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Paleoclimate reconstructions from the Holocene are important for defining baseline conditions in order to interpret and contextualize the effects of modern climate change. Such records are particularly lacking for Siberia, a region that represents ~ 50% of the Arctic. In addition, the majority of proxy-based paleoclimate reconstructions for the Holocene represent mean annual conditions, and few quantify winter temperature, which is particularly important for predicting the effects of global warming in Arctic environments. Here we provide the first quantitative proxy reconstruction of precipitation and temperature for both summer and winter for 3000 years ago via novel high-resolution intra-annual carbon and oxygen isotope measurements across annual growth rings of fossil wood mummified within the permafrost of far northeastern Siberia. We found that the site experienced greater precipitation year-round (~ 10% increase in summer and ~ 30% increase in winter), cooler summer temperatures, and warmer winter temperatures, compared with today. Our findings indicate that warmer winter temperatures (+ 3.0 °C above early twentieth century values) in the Arctic 3000 years ago drove higher mean annual temperature by up to 1 °C, despite the existence of cooler summers, a similar phenomenon to what is observed within today's Arctic environments, and past intervals of extreme global warmth.
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RATIONALE: Position-specific 13 C/12 C ratios within amino acids remain largely unexplored in environmental samples due to methodological limitations. We hypothesized that natural-abundance isotope patterns in serine may serve as a proxy for plant metabolic fluxes including photorespiration. Here we describe an Orbitrap method optimized for the position-specific carbon isotope analysis of serine to test our hypothesis and discuss the generalizability of this method to other amino acids. METHODS: Position-specific carbon isotope ratios of serine were measured using a Thermo Scientific™ Q Exactive™ GC Orbitrap™. Amino acids were hydrolyzed from Arabidopsis biomass, purified from potential matrix interferences, and derivatized alongside standards. Derivatized serine (N,O-bis(trifluoroacetyl)methyl ester) was isolated using gas chromatography, trapped in a reservoir, and purged into the electron ionization source over tens of minutes, producing fragment ions containing different combinations of atoms from the serine-derivative molecule. The 13 C/12 C ratios of fragments with monoisotopic masses of 110.0217, 138.0166, and 165.0037 Da were monitored in the mass analyzer and used to calculate position-specific δ13 C values relative to a working standard. RESULTS: This methodology constrains position-specific δ13 C values for nanomole amounts of serine isolated from chemically complex mixtures. The δ13 C values of fragment ions of serine were characterized with ≤1 precisions, leading to propagated standard errors of 0.7-5 for each carbon position. Position-specific δ13 C values differed by up to ca 28 ± 5 between serine molecules hydrolyzed from plants grown under contrasting pCO2 , selected to promote different fluxes through photosynthesis and photorespiration. The method was validated using pure serine standards characterized offline. CONCLUSIONS: This study presents the first Orbitrap-based measurements of natural-abundance, position-specific carbon isotope variation in an amino acid isolated from a biological matrix. We present a method for the precise characterization of isotope ratios in serine and propose applications probing metabolism in plants. We discuss the potential for extending these approaches to other amino acids, paving the way for novel applications.
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Aminoácidos , Serina , Aminas/análisis , Aminoácidos/química , Isótopos de Carbono/análisis , Cromatografía de Gases y Espectrometría de Masas/métodosRESUMEN
The δ13C value of blood is a novel proposed biomarker of added sugars (AS) intake. AS prediction equations using either a single- (δ13C) or dual-isotope model (δ13C and δ15N) were previously developed in an adult population with high AS intake living in southwest Virginia (reference group). The purpose of this investigation was to test the δ13C single- and δ13C and δ15N dual-isotope prediction equations for AS intake in adults with a lower mean AS intake and different demographic characteristics (test group). The blood samples for the reference (n = 257 for single-isotope, n = 115 for dual-isotope) and test groups (n = 56) were analyzed for δ13C and δ15N values using natural abundance stable isotope mass spectrometry and were compared to reported dietary AS intake. When the δ13C single-isotope equation was applied to the test group, predicted AS intake was not significantly different from reported AS intake (mean difference ± standard error = -3.6 ± 5.5 g, Z = -0.55, p = 0.51). When testing the dual-isotope equation, predicted AS was different from reported AS intake (mean difference ± SEM = 13.0 ± 5.4 g, Z = -2.95, p = 0.003). δ13C value was able to predict AS intake using a blood sample within this population subset. The single-isotope prediction equation may be an alternative method to assess AS intake and is more objective, cost-feasible, and efficient than traditional dietary assessment methods. However, more research is needed to assess this biomarker with rigorous study designs such as controlled feeding.
