Metabolomics to understand placental biology: Where are we now?

Metabolomics, the application of analytical chemistry methodologies to survey the chemical composition of a biological system, is used to globally profile and compare metabolites in one or more groups of samples. Given that metabolites are the terminal end-products of cellular metabolic processes, or 'phenotype' of a cell, tissue, or organism, metabolomics is valuable to the study of the maternal-fetal interface as it has the potential to reveal nuanced complexities of a biological system as well as differences over time or between individuals. The placenta acts as the primary site of maternal-fetal exchange, the success of which is paramount to growth and development of offspring during pregnancy and beyond. Although the study of metabolomics has proven moderately useful for the screening, diagnosis, and understanding of the pathophysiology of pregnancy complications, the placental metabolome in the context of a healthy pregnancy remains poorly characterized and understood. Herein, we discuss the technical aspects of metabolomics and review the current literature describing the placental metabolome in human and animal models, in the context of health and disease. Finally, we highlight areas for future opportunities in the emerging field of placental metabolomics.

Enzymes enhance degradation of the fiber-starch-protein matrix of distillers dried grains with solubles as revealed by a porcine in vitro fermentation model and microscopy.

Effects of treating corn and wheat distillers dried grains with solubles (DDGS) with a multicarbohydrase alone or in combination with a protease on porcine in vitro fermentation characteristics and the matrix structure of the DGGS before and after the fermentation were studied. Three DDGS samples (wheat DDGS sample 1 [wDDGS1], wheat DDGS sample 2 [wDDGS2], and corn DDGS [cDDGS]) were predigested with pepsin and pancreatin. Residues were then subjected to in vitro fermentation using buffered mineral solution inoculated with fresh pig feces without or with a multicarbohydrase alone or in combination with protease in a 3 × 3 factorial arrangement. Accumulated gas production was measured for up to 72 h. Concentration of VFA was measured in fermented solutions. The matrix of native DDGS and their residues after fermentation was analyzed using confocal laser scanning microscopy and scanning electron microscopy to determine internal and external structures, respectively. On a DM basis, wDDGS1, wDDGS2, and cDDGS contained 35.5, 43.4, and 29.0% CP; 2.23, 0.51, and 6.40% starch; 0.82, 0.80, and 0.89% available Lys; and 24.8, 22.5, and 23.0% total nonstarch polysaccharides, respectively. The in vitro digestibility of DM for wDDGS1, wDDGS2, and cDDGS was 67.7, 72.1, and 59.6%, respectively. The cDDGS had greater ( < 0.05) total gas and VFA production than both wheat DDGS. The wDDGS2 had lower ( < 0.05) total gas production than wDDGS1. Multicarbohydrase increased ( < 0.05) total gas production for cDDGS and total VFA production for wDGGS1 but did not increase gas or VFA production for wDDGS2. Addition of protease with multicarbohydrase to DDGS reduced ( < 0.05) total gas and VFA productions and increased ( < 0.05) branched-chain VFA regardless of DDGS type. Confocal laser scanning microscopy and scanning electron microscopy revealed that DDGS were mainly aggregates of resistant and nonfermentable starchy and nonstarchy complexes formed during DDGS production. After in vitro fermentation with porcine fecal inoculum, particles of enzyme-treated DDGS were generally smaller than those of the untreated DDGS. In conclusion, cDDGS had a more porous matrix that was more fermentable than the wheat DDGS. The wDDGS2 was less fermentable than wDDGS1. Multicarbohydrase increased fermentability of cDDGS and wDDGS1 but not wDDGS2, indicating that its efficacy in DDGS is dependent on matrix porosity and DDGS source. Protease hindered efficacy of multicarbohydrase.

LPS from bovine serum albumin drives TNF-α release during ex-vivo placenta perfusion experiments, contaminates the perfusion system but can be effectively removed by oxidative cleaning.

