{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T02:44:42Z","timestamp":1760237082961,"version":"build-2065373602"},"reference-count":35,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2020,3,2]],"date-time":"2020-03-02T00:00:00Z","timestamp":1583107200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Medical Biotechnology (CEMB), S&amp;T Postgraduate Education and Research Development Office (PERDO), Office of Higher Education Commission (OHEC), Thailand","award":["SD-60-003-04"],"award-info":[{"award-number":["SD-60-003-04"]}]},{"name":"RGJ Ph.D. Programme Thailand Research Fund","award":["PHD\/1086\/2557"],"award-info":[{"award-number":["PHD\/1086\/2557"]}]},{"name":"Faculty of Science, Mahidol University and the Thailand Research Fund","award":["IRG5980001"],"award-info":[{"award-number":["IRG5980001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Calcium pyrophosphate deposition disease (CPPD) is a crystal induced inflammation in joints, and causes severe pain in elderly people. The accumulation of pyrophosphate (PPi) in synovial fluid (SF) results from several enzymatic reactions, especially the highly activated e-NPPs, which catalyze the conversion of ATP to PPi. This study demonstrates the detection of relative catalytic activity of 3 enzymes\u2014ecto-nucleotide pyrophosphatase\/phosphodiesterases (e-NPPs), tissue nonspecific alkaline phosphatase (TNAP), and ecto-nucleoside triphosphate diphosphohydrolases (e-NTPDases)\u2014using a single molecular sensor called Kyoto Green. Kyoto Green exhibits excellent performance in sensing the catalytic activity of the commercial representatives of the e-NPPs, TNAP, and e-NTPDases, which are ENPP1, PPase, and apyrase, respectively, in both single-enzyme and multi-enzyme assays. Analysis of SF enzymes in 19 SF samples from human and swine revealed moderate activity of e-NPPs, high activity of e-NTPDases, and low activity of TNAP. Our newly developed method for analysis of multiple enzymatic activities using Kyoto Green in biological SF will assist improvement in accuracy of the CPPD prognosis\/diagnosis, which will minimize unnecessary medical procedures.<\/jats:p>","DOI":"10.3390\/molecules25051116","type":"journal-article","created":{"date-parts":[[2020,3,4]],"date-time":"2020-03-04T03:24:20Z","timestamp":1583292260000},"page":"1116","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Fluorescence Differentiation of ATP-Related Multiple Enzymatic Activities in Synovial Fluid as a Marker of Calcium Pyrophosphate Deposition Disease Using Kyoto Green"],"prefix":"10.3390","volume":"25","author":[{"given":"Nattha","family":"Yongwattana","sequence":"first","affiliation":[{"name":"Department of Biotechnology, Faculty of Science, Mahidol University, 272 Rama 6 Road, Bangkok 10400, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nutsara","family":"Mekjinda","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Faculty of Science, Mahidol University, 272 Rama 6 Road, Bangkok 10400, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tulyapruek","family":"Tawonsawatruk","sequence":"additional","affiliation":[{"name":"Department of Orthopedics, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, 270 Rama 6 Road, Bangkok 10400, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Itaru","family":"Hamachi","sequence":"additional","affiliation":[{"name":"Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Kyoto 615-8510, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Akio","family":"Ojida","sequence":"additional","affiliation":[{"name":"Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8269-4602","authenticated-orcid":false,"given":"Jirarut","family":"Wongkongkatep","sequence":"additional","affiliation":[{"name":"Department of Biotechnology, Faculty of Science, Mahidol University, 272 Rama 6 Road, Bangkok 10400, Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,3,2]]},"reference":[{"key":"ref_1","first-page":"633","article-title":"The prevalence of chondrocalcinosis in the human knee joint. 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