Abstract, title and reference: ● Is the aim clear? In this study author has aimed to investigate how oxytocin administration and classically conditioned oxytocin influence brain responses. ● Is it clear what the study found and how they did it? This study was done on females (n=88) and divided into three groups: oxytocin administration, conditioned oxytocin, or placebo. Participants in the conditioned group received 24 IU of oxytocin together with a conditioned stimulus (CS).The oxytocin administration group received 24 IU of oxytocin and the placebo group received placebo during all days. On the last evocation day, fMRI scanning was performed for all participants during three tasks previously shown to be affected by oxytocin: presentation of emotional faces, crying baby sounds and heat pain. ● Is the title informative and relevant? Yes, the title is informative and relevant. ● Are the references: ● Relevant? All references cited in this manuscript are relevant. ● Recent? Authors has cited only few recent references. However, lot of recent literature is available. ● Referenced correctly? In few references author has not mentioned volume, issue number, and page no. (References 1 and 24). ● Are appropriate key studies included? Yes. Introduction/background: ● Is it clear what is already known about this topic? Yes, author has beautifully explained pharmacological conditioning with oxytocin. ● Is the research question clearly outlined? Yes, author has tried to compare the conditioned oxytocin group would demonstrate similar brain activation patterns as the group that received exogenous oxytocin in selected area of brain. ● Is the research question justified given what is already known about the topic? Research question justified somewhat. Author should also focus on regional expression of oxytocin receptor by intranasal delivery in the brain. Therefore it would be better perspective for treatment of neuropsychiatric disorder. Methods: Is the process of subject selection clear? Subject selection process is descent. ● Are the variables defined and measured appropriately? Author has defined variable and method of data analysis is also appropriate as well as study has also ethically approved and done as per guideline of NIH. ● Are the study methods valid and reliable? Yes, In this study Group differences in age, BMI, and baseline oxytocin salivary levels from three evocation days were examined using one-way analysis of variance (ANOVA). Image preprocessing and analyses. The data were pre-processed and analysed with FSL software Version 5.0.10 (FMRIB’s Software Library, www.fmrib.ox.ac.uk/fsl. The statistical tests were corrected for multiple comparisons with the threshold-free cluster enhancement (TFCE). A TFCE corrected statistical threshold of p < 0.05 was chosen. ● Is there enough detail in order to replicate the study? Yes, there is enough detail to replicate the study. Results: ● Is the data presented in an appropriate way? Yes, the authors has presented data systematically. ● Tables and figures relevant and clearly presented? Author has clearly presented results in table and figures. ● Appropriate units, rounding, and number of decimals? Results has presented in appropriate SI unit and two decimals places. ● Titles, columns, and rows labelled correctly and clearly? ● Categories grouped appropriately? Yes, author has categorized appropriately i.e., Placebo group (n = 25) Oxytocin group (n = 23) Conditioned group (n = 25) ● Does the text in the results add to the data or is it repetitive? Yes, the text in the results add to the data. ● Are you clear about what is a statistically significant result? In this study author has studied differences in the brain activation between the oxytocin administration and placebo groups were demonstrated in the right amygdala and in two clusters in superior temporal gyrus for the emotional faces task only, while brain activation of the conditioned oxytocin group was in between that of the oxytocin administration and the placebo group but did not significantly differ from either group. ● Are you clear about what is a practically meaningful result? Yes Discussion and Conclusion: ● Are the results discussed from multiple angles and placed into context without being overinterpreted? Authors has discussed results very well. ● Do the conclusions answer the aims of the study? Not so clear. ● Are the conclusions supported by references or results? Yes ● Are the limitations of the study fatal or are they opportunities to inform future research? For future studies, it is advised that the effects of oxytocin conditioning are studied on the first evocation day, and possibly immediately following the placebo administration with repeated measurements across time to find the peak of the conditioned response. Changing hormonal levels with a behavioral manipulation can have important clinical implications especially for disorders related to dysfunction of the endocrine system.

Overall: ● Was the study design appropriate to answer the aim? Yes ● What did this study add to what was already known on this topic? It was already known that, exogenously administered oxytocin modifies brain responses to emotional and aversive visual stimuli, motivational tasks, involving trust, empathy, reward, and pain. Particularly, oxytocin has been shown to reduce amygdala activation in response to aversive and painful stimuli, which can be an underlying mechanism of stress reducing and analgesic effects of oxytocin described in previous research. Considering these positive physiological and psychological effects of oxytocin, pharmacological conditioning of oxytocin might have important clinical implications both in somatic and mental health. In this randomized placebo-controlled trial, author has investigated the effects of oxytocin administration and conditioned oxytocin in comparison to a placebo-control group on brain activation in response to fMRI tasks that have previously shown to be affected by exogenous oxytocin administration: presentation of emotional faces, presentation of crying baby sounds and thermal pain stimulation. ● What were the major flaws of this article? Author should also focus on regional expression of oxytocin receptor by intranasal delivery in the brain. Therefore it would be better perspective for treatment of neuropsychiatric disorder. ● Is the article consistent within itself? Yes