A genome-wide metabolic QTL analysis in Europeans implicates two loci shaped by recent positive selection.

We have performed a metabolite quantitative trait locus (mQTL) study of the (1)H nuclear magnetic resonance spectroscopy ((1)H NMR) metabolome in humans, building on recent targeted knowledge of genetic drivers of metabolic regulation. Urine and plasma samples were collected from two cohorts of individuals of European descent, with one cohort comprised of female twins donating samples longitudinally. Sample metabolite concentrations were quantified by (1)H NMR and tested for association with genome-wide single-nucleotide polymorphisms (SNPs). Four metabolites' concentrations exhibited significant, replicable association with SNP variation (8.6×10(-11)

Objective grading of rib eye traits using the Q-FOM™ camera in Australian beef carcasses.

The objective of this study was to develop calibration models against rib eye traits and independently validate the precision, accuracy, and repeatability of the Frontmatec Q-FOM™ Beef grading camera in Australian carcasses. This study compiled 12 different research datasets acquired from commercial processing facilities and were comprised of a diverse range of carcass phenotypes, graded by industry identified expert Meat Standards Australia (MSA) graders and sampled for chemical intramuscular fat (IMF%). Calibration performance was maintained when the device was independently validated. For continuous traits, the Q-FOM™ demonstrated precise (root mean squared error of prediction, RMSEP) and accurate (coefficient of determination, R2) prediction of eye muscle area (EMA) (R2 = 0.89, RMSEP = 4.3 cm2, slope = 0.96, bias = 0.7), MSA marbling (R2 = 0.95, RMSEP = 47.2, slope = 0.98, bias = -12.8) and chemical IMF% (R2 = 0.94, RMSEP = 1.56%, slope = 0.96, bias = 0.64). For categorical traits, the Q-FOM™ predicted 61%, 64.3% and 60.8% of AUS-MEAT marbling, meat colour and fat colour scores equivalent, and 95% within ±1 classes of expert grader scores. The Q-FOM™ also demonstrated very high repeatability and reproducibility across all traits.

Human metabolic profiles are stably controlled by genetic and environmental variation.

¹H Nuclear Magnetic Resonance spectroscopy (¹H NMR) is increasingly used to measure metabolite concentrations in sets of biological samples for top-down systems biology and molecular epidemiology. For such purposes, knowledge of the sources of human variation in metabolite concentrations is valuable, but currently sparse. We conducted and analysed a study to create such a resource. In our unique design, identical and non-identical twin pairs donated plasma and urine samples longitudinally. We acquired ¹H NMR spectra on the samples, and statistically decomposed variation in metabolite concentration into familial (genetic and common-environmental), individual-environmental, and longitudinally unstable components. We estimate that stable variation, comprising familial and individual-environmental factors, accounts on average for 60% (plasma) and 47% (urine) of biological variation in ¹H NMR-detectable metabolite concentrations. Clinically predictive metabolic variation is likely nested within this stable component, so our results have implications for the effective design of biomarker-discovery studies. We provide a power-calculation method which reveals that sample sizes of a few thousand should offer sufficient statistical precision to detect ¹H NMR-based biomarkers quantifying predisposition to disease.

Optimization of human plasma 1H NMR spectroscopic data processing for high-throughput metabolic phenotyping studies and detection of insulin resistance related to type 2 diabetes.

Optimizing NMR experimental parameters for high-throughput metabolic phenotyping requires careful examination of the total biochemical information obtainable from (1)H NMR data, which includes concentration and molecular dynamics information. Here we have applied two different types of mathematical transformation (calculation of the first derivative of the NMR spectrum and Gaussian shaping of the free-induction decay) to attenuate broad spectral features from macromolecules and enhance the signals of small molecules. By application of chemometric methods such as principal component analysis (PCA), orthogonal projections to latent structures discriminant analysis (O-PLS-DA) and statistical spectroscopic tools such as statistical total correlation spectroscopy (STOCSY), we show that these methods successfully identify the same potential biomarkers as spin-echo (1)H NMR spectra in which broad lines are suppressed via T2 relaxation editing. Finally, we applied these methods for identification of the metabolic phenotype of patients with type 2 diabetes. This "virtual" relaxation-edited spectroscopy (RESY) approach can be particularly useful for high-throughput screening of complex mixtures such as human plasma and may be useful for extraction of latent biochemical information from legacy or archived NMR data sets for which only standard 1D data sets exist.