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Isótopos de Carbono/sangre , Azúcares de la Dieta/análisis , Ingestión de Alimentos/fisiología , Modelos Teóricos , Evaluación Nutricional , Adulto , Biomarcadores/sangre , Estudios de Factibilidad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Isótopos de Nitrógeno/sangre , Valor Predictivo de las Pruebas , Ensayos Clínicos Controlados Aleatorios como Asunto , Autoinforme , VirginiaRESUMEN
BACKGROUND: Radioactive or stable isotopic labeling of metabolites is a strategy that is routinely used to map the cellular fate of a selected labeled metabolite after it is added to cell culture or to the circulation of an animal. However, a labeled metabolite can be enzymatically changed in cellular metabolism, complicating the use of this experimental strategy to understand how a labeled metabolite moves between organs. These methods are also technically demanding, expensive and potentially toxic. To allow quantification of the bulk movement of metabolites between organs, we have developed a novel application of stable isotope ratio mass spectrometry (IRMS). RESULTS: We exploit natural differences in 13C/12C ratios of plant nutrients for a low-cost and non-toxic carbon labeling, allowing a measurement of bulk carbon transfer between organs in vivo. IRMS measurements were found to be sufficiently sensitive to measure organs from individual Drosophila melanogaster larvae, giving robust measurements down to 2.5 µg per sample. We apply the method to determine if carbon incorporated into a growing solid tumor is ultimately derived from food or host tissues. CONCLUSION: Measuring tumor growth in a D. melanogaster larvae tumor model reveals that these tumors derive a majority of carbon from host sources. We believe the low cost and non-toxic nature of this methodology gives it broad applicability to study carbon flows between organs also in other animals and for a range of other biological questions.
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Neoplasias , Animales , Carbono , Isótopos de Carbono , Drosophila melanogaster , Espectrometría de MasasRESUMEN
BACKGROUND: Consumption of added sugars (AS) and sugar-sweetened beverages (SSB) may adversely affect adolescents' weight and cardiovascular disease risk. Reliance on self-reported dietary assessment methods is a common research limitation, which could be overcome by dietary intake biomarkers. AIM: The investigation was a proof-of-concept study to evaluate the proposed carbon isotope ratio (δ13C) biomarker of AS intake in adolescents, using a controlled feeding design. METHODS: Participants (n = 33, age 15.3 years, 53% female) underwent two seven-day controlled feeding periods in a randomly assigned order. Diets were matched in composition except for AS content (5% or 25% of total energy). Fasting fingerstick blood samples were collected daily during each diet period. RESULTS: Fingerstick δ13C values changed from day 1 to 8 by -0.05 ± 0.071 on 5% AS, and +0.03 ± 0.083 on 25% AS (p ≤ 0.001). Reliability was demonstrated between day 7 and 8 δ13C values on the 5% (ICC = 0.996, p ≤ 0.001) and 25% (ICC = 0.997, p ≤ 0.001) AS diets. CONCLUSIONS: Larger scale investigations are warranted to determine if this technique could be applied to population-level research in order to help assess the effectiveness of interventions aimed at reducing the consumption of AS or SSB intake.
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Isótopos de Carbono/sangre , Dieta/métodos , Azúcares de la Dieta/sangre , Azúcares de la Dieta/farmacología , Adolescente , Biomarcadores/sangre , Femenino , Humanos , Masculino , Reproducibilidad de los Resultados , TiempoRESUMEN
RATIONALE: Stable isotope analysis (SIA), a potential method of verifying the geographic origin and production method of dairy products, has not been applied to United States (U.S.) dairy samples on a national scale. To determine the potential of carbon and nitrogen SIA in authenticity assessment of U.S. dairy products, we analyzed a geographically representative collection of conventional milk samples to determine isotopic variations with (1) Purchase Location and (2) Macronutrient Content. METHODS: A total of 136 milk samples spanning five commercially available varieties (3.25% [i.e., 'whole'], 2%, 1%, 0% [i.e., 'skim'] and 1% chocolate) were collected from randomly selected counties across the U.S. as part of the United States Department of Agriculture's (USDA's) National Food and Nutrient Analysis program. δ13 C and δ15 N values of bulk samples determined via elemental analysis/isotope ratio mass spectrometry (EA/IRMS) were used to assess the contribution of fat content, added sugar content and census-designated region of collection to isotopic variations within the dataset. RESULTS: There was a negative linear relationship between fat content and δ13 C values, with average milk δ13 C values decreasing by 0.33 for each 8.75% increase in dry weight (1% wet weight) fat content. The average δ13 C value of flavored 1% chocolate milk samples, which contain an additional 12 g of added sugar, was 2.05 higher than that of 1% unflavored milk (-16.47 for chocolate milk vs -18.52 for unflavored milk). When controlling for macronutrient content, milk samples collected in West region supermarkets possessed significantly lower δ13 C values than samples collected from Midwest, South, and Northeast regions. δ15 N values did not vary with macronutrient content or region of collection. CONCLUSIONS: Carbon stable isotope ratios in U.S. milk samples varied with macronutrient content and region of purchase, suggesting that SIA can provide insight into production processes within the U.S. dairy industry, with potential applications in national food adulteration and authentication efforts.
