Amino acid intake and plasma concentrations and their interplay with gut microbiota in vegans and omnivores in Germany

Purpose It has been estimated that most vegans meet the total protein requirements, but whether this is also true for individual essential amino acids (AAs) is unclear. Furthermore, a shift in protein intake is suggested to alter microbiota composition, but this association is unknown in terms of veganism or individual AAs. This cross-sectional study compared vegans and omnivores regarding dietary intake and plasma concentration of AAs. The prevalence of insufficient intake of essential AAs among vegans was determined using estimated average requirements (EAR) of WHO. Moreover, correlations between AAs intake and gut microbiota were investigated. Methods Data of 36 vegans and 36 omnivores (30–60 years) were analysed. AA intake, AA plasma concentrations and gut microbiota were ascertained by three-day weighed food protocols, gas/liquid chromatography-tandem mass spectrometry and 16S rRNA sequencing, respectively. Results At almost the same energy intake, the intake of 9 AAs in vegans was significantly lower than in omnivores, with median differences of − 27.0% to − 51.9%. However, only one female vegan showed total protein and lysine intake below the EAR. Vegans showed lower lysine (− 25.0%), but higher glycine (+ 25.4%) and glutamate (+ 13.1%) plasma concentrations than omnivores. Correlation patterns between AA intake and bacterial microbiota differed between vegans and omnivores. In vegans 19 species and in omnivores 5 species showed correlations with AA intake. Conclusion Vegans consumed apparently sufficient but lower AAs than omnivores. In addition, the different AAs intake seems to influence the microbiota composition. The use of short-term dietary data without considering usual intake limits these findings.

Red Light Regulates the Metabolite Biosynthesis in the Leaves of “Huangjinya” Through Amino Acid and Phenylpropanoid Metabolisms

“Huangjinya” is a light-sensitive albino variety and is widely cultivated in China. It has been proved that red light could promote the vegetable growth of plants. However, the mechanism of “Huangjinya” in response to a red light is unclear. This study used high-throughput sequencing technology to analyze the transcriptome of tender shoots of “Huangjinya” under the white and red light supplement conditions. At the same time, liquid chromatography tandem mass spectrometry (LC-MS) was used to analyze metabolite changes under different light conditions. Transcriptome analysis revealed that a total of 174 differentially expressed genes (DEGs) were identified after the red light supplement. Kyoto encyclopedia of genes and genomes (KEGG) classification indicated that amino acid metabolism enriched the most DEGs. In addition, two phenylpropanoid metabolism-related genes and five glutathione S-transferase genes (CsGSTs) were found to be expressed differently. Metabolome analysis revealed that 193 differential metabolites were obtained. Being the same as transcriptome analysis, most differential metabolites were enriched in amino acids, sweet and umami tasting amino acids were increased, and bitter-tasting amino acids were decreased after the red light supplement. In summary, red light supplementary treatment may be propitious to the quality of “Huangjinya” due to its regulatory effect on amino acid metabolism. Also, CsGSTs involved phenylpropanoid metabolism contributed to tea quality changes in “Huangjinya.”

Simultaneous Quantification of Chloramphenicol, Thiamphenicol, Florfenicol, and Florfenicol Amine in Animal and Aquaculture Products Using Liquid Chromatography-Tandem Mass Spectrometry

The accumulation of antimicrobial residues in edible animal products and aquaculture products could pose health concerns to unsuspecting consumers. Hence, this study aimed to develop a validated method for simultaneous quantification of chloramphenicol (CAP), thiamphenicol (TAP), florfenicol (FF), and florfenicol amine (FFA) in beef, pork, chicken, shrimp, eel, and flatfish using a quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction method coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Primary-secondary amine (PSA) and MgSO4 were used for sample purification. The analytes were separated on a reversed-phase analytical column. The coefficients of determination for the linear matrix-matched calibration curves were ≥0.9941. Recovery rates ranged between 64.26 and 116.51% for the four analytes with relative standard deviations (RSDs) ≤ 18.05%. The calculated limits of detection (LODs) and limits of quantification (LOQs) were 0.005–3.1 and 0.02–10.4 μg/kg, respectively. The developed method was successfully applied for monitoring samples obtained from local markets in Seoul, Republic of Korea. The target residues were not detected in any tested matrix. The designed method was versatile, sensitive, and proved suitable for quantifying residues in animal-derived products.

Efficient degradation of various emerging pollutants by wild type and evolved fungal DyP4 peroxidases

The accumulation of emerging pollutants in the environment remains a major concern as evidenced by the increasing number of reports citing their potential risk on environment and health. Hence, removal strategies of such pollutants remain an active area of investigation. One way through which emerging pollutants can be eliminated from the environment is by enzyme-mediated bioremediation. Enzyme-based degradation can be further enhanced via advanced protein engineering approaches. In the present study a sensitive and robust bioanalytical liquid chromatography-tandem mass spectrometry (LCMSMS)-based approach was used to investigate the ability of a fungal dye decolorizing peroxidase 4 (DyP4) and two of its evolved variants—that were previously shown to be H2O2 tolerant—to degrade a panel of 15 different emerging pollutants. Additionally, the role of a redox mediator was examined in these enzymatic degradation reactions. Our results show that three emerging pollutants (2-mercaptobenzothiazole (MBT), paracetamol, and furosemide) were efficiently degraded by DyP4. Addition of the redox mediator had a synergistic effect as it enabled complete degradation of three more emerging pollutants (methyl paraben, sulfamethoxazole and salicylic acid) and dramatically reduced the time needed for the complete degradation of MBT, paracetamol, and furosemide. Further investigation was carried out using pure MBT to study its degradation by DyP4. Five potential transformation products were generated during the enzymatic degradation of MBT, which were previously reported to be produced during different bioremediation approaches. The current study provides the first instance of the application of fungal DyP4 peroxidases in bioremediation of emerging pollutants.

