of food and is greatest during food shortages.

A prospective study of one-carbon metabolism biomarkers and risk of renal cell carcinoma.

OBJECTIVE: Previous studies have found associations between one-carbon metabolism factors and risk of several cancers, but little is known regarding renal cell carcinoma (RCC). We conducted a nested case-control study within the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study, a prospective study of Finnish male smokers aged 50-69 at baseline. METHODS: Prediagnostic folate, vitamin B(6), vitamin B(12), cysteine, riboflavin, and homocysteine concentrations were measured in fasting serum from 224 incident RCC cases and 224 controls (matched on age and date of serum collection). Conditional logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs), adjusted for potential confounders. RESULTS: Serum folate tended to be inversely associated with RCC, compared to the first quartile, the odds ratios (95% CI) for subsequent quartiles were 0.62 (0.35-1.08), 0.52 (0.29-0.93), and 0.67 (0.37-1.20) (P-trend = 0.19). When modeled as a threshold effect, subjects in the lowest serum folate quartile (