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Background: Reliance on self-reported dietary intake methods is a commonly cited research limitation, and dietary misreporting is a particular problem in children and adolescents. Objective indicators of dietary intake, such as dietary biomarkers, are needed to overcome this research limitation. The added sugar (AS) biomarker δ13C, which measures the relative abundance of 13C to 12C, has demonstrated preliminary validity in adults. Objective: The purpose of this investigation was to determine the comparative validity, test-retest reliability, and sensitivity of the δ13C biomarker to detect AS and sugar-sweetened beverage (SSB) intake using fingerstick blood samples in children and adolescents. Methods: Children (aged 6-11 y, n = 126, 56% male, mean ± SD age: 9 ± 2 y) and adolescents (aged 12-18 y, n = 200, 44% male, mean ± SD age: 15 ± 2 y) completed 4 testing sessions within a 3-wk period. Participants' height, weight, demographic characteristics, and health history were determined at the first session; 24-h recalls were obtained at each visit and fingerstick blood samples were collected at visits 1 and 3. Samples were analyzed for δ13C value using natural abundance stable isotope mass spectrometry. δ13C value was compared with dietary outcomes in the full sample, and in child and adolescent subgroups. t Tests and correlational analyses were used to assess biomarker validity and reliability, whereas logistic regression and area under the receiver-operator characteristic curve (AUC) were used to evaluate sensitivity. Results: Reported mean ± SD AS consumption was 82.2 ± 35.8 g/d and 329 ± 143 kcal/d, and SSB consumption was 222 ± 243 mL/d and 98 ± 103 kcal/d. Mean δ13C value was -19.65 ± 0.69, and was lower in children than in adolescents (-19.80 ± 0.67 compared with -19.56 ± 0.67, P = 0.002). δ13C values were similar across sessions (visit 1: -19.66 ± 0.68; visit 3: -19.64 ± 0.68; r = 0.99, P < 0.001) and were associated (P < 0.001) with intake of total AS (grams, kilocalories: r = 0.29) and SSB (milliliters, kilocalories: r = 0.35). The biomarker was able to better discriminate between high and low SSB consumers than high and low AS consumers, as demonstrated by the AUC (0.75 and 0.62, respectively). Conclusions: The δ13C biomarker is a promising, minimally invasive, objective biomarker of SSB intake in children and adolescents. Further evaluation using controlled feeding designs is warranted. Registered at clinicaltrials.gov as NCT02455388.
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Bebidas , Biomarcadores/sangre , Isótopos de Carbono/sangre , Adolescente , Índice de Masa Corporal , Niño , Estudios Transversales , Dieta , Azúcares de la Dieta/administración & dosificación , Metabolismo Energético , Femenino , Humanos , Masculino , Evaluación Nutricional , Edulcorantes Nutritivos/administración & dosificación , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Encuestas y CuestionariosRESUMEN
The role of environmental change in the late Pleistocene megafaunal extinctions remains a key question, owing in part to uncertainty about landscape changes at continental scales. We investigated the influence of environmental changes on megaherbivores using bone collagen nitrogen isotopes (n = 684, 63 new) as a proxy for moisture levels in the rangelands that sustained late Pleistocene grazers. An increase in landscape moisture in Europe, Siberia and the Americas during the Last Glacial-Interglacial Transition (LGIT; ~25-10 kyr bp) directly affected megaherbivore ecology on four continents, and was associated with a key period of population decline and extinction. In all regions, the period of greatest moisture coincided with regional deglaciation and preceded the widespread formation of wetland environments. Moisture-driven environmental changes appear to have played an important part in the late Quaternary megafaunal extinctions through alteration of environments such as rangelands, which supported a large biomass of specialist grazers. On a continental scale, LGIT moisture changes manifested differently according to regional climate and geography, and the stable presence of grasslands surrounding the central forested belt of Africa during this period helps to explain why proportionally fewer African megafauna became extinct during the late Pleistocene.