Anti-Colon Cancer Activity of Novel Peptides Isolated from In Vitro Digestion of Quinoa Protein in Caco-2 Cells

Quinoa peptides are the bioactive components obtained from quinoa protein digestion, which have been proved to possess various biological activities. However, there are few studies on the anticancer activity of quinoa peptides, and the mechanism has not been clarified. In this study, the novel quinoa peptides were obtained from quinoa protein hydrolysate and identified by liquid chromatography–tandem mass spectrometry (LC–MS/MS). The anticancer activity of these peptides was predicted by PeptideRanker and evaluated using an antiproliferative assay in colon cancer Caco-2 cells. Combined with the result of histone deacetylase 1 (HDAC1) inhibitory activity assay, the highly anticancer activity peptides FHPFPR, NWFPLPR, and HYNPYFPG were screened and further investigated. Molecular docking was used to analyze the binding site between peptides and HDAC1, and results showed that three peptides were bound in the active pocket of HDAC1. Moreover, real-time quantitative polymerase chain reaction (RT-qPCR), and Western blot showed that the expression of HDAC1, NFκB, IL-6, IL-8, Bcl-2 was significantly decreased, whereas caspase3 expression showed a remarkable evaluation. In conclusion, quinoa peptides may have the potential to protect against cancer development by inhibiting HDAC1 activity and regulating the expression of the cancer-related genes, which indicates that these peptides could be explored as functional foods to alleviate colon cancer.

Next-generation biomonitoring of the early-life chemical exposome in neonatal and infant development

Exposure to man-made and natural chemicals is a major, yet not sufficiently considered, environmental risk factor in the etiology of chronic diseases. Current human biomonitoring approaches typically measure a limited number of exposures rather than investigating complex mixtures. The latter would be fundamental and necessary for a holistic assessment of chemical exposure in exposome-wide association studies. In this work, an highly-sensitive liquid chromatography-tandem mass spectrometry approach was developed and thoroughly-validated. The assay enables the simultaneous and targeted assessment of more than 80 highly-diverse xenobiotics in the investigated body fluids of urine, serum/plasma, and breast milk; the detection limit for most toxicants are in the pg-ng/mL range. In the plasma of extremely-premature infants (gestational age <28 weeks, birth weight <1 kg) a total of 27 different xenobiotics are identified; including severe contamination with synthetic plasticizers, perfluorinated alkylated substances and parabens. In an independent sample set of breast milk that was longitudinally collected over the first 211 days post-partum, a total of 29 analytes is detected, including the first-ever identification of pyrrolizidine- and tropane alkaloids in this matrix. Based on the generated data, a preliminary estimation of daily toxicant intake via breast milk is conducted. In conclusion, our proof-of-principle experiments show significant early-life co-exposure to multiple toxicants, and demonstrate the method’s applicability in future large-scale exposomics-type cohort studies in vulnerable populations.

Multiple neonicotinoids in children’s cerebro-spinal fluid, plasma, and urine

Background Neonicotinoids (NN) are selective neurotoxic pesticides that bind to insect but also mammal nicotinic acetycholine receptors (nAChRs). As the most widely used class of insecticides worldwide, they are ubiquitously found in the environment, wildlife, and foods, and thus of special concern for their impacts on the environment and human health. nAChRs are vital to proper brain organization during the prenatal period and play important roles in various motor, emotional, and cognitive functions. Little is known on children’s contamination by NN. In a pilot study we tested the hypothesis that children’s cerebro-spinal fluid (CSF) can be contaminated by NN. Methods NN were analysed in leftover CSF, blood, and urine samples from children treated for leukaemias and lymphomas and undergoing therapeutic lumbar punctions. We monitored all neonicotinoids approved on the global market and some of their most common metabolites by ultra-high performance liquid chromatography-tandem mass spectrometry. Results From August to December 2020, 14 children were consecutively included in the study. Median age was 8 years (range 3–18). All CSF and plasma samples were positive for at least one NN. Nine (64%) CSF samples and 13 (93%) plasma samples contained more than one NN. Thirteen (93%) CSF samples had N-desmethyl-acetamiprid (median concentration 0.0123, range 0.0024–0.1068 ng/mL), the major metabolite of acetamiprid. All but one urine samples were positive for ≥ one NN. A statistically significant linear relationship was found between plasma/urine and CSF N-desmethyl-acetamiprid concentrations. Conclusions We have developed a reliable analytical method that revealed multiple NN and/or their metabolites in children’s CSF, plasma, and urine. Our data suggest that contamination by multiple NN is not only an environmental hazard for non-target insects such as bees but also potentially for children.