INTRODUCTION: The dual ex-vivo perfusion of human placental tissue is useful to study inflammatory pathways. We found significant TNF-α release in negative controls similar in concentration to lipopolysaccharide (LPS) stimulated placentas. The aim of the current study was to (i) identify sources driving TNF-α release and (ii) develop an approach to control for it. METHOD: (i) To determine sources leading to TNF-α release, solutions frequently circulated through the perfusion system and perfusion media with different bovine serum albumin (BSA) quality were exposed to mouse macrophage cell lines (RAW264.7) and subsequently measured for TNF-α expression. (ii) To assess memory effects and validate cleaning procedures, sham perfusion experiments were conducted either in the presence or absence of exogenous LPS, in new tubing that was contaminated, cleaned and analyzed for the effectiveness of LPS removal. Oxidative and acid-base cleaning were tested for their effectiveness to reduce LPS contamination. RESULTS: TNF-α release, observed in negative control experiments, was attributed to the use of LPS-contaminated BSA as well as inadequate cleaning of the perfusion system. Once introduced in the perfusion system, LPS accumulated and created a memory effect. Oxidative but not acid-base depyrogenation effectively reduced LPS levels to concentrations that were in accordance with FDA guidelines (<0.5 EU/mL) for medical equipment redeemed appropriate for re-use. DISCUSSION: LPS contamination of the placenta perfusion model could have confounding effects on experimental outcomes leading to misinterpretation of data. To circumvent LPS contamination LPS-free BSA and oxidative depyrogenation cleaning techniques should be implemented in future placental perfusion studies.

Use of insecticide-treated bednets in an urban coastal area of Puducherry, India: a cross-sectional study.

Use of Insecticide-Treated Bednets (ITNs) has been shown to reduce the incidence of mosquito-borne diseases. However, the impact of ITNs depends on its community acceptance. We studied the ITN usage and factors influencing it, following the distribution of one ITN to each family in an urban area of Puducherry. Around 93.6% of the 157 respondents surveyed were aware of mosquito-borne diseases. Coils and vaporizers were used in 91.1% of the families. Around two-fifths (41.3%) of the 116 families who received the ITNs used it regularly with another 5.1% using it irregularly. Majority of the users (85%) reported reduced mosquito bites and pleasant sleep after use. Small size of ITNs was the most common reason for non-use (46.3%). Families not using normal bed nets at the time of distribution of ITNs were unlikely to use ITNs at present (odds ratio = 5.22, P < 0.001). Therefore, ITN size should be increased and distribution in urban settings should accompany behavior change communication.

Tocol composition and supercritical carbon dioxide extraction of lipids from barley pearling flour.

Samples of whole grain and 35% pearling flour of 20 different barley varieties grown in Alberta were analyzed for their lipid contents. Total lipid contents of whole grains were within 1.9% to 3.0% (w/w), whereas those of the 35% pearling flour were 4.3% to 7.9%. Lipids of 35% pearling flour fraction of Tercel barley were extracted using supercritical carbon dioxide (SC-CO2 ) at different pressures (24, 45, and 58 MPa) and temperatures (40 and 60 °C) for 3 h. Lipid recoveries of 73% to 97% were achieved using SC-CO2 extraction under different operational conditions. Tocol contents and compositions of whole grain, 35% pearling flour, and SC-CO2 extracts were analyzed using HPLC. Tocol content of the whole grain was 53.8 to 124.9 µg/g and that of the pearling flour was 195 to 363 µg/g of flour. The hulless barley varieties were higher in tocols, with waxy, double waxy and Tercel varieties having the highest levels (P < 0.05). The ratios of total tocotrienols to total tocopherols varied within 1.6 to 3.9 range. Tocol concentrations of SC-CO2 extract fractions varied from 1171 to 4391 µg/g extract depending on the operational conditions. Barley oil is a good natural source of different tocol isomers rich in tocotrienols.

Barley-derived ß-glucans increases gut permeability, ex vivo epithelial cell binding to E. coli, and naive T-cell proportions in weanling pigs.