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OBJECTIVE: Controversy exists surrounding the health effects of added sugar (AS) and sugar-sweetened beverage (SSB) intakes, primarily due to a reliance on self-reported dietary intake. The purpose of the current investigation was to determine if a 6-month intervention targeting reduced SSB intake would impact δ13C AS intake biomarker values. DESIGN: A randomized controlled intervention trial. At baseline and at 6 months, participants underwent assessments of anthropometrics and dietary intake. Fasting fingerstick blood samples were obtained and analysed for δ13C value using natural abundance stable isotope MS. Statistical analysis included descriptive statistics, correlational analyses and multilevel mixed-effects linear regression analysis using an intention-to-treat approach. SETTING: Rural Southwest Virginia, USA. SUBJECTS: Adults aged ≥18 years who consumed ≥200 kcal SSB/d (≥837 kJ/d) were randomly assigned to either the intervention (n 155) or a matched-contact group (n 146). Participants (mean age 42·1 (sd 13·4) years) were primarily female and overweight (21·5 %) or obese (57·0 %). RESULTS: A significant group by time difference in δ13C value was detected (P<0·001), with mean (sd) δ13C value decreasing in the intervention group (pre: -18·92 (0·65) , post: -18·97 (0·65) ) and no change in the comparison group (pre: -18·94 (0·72) , post: -18·92 (0·73) ). Significant group differences in weight and BMI change were also detected. Changes in biomarker δ13C values were consistent with changes in self-reported AS and SSB intakes. CONCLUSIONS: The δ13C sugar intake biomarker assessed using fingerstick blood samples shows promise as an objective indicator of AS and SSB intakes which could be feasibly included in community-based research trials.
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Bebidas/análisis , Biomarcadores/sangre , Azúcares de la Dieta/análisis , Edulcorantes Nutritivos/análisis , Obesidad/sangre , Sobrepeso/sangre , Adulto , Índice de Masa Corporal , Isótopos de Carbono/análisis , Dieta , Azúcares de la Dieta/administración & dosificación , Femenino , Humanos , Masculino , Persona de Mediana Edad , Edulcorantes Nutritivos/administración & dosificación , Población Rural , VirginiaRESUMEN
OBJECTIVE: The δ 13C value of human blood is an emerging novel biomarker of added sugar (AS) intake for adults. However, no free-living, community-based assessments of comparative validity of this biomarker have been conducted. The purpose of the present investigation was to determine if Healthy Eating Index-2010 (HEI-2010) score, SoFAAS score (HEI-2010 sub-component for solid fat, alcohol and AS), AS and sugar-sweetened beverage (SSB) intakes were associated with δ 13C value of fingerstick blood in a community-based sample of adults, while controlling for relevant demographics. DESIGN: A cross-sectional analysis of data obtained from assessments of BMI, dietary intake using 24 h recalls and a fingerstick blood sample was completed. Statistical analyses included descriptive statistics, multiple linear regression and one-way ANOVA. SETTING: Rural Southwest Virginia, U.S.A. SUBJECTS: Adults (n 216) aged >18 years who consumed at least 837 kJ/d (200 kcal/d) from SSB. RESULTS: This sample of adult participants with low socio-economic status demonstrated a mean HEI-2010 score of 43.4 (sd 12.2), mean SoFAAS score of 10.2 (sd 5.7), mean AS intake of 93 (sd 65) g/d and mean blood δ 13C value of -18.88 (sd 0.7) . In four separate regression models, HEI-2010 (R 2=0.16), SoFAAS (R 2=0.19), AS (R 2=0.15) and SSB (R 2=0.14) predicted δ 13C value (all P≤0.001). Age was also predictive of δ 13C value, but not sex or race. CONCLUSIONS: These findings suggest that fingerstick δ 13C value has the potential to be a minimally invasive method for assessing AS and SSB intake and overall dietary quality in community-based settings. Strengths, limitations and future areas of research for using an objective δ 13C biomarker in diet-related public health studies are discussed.
