Application on infrared spectroscopy for the analysis of total phenolic compounds in fruits.

Recent studies have demonstrated the metabolic benefits of phenolic compounds on human health. However, traditional analytical methods used for quantification of total phenolic compounds are time-consuming, laborious, require a high volume of reagents, mostly toxic substances, and involve several steps that can result in systematic and instrumental errors. Spectroscopic techniques have been used as alternatives to these methods for the determination of bioactive compounds directly in the food matrix by minimal sample preparation, without using toxic reagents. Therefore, this overview presents the advantages of nondestructive methods focusing on infrared spectroscopy (IR), for the quantification of total phenolic compounds in fruits. In addition, the main difficulties in applying these spectroscopic techniques are presented, as well as a comparison between the quantification of total phenolic compounds by traditional and IR methods. This review concludes by focusing on model building, highlighting that IR data are mainly processed using the partial least-squares (PLS) regression method to predict total phenolic content. The development of portable and inexpensive IR instruments, combined with multivariate data processing, could give to the consumers a straightforward technology to evaluate the total phenolic content of fruits prior to purchase.

Rapid analyses of oil and fat content in agri-food products using continuous wave free precession time domain NMR.

Time-domain nuclear magnetic resonance (TD-NMR) is one of the most popular solutions for quality control in the food industry. Despite the recognized success of TD-NMR in quality control and quality assurance, the speed by which samples can be characterized by TD-NMR techniques is still a concern, primarily when considering online or high-throughput applications. Therefore, to enhance the speed of TD-NMR analysis, we developed rapid methods based on steady-state free precession of nuclear spins, which we denoted continuous wave free precession (CWFP). CWFP substantially increases the sensitivity of TD-NMR compared with free induction decay or spin-echo detection, which are traditionally used. The objective of this paper was to present the physical background of CWFP and review its recent developments and applications in fat and oil quantifications in agri-food products.

Serum metabolomic fingerprints of lambs fed chitosan and its association with performance and meat quality traits.

Chitosan (CHI) is a natural biopolymer with antimicrobial, anti-inflammatory, antioxidant and digestive modulatory effects, which can be used in the ruminant diet to replace antibiotics. The aim of this study was to evaluate the effects of CHI on lamb growth traits, nutrients digestibility, muscle and fatty deposition, meat fatty acid (FA) profile, meat quality traits and serum metabolome. Thirty 30-month-old male lambs, half Suffolk and half Dorper, with an average BW of 21.65 ± 0.86 kg, were fed in a feedlot system for a total of 70 days. The lambs were separated into two groups according to the diet: the control (CON) group which received the basal diet and the CHI group which received the basal diet with the addition of CHI as 2 g/kg of DM in the diet. Lambs supplemented with CHI had a greater (P < 0.05) final BW, DM intake, final body metabolic weight (P < 0.05) and lower residual feed intake than the CON group. Animals fed CHI had a greater (P < 0.05) starch digestibility at 14 and 28 days, average daily gain at 14, 42 and 56 days, greater feed efficiency at 28 days and feed conversation at 14 and 42 days in feedlot. Most of the carcass traits were not affected (P > 0.05) by the treatment; however, the CHI supplementation improved (P < 0.05) dressing and longissimus muscle area. The treatments had no effect (P > 0.05) on the meat colour and other quality measurements. Meat from the CHI-fed lambs had a greater concentration (P < 0.05) of oleic-cis-9 acid, linoleic acid, linolenic-trans-6 acid, arachidonic acid and eicosapentaenoic acid. According to the variable importance in projection score, the most important metabolites to differentiate between the CON and the CHI group were hippurate, acetate, hypoxanthine, arginine, malonate, creatine, choline, myo-inositol, 2-oxoglutarate, alanine, glycerol, carnosine, histidine, glutamate and 3-hydroxyisobutyrate. Similarly, fold change (FC) analysis highlighted succinate (FC = 1.53), arginine (FC = 1.51), hippurate (FC = 0.68), myo-inositol (FC = 1.48), hypoxanthine (FC = 1.45), acetate (FC = 0.73) and malonate (FC = 1.35) as metabolites significantly different between groups. In conclusion, the present data showed that CHI changes the muscle metabolism improving muscle mass deposition, the lamb's performance and carcass dressing. In addition, CHI led to an alteration in the FA metabolism, changes in the meat FA profile and improvements in meat quality.

Performance, carcass traits and serum metabolomic profile of Nellore males with different genetic potential for post-weaning growth.

