A fluorometric microplate assay has been developed to determine Escherichia (E.) coli adhesion to uroepithelial cells (UEC). P-fimbriated E. coli were labeled with BacLight Green and preincubated 30 min with human urine or standard. Fluorescent-E. coli were added to UEC in mircoplates at a 400:1 ratio, incubated 1 h, and washed, and the fluorescence intensity was measured. Specific labeling and adherence were confirmed by flow cytometry. A myricetin (1) standard curve (0-30 μg/mL) was developed; the lower limit of detection was 0.1 μg/mL, and half-maximal inhibitory concentration was 0.88 μg/mL (intra- and interassay coefficients of variance were <10% and <15%, respectively). Vaccinium macrocarpon (cranberry) extracts, quercetin (2), and procyanidins B1 (3), B2 (4), and C1 (5) showed similar inhibition. Antiadhesion activity of urine samples from subjects (n = 12) consuming placebo or V. macrocarpon beverage determined using this assay was positively correlated (R(2) = 0.78; p < 0.01) with a radiolabeled-E. coli assay.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1021/np400781y | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Mitochondrial Membrane Potential (ΔΨ) Fluctuations Associated with the Metabolic States of Mitochondria.
Methods Mol Biol
November 2024
Department of Life Sciences, University of Coimbra, Coimbra, Portugal.
The proton electrochemical gradient generated by the respiratory chain activity accounts for over 90% of the available respiratory energy and, as such, its evaluation and accurate measurement regarding total values and fluctuations are an invaluable component of the understanding of mitochondrial function. Consequently, alterations in electric potential across the inner mitochondrial membrane generated by differential protonic accumulation and transport are known as the mitochondrial membrane potential, or Δψ, and are reflective of the functional metabolic status of mitochondria. There are several experimental approaches to measure Δψ, ranging from fluorometric evaluations to electrochemical probes.
View Article and Find Full Text PDFEnhancing peroxidase activity of NiCoO nanoenzyme by Mn doping for catalysis of CRISPR/Cas13a-mediated non-coding RNA detection.
Int J Biol Macromol
December 2024
College of Food Science and Technology, Hebei Agricultural University, Baoding 071001, China; College of Science, Hebei Agricultural University, Baoding 071001, China. Electronic address:
CRISPR/Cas13a with precise and controllable programming of endonuclease activity has been served as powerful tool for RNA sensing. Although with high sensitivity, existing CRISPR/Cas13a-based biosensors need complex amplification procedure or special equipment that limited quantification capability. Here, Mn-doped NiCoO (Mn/NiCoO) nanozyme with enhanced peroxidase activity was synthesized and combined with CRISPR/Cas13a-based reaction to develop a simple, sensitive and universal biosensor for RNA detection, which is achieved through target recognition that activates Cas enzymes to cleave RNA reporter for inhibiting Mn/NiCoO nanozyme to assemble on microplate.
View Article and Find Full Text PDFFluorometric Methods to Measure Bioavailable and Total Heme.
Methods Mol Biol
July 2024
School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA.
Heme b (iron protoporphyrin IX) is an essential but potentially cytotoxic cofactor, signaling molecule, and nutritional source of iron. Its importance in cell biology and metabolism is underscored by the fact that numerous diseases, including various cancers, neurodegenerative disorders, infectious diseases, anemias, and porphyrias, are associated with the dysregulation of heme synthesis, degradation, trafficking, and/or transport. Consequently, methods to measure, image, and quantify heme in cells are required to better understand the physiology and pathophysiology of heme.
View Article and Find Full Text PDFHigh-Throughput Fluorometric Assay For Quantifying Polysorbate In Biopharmaceutical Products Using Micelle Activated Fluorescence Probe N-Phenyl-1-Naphthylamine.
Pharm Res
July 2024
Quantitative Biosciences, MRL, Merck & Co., Inc., 126 E Lincoln Ave, Rahway, NJ, 07065, USA.
Purpose: Polysorbates are among the most used surfactants in biopharmaceutical products containing proteins. Our work aims to develop a high-throughput fluorometric assay to further diversify the analytical toolbox for quantification of PSs.
Method: The assay leverages the micelle activated fluorescence signal from N-Phenyl-1-Naphthylamine (NPN).
Colorimetric and fluorometric determination of uric acid by a suspension-based assay using enzyme-immobilized micro-sized particles.
Anal Sci
May 2024
Department of Biomedical Analysis, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo, 192-0392, Japan.
Article Synopsis
- Daily monitoring of serum uric acid is crucial for personalized treatment in hyperuricemic patients.
- A new suspension-based assay has been developed, utilizing uricase and horseradish peroxidase to measure uric acid by producing a red color from resorufin.
- The method offers improved visibility for colorimetric analysis and allows for quantitative measurement of uric acid using a 96-well fluorescent microplate reader, with a detection limit of 2.2 × 10 μg/mL.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!