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Bebidas/análisis , Biomarcadores/sangre , Sacarosa en la Dieta/sangre , Población Rural , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Índice de Masa Corporal , Isótopos de Carbono , Estudios Transversales , Sacarosa en la Dieta/administración & dosificación , Ingestión de Energía , Femenino , Humanos , Modelos Lineales , Masculino , Recuerdo Mental , Persona de Mediana Edad , Salud Pública , Autoinforme , Factores Socioeconómicos , Virginia , Adulto JovenRESUMEN
RATIONALE: Isotope ratio mass spectrometry (IRMS) is used extensively to reconstruct general attributes of prehistoric and modern diets in both humans and animals. In order to apply these methods to the accurate determination of specific intakes of foods/nutrients of interest, the isotopic signature of individually consumed foods must be constrained. For example, 86% of the calories consumed in the USA are derived from processed and prepared foods, but the relationship between the stable isotope composition of raw ingredients and the resulting products has not been characterized. METHODS: To examine the effect of common cooking techniques on the stable isotope composition of grain-based food items, we prepared yeast buns and sugar cookies from standardized recipes and measured bulk δ(13) C and δ(15) N values of samples collected throughout a 75 min fermentation process (buns) and before and after baking at 190°C (buns and cookies). Simple isotope mixing models were used to determine if the isotopic signatures of 13 multi-ingredient foods could be estimated from the isotopic signatures of their constituent raw ingredients. RESULTS: No variations in δ(13) C or δ(15) N values were detected between pre- and post-baked yeast buns (pre: -24.78/2.61, post: -24.75/2.74), beet-sugar cookies (pre: -24.48/3.84, post: -24.47/3.57), and cane-sugar cookies (pre: -19.07/2.97, post: -19.02/3.21), or throughout a 75 min fermentation process in yeast buns. Using isotopic mass balance equations, the δ(13) C/δ(15) N values of multi-ingredient foods were estimated from the isotopic composition of constituent raw ingredients to within 0.14 ± 0.13/0.24 ± 0.17 for gravimetrically measured recipes and 0.40 ± 0.38/0.58 ± 0.53 for volumetrically measured recipes. CONCLUSIONS: Two common food preparation techniques, baking and fermentation, do not substantially affect the carbon or nitrogen isotopic signature of grain-based foods. Mass-balance equations can be used to accurately estimate the isotopic signature of multi-ingredient food items for which quantitative ingredient information is available.
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Culinaria , Grano Comestible/química , Fermentación , Isótopos de Carbono/análisis , Culinaria/métodos , Isótopos de Nitrógeno/análisisRESUMEN
RATIONALE: Throughout at least the next century, CO(2) fertilization and environmental stresses (e.g. nutrient, moisture, insect herbivory) are predicted to affect yields of economically important crop species. Stable isotopes of carbon are used to study plant stresses, which affect yields, but a growth chamber design that can be used to isolate the effects of environmental stresses on crop-sized species through precise maintenance of pCO(2) levels and the δ(13)C values of atmospheric CO(2) (δ(13) C(CO2)) is lacking. METHODS: We designed and built low-cost plant growth chambers for growing staple crop species under precise pCO(2) and δ(13) C(CO2) conditions. Over the course of 14 hours, we assessed for pCO(2) stability at two targeted levels (ambient, ~400 ppm; and 2×, ~800 ppm) and measured the δ(13) C(CO2) value within the two chambers using a stable isotope ratio mass spectrometer. We also compared the temperature and relative humidity conditions within the two growth chambers, and in the ambient, outside air. RESULTS: Across our experimental period, we achieved δ(13) C(CO2) stability (ambient: -8.05 ± 0.17; elevated: -12.99 ± 0.29) that showed nearly half the variability of any previously reported values for other chamber designs. The stability of the pCO(2) conditions (ambient: 406 ± 3 ppm; elevated: 793 ± 54 ppm) was comparable with that in previous studies, but our design provided ~8 times more growing space than previous chamber designs. We also measured nearly identical temperature and relative humidity conditions for the two chambers throughout the experiment. CONCLUSIONS: Our growth chamber design marks a significant improvement in our ability to test for plant stress across a range of future pCO(2) scenarios. Through significant improvement in δ(13) C(CO2) stability and increased chamber size, small changes in carbon isotope fractionation can be used to assess stress in crop species under specific environmental (temperature, relative humidity, pCO(2)) conditions.