Weaning in young animals is associated with an increased incidence of gastrointestinal infections. ß-glucans exert numerous physiological effects, including altering immune function. The objective of this study was to determine the effects of feeding barley (Hordeum vulgare L.)-derived ß-glucans on immune and intestinal function in weanling pigs (Sus scrofa). Thirty-one individually-housed Dutch Landrace pigs (21 d; initial BW, 6,298 ± 755 g) were weaned and fed a wheat-based diet (control) or a low (Lo-BG), medium (Med-BG), or high ß-glucan-containing barley-based diet (Hi-BG) for 2 wk with 7 or 8 pigs/treatment. Intestinal segments were analyzed for permeability using Ussing chambers and K88 Escherichia coli adhesion to enterocytes was assessed ex vivo. Immune cells from mesenteric lymph nodes, peripheral blood, and Peyer's patches were analyzed for lymphocyte subsets by indirect immunofluorescence and the ability to respond ex vivo to mitogens by (3)H-thymidine incorporation. Hematology and neutrophil function were determined by flow cytometry. Neutrophil burst, size, and granularity, lymphocyte proliferation, and B-cell distribution in peripheral blood lymphocytes, Peyer's patches, and mesenteric lymph nodes were not affected by ß-glucans content of the diet. The ß-glucans content of the diet altered blood concentrations of erythrocytes and leukocytes, CD4, CD45RA, and CD8 blood cells (P < 0.05). In addition, feeding ß-glucan resulted in increased (P < 0.05) percentage CD45RA positive cells in peripheral blood lymphocytes, Peyer's patches, and mesenteric lymph nodes. Mannitol permeability and tissue conductance were increased (P < 0.05) in Hi-BG fed pigs compared with control pigs. Percentage maximum K88-E.coli binding was increased in proportion to the ß-glucan content of the diet (P < 0.05). Although ß-glucan feeding during the weaning period increased blood lymphocytes and the proportion of naïve T-cells, it also increased E. coli-enterocyte binding and intestinal permeability. ß-glucan may alter immune and intestinal function of weaning pigs.

Identification of Parapenaeopsis stylifera, a new host for Epipenaeon ingens.

Epipenaeon ingens (Isopoda: Bopyridae) is reported for the first time from Parapenaeopsis stylifera (Decapoda: Penaeidae) in South India. According to the month the parasitic prevalence ranged from 0.0 to 2.9%. A characteristic bulge of the branchial chamber growth retardation and degenerated sex organs were observed in the infested shrimp.

Comparison of MALDI-TOF mass spectrometric to enzyme colorimetric quantification of glucose from enzyme-hydrolyzed starch.

Successful quantification of the glucose produced by enzyme hydrolysis of starch was achieved by a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) protocol, using sorbitol as an internal standard. The starch contents measured by MALDI-TOF MS of corn starch, fiber-enriched oat flour derivatives, oat and barley flours, and barley flour/corn starch composites were evaluated in comparison to a widely accepted and validated method of starch determination, which relies on enzyme colorimetry (EC). The average starch content measured in a series of corn starch samples of different masses was 93 and 101% for EC and MALDI-TOF MS, respectively, values that represent the estimated purity of the sample. There was an agreement of 99% between the starch contents determined by the two analytical methods for complex flour-derived samples. Starch values estimated by MALDI-TOF MS consistently showed a greater degree of variability than those determined by EC, but this limitation was readily compensated by rapid acquisition of multiple mass spectra. This study is the first to report the quantification of glucose by MALDI-TOF MS, and it offers new perspectives into the potential utility of MALDI-TOF MS as a definitive tool for monosaccharide analysis and rapid starch determination in complex samples.

Analysis of phenolic acids in barley by high-performance liquid chromatography.

Phenolic acids from 30 barley varieties (combination of hulled/hulless/two-row/six-row/regular/waxy) were investigated by HPLC following four different sample treatments: (a) simple hot water extraction, (b) extraction after acid hydrolysis, (c) acid plus alpha-amylase hydrolysis, and (d) acid plus alpha-amylase plus cellulase hydrolysis treatments. The benzoic acid (p-hydroxybenzoic, vanillic, and protocatechuic acids) and cinnamic acid derivatives (coumaric, caffeic, ferulic, and chlorogenic acids) were identified, and some of the phenolic acids were quantified after each above-mentioned treatment. The data indicated that a combination of sequential acid, alpha-amylase, and cellulase hydrolysis treatments might be applicable for release of more phenolic acids from barley.

