A nested case–control study was conducted using the antibody levels detected by AlphaLISA. This study was nested within the Japan Public Health Center (JPHC)-based Prospective Study [37, 38], which stored the plasma samples of approximately 30,000 Japanese individuals aged 40–69 years at a baseline period of 1990–1994. We used the samples of 202 incidental cases of acute ischemic stroke developed between the baseline and 2008 and those of 202 controls. The age (within 2 years), sex, blood sampling date (within 3 months), time since last meal (within 4 h), and study location (Public Health Center area) of these controls were matched with those of the cases. The odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using a conditional logistic regression model. We informed the study participants of the objectives and methods of the study; those who answered the questionnaire and donated blood indicated that they gave informed consent to participate. The ethics committee of the National Cancer Center, Osaka University, and Tsukuba University approved this study.

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