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BACKGROUND: An objective measure of added sugar (AS) and sugar-sweetened beverage (SSB) intake is needed. The δ(13)C value of finger-stick blood is a novel validated biomarker of AS/SSB intake; however, nonsweetener corn products and animal protein also carry a δ(13)C value similar to AS sources, which may affect blood δ(13)C values. The δ(15)N value of blood has been proposed as a "correction factor" for animal protein intake. OBJECTIVES: The objectives were to 1) identify foods associated with δ(13)C and δ(15)N blood values, 2) determine the contribution of nonsweetener corn to the diet relative to AS intake, and 3) determine if the dual-isotope model (δ(13)C and δ(15)N) is a better predictor of AS/SSB intake than δ(13)C alone. METHODS: A cross-sectional sample of southwest Virginian adults (n = 257; aged 42 ± 15 y; 74% overweight/obese) underwent dietary intake assessments and provided finger-stick blood samples, which were analyzed for δ(13)C and δ(15)N values by using natural abundance stable isotope mass spectrometry. Statistical analyses included ANOVAs, paired-samples t tests, and multiple linear regressions. RESULTS: The mean ± SD daily AS intake was 88 ± 59 g and nonsweetener corn intake was 13 ± 13 g. The mean δ(13)C value was -19.1 ± 0.9, which was significantly correlated with AS and SSB intakes (r = 0.32 and 0.39, respectively; P ≤ 0.01). The δ(13)C value and nonsweetener corn intake and the δ(15)N value and animal protein intake were not correlated. AS intake was significantly greater than nonsweetener corn intake (mean difference = 76.2 ± 57.2 g; P ≤ 0.001). The δ(13)C value was predictive of AS/SSB intake (ß range: 0.28-0.35; P ≤ 0.01); however, δ(15)N was not predictive and minimal increases in R(2) values were observed when the δ(15)N value was added to the model. CONCLUSIONS: The data do not provide evidence that the dual-isotope method is superior for predicting AS/SSB intakes within a southwest Virginian population. Our results support the potential of the δ(13)C value of finger-stick blood to serve as an objective measure of AS/SSB intake. This trial was registered at clinicaltrials.gov as NCT02193009.
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Carbohidratos/administración & dosificación , Isótopos de Carbono/sangre , Conducta Alimentaria , Isótopos de Nitrógeno/sangre , Edulcorantes/administración & dosificación , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Bebidas , Biomarcadores/sangre , Índice de Masa Corporal , Estudios Transversales , Dieta , Proteínas en la Dieta/administración & dosificación , Ingestión de Energía , Humanos , Persona de Mediana Edad , Modelos Teóricos , Evaluación Nutricional , Obesidad/sangre , Sobrepeso/sangre , Virginia , Adulto JovenRESUMEN
Serum carbon isotope values [13C-to-12C serum carbon isotope ratio (δ(13)C)], which reflect consumption of corn- and cane-based foods, differ between persons consuming high and low amounts of sugar-sweetened beverages (SSBs). In this study, we determined whether serum δ(13)C changes in response to change in SSB intake during an 18-mo behavioral intervention trial. Data were from a subset of 144 participants from the PREMIER trial, a completed behavioral intervention (Maryland, 1998-2004). SSB intake was assessed using 2 24-h dietary recall interviews. Blinded serum samples were assayed for δ(13)C by natural abundance stable isotope mass spectroscopy. Multiple linear regression models with generalized estimating equations and robust variance estimation were used. At baseline, mean SSB intake was 13.8 ± 14.2 fl oz/d, and mean δ(13)C serum value was -19.3 ± 0.6 units per mil (designated ). A reduction of 12 oz (355 mL)/d SSB (equivalent to 1 can of soda per day) was associated with 0.17 (95% CI: 0.08, 0.25 P < 0.0001) reduction in serum δ(13)C values over 18 mo (equivalent to a 1% reduction in δ(13)C from baseline). After adjusting for potential confounders, a reduction of 12 oz/d SSB (equivalent to 1 can of soda per day), over an 18-mo period, was associated with 0.12 (95% CI: 0.01, 0.22 P = 0.025) reduction in serum δ(13)C. These findings suggest that serum δ(13)C can be used as a measure of dietary changes in SSB intake.