The BW has been largely used as a selection criterion in genetic selection programmes; however, increases in BW can affect animal metabolism and metabolites. The knowledge of how genetic potential for growth affects the metabolites can give a footprint of growth metabolism. This research aimed to evaluate the effect of genetic potential for post-weaning growth (GG) on performance, carcass traits and serum metabolome of non-castrated Nellore males during the finishing phase. Forty-eight Nellore non-castrated males, with divergent potential for post-weaning growth, were selected and divided into two groups: high potential for post-weaning growth (HG; n = 24) and low potential for post-weaning growth (LG; n = 24). Animals were kept and fed for 90 days where performance and ultrasound carcass traits were evaluated. Blood samples were collected at the beginning and end of feeding period to analyse serum metabolites concentration. The hot carcass weight and dressing percentage were recorded at slaughter. The feedlot performance and carcass traits were not affected by genetic potential. The HG animals had a lower glucose (P = 0.039), glutamate (P = 0.038), glutamine (P = 0.004), greater betaine (P = 0.039) and pyruvate (P = 0.039) compared to the LG group at the beginning of feedlot. In addition, higher creatine phosphate concentrations were observed at the beginning of feeding period, compared to final, for both groups (P = 0.039). In conclusion, the genetic potential for post-weaning growth does not affect performance and carcass traits during the finishing period. Differences in metabolite concentrations can be better found at the beginning of feedlot, providing a footprint of growth metabolism, but similar metabolite concentration at the end of finishing period.

NMR studies of the structure and dynamics of membrane-bound bacteriophage Pf1 coat protein.

Filamentous bacteriophage coat protein undergoes a remarkable structural transition during the viral assembly process as it is transferred from the membrane environment of the cell, where it spans the phospholipid bilayer, to the newly extruded virus particles. Nuclear magnetic resonance (NMR) studies show the membrane-bound form of the 46-residue Pf1 coat protein to be surprisingly complex with five distinct regions. The secondary structure consists of a long hydrophobic helix (residues 19 to 42) that spans the bilayer and a short amphipathic helix (residues 6 to 13) parallel to the plane of the bilayer. The NH2-terminus (residues 1 to 5), the COOH-terminus (residues 43 to 46), and residues 14 to 18 connecting the two helices are mobile. By comparing the structure and dynamics of the membrane-bound coat protein with that of the viral form as determined by NMR and neutron diffraction, essential features of assembly process can be identified.

Identification of free fatty acids in maize protein bodies and purified alpha zeins by (13)C and (1)H nuclear magnetic resonance.

Zeins, the maize storage proteins, are the most abundant proteins in the corn endosperm, and are synthesized on the rough endoplasmatic reticulum and deposited in discrete organelles called protein bodies. Several authors, using circular dichroism and optical rotatory dispersion, have concluded that these proteins have a high alpha-helical content in alcoholic solution. In this work we have studied these proteins, within the protein bodies themselves and after extraction from the corn grains with 70% ethanol, using NMR (nuclear magnetic resonance) spectroscopy. We conclusively demonstrate the presence of free fatty acids within both the protein bodies and also in the alcohol extracted alpha zeins. We present evidence for a direct interaction between the free fatty acids and the alpha zein proteins within the protein body and suggest possible mechanisms by which such an association has arisen during the evolution of the maize endosperm.

In situ analysis of copper electrodeposition reaction using unilateral NMR sensor.

The uses of high-resolution NMR spectroscopy and imaging (MRI) to study electrochemical reactions in situ have greatly increased in the last decade. However, most of these applications are limited to specialized NMR laboratories and not feasible for routine analysis. Recently we have shown that a bench top, time domain NMR spectrometer can be used to monitor in situ copper electrodeposition reaction and the effect of Lorentz force in the reaction rate. However these spectrometers limit the cell size to the magnet gap and cannot be used with standard electrochemical cells. In this paper we are demonstrating that unilateral NMR sensor (UNMR), which does not limit sample size/volume, can be used to monitor electrodeposition of paramagnetic ions in situ. The copper electrodeposition reaction was monitored remotely and in situ, placing the electrochemical cell on top of the UNMR sensor. The Cu(2+) concentration was measured during three hours of the electrodeposition reactions, by using the transverse relaxation rate (R2) determined with the Carr-Purcell-Meiboom-Gill pulse sequence. The reaction rate increased fourfold when the reaction was performed in the presence of a magnetic field (in situ), in comparison to the reactions in the absence of the magnetic field (ex situ). The increase of reaction rate, in the presence of the UNMR magnet, was related to the magneto hydrodynamic force (FB) and magnetic field gradient force (F∇B). F∇B was calculated to be one order of magnitude stronger than FB. The UNMR sensor has several advantages for in situ measurements when compared to standard NMR spectrometers. It is a low cost, portable, open system, which does not limit sample size/volume and can be easily be adapted to standard electrochemical cells or large industrial reactors.

Mobility and free radical concentration effects in proton-electron double-resonance imaging.

The dependence of the enhancement of proton-electron double-resonance images upon the mobility of the proton bearing molecules, of the concentration of free radicals, and of the pulsed saturating RF power is studied in a magnetic field of 16 mT. The data exhibit a behavior which, in the potentially interesting region of small free radical concentration, may differ substantially from the high-concentration regime depending upon experimental conditions. The results permit a clearer understanding of the factors determining enhancement and contrast in images obtained by dynamic nuclear polarization.

Flow sensitivity and coherence in steady-state free spin precession.

