A Combined Metabolomic and Proteomic Analysis of Gestational Diabetes Mellitus.

The aim of this pilot study was to apply a novel combined metabolomic and proteomic approach in analysis of gestational diabetes mellitus. The investigation was performed with plasma samples derived from pregnant women with diagnosed gestational diabetes mellitus (n = 18) and a matched control group (n = 13). The mass spectrometry-based analyses allowed to determine 42 free amino acids and low molecular-weight peptide profiles. Different expressions of several peptides and altered amino acid profiles were observed in the analyzed groups. The combination of proteomic and metabolomic data allowed obtaining the model with a high discriminatory power, where amino acids ethanolamine, L-citrulline, L-asparagine, and peptide ions with m/z 1488.59; 4111.89 and 2913.15 had the highest contribution to the model. The sensitivity (94.44%) and specificity (84.62%), as well as the total group membership classification value (90.32%) calculated from the post hoc classification matrix of a joint model were the highest when compared with a single analysis of either amino acid levels or peptide ion intensities. The obtained results indicated a high potential of integration of proteomic and metabolomics analysis regardless the sample size. This promising approach together with clinical evaluation of the subjects can also be used in the study of other diseases.

The application of fuzzy statistics and linear discriminant analysis as criteria for optimizing the preparation of plasma for matrix-assisted laser desorption/ionization mass spectrometry peptide profiling.

An alternative bioinformatics approach based on fuzzy theory statistics and linear discriminant analysis is proposed for the interpretation of MALDI MS spectra in peptide profiling. When applied, the methodology enables the establishment of a reproducible plasma preparation protocol appropriate for the evaluation of small data sets. The samples were collected from pregnant women affected by gestational diabetes mellitus (GDM), n=18 and control group, n=13. The following pre-treatment sets were tested: pipette tips with C18 stationary phase (ZipTip, Millipore and Omix, Agilent) and magnetic bead-based weak cation exchange chromatography kit (MB WCX, Bruker Daltonics). The spectra were recorded using a MALDI TOF mass spectrometer (UltrafleXtreme, Bruker Daltonics) for a mass range of m/z from 1000 to 10,000. The significant features were selected using the wrapper selection method, and two classification systems were tested: discriminant analysis (DA) and fuzzy inference system (FIS). ClinProTools software was employed to compare the usefulness of the proposed methodology. The study showed that the optimum results for MS spectra were obtained after the use of the ZipTip as pre-treatment method in plasma preparation. Chemometric analysis allowed the differentiation of the GDM group from the control with a high degree of accuracy: 0.7333 (DA) and 0.8065 (FIS).