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Bebidas/análisis , Isótopos de Carbono/sangre , Sacarosa en la Dieta/administración & dosificación , Edulcorantes/administración & dosificación , Adulto , Anciano , Índice de Masa Corporal , Dieta , Ingestión de Energía , Femenino , Humanos , Masculino , Maryland , Persona de Mediana EdadRESUMEN
RATIONALE: Workers have shown a correlation between temperature and the pollen δ(13)C value, and therefore suggested using pollen δ(13)C values to reconstruct paleotemperature. To evaluate the potential for pollen δ(13)C values to be used as a paleotemperature proxy, it is essential to quantify the variability in pollen δ(13)C values and to evaluate the effect of temperature on pollen δ(13)C values, in isolation, under controlled environmental conditions. METHODS: Pollen was isolated from 146 Hibiscus flowers from 26 plants within a single climate environment to evaluate isotopic variability in pollen δ(13)C values. The nearest leaf (n = 82) and flower phloem (n = 30) were also sampled to measure the δ(13)C variability in carbon providing the raw material for new growth. To evaluate the correlation between temperature and pollen δ(13)C values, we isolated pollen from 89 Brassica rapa plants grown in controlled growth chambers with temperatures ranging from 17 to 32°C. RESULTS: The range in pollen δ(13)C values collected from different flowers on the same Hibiscus plant was large (average = 1.6), and could be as much as 3.2. This amount of variability was similar to that seen between flower-adjacent leaves, and phloem extracted from styles of individual flowers. In controlled growth chamber experiments, we saw no correlation between temperature and the pollen (R(2) = 0.005) or leaf (R(2) = 0.10) δ(13)C values. CONCLUSIONS: We measured large variability in pollen δ(13)C values. When temperature was isolated from other environmental parameters, temperature did not correlate with the pollen δ(13)C value. These results complicate the supposed relationship between temperature and pollen δ(13)C values and caution against using nanogram isotope analytical techniques for characterizing whole-plant individuals.
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Isótopos de Carbono/análisis , Clima , Polen/química , Brassica rapa/química , Brassica rapa/metabolismo , Isótopos de Carbono/metabolismo , Flores/química , Hibiscus/química , Hibiscus/metabolismo , Humedad , Paleontología , Floema/química , Hojas de la Planta/química , Polen/metabolismo , TemperaturaRESUMEN
RATIONALE: Accurate online analysis of the δ(18)O values of nitrogen-bearing organic compounds is of interest to several emergent fields, including ecology, forensics and paleontology. During online analysis, high-temperature conversion (HTC) of nitrogen-bearing organics produces N(2) gas which creates isobaric interference with the isotopic measurement. Specifically, N(2) reacts with trace amounts of oxygen in the mass spectrometer source to form (14)N(16)O (m/z 30), which prevents accurate evaluation of the sample (12)C(18)O peak (m/z 30). METHODS: We present an alternative system to the conventional HTC, which uses a nickel-catalyzed ("NiCat") reduction furnace to convert HTC-produced CO into CO(2), allowing for δ(18)O measurement using signal intensities at m/z 44 and 46. RESULTS: This system yields identical δ(18)O values for nitrogen-doped and undoped sucrose and cellulose compounds up to molar yield ratios of N(2):CO = 0.22. In contrast, our conventional HTC system configured to factory recommendations with the stock gas chromatography (GC) column produced a discrepancy of ~5 between nitrogen-doped and undoped samples. CONCLUSIONS: Because of its ability to eliminate isobaric interference, the NiCat system is a viable alternative to conventional HTC for δ(18)O measurement, and can be constructed from relatively inexpensive and readily available materials. As an additional advantage, the CO(2) analyte produced by NiCat may be cryofocused, to allow for oxygen-isotope determinations on very small amounts of sample substrate.
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A reliance on self-reported dietary intake measures is a common research limitation, thus the need for dietary biomarkers. Added-sugar intake may play a role in the development and progression of obesity and related comorbidities; common sweeteners include corn and sugar cane derivatives. These plants contain a high amount of ¹³C, a naturally occurring stable carbon isotope. Consumption of these sweeteners, of which sugar-sweetened beverages are the primary dietary source, might be reflected in the δ¹³C value of blood. Fingerstick blood represents an ideal substrate for bioassay because of its ease of acquisition. The objective of this investigation was to determine if the δ¹³C value of fingerstick blood is a potential biomarker of added-sugar and sugar-sweetened beverage intake. Individuals aged 21 years and older (n = 60) were recruited to attend three laboratory visits; assessments completed at each visit depended upon a randomly assigned sequence (sequence one or two). The initial visit included assessment of height, weight, and dietary intake (sequence one: beverage intake questionnaire, sequence two: 4-day food intake record). Sequence one participants completed a food intake record at visit two, and nonfasting blood samples were obtained via routine fingersticks at visits one and three. Sequence two participants completed a beverage intake questionnaire at visit two, and provided fingerstick blood samples at visits two and three. Samples were analyzed for δ¹³C value using natural abundance stable isotope mass spectrometry. δ¹³C value was compared to dietary outcomes in all participants, as well as among those in the highest and lowest tertile of added-sugar intake. Reported mean added-sugar consumption was 66 ± 5 g/day, and sugar-sweetened beverage consumption was 330 ± 53 g/day and 134 ± 25 kcal/day. Mean fingerstick δ¹³C value was -19.94 ± 0.10, which differed by body mass index status. δ¹³C value was associated (all P < 0.05) with intake of total added sugars (g, r = 0.37; kcal, r = 0.37), soft drinks (g, r = 0.26; kcal, r = 0.27), and total sugar-sweetened beverage (g, r = 0.28; kcal, r = 0.35). The δ¹³C value in the lowest and the highest added-sugar intake tertiles were significantly different (mean difference = -0.48; P = 0.028). Although there are several potential dietary sources for blood carbon, the δ¹³C value of fingerstick blood shows promise as a noninvasive biomarker of added-sugar and sugar-sweetened beverage intake based on these findings.
