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Development and validation of a sensitive, high-throughput bioassay for the adhesion of radiolabeled E. coli to uroepithelial cells in vitro.
J Nat Prod. 2013 Sep 27; 76(9):1605-11.JN

Abstract

Vaccinium macrocarpon (cranberry) products have been used to prevent uropathogenic Escherichia (E.) coli adherence to uroepithelial cells (UEC) and may help reduce risk of urinary tract infection. Reported herein are the development and validation of an assay to assess antiadhesion activity of V. macrocarpon extracts and human urine. P-fimbriated E. coli (CFT073) was labeled with ³H-uridine, then co-incubated with HTB-4 UEC at a 400:1 ratio. V. macrocarpon extracts (0-17 mg proanthocyanidins/mL) were added to ³H-labeled E. coli before co-incubating with UEC. The assay yielded a sensitive inhibition curve: the lower limit of detection and half-maximal inhibitory concentration were 0.43 and 1.59 mg proanthocyanidins/mL for V. macrocarpon extract CEP 55; intra- and interassay coefficients of variance were <10% and <15%, respectively. V. macrocarpon extract CEP 3283 showed identical adhesion inhibition. Serial dilutions of urine from human participants who consumed V. macrocarpon beverages showed a linear decrease in antiadhesion activity. Antiadhesion assays conducted with urine from a human intervention study also showed good agreement with results obtained using the hemagglutination assay. Therefore, a sensitive, high-throughput, biologically relevant antiadhesion assay using ³H-E. coli co-incubated with UEC is reported, which can be used for studying the action of V. macrocarpon bioactives.

Authors+Show Affiliations

School of Food Science, Washington State University , Pullman, Washington 99164-6376, United States.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23964569

Citation

Mathison, Bridget D., et al. "Development and Validation of a Sensitive, High-throughput Bioassay for the Adhesion of Radiolabeled E. Coli to Uroepithelial Cells in Vitro." Journal of Natural Products, vol. 76, no. 9, 2013, pp. 1605-11.
Mathison BD, Kimble LL, Kaspar KL, et al. Development and validation of a sensitive, high-throughput bioassay for the adhesion of radiolabeled E. coli to uroepithelial cells in vitro. J Nat Prod. 2013;76(9):1605-11.
Mathison, B. D., Kimble, L. L., Kaspar, K. L., Khoo, C., & Chew, B. P. (2013). Development and validation of a sensitive, high-throughput bioassay for the adhesion of radiolabeled E. coli to uroepithelial cells in vitro. Journal of Natural Products, 76(9), 1605-11. https://doi.org/10.1021/np400264b
Mathison BD, et al. Development and Validation of a Sensitive, High-throughput Bioassay for the Adhesion of Radiolabeled E. Coli to Uroepithelial Cells in Vitro. J Nat Prod. 2013 Sep 27;76(9):1605-11. PubMed PMID: 23964569.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development and validation of a sensitive, high-throughput bioassay for the adhesion of radiolabeled E. coli to uroepithelial cells in vitro. AU - Mathison,Bridget D, AU - Kimble,Lindsey L, AU - Kaspar,Kerrie L, AU - Khoo,Christina, AU - Chew,Boon P, Y1 - 2013/08/21/ PY - 2013/8/23/entrez PY - 2013/8/24/pubmed PY - 2013/12/18/medline SP - 1605 EP - 11 JF - Journal of natural products JO - J. Nat. Prod. VL - 76 IS - 9 N2 - Vaccinium macrocarpon (cranberry) products have been used to prevent uropathogenic Escherichia (E.) coli adherence to uroepithelial cells (UEC) and may help reduce risk of urinary tract infection. Reported herein are the development and validation of an assay to assess antiadhesion activity of V. macrocarpon extracts and human urine. P-fimbriated E. coli (CFT073) was labeled with ³H-uridine, then co-incubated with HTB-4 UEC at a 400:1 ratio. V. macrocarpon extracts (0-17 mg proanthocyanidins/mL) were added to ³H-labeled E. coli before co-incubating with UEC. The assay yielded a sensitive inhibition curve: the lower limit of detection and half-maximal inhibitory concentration were 0.43 and 1.59 mg proanthocyanidins/mL for V. macrocarpon extract CEP 55; intra- and interassay coefficients of variance were <10% and <15%, respectively. V. macrocarpon extract CEP 3283 showed identical adhesion inhibition. Serial dilutions of urine from human participants who consumed V. macrocarpon beverages showed a linear decrease in antiadhesion activity. Antiadhesion assays conducted with urine from a human intervention study also showed good agreement with results obtained using the hemagglutination assay. Therefore, a sensitive, high-throughput, biologically relevant antiadhesion assay using ³H-E. coli co-incubated with UEC is reported, which can be used for studying the action of V. macrocarpon bioactives. SN - 1520-6025 UR - https://www.unboundmedicine.com/medline/citation/23964569/Development_and_validation_of_a_sensitive_high_throughput_bioassay_for_the_adhesion_of_radiolabeled_E__coli_to_uroepithelial_cells_in_vitro_ L2 - https://dx.doi.org/10.1021/np400264b DB - PRIME DP - Unbound Medicine ER -