The study explores the sex-specific anxiolytic effects of linalool and β-myrcene, demonstrating differences in efficacy based on exposure duration and terpene combinations with cannabidiol (CBD). While the findings are compelling, one potential issue is the limited exploration of baseline variability in male elevated plus maze (EPM) performance, which the authors acknowledge as a limitation. Given that baseline variability can affect treatment outcomes, could the authors provide further clarification on how this variability was controlled beyond the within-subjects design? Additionally, the study focuses on a single anxiety assay (EPM), raising concerns about whether the findings generalize across other validated models for anxiety. Would incorporating complementary assays strengthen the robustness of these conclusions?
– The primary experiments (Figure 1) employ a repeated-measures design where animals are tested with both vehicle and a terpene, spaced two weeks apart. This is a significant flaw for assessing the acute effects of a drug. The “semi-within subjects” design does not control for order effects, and the data shows drastically different baseline (vehicle) performance between sessions for both males and females. How can the authors rule out that the observed effects are due to habituation, learning, or a practice effect on the EPM rather than a genuine drug effect, especially given the statement that animals were only tested a maximum of “more than twice” in a different experiment?
– In Figure 1, panels G and H are described in the main text (Section 3.1, fourth paragraph) as showing the effects of β-myrcene on locomotor activity. However, the figure legend for panels G and H incorrectly states the data were collected “after 30 min of linalool vapor pulls.” This is a direct error in the figure legend. Does this error affect the interpretation or reproducibility of the reported findings?
– The authors repeatedly describe the effects of the terpenes as “antipodal” between the sexes (e.g., reducing distance in females but enhancing it in males for linalool). However, the baseline (vehicle) values for males and females in these experiments are not comparable. The observed “reduction” in males might simply be a regression to the mean or a loss of a practice effect, as the males show exceptionally high vehicle activity in the second session. How can a change from an already elevated baseline in males be equated to a true “antipodal” effect?