Investigating the interaction between dietary polyphenols, the SARS CoV-2 spike protein and the ACE-2 receptor.

The global spread of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has called for an urgent need for the identification of compounds able to control, prevent or slow down the global pandemic. Several dietary polyphenols were assayed against binding to the SARS CoV-2 S1 spike protein and the human ACE-2 receptor, the target of the SARS CoV-2 virus using nano differential scanning fluorimetry, suggesting interaction of dietary polyphenols with both proteins. Following this initial screening the two dietary polyphenols with the strongest affinity were evaluated in a second functional binding assay. The assay was based on the thermophoresis of a fluorescently labelled spike protein and the ACE-2 receptor in the presence of dietary concentrations of the polyphenol in question. It could be experimentally shown that 5-caffeoyl quinic acid and epicatechin reduce the binding constant between SARS CoV-2 spike protein of the alpha variant and the ACE-2 receptor by a factor of ten. The finding could as well be applied to black tea and a coffee beverage with dietary 5-CQA concentrations for the alpha variant Spike protein. Hence it can be speculated that a cup of coffee reduces binding of the virus to its human target, therefore reducing the likelihood of infection with SARS CoV-2, acting as a virus entry-inhibitor.

LC-MS based metabolomic approach for the efficient identification and relative quantification of bioavailable cocoa phenolics in human urine.

This study was designed to investigate the rate and extent of urinary excretion of cocoa phenolic metabolites after human intake using metabolomics approach. In this context, a feeding trial was conducted where urine samples were collected at different time points over 48-h period. Several biomarkers were highlighted in LC-MS based chemometrics using principal component (PCA) and partial least squares discriminant analysis (PLS-DA), which revealed the presence of both epicatechin and gut microbial phenyl-γ-valerolactones (PVLs) conjugated analogues. The presences of these metabolites segregated and grouped the samples based on cocoa and non-cocoa ingestion. Furthermore, semi quantification of major bioavailable metabolites was performed to determine the interindividual differences and assess the relative bioavailability of cocoa compounds in the human body. Our approach presented here is unique in displaying a combination of LC-MS based chemometrics visualization strategies, which revealed and identified significant biomarkers that could reduce the problems associated with data screening complexity.

LC-MS/MS based molecular networking approach for the identification of cocoa phenolic metabolites in human urine.

Dietary phenolic compounds are often transformed by gut microbiota prior to absorption. This transformation may modify their structures, producing novel gut flora metabolites associated with numerous health benefits. Traditional mass spectrometry (MS) based approaches for assessing dietary exposure of cocotea (cocoa, coffee and tea) products provided very little information about the modification and fate of dietary phenolics after ingestion, mainly due to limitation of complex sample nature and their data analyses. Mass spectrometry techniques are well-suited to a high-throughput characterization of natural products, however, analyzing MS based data of complex biological matrix is still considered a challenge. In order to overcome such limitations and simplify the analysis of complex MS data, a cocotea based human trial was conducted where MS based molecular networking approach was implemented. To demonstrate the utility of this approach in one of the specific cocotea diets, we have applied it to a diverse collection of human (n = 15) urine samples, who consumed cocoa rich in polyphenols over a 48-h period. This approach illustrated the power of the new strategy, allowing the rapid identification of new analogues of cocoa metabolites after human consumption. Analysis of human urine samples after cocoa consumption revealed (by assignment of unknown metabolites based on the network similarities) that monomeric flavanols are mainly absorbed and transformed directly into their glucuronide and sulfated moieties. Subsequently, the hydroxy and methoxy phenyl-g-velerolactone as well as their smaller metabolites (such as hydroxyphenyl valeric acids, hydroxy and methoxy phenyl propionic acids and their derivates) are indicative of bacterial metabolism of cocoa major flavanols. For the first time, our study exemplifies and highlight the implementation of MS based molecular networking approach in illustrating the tracking of various structural motifs of ingested cocoa phenolics in human based study.

Changes in the fucoxanthin production and protein profiles in Cylindrotheca closterium in response to blue light-emitting diode light.

BACKGROUND: Marine diatoms have a higher fucoxanthin content in comparison to macroalgae. Fucoxanthin features many potent bioactive properties, particularly anti-obesity properties. Despite the great potential for harvesting larger amounts of fucoxanthin, the impacts of light quality (light source, intensity, and photoperiod) on fucoxanthin production and the essential proteins involved in fucoxanthin biosynthesis in marine diatoms remain unclear. RESULTS: In the present study, Cylindrotheca closterium was selected from four different species of diatoms based on its high fucoxanthin content and productivity. Optimal light conditions (light source, intensity, and regime) were determined by a "Design of Experiment" approach (software MODDE Pro 11 was used). The model indicated that an 18/6 light/darkness regime increased fucoxanthin productivity remarkably as opposed to a 12/12 or 24/0 regime. Eventually, blue light-emitting diode light, as an alternative to fluorescent light, at 100 µmol/m2/s and 18/6 light/darkness regime yielded maximum fucoxanthin productivity and minimal energy consumption. The fucoxanthin production of C. closterium under the predicted optimal light conditions was assessed both in bottle and bag photobioreactors (PBRs). The high fucoxanthin content (25.5 mg/g) obtained from bag PBRs demonstrated the feasibility of large-scale production. The proteomes of C. closterium under the most favorable and unfavorable fucoxanthin biosynthesis light/darkness regimes (18/6 and 24/0, respectively) were compared to identify the essential proteins associated with fucoxanthin accumulation by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry. Six proteins that were up-regulated in the 18/6 regime but down-regulated in the 24/0 were identified as important chloroplastic proteins involved in photosynthesis, energy metabolism, and cellular processes. CONCLUSIONS: Blue light-emitting diode light at 100 µmol/m2/s and 18/6 light/darkness regime induced maximum fucoxanthin productivity in C. closterium and minimized energy consumption. The high fucoxanthin production in the bag photobioreactor under optimal light conditions demonstrated the possibility of commercialization. Proteomics suggests that fucoxanthin biosynthesis is intimately associated with the photosynthetic efficiency of the diatom, providing another technical and bioengineering outlook on fucoxanthin enhancement.

