Emulsifying properties of hemp proteins: Effect of isolation technique

peer-reviewedHemp protein was isolated from hemp seed meal using two different isolation procedures: alkali extraction/isoelectric precipitation (HPI) and micellization (HMI). The ability of these proteins to form and stabilize 10% (w/w) sunflower oil-in-water emulsions (at pH = 3.0) was studied at three different concentrations, 0.25, 0.75 and 1.5% (w/w), by monitoring emulsion droplet size distribution, microstructural and morphological properties, rheological behaviour and stability against flocculation, coalescence and creaming. In addition, hemp proteins were analysed for water solubility, denaturation degree and surface/interfacial activity. HMI protein, which was found to be less denatured after isolation, exhibited higher solubility and slightly higher surface/interfacial activity than HPI protein. HMI emulsions possessed a smaller volume mean droplet diameter (d4,3 = 1.92–3.42 μm in 2% SDS) than HPI emulsions (d4,3 = 2.25–15.77 μm in 2% SDS). While HMI stabilized emulsions were characterized with individual droplets covered by protein film, both confocal laser scanning microscopy and flocculation indices indicated occurrence of bridging flocculation in HPI stabilized emulsions. Protein aggregation, which induced flocculation of the droplets, contributed to higher apparent viscosity of HPI stabilized emulsions compared to HMI stabilized emulsions. Interestingly, emulsions stabilized with 1.5% (w/w) HPI exhibited much better creaming and coalescence stability than other emulsions due to the formation of a weak transient network of floccules and higher continuous phase viscosity which both suppressed the movement of the droplets

Analysis of oxylipins in human plasma: Comparison ofultrahigh-performance liquid chromatography andultrahigh-performance supercritical fluid chromatography coupled tomass spectrometry

tThe potential of ultrahigh-performance liquid chromatography–mass spectrometry (UHPLC/MS) andultrahigh-performance supercritical fluid chromatography (UHPSFC) coupled to negative-ion electro-spray ionization mass spectrometry (ESI–MS) for the analysis of 46 oxylipins and 2 fatty acid standardsis compared in terms of their chromatographic resolution with the emphasis on distinguishing isobaricinterferences and the method sensitivity. UHPLC provides the baseline separation of 24 isobaric oxylipinswithin 13 min, while UHPSFC enables the separation of only 20 isobaric oxylipins within 8 min. More-over, the UHPLC/ESI–MS method provides an average improvement of sensitivity by 3.5-fold. A similartrend is observed in the analysis of human plasma samples, but lower ion suppression effects causedby lysophospholipids (LPL) are observed in case of UHPSFC/ESI–MS due to better separation of LPL. Bothmethods are fully applicable for the analysis of oxylipins, but UHPLC/ESI–MS method is preferred dueto better separation and higher sensitivity, which results in the identification of 31 oxylipins in humanplasma based on available standards and additional tentative 20 identifications based on accurate m/zvalues and the fragmentation behavior known from the literature

Analysis of maternal polymorphisms in arsenic (+3 oxidation state)-methyltransferase AS3MT and fetal sex in relation to arsenic metabolism and infant birth outcomes: Implications for risk analysis

Arsenic (+3 oxidation state) methyltransferase (AS3MT) is the key enzyme in the metabolism of inorganic arsenic (iAs). Polymorphisms of AS3MT influence adverse health effects in adults, but little is known about their role in iAs metabolism in pregnant women and infants. The relationships between seven single nucleotide polymorphisms (SNPs) in AS3MT and urinary concentrations of iAs and its methylated metabolites were assessed in mother-infant pairs of the Biomarkers of Exposure to ARsenic (BEAR) cohort. Maternal alleles for five of the seven SNPs (rs7085104, rs3740400, rs3740393, rs3740390, and rs1046778) were associated with urinary concentrations of iAs metabolites, and alleles for one SNP (rs3740393) were associated with birth outcomes/measures. These associations were strongly dependent upon the male sex of the fetus but independent of fetal genotype for AS3MT. These data highlight a potential sex-dependence of the relationships among maternal genotype, iAs metabolism and infant health outcomes

Epigenetic Changes in Individuals with Arsenicosis

Inorganic arsenic (iAs) is an environmental toxicant currently poisoning millions of people worldwide, and chronically exposed individuals are susceptible to arsenicosis or arsenic poisoning. Using a state-of-the-art technique to map the methylomes of our study subjects, we identified a large interactome of hypermethylated genes that are enriched for their involvement in arsenic-associated diseases, such as cancer, heart disease, and diabetes. Notably, we have uncovered an arsenic-induced tumor suppressorome, a complex of 17 tumor suppressors known to be silenced in human cancers. This finding represents a pivotal clue in unraveling a possible epigenetic mode of arsenic-induced disease

Neonatal Metabolomic Profiles Related to Prenatal Arsenic Exposure

Prenatal inorganic arsenic (iAs) exposure is associated with health effects evident at birth and later in life. An understanding of the relationship between prenatal iAs exposure and alterations in the neonatal metabolome could reveal critical molecular modifications, potentially underpinning disease etiologies. In this study, nuclear magnetic resonance (NMR) spectroscopy-based metabolomic analysis was used to identify metabolites in neonate cord serum associated with prenatal iAs exposure in participants from the Biomarkers of Exposure to ARsenic (BEAR) pregnancy cohort, in GoÌmez Palacio, Mexico. Through multivariable linear regression, ten cord serum metabolites were identified as significantly associated with total urinary iAs and/or iAs metabolites, measured as %iAs, %monomethylated arsenicals (MMAs), and %dimethylated arsenicals (DMAs). A total of 17 metabolites were identified as significantly associated with total iAs and/or iAs metabolites in cord serum. These metabolites are indicative of changes in important biochemical pathways such as vitamin metabolism, the citric acid (TCA) cycle, and amino acid metabolism. These data highlight that maternal biotransformation of iAs and neonatal levels of iAs and its metabolites are associated with differences in neonate cord metabolomic profiles. The results demonstrate the potential utility of metabolites as biomarkers/indicators of in utero environmental exposure

Prenatal Arsenic Exposure and the Epigenome: Identifying Sites of 5-methylcytosine Alterations that Predict Functional Changes in Gene Expression in Newborn Cord Blood and Subsequent Birth Outcomes

Prenatal exposure to inorganic arsenic (iAs) is detrimental to the health of newborns and increases the risk of disease development later in life. Here we examined a subset of newborn cord blood leukocyte samples collected from subjects enrolled in the Biomarkers of Exposure to ARsenic (BEAR) pregnancy cohort in Gómez Palacio, Mexico, who were exposed to a range of drinking water arsenic concentrations (0.456–236 µg/l). Changes in iAs-associated DNA 5-methylcytosine methylation were assessed across 424 935 CpG sites representing 18 761 genes and compared with corresponding mRNA expression levels and birth outcomes. In the context of arsenic exposure, a total of 2919 genes were identified with iAs-associated differences in DNA methylation. Site-specific analyses identified DNA methylation changes that were most predictive of gene expression levels where CpG methylation within CpG islands positioned within the first exon, the 5′ untranslated region and 200 bp upstream of the transcription start site yielded the most significant association with gene expression levels. A set of 16 genes was identified with correlated iAs-associated changes in DNA methylation and mRNA expression and all were highly enriched for binding sites of the early growth response (EGR) and CCCTC-binding factor (CTCF) transcription factors. Furthermore, DNA methylation levels of 7 of these genes were associated with differences in birth outcomes including gestational age and head circumference.These data highlight the complex interplay between DNA methylation, functional changes in gene expression and health outcomes and underscore the need for functional analyses coupled to epigenetic assessments