Steady-state free precession (SSFP) of nuclear spins in the presence of a magnetic field gradient is known to be very sensitive to flow. We present a theoretical and experimental study of flow sensitivity in a regime where the spacing of the radio-frequency pulses is extremely short compared with the free induction decay time and the relaxation times. Under these rather drastic conditions, a truly continuous wave free precession (CWFP) regime is established, in which, unlike standard SSFP, a large degree of coherence is preserved. This leads to a quite different flow sensitivity, which is significant even when very small magnetic field gradients are present. The unspoiled coherence is predicted to cause different flow effects which we confirmed experimentally. Tailored flow sensitivity can be achieved by adjusting the frequency offset from resonance, which plays a dominant role in the CWFP regime.

Preliminary Results from Clinical Application of a Natural Oral Contrast Agent in Magnetic Resonance Imaging (MRI) of the Gastrointestinal (GI) System.

A natural magnetic resonance imaging (MRI) contrast based on the pulp of a fruit "Euterpe Olerácea", popularly known as Açaí, was investigated. T1 and T2 contrasted images shown the effects of the contrast agent increasing the ability to visualize the contour of segments of the gastrointestinal tract.

Dynamics of fd coat protein in the bacteriophage.

The dynamics of the coat protein in fd bacteriophage are described with solid-state 15N and 2H NMR experiments. The virus particles and the coat protein subunits are immobile on the time scales of the 15N chemical shift anisotropy (10(3) Hz) and 2H quadrupole (10(6) Hz) interactions. Previously we have shown that the Trp-26 side chain is immobile, that the two Tyr and three Phe side chains undergo only rapid twofold jump motions about their C beta-C gamma bond axis [Gall, C. M., Cross, T. A., DiVerdi, J. A., & Opella, S. J. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 101-105], and that most of the backbone peptide linkages are highly constrained but do undergo rapid small amplitude motions [Cross, T. A., & Opella, S. J. (1982) J. Mol. Biol. 159, 543-549] in the coat protein subunits in the virus particles. In this paper, we demonstrate that the four N-terminal residues of the coat protein subunits are highly mobile, since both backbone and side-chain sites of these residues undergo large amplitude motions that are rapid on the time scales of the solid-state NMR experiments. In addition, the dynamics of the methyl-containing aliphatic residues Ala, Leu, Val, Thr, and Met are analyzed. Large amplitude jump motions are observed in nearly all of these side chains even though, with the exception of the N-terminal residue Ala-1, their backbone peptide linkages are highly constrained. The established information about the dynamics of the structural form of fd coat protein in the virus particle is summarized qualitatively.(ABSTRACT TRUNCATED AT 250 WORDS)

Interaction of two prolamins with 1-13C oleic acid by 13C NMR.

In this paper, we analyzed the interaction of Z19 prolamin from a BR451 maize variety and pennisetin from a BRS1501 pearl millet variety with 1-(13)C-enriched oleic acid (OA) by (13)C NMR in solution. In both proteins, we identified the presence of free fatty acids by NMR in solid state and solution. The interactions were analyzed at the protein/OA molar ratios of 1:1 and 1:4. In the Z19/OA 1:1 mixture in 70% ethanol and 30% D(2)O, the chemical shift of OA C1 was 182.9 ppm, about 3 ppm above that of the pure OA in the same solvent. In contrast, upon addition of OA to the pennisetin (1:1), the chemical-shift value slightly decreased by less than 1 ppm. The chemical-shift titration curve of OA C1 in an apparent pH range of 5.5-7.3 shifted by approximately 0.3 pH units toward higher pH values in the pennisetin/OA 1:1 complex relative to the pure OA. The results obtained for the pennisetin/OA 1:4 mixture were similar to the complexes at a 1:1 molar ratio. A significant difference was observed between the 1:1 and 1:4 curves for Z19. The titration curve for Z19/OA 1:1 suggested specific binding at the sites with electrostatic interaction.

Protein structure in KBr pellets by infrared spectroscopy.

In this work we analyzed the secondary structure of 13 globular proteins in KBr pellet through Fourier transform infrared spectroscopy (FTIR). The quantification was based in singular value decomposition (SVD) theory, a pattern recognition method. The results show better correlation for alpha helix (0.90) and beta sheet (0.84) in amide I band, similar to the results obtained for proteins in solution. These results show that the protein secondary structure is conserved in solid state, in opposition to the results observed by FTIR using resolution enhancement techniques. The SVD analysis also show that in KBr pellets the protein secondary structures have absorbances in different wavenumbers when compared to those in solution. In this way, the use of KBr pellet and the pattern recognition method can be an ideal method to analyze protein secondary structure by FTIR.

Dynamics of fd coat protein in lipid bilayers.

The dynamics of backbone and side-chain sites of the membrane-bound form of fd coat protein are described with solid-state 2H and 15N NMR experiments. The samples were isotopically labeled coat protein in phospholipid bilayers in excess water. The protein itself is immobile and does not undergo rapid rotation within the bilayer. Like the structural form of the protein, the membrane-bound form has four mobile residues at the N-terminus. The membrane-bound form differs from the structural form in having several mobile residues at the C-terminus. Many of the side chains of residues with immobile backbone sites undergo large amplitude jump motions. The dynamics are generally similar in both the structural and membrane-bound forms of the protein.