Tea and coffee time with bacteria - Investigation of uptake of key coffee and tea phenolics by wild type E. coli.

Dietary phenolic compounds are often transformed by gut microbiota prior to absorption. This transformation may modulate their biological activities. Many fundamental questions still need to be addressed to understand how the gut microbiota-diet interactions affect human health. Herein, a UHPLC-QTOF mass spectrometry-based method for the quantification of uptake and determination of intracellular bacterial concentrations of dietary phenolics from coffee and tea was developed. Quantitative uptake data for selected single purified phenolics were determined. The specific uptake from mixtures containing up to four dietary relevant compounds was investigated to assess changes of uptake parameters in a mixture model system. Indeed, perturbation of bacteria by several compounds alters uptake parameter in particular tmax. Finally, model bacteria were dosed with complex dietary mixtures such as diluted tea or coffee extracts. The uptake kinetics of the twenty most abundant phenolics was quantified and the findings are discussed. For the first time, quantitative data on in-vitro uptake of dietary phenolics from food matrices were obtained indicating a time-dependent differential uptake of nutritional compounds.

Determination of hydroxycinnamic acids present in Rhododendron species.

Hydroxycinnamates including free hydroxycinnamic acids and their chlorogenic acid derivatives and glycosides have been profiled in leaf extracts of 98 Rhododendron species using LC-MS techniques. In total, 69 hydroxycinnamic acid derivatives were identified in the leaves of 98 Rhododendron species. Some derivatives serve as unique phytochemical marker for a single species, whereas other compounds are limited to certain subgenera. The distribution of compounds among six different subgenera of Rhododendron was studied using PCA and PLS-DA. This contribution presents data that provide unique metabolomic insight in the distribution of a class of secondary metabolites within a large selection of species from the botanically diverse plant genus Rhododendron.

Metabolome Comparison of Bioactive and Inactive Rhododendron Extracts and Identification of an Antibacterial Cannabinoid(s) from Rhododendron collettianum.

INTRODUCTION: The science of metabolomics offers the possibility to measure full secondary plant metabolomes with limited experimental effort to allow identification of metabolome differences using principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) of liquid chromatography mass spectrometry (LC-MS) data. OBJECTIVE: To demonstrate a bioinformatics driven hypothesis generator for identification of biologically active compounds in plant crude extracts, which is validated by activity guided fractionation. METHODOLOGY: Crude extracts of Rhododendron leaves were tested for their antibacterial activity using agar diffusion and minimum inhibitory concentration assays. Extracts were profiled by LC-MS. PCA and PLS-DA were used for differentiation of bioactive and inactive extracts and their metabolites. Preparative-high performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR) spectroscopy were used for separation and structure elucidation of pure compound(s) respectively. RESULTS: An antibacterial Rhododendron collettianum was compared to a series of inactive extracts. Three metabolites were found to distinguish R. collettianum from other species indicating the ability of PCA and PLS-DA to suggest potential bioactive substances. An activity-guided fractionation of R. collettianum extracts was carried out and cannabiorcichromenic acid (CCA) was identified as antibacterial compound thereby validating the PCA and PLS-DA generated hypothesis. Four mammalian cell lines were used to estimate possible cytotoxicity of CCA. CONCLUSION: It was shown that bioinformatics tools facilitate early stage identification of a biologically active compound(s) using LC-MS data, which reduce complexity and number of separation steps in bioactive screening. Copyright © 2017 John Wiley & Sons, Ltd.

Bioactivity in Rhododendron: A Systemic Analysis of Antimicrobial and Cytotoxic Activities and Their Phylogenetic and Phytochemical Origins.

The exceptional diversity of the genus Rhododendron has a strong potential for identification, characterization, and production of bioactive lead compounds for health purposes. A particularly relevant field of application is the search for new antibiotics. Here, we present a comparative analysis of nearly 90 Rhododendron species targeted toward the search for such candidate substances. Through a combination of phytochemical profiles with antimicrobial susceptibility and cytotoxicity, complemented by phylogenetic analyses, we identify seven potentially antimicrobial active but non-cytotoxic compounds in terms of mass-to-charge ratios and retention times. Exemplary bioactivity-guided fractionation for a promising Rhododendron species experimentally supports in fact one of these candidate lead compounds. By combining categorical correlation analysis with Boolean operations, we have been able to investigate the origin of bioactive effects in further detail. Intriguingly, we discovered clear indications of systems effects (synergistic interactions and functional redundancies of compounds) in the manifestation of antimicrobial activities in this plant genus.