Development and Validation of a Sensitive, High-Throughput Bioassay for the Adhesion of Radiolabeled E. coli to Uroepithelial Cells in Vitro
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. ma...
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| Veröffentlicht in: | Journal of natural products (Washington, D.C.) D.C.), 2013-09, Vol.76 (9), p.1605-1611 |
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| creator | Mathison, Bridget D Kimble, Lindsey L Kaspar, Kerrie L Khoo, Christina Chew, Boon P |
| description | 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 |
| doi_str_mv | 10.1021/np400264b |
| format | Article |
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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.</description><identifier>ISSN: 0163-3864</identifier><identifier>EISSN: 1520-6025</identifier><identifier>DOI: 10.1021/np400264b</identifier><identifier>PMID: 23964569</identifier><language>eng</language><publisher>United States: American Chemical Society and American Society of Pharmacognosy</publisher><subject>Bacterial Adhesion - drug effects ; Biological Assay - methods ; Escherichia ; Escherichia coli ; Humans ; Proanthocyanidins - analysis ; Proanthocyanidins - chemistry ; Proanthocyanidins - isolation & purification ; Proanthocyanidins - pharmacology ; Urinary Tract Infections - prevention & control ; Urinary Tract Infections - urine ; Uropathogenic Escherichia coli - drug effects ; Urothelium - drug effects ; Vaccinium macrocarpon ; Vaccinium macrocarpon - chemistry</subject><ispartof>Journal of natural products (Washington, D.C.), 2013-09, Vol.76 (9), p.1605-1611</ispartof><rights>Copyright © 2013 American Chemical Society and American Society of Pharmacognosy</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a348t-973669ca1211bac4cab15c938e4b8790b2782632382ea50f2163186a8e1a8fa43</citedby><cites>FETCH-LOGICAL-a348t-973669ca1211bac4cab15c938e4b8790b2782632382ea50f2163186a8e1a8fa43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/np400264b$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/np400264b$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>315,781,785,2766,27078,27926,27927,56740,56790</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23964569$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Mathison, Bridget D</creatorcontrib><creatorcontrib>Kimble, Lindsey L</creatorcontrib><creatorcontrib>Kaspar, Kerrie L</creatorcontrib><creatorcontrib>Khoo, Christina</creatorcontrib><creatorcontrib>Chew, Boon P</creatorcontrib><title>Development and Validation of a Sensitive, High-Throughput Bioassay for the Adhesion of Radiolabeled E. coli to Uroepithelial Cells in Vitro</title><title>Journal of natural products (Washington, D.C.)</title><addtitle>J. Nat. Prod</addtitle><description>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.</description><subject>Bacterial Adhesion - drug effects</subject><subject>Biological Assay - methods</subject><subject>Escherichia</subject><subject>Escherichia coli</subject><subject>Humans</subject><subject>Proanthocyanidins - analysis</subject><subject>Proanthocyanidins - chemistry</subject><subject>Proanthocyanidins - isolation & purification</subject><subject>Proanthocyanidins - pharmacology</subject><subject>Urinary Tract Infections - prevention & control</subject><subject>Urinary Tract Infections - urine</subject><subject>Uropathogenic Escherichia coli - drug effects</subject><subject>Urothelium - drug effects</subject><subject>Vaccinium macrocarpon</subject><subject>Vaccinium macrocarpon - chemistry</subject><issn>0163-3864</issn><issn>1520-6025</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0c9O3DAQBnCrKipb6IEXQL5UaqUG_H-dI93SgoSERIFrNEkmxMgbB9tB4h146AbtwqkSp7n85rM1HyEHnB1xJvjxMCrGhFH1B7LgWrDCMKE_kgXjRhbSGrVLPqd0zxiTrNSfyK6QpVHalAvy_Asf0YdxjUOmMLT0FrxrIbsw0NBRoH9xSC67R_xBz9xdX1z3MUx3_Thl-tMFSAmeaBcizT3Sk7bHtN28gtYFDzV6bOnpEW2CdzQHehMDjm7W3oGnK_Q-UTfQW5dj2Cc7HfiEX7Zzj9z8Pr1enRUXl3_OVycXBUhlc1EupTFlA1xwXkOjGqi5bkppUdV2WbJaLK0wUkgrEDTrxHwGbg1Y5GA7UHKPfNvkjjE8TJhytXapmb8CA4YpVVwrxTi3SrxPlVzq-U1dzvT7hjYxpBSxq8bo1hCfKs6ql56qt55me7iNneo1tm_ytZgZfN0AaFJ1H6Y4zAf5T9A_PZyY8w</recordid><startdate>20130927</startdate><enddate>20130927</enddate><creator>Mathison, Bridget D</creator><creator>Kimble, Lindsey L</creator><creator>Kaspar, Kerrie L</creator><creator>Khoo, Christina</creator><creator>Chew, Boon P</creator><general>American Chemical Society and American Society of Pharmacognosy</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>20130927</creationdate><title>Development and Validation of a Sensitive, High-Throughput Bioassay for the Adhesion of Radiolabeled E. coli to Uroepithelial Cells in Vitro</title><author>Mathison, Bridget D ; Kimble, Lindsey L ; Kaspar, Kerrie L ; Khoo, Christina ; Chew, Boon P</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a348t-973669ca1211bac4cab15c938e4b8790b2782632382ea50f2163186a8e1a8fa43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Bacterial Adhesion - drug effects</topic><topic>Biological Assay - methods</topic><topic>Escherichia</topic><topic>Escherichia coli</topic><topic>Humans</topic><topic>Proanthocyanidins - analysis</topic><topic>Proanthocyanidins - chemistry</topic><topic>Proanthocyanidins - isolation & purification</topic><topic>Proanthocyanidins - pharmacology</topic><topic>Urinary Tract Infections - prevention & control</topic><topic>Urinary Tract Infections - urine</topic><topic>Uropathogenic Escherichia coli - drug effects</topic><topic>Urothelium - drug effects</topic><topic>Vaccinium macrocarpon</topic><topic>Vaccinium macrocarpon - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mathison, Bridget D</creatorcontrib><creatorcontrib>Kimble, Lindsey L</creatorcontrib><creatorcontrib>Kaspar, Kerrie L</creatorcontrib><creatorcontrib>Khoo, Christina</creatorcontrib><creatorcontrib>Chew, Boon P</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Journal of natural products (Washington, D.C.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Mathison, Bridget D</au><au>Kimble, Lindsey L</au><au>Kaspar, Kerrie L</au><au>Khoo, Christina</au><au>Chew, Boon P</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development and Validation of a Sensitive, High-Throughput Bioassay for the Adhesion of Radiolabeled E. coli to Uroepithelial Cells in Vitro</atitle><jtitle>Journal of natural products (Washington, D.C.)</jtitle><addtitle>J. Nat. Prod</addtitle><date>2013-09-27</date><risdate>2013</risdate><volume>76</volume><issue>9</issue><spage>1605</spage><epage>1611</epage><pages>1605-1611</pages><issn>0163-3864</issn><eissn>1520-6025</eissn><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.</abstract><cop>United States</cop><pub>American Chemical Society and American Society of Pharmacognosy</pub><pmid>23964569</pmid><doi>10.1021/np400264b</doi><tpages>7</tpages></addata></record> |
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| ispartof | Journal of natural products (Washington, D.C.), 2013-09, Vol.76 (9), p.1605-1611 |
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| subjects | Bacterial Adhesion - drug effects Biological Assay - methods Escherichia Escherichia coli Humans Proanthocyanidins - analysis Proanthocyanidins - chemistry Proanthocyanidins - isolation & purification Proanthocyanidins - pharmacology Urinary Tract Infections - prevention & control Urinary Tract Infections - urine Uropathogenic Escherichia coli - drug effects Urothelium - drug effects Vaccinium macrocarpon Vaccinium macrocarpon - chemistry |
| title | Development and Validation of a Sensitive, High-Throughput Bioassay for the Adhesion of Radiolabeled E. coli to Uroepithelial Cells in Vitro |
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