Ee times (3S). In the second stage, the second player decides how much (B) out of 3S to second back to the first player. At the end, the first player receives (20 ?S + B) being the amount he/she keeps plus the amount the second player sends back while the second player receives three times the amount sent deducting the amount sent back (3S ?B). We use the strategic method [24], in which the second player states his/her response to each of 21 possible choices from the first player. Every participant plays both roles of first and second players without any feedback. At the payment stage with real money, we randomly determine the specific role ?first or second mover ?for each pair of subjects. The amount sent by first player is used as a measure for trust while the average return amount from the second player is a measure for trustworthiness. As trust is an inherently risky behavior viz., the trustee may not reciprocate with an act of trustworthiness, we aim 
to test whether OT is specific to trust in the social interactions or alternatively plasma OT indexes risk in general. We therefore include a risk task using portfolio choice design [25]. In this risk task, subjects 
are endowed with SGD 20, and decide how much to FCCP web invest on an experimental stock. For the amount invested, there is 50 chance that it will become 2.5 times, and 50 chance that it will become zero. This design enables us to observe different levels of risk aversion for each subject, and allows ascertainment of the specificity of plasma OT levels towards trust without including a confound i.e. a possible connection between OT and risk attitude.Assay ProceduresBlood samples for oxytocin assay were collected from the antecubital vein into pre-chilled 5 ml EDTA tubes with 250 KIU of apoprotinin, and refrigerated until processing. Plasma was isolated by centrifugation at 1800 g, 15 minutes, 4uC, and stored in aliquots at 270uC. Oxytocin immunoreactivity levels were quantified in duplicates using a commercial oxytocin ELISA kit (Enzo Life Sciences, NY, USA, formerly Assays Designs, MI, USA), as recommended in previous publications [11]. Thawed samples on ice were diluted 1:2 23727046 times in assay buffer and assayed according to manufacturer’s instructions. The oxytocin assay had a sensitivity of 11.7 pg/ml, and inter- and intra-assay coefficient of variations below 15 . Currently there are differences in opinions surrounding the measurement of oxytocin and particularly concerning the requirement of sample extraction. The commercially available oxytocin EIA kit from Enzo Life (formerly Assay Designs), which has been validated by for linearity, cross reactivity, matrix effects, accuracy, precision and recovery [26], was used in the current study. The experience of some investigators suggests that extraction of the samples leads to significant loss of measureable oxytocin. Importantly, the oxytocin data from non-extracted samples makes biological sense as compared to those from extracted samples, which often gave rise to non-detectable levelsEthics StatementThe study is approved by the Internal Review Board at the National University of Singapore. Each participant provides written informed consent (as outlined in the PLoS consent form) to participate in the experiment at the beginning of the study.Materials and Methods SubjectsWe recruited 1,158 (584 females; age, mean 21.26 S.D. 1.5) Han Chinese 3PO undergraduate students at the National University of Singapore to participate in a study of t.Ee times (3S). In the second stage, the second player decides how much (B) out of 3S to second back to the first player. At the end, the first player receives (20 ?S + B) being the amount he/she keeps plus the amount the second player sends back while the second player receives three times the amount sent deducting the amount sent back (3S ?B). We use the strategic method [24], in which the second player states his/her response to each of 21 possible choices from the first player. Every participant plays both roles of first and second players without any feedback. At the payment stage with real money, we randomly determine the specific role ?first or second mover ?for each pair of subjects. The amount sent by first player is used as a measure for trust while the average return amount from the second player is a measure for trustworthiness. As trust is an inherently risky behavior viz., the trustee may not reciprocate with an act of trustworthiness, we aim to test whether OT is specific to trust in the social interactions or alternatively plasma OT indexes risk in general. We therefore include a risk task using portfolio choice design [25]. In this risk task, subjects are endowed with SGD 20, and decide how much to invest on an experimental stock. For the amount invested, there is 50 chance that it will become 2.5 times, and 50 chance that it will become zero. This design enables us to observe different levels of risk aversion for each subject, and allows ascertainment of the specificity of plasma OT levels towards trust without including a confound i.e. a possible connection between OT and risk attitude.Assay ProceduresBlood samples for oxytocin assay were collected from the antecubital vein into pre-chilled 5 ml EDTA tubes with 250 KIU of apoprotinin, and refrigerated until processing. Plasma was isolated by centrifugation at 1800 g, 15 minutes, 4uC, and stored in aliquots at 270uC. Oxytocin immunoreactivity levels were quantified in duplicates using a commercial oxytocin ELISA kit (Enzo Life Sciences, NY, USA, formerly Assays Designs, MI, USA), as recommended in previous publications [11]. Thawed samples on ice were diluted 1:2 23727046 times in assay buffer and assayed according to manufacturer’s instructions. The oxytocin assay had a sensitivity of 11.7 pg/ml, and inter- and intra-assay coefficient of variations below 15 . Currently there are differences in opinions surrounding the measurement of oxytocin and particularly concerning the requirement of sample extraction. The commercially available oxytocin EIA kit from Enzo Life (formerly Assay Designs), which has been validated by for linearity, cross reactivity, matrix effects, accuracy, precision and recovery [26], was used in the current study. The experience of some investigators suggests that extraction of the samples leads to significant loss of measureable oxytocin. Importantly, the oxytocin data from non-extracted samples makes biological sense as compared to those from extracted samples, which often gave rise to non-detectable levelsEthics StatementThe study is approved by the Internal Review Board at the National University of Singapore. Each participant provides written informed consent (as outlined in the PLoS consent form) to participate in the experiment at the beginning of the study.Materials and Methods SubjectsWe recruited 1,158 (584 females; age, mean 21.26 S.D. 1.5) Han Chinese undergraduate students at the National University of Singapore to participate in a study of t.