Assessment of the effect of high or low protein diet on the human urine metabolome as measured by NMR.

The objective of this study was to identify urinary metabolite profiles that discriminate between high and low intake of dietary protein during a dietary intervention. Seventy-seven overweight, non-diabetic subjects followed an 8-week low-calorie diet (LCD) and were then randomly assigned to a high (HP) or low (LP) protein diet for 6 months. Twenty-four hours urine samples were collected at baseline (prior to the 8-week LCD) and after dietary intervention; at months 1, 3 and 6, respectively. Metabolite profiling was performed by (1)H NMR and chemometrics. Using partial least squares regression (PLS), it was possible to develop excellent prediction models for urinary nitrogen (root mean square error of cross validation (RMSECV) = 1.63 mmol/L; r = 0.89) and urinary creatinine (RMSECV = 0.66 mmol/L; r = 0.98). The obtained high correlations firmly establish the validity of the metabolomic approach since urinary nitrogen is a well established biomarker for daily protein consumption. The models showed that trimethylamine-N-oxide (TMAO) is correlated to urinary nitrogen. Furthermore, urinary creatine was found to be increased by the HP diet whereas citric acid was increased by the LP diet. Despite large variations in individual dietary intake, differentiated metabolite profiles were observed at the dietary group-level.

Integration of clinical chemistry, expression, and metabolite data leads to better toxicological class separation.

A large number of databases are currently being implemented within toxicology aiming to integrate diverse biological data, such as clinical chemistry, expression, and other types of data. However, for these endeavors to be successful, tools for integration, visualization, and interpretation are needed. This paper presents a method for data integration using a hierarchical model based on either principal component analysis or partial least squares discriminant analysis of clinical chemistry, expression, and nuclear magnetic resonance data using a toxicological study as case. The study includes the three toxicants alpha-naphthyl-isothiocyanate, dimethylnitrosamine, and N-methylformamide administered to rats. Improved predictive ability of the different classes is seen, suggesting that this approach is a suitable method for data integration and visualization of biological data. Furthermore, the method allows for correlation of biological parameters between the different data types, which could lead to an improvement in biological interpretation.

Metabonomics in diabetes research.

Metabonomics has been defined as "quantitative measurement of the dynamic multiparametric metabolic response of living systems to pathophysiological stimuli or genetic modification" and can provide information on disease processes, drug toxicity, and gene function. In this approach many samples of biological origin (biofluids such as urine or plasma) are analyzed using techniques that produce simultaneous detection. A variety of analytical metabolic profiling tools are used routinely, are also currently under development, and include proton nuclear magnetic resonance spectroscopy and mass spectrometry with a prior online separation step such as high-performance liquid chromatography, ultra-performance liquid chromatography, or gas chromatography. Data generated by these analytical techniques are often combined with multivariate data analysis, i.e., pattern recognition, for respectively generating and interpreting the metabolic profiles of the investigated samples. Metabonomics has gained great prominence in diabetes research within the last few years and has already been applied to understand the metabolism in a range of animal models and, more recently, attempts have been done to process complex metabolic data sets from clinical studies. A future hope for the metabonomic approach is the identification of biomarkers that are able to highlight individuals likely to suffer from diabetes and enable early diagnosis of the disease or the identification of those at risk. This review summarizes the technologies currently being used in metabonomics, as well as the studies reported related to diabetes prior to a description of the general objective of the research plan of the metabonomics part of the European Union project, Molecular Phenotyping to Accelerate Genomic Epidemiology.