Asunto(s)
Bebidas , Isótopos de Carbono/sangre , Sacarosa en la Dieta/administración & dosificación , Sacarosa en la Dieta/sangre , Edulcorantes/administración & dosificación , Adulto , Anciano , Anciano de 80 o más Años , Bebidas/estadística & datos numéricos , Biomarcadores/sangre , Registros de Dieta , Femenino , Humanos , Masculino , Persona de Mediana Edad , Obesidad/sangre , Obesidad/etiología , Obesidad/prevención & control , Autorrevelación , Encuestas y Cuestionarios , Edulcorantes/metabolismo , Adulto JovenRESUMEN
Gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) is increasingly applied to food and metabolic studies for stable isotope analysis (δ(13) C), with the quantification of analyte concentration often obtained via a second alternative method. We describe a rapid direct transesterification of triacylglycerides (TAGs) for fatty acid methyl ester (FAME) analysis by GC-C-IRMS demonstrating robust simultaneous quantification of amount of analyte (mean r(2) =0.99, accuracy ±2% for 37 FAMEs) and δ(13) C (±0.13) in a single analytical run. The maximum FAME yield and optimal δ(13) C values are obtained by derivatizing with 10% (v/v) acetyl chloride in methanol for 1 h, while lower levels of acetyl chloride and shorter reaction times skewed the δ(13) C values by as much as 0.80. A Bland-Altman evaluation of the GC-C-IRMS measurements resulted in excellent agreement for pure oils (±0.08) and oils extracted from French fries (±0.49), demonstrating reliable simultaneous quantification of FAME concentration and δ(13) C values. Thus, we conclude that for studies requiring both the quantification of analyte and δ(13) C data, such as authentication or metabolic flux studies, GC-C-IRMS can be used as the sole analytical method.
RESUMEN
Although the advantages of online δ(18)O analysis of organic compounds make its broad application desirable, researchers have encountered NO(+) isobaric interference with CO(+) at m/z 30 (e.g. (14)N(16)O(+), (12)C(18)O(+)) when analyzing nitrogenous substrates. If the δ(18)O value of inter-laboratory standards for substrates with high N:O value could be confirmed offline, these materials could be analyzed periodically and used to evaluate δ(18)O data produced online for nitrogenous unknowns. To this end, we present an offline method based on modifications of the methods of Schimmelmann and Deniro (Anal. Chem. 1985; 57: 2644) and Sauer and Sternberg (Anal. Chem. 1994; 66: 2409), whereby all the N(2) from the gas products of a chlorinated pyrolysis was eliminated, resulting in purified CO(2) for analysis via a dual-inlet isotope ratio mass spectrometry system. We evaluated our method by comparing observed δ(18)O values with previously published or inter-laboratory calibrated δ(18)O values for five nitrogen-free working reference materials; finding isotopic agreement to within ±0.2 for SIGMA® cellulose, IAEA-CH3 cellulose (C(6)H(10)O(5)) and IAEA-CH6 sucrose (C(12)H(22)O(11)), and within ±1.8 for IAEA-601 and IAEA-602 benzoic acids (C(7)H(6)O(2)). We also compared the δ(18)O values of IAEA-CH3 cellulose and IAEA-CH6 sucrose that was nitrogen-'doped' with adenine (C(5)H(5)N(5)), imidazole (C(3)H(4)N(2)) and 2-aminopyrimidine (C(4)H(5)N(3)) with the undoped δ(18)O values for the same substrates; yielding isotopic agreement to within ±0.7. Finally, we provide an independent analysis of the δ(18)O value of IAEA-600 caffeine (C(8)H(10)N(4)O(2)), previously characterized using online systems exclusively, and discuss the reasons for an average 1.4 enrichment in δ(18)O observed offline relative to the consensus online δ(18)O value.