scholarly journals Development of a High-Throughput Method to Study the Inhibitory Effect of Phytochemicals on Trimethylamine Formation

Nutrients ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 1466
Author(s):  
Lisard Iglesias-Carres ◽  
Lauren A. Essenmacher ◽  
Kathryn C. Racine ◽  
Andrew P. Neilson
Keyword(s):  
Chlorogenic Acid ◽  
Gallic Acid ◽  
High Throughput ◽  
Anaerobic Fermentation ◽  
Inhibitory Effect ◽  
Healthy Human ◽  
High Throughput Method ◽  
Lyase Activity ◽  
Inhibitory Potential ◽  
Potential Maximum

Choline is metabolized by the gut microbiota into trimethylamine (TMA), the precursor of pro-atherosclerotic molecule trimethylamine N-oxide (TMAO). A reduction in TMA formation has shown cardioprotective effects, and some phytochemicals may reduce TMA formation. This study aimed to develop an optimized, high-throughput anaerobic fermentation methodology to study the inhibition of choline microbial metabolism into TMA by phenolic compounds with healthy human fecal starter. Optimal fermentation conditions were: 20% fecal slurry (1:10 in PBS), 100 µM choline, and 12 h fermentation. Additionally, 10 mM of 3,3-dimethyl-1-butanol (DMB) was defined as a positive TMA production inhibitor, achieving a ~50% reduction in TMA production. Gallic acid and chlorogenic acid reported higher TMA inhibitory potential (maximum of 80–90% TMA production inhibition), with IC50 around 5 mM. Neither DMB nor gallic acid or chlorogenic acid reduced TMA production through cytotoxic effects, indicating mechanisms such as altered TMA-lyase activity or expression.

scholarly journals Development of a High Throughput Method to Study the Inhibitory Effect of Phytochemicals on Trimethylamine Formation

2021 ◽  
Author(s):  
Lisard Iglesias-Carres ◽  
Lauren A. Essenmacher ◽  
Kathryn C. Racine ◽  
Andrew P. Neilson
Keyword(s):  
Chlorogenic Acid ◽  
Gallic Acid ◽  
High Throughput ◽  
Anaerobic Fermentation ◽  
Inhibitory Effect ◽  
Healthy Human ◽  
High Throughput Method ◽  
Lyase Activity ◽  
Inhibitory Potential ◽  
Potential Maximum

Choline is metabolized by the gut microbiota into trimethylamine (TMA), the precursor of pro-atherosclerotic molecule trimethylamine N-oxide (TMAO). Reduction of TMA formation has been shown to provide to cardioprotective effects, and some phytochemicals may produce such reduction. This study aimed to develop an optimized, high-throughput anaerobic fermentation methodology to study inhibition of choline microbial metabolism into TMA by phenolic compounds with healthy human fecal starter. Optimal fermentation conditions were: 20 % fecal slurry (1:10 in PBS), 100 M choline, and 12 h fermentation. Also, 10 mM of 3,3-dimethyl-1-butanol (DMB) was defined as a positive TMA production inhibitor, achieving a ~50 % reduction in TMA production. Gallic acid and chlorogenic acid reported higher TMA inhibitory potential (maximum of 80 -90 % in. TMA production inhibition), with IC50 around 5 mM. Nor DMB neither gallic acid and chlorogenic acid reduced TMA production through cytotoxic effects, indicating mechanisms such as altered TMA lyase activity or expression.

scholarly journals Development of a High Throughput In Vitro Fecal Fermentation Method to Screen for TMA Production Inhibitors

2021 ◽  
Vol 5 (Supplement_2) ◽  
pp. 1160-1160
Author(s):  
Lisard Iglesias-Carres ◽  
Lauren Essenmacher ◽  
Kathryn Racine ◽  
Andrew Neilson
Keyword(s):  
Chlorogenic Acid ◽  
Gallic Acid ◽  
Cell Density ◽  
High Throughput ◽  
Positive Control ◽  
In Vivo Studies ◽  
Fermentation Conditions ◽  
Fermentation Method

