Determination of Soybean Oil, Protein and Amino Acid Residues in Soybean Seeds by High Resolution Nuclear Magnetic Resonance (NMRS) and Near Infrared (NIRS)

A detailed account is presented of our high resolution nuclear magnetic resonance (HR-NMR) and near infrared (NIR) calibration models, methodologies and validation procedures, together with a large number of composition analyses for soybean seeds. NIR calibrations were developed based on both HR-NMR and analytical chemistry reference data for oil and twelve amino acid residues in mature soybeans and soybean embryos. This is our first report of HR-NMR determinations of amino acid profiles of proteins from whole soybean seeds, without protein extraction from the seed. It was found that the best results for both oil and protein calibrations were obtained with a Partial Least Squares Regression (PLS-1) analysis of our extensive NIR spectral data, acquired with either a DA7000 Dual Diode Array (Si and InGaAs detectors) instrument or with several Fourier Transform NIR (FT-NIR) spectrometers equipped with an integrating sphere/InGaAs detector accessory. In order to extend the bulk soybean samples calibration models to the analysis of single soybean seeds, we have analized in detail the component NIR spectra of all major soybean constituents through spectral deconvolutions for bulk, single and powdered soybean seeds. Baseline variations and light scattering effects in the NIR spectra were corrected, respectively, by calculating the first-order derivatives of the spectra and the Multiplicative Scattering Correction (MSC). The single soybean seed NIR spectra are broadly similar to those of bulk whole soybeans, with the exception of minor peaks in single soybean NIR spectra in the region from 950 to 1,000 nm. Based on previous experience with bulk soybean NIR calibrations, the PLS-1 calibration model was selected for protein, oil and moisture calibrations that we developed for single soybean seed analysis. In order to improve the reliability and robustness of our calibrations with the PLS-1 model we employed standard samples with a wide range of soybean constituent compositions: from 34% to 55% for protein, from 11% to 22% for oil and from 2% to 16% for moisture. Such calibrations are characterized by low standard errors and high degrees of correlation for all major soybean constituents. Morever, we obtained highly resolved NIR chemical images for selected regions of mature soybean embryos that allow for the quantitation of oil and protein components. Recent developments in high-resolution FT-NIR microspectroscopy extend the NIR sensitivity range to the picogram level, with submicron spatial resolution in the component distribution throughout intact soybean seeds and embryos. Such developments are potentially important for biotechnology applications that require rapid and ultra- sensitive analyses, such as those concerned with high-content microarrays in Genomics and Proteomics research. Other important applications of FT-NIR microspectroscopy are envisaged in biomedical research aimed at cancer prevention, the early detection of tumors by NIR-fluorescence, and identification of single cancer cells, or single virus particles in vivo by super-resolution microscopy/ microspectroscopy

On Verifying Complex Properties using Symbolic Shape Analysis

One of the main challenges in the verification of software systems is the analysis of unbounded data structures with dynamic memory allocation, such as linked data structures and arrays. We describe Bohne, a new analysis for verifying data structures. Bohne verifies data structure operations and shows that 1) the operations preserve data structure invariants and 2) the operations satisfy their specifications expressed in terms of changes to the set of objects stored in the data structure. During the analysis, Bohne infers loop invariants in the form of disjunctions of universally quantified Boolean combinations of formulas. To synthesize loop invariants of this form, Bohne uses a combination of decision procedures for Monadic Second-Order Logic over trees, SMT-LIB decision procedures (currently CVC Lite), and an automated reasoner within the Isabelle interactive theorem prover. This architecture shows that synthesized loop invariants can serve as a useful communication mechanism between different decision procedures. Using Bohne, we have verified operations on data structures such as linked lists with iterators and back pointers, trees with and without parent pointers, two-level skip lists, array data structures, and sorted lists. We have deployed Bohne in the Hob and Jahob data structure analysis systems, enabling us to combine Bohne with analyses of data structure clients and apply it in the context of larger programs. This report describes the Bohne algorithm as well as techniques that Bohne uses to reduce the ammount of annotations and the running time of the analysis