Pearling of hull-less barley: product composition and gel color of pearled barley flours as affected by the degree of pearling.

Barley grains from two hull-less varieties, Phoenix and Candle, were pearled to various degrees (10-80%). The composition (starch, protein, beta-glucan, lipid, and ash) of pearled grain (PG) and pearling flour (PF) was determined. Effect of pearling on Hunter L, a, and b color parameters of uncooked and cooked (gel) barley flour (milled from PG) was investigated over a 3 day storage at 4 degrees C.

MALDI-MS and HPLC quantification of oligosaccharides of lichenase-hydrolyzed water-soluble beta-glucan from ten barley varieties.

This study is the first to apply matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) to both qualitative and quantitative analyses of oligosaccharides of lichenase-hydrolyzed water-soluble beta-glucan from barley. Compared to high-performance liquid chromatography (HPLC) with an evaporative light-scattering detector, MALDI-MS is a rapid technique with high accuracy and sensitivity and could be used to assess primary structural features of water-soluble beta-glucan from different barley varieties.

Volatiles from roasted byproducts of the poultry-processing industry.

Volatiles of roasted chicken breast muscle and byproducts, such as backbones, breastbones, spent bones, and skin, were investigated. Total volatile concentrations ranged from 2030 ppb in the roasted backbones to 4049 ppb in the roasted skin. The major classes of volatile compounds detected in roasted samples were aldehydes (648-1532 ppb) and alcohols (336-1006 ppb). Nitrogen- and/or sulfur-containing compounds were also detected in appreciable quantities (161-706 ppb) in all samples. For all samples, hexanal and 2-methyl-2-buten-1-ol were dominant among the aldehydes and alcohols, respectively. Among the nitrogen- and sulfur-containing compounds, Maillard reaction products, such as tetrahydropyridazines, piperidines, and thiazoles, were the major contributors to the total volatile content in all samples. The composition of volatiles observed in roasted byproducts was markedly different from that of the roasted breast muscle. Therefore, the blending of the byproducts in appropriate proportions or blending of volatile flavor extracts from different byproducts may be necessary to obtain an aroma that mimics roasted chicken aroma.

Effect of heat-moisture treatment on the structure and physicochemical properties of cereal, legume, and tuber starches.

Native wheat, oat, lentil, yam, and potato starches were heat treated at 100 degrees C for 16 h at moisture contents between 10 and 30%. The heat treatment did not change granule size and shape. In oat starch, granules were less compactly packed after heat treatment. The X-ray diffraction intensities increased in wheat, oat, and lentil starches, but decreased in potato and yam. The X-ray patterns of wheat and oat starches remained unchanged, while those of lentil, potato, and yam starches became more cereal-like. In all starches, the swelling factor and amylose leaching decreased, being more pronounced in potato. Heat treatment induced complex formation between amylose and native lipids. Differential scanning calorimetry of the heat-treated samples showed a broadening of the gelatinization-temperature range and a shifting of the endothermal transition towards higher temperatures. These changes were more pronounced in potato starch. The gelatinization enthalpy of wheat, oat, and lentil starches remained unchanged, but those of potato and yam starches decreased on heat treatment. Heat treatment increased the 95 degrees C viscosity of wheat starch, but decreased those of oat lentil, potato, and yam starches. In all starches, thermal and shear stability increased after heat treatment. Acid hydrolysis decreased on heat treatment of wheat and lentil starches, but increased in oat, potato, and yam starches. However, in potato and yam starches, the foregoing trend was evident only during the first seven days of hydrolysis. Thereafter, acid hydrolysis was more pronounced in native than in heat-treated starches. The susceptibility towards hydrolysis by porcine pancreatic alpha amylase decreased on heat treatment of wheat and lentil starches, whereas increases were observed for oat, potato, and yam starches. The results indicated that the extent of starch-chain associations within the amorphous regions and the degree of crystalline order are altered during heat-moisture treatment. The magnitude of these changes were found to be dependent upon the moisture content during heat treatment and on the starch source.