Mice deficient in the mitochondrial branched-chain aminotransferase (BCATm) respond with delayed tumour growth to a challenge with EL-4 lymphoma.

BACKGROUND: The mitochondrial branched-chain aminotransferase (BCATm) is a recently discovered cancer marker with a poorly defined role in tumour progression. METHODS: To understand how a loss of function of BCATm affects cancer, the global knockout mouse BCATmKO was challenged with EL-4 lymphoma under different diet compositions with varying amounts of branched-chain amino acids (BCAAs). Next, the growth and metabolism of EL-4 cells were studied in the presence of different leucine concentrations in the growth medium. RESULTS: BCATmKO mice experienced delayed tumour growth when fed standard rodent chow or a normal BCAA diet. Tumour suppression correlated with 37.6- and 18.9-fold increases in plasma and tumour BCAAs, 37.5% and 30.4% decreases in tumour glutamine and alanine, and a 3.5-fold increase in the phosphorylation of tumour AMPK in BCATmKO mice on standard rodent chow. Similar results were obtained with a normal but not with a choice BCAA diet. CONCLUSIONS: Global deletion of BCATm caused a dramatic build-up of BCAAs, which could not be utilised for energy or amino acid synthesis, ultimately delaying the growth of lymphoma tumours. Furthermore, physiological, but not high, leucine concentrations promoted the growth of EL-4 cells. BCATm and BCAA metabolism were identified as attractive targets for anti-lymphoma therapy.

Carbon and nitrogen stable isotopes in U.S. milk: Insight into production process.

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.

A Dual-Carbon-and-Nitrogen Stable Isotope Ratio Model Is Not Superior to a Single-Carbon Stable Isotope Ratio Model for Predicting Added Sugar Intake in Southwest Virginian Adults.

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.

Effect of baking and fermentation on the stable carbon and nitrogen isotope ratios of grain-based food.

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.