Abstract Objectives Choline is metabolized by gut bacteria to trimethylamine (TMA), which is further metabolized by the host into trimethylamine N-oxide (TMAO). There is significant interest in reducing TMAO formation to reduce atherosclerosis risk. Our objective was to develop an in vitro fermentation methodology to screen for bioactives able to reduce TMA formation. Methods Choline (5–100 µM) fermentation was optimized under anaerobic conditions at 37°C with the presence of human fecal slurry (OpenBiome) diluted 1:10 in PBS 1X (5–45%) over 36 h. Common dietary phenolics (gallic acid and chlorogenic acid, 0.1–10 mM) were evaluated as TMA production inhibitors under optimal fermentation conditions. 3,3-dimethyl-1-butanol (DMB, 10 mM) was used as a positive control. TMA and choline levels were monitored and analyzed by UPLC-ESI-MS/MS. Cell density (O.D. at 600 nm) was evaluated to account potential cytotoxicity. Results TMA kinetic production curves from choline at >25 µM were statistically different from background (no choline added), suggesting its potential to be used to assay inhibition of TMA production. Fecal slurry concentration of 5% did not reach a TMA plateau within 36 h of fermentation, while the kinetics reported by 45% were fast, reaching a plateau at 12 h, suggesting the need for intermediate fecal concentrations. Optimal fermentation conditions were 100 µM choline and 20%. fecal slurry Under those conditions, exogenous choline was consumed within the first 12 h of fermentation, during which TMA formation plateaued. Under optimal conditions plus gallic acid or chlorogenic acid, TMA formation was significantly reduced, reaching 50% of inhibitor-free control at concentrations >5 mM at 8 h. Also, >2 mM reported higher inhibition potential than DMB 10 mM. Of note, no reductions in cell density were reported due to treatment administration, suggesting a lack of cytotoxicity. Conclusions Our results suggest that gallic acid and chlorogenic acid are promising compounds for in vivo studies. Moreover, our fermentation method can be used to screen for TMA production inhibitors in a high-throughput fashion. Funding Sources Supported by through startup funding from North Carolina State University and the Hatch Program of the National Institute of Food and Agriculture, U.S. Department of Agriculture.

scholarly journals Measuring Sulforaphane and Its Metabolites in Human Plasma: A High Throughput Method

Molecules ◽  
2020 ◽  
Vol 25 (4) ◽  
pp. 829 ◽  
Author(s):  
Annie Langston-Cox ◽  
Dovile Anderson ◽  
Darren J. Creek ◽  
Kirsten Palmer ◽  
Euan M. Wallace ◽  
...  
Keyword(s):  
Human Plasma ◽  
High Throughput ◽  
Clinical Investigation ◽  
Healthy Human ◽  
Potential Health ◽  
Low Concentrations ◽  
High Throughput Method ◽  
Standard Calibration ◽  
Internal Standardization ◽  
Cost Efficient

(1) Background: There is increasing understanding of the potential health benefits of cruciferous vegetables. In particular sulforaphane (SFN), found in broccoli, and its metabolites sulforaphane-glutathione (SFN-GSH), sulforaphane-cysteine (SFN-Cys), sulforaphane cysteine-glycine (SFN-CG) and sulforaphane-N-acetyl-cysteine (SFN-NAC) have potent antioxidant effects that may offer therapeutic value. Clinical investigation of sulforaphane as a therapeutic antioxidant requires a sensitive and high throughput process for quantification of sulforaphane and metabolites; (2) Methods: We collected plasma samples from healthy human volunteers before and for eight hours after consumption of a commercial broccoli extract supplement rich in sulforaphane. A rapid and sensitive method for quantification of sulforaphane and its metabolites in human plasma using Liquid Chromatography–Mass Spectrometry (LC–MS) has been developed; (3) Results: The LC–MS analytical method was validated at concentrations ranging between 3.9 nM and 1000 nM for SFN-GSH, SFN-CG, SFN-Cys and SFN-NAC and between 7.8 nM and 1000 nM in human plasma for SFN. The method displayed good accuracy (1.85%–14.8% bias) and reproducibility (below 9.53 %RSD) including low concentrations 3.9 nM and 7.8 nM. Four SFN metabolites quantitation was achieved using external standard calibration and in SFN quantitation, SFN-d8 internal standardization was used. The reported method can accurately quantify sulforaphane and its metabolites at low concentrations in plasma; (4) Conclusions: We have established a time- and cost-efficient method of measuring sulforaphane and its metabolites in human plasma suitable for high throughput application to clinical trials.

