scholarly journals A user-friendly, high-throughput tool for the precise fluorescent quantification of deoxyribonucleoside triphosphates from biological samples

2020 ◽  
Vol 48 (8) ◽  
pp. e45-e45 ◽  
Author(s):  
Judit Eszter Szabó ◽  
Éva Viola Surányi ◽  
Bence Sándor Mébold ◽  
Tamás Trombitás ◽  
Mihály Cserepes ◽  
...  
Keyword(s):  
High Throughput ◽  
Biological Samples ◽  
Cell Cycle Control ◽  
Taq Polymerase ◽  
Replication Arrest ◽  
Physiological Processes ◽  
Analysis Process ◽  
High Throughput Method ◽  
Inexperienced User ◽  
User Friendly

Abstract Cells maintain a fine-tuned, dynamic concentration balance in the pool of deoxyribonucleoside 5′-triphosphates (dNTPs). This balance is essential for physiological processes including cell cycle control or antiviral defense. Its perturbation results in increased mutation frequencies, replication arrest and may promote cancer development. An easily accessible and relatively high-throughput method would greatly accelerate the exploration of the diversified consequences of dNTP imbalances. The dNTP incorporation based, fluorescent TaqMan-like assay published by Wilson et al. has the aforementioned advantages over mass spectrometry, radioactive or chromatography based dNTP quantification methods. Nevertheless, the assay failed to produce reliable data in several biological samples. Therefore, we applied enzyme kinetics analysis on the fluorescent dNTP incorporation curves and found that the Taq polymerase exhibits a dNTP independent exonuclease activity that decouples signal generation from dNTP incorporation. Furthermore, we found that both polymerization and exonuclease activities are unpredictably inhibited by the sample matrix. To resolve these issues, we established a kinetics based data analysis method which identifies the signal generated by dNTP incorporation. We automated the analysis process in the nucleoTIDY software which enables even the inexperienced user to calculate the final and accurate dNTP amounts in a 96-well-plate setup within minutes.

A high-throughput method for the determination of Os concentrations and isotope ratio measurements in small-size biological samples

2013 ◽  
Vol 28 (10) ◽  
pp. 1591 ◽  
Author(s):  
Nicola Pallavicini ◽  
Frauke Ecke ◽  
Emma Engström ◽  
Douglas C. Baxter ◽  
Ilia Rodushkin
Keyword(s):  
High Throughput ◽  
Biological Samples ◽  
Isotope Ratio ◽  
High Throughput Method ◽  
Isotope Ratio Measurements

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 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 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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