A Fluorescent High Throughput Method To Identify Potential Skin Sensitizers

Planta Medica ◽  
2016 ◽  
Vol 82 (05) ◽  
Author(s):  
C Avonto ◽  
AG Chittiboyina ◽  
D Rua ◽  
IA Khan
Keyword(s):  
High Throughput ◽  
High Throughput Method

scholarly journals High Throughput Centrifugal Electrospinning of Polyacrylonitrile Nanofibers for Carbon Fiber Nonwovens

Polymers ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1313
Author(s):  
Andreas Hoffmann ◽  
Alexander J. C. Kuehne
Keyword(s):  
High Throughput ◽  
Carbon Nanofiber ◽  
Fiber Diameter ◽  
Carbon Precursor ◽  
Production Methods ◽  
High Throughput Method ◽  
Radial Forces ◽  
Fiber Deposition ◽  
The Impact ◽  
Pan Fibers

Carbon nanofiber nonwovens are promising materials for electrode or filtration applications; however, their utilization is obviated by a lack of high throughput production methods. In this study, we utilize a highly effective high-throughput method for the fabrication of polyacrylonitrile (PAN) nanofibers as a nonwoven on a dedicated substrate. The method employs rotational-, air pressure- and electrostatic forces to produce fibers from the inner edge of a rotating bell towards a flat collector. We investigate the impact of all above-mentioned forces on the fiber diameter, morphology, and bundling of the carbon-precursor PAN fibers. The interplay of radial forces with collector-facing forces has an influence on the uniformity of fiber deposition. Finally, the obtained PAN nanofibers are converted to carbon nonwovens by thermal treatment.

scholarly journals A neural network-based algorithm for high-throughput characterisation of viscoelastic properties of flowing microcapsules

Soft Matter ◽  
2021 ◽  
Author(s):  
Tao Lin ◽  
Zhen Wang ◽  
Wen Wang ◽  
Yi Sui
Keyword(s):  
Neural Network ◽  
High Throughput ◽  
Viscoelastic Properties ◽  
Dynamic Deformation ◽  
Membrane Elasticity ◽  
Hybrid Neural Network ◽  
High Throughput Method

We have developed a high-throughput method, by combining a hybrid neural network with a mechanistic capsule model, to predict membrane elasticity and viscosity of microcapsules from their dynamic deformation in a branched microchannel.

scholarly journals High-Throughput Chlorophyll and Carotenoid Profiling Reveals Positive Associations with Sugar and Apocarotenoid Volatile Content in Fruits of Tomato Varieties in Modern and Wild Accessions

Metabolites ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 398
Author(s):  
Yusuke Aono ◽  
Yonathan Asikin ◽  
Ning Wang ◽  
Denise Tieman ◽  
Harry Klee ◽  
...  
Keyword(s):  
High Throughput ◽  
Fruit Color ◽  
Wild Relatives ◽  
Aroma Compounds ◽  
Soluble Solids ◽  
Sugar Production ◽  
Volatile Content ◽  
Factors Influencing ◽  
High Throughput Method ◽  
Wild Accessions

Flavor and nutritional quality has been negatively impacted during the course of domestication and improvement of the cultivated tomato (Solanum lycopersicum). Recent emphasis on consumers has emphasized breeding strategies that focus on flavor-associated chemicals, including sugars, acids, and aroma compounds. Carotenoids indirectly affect flavor as precursors of aroma compounds, while chlorophylls contribute to sugar production through photosynthesis. However, the relationships between these pigments and flavor content are still unclear. In this study, we developed a simple and high-throughput method to quantify chlorophylls and carotenoids. This method was applied to over one hundred tomato varieties, including S. lycopersicum and its wild relatives (S. l. var. cerasiforme and S. pimpinellifolium), for quantification of these pigments in fruits. The results obtained by integrating data of the pigments, soluble solids, sugars, and aroma compounds indicate that (i) chlorophyll-abundant varieties have relatively higher sugar accumulations and (ii) prolycopene is associated with an abundance of linear carotenoid-derived aroma compounds in one of the orange-fruited varieties, “Dixie Golden Giant”. Our results suggest the importance of these pigments not only as components of fruit color but also as factors influencing flavor traits, such as sugars and aroma.

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