Re: FEBSOPEN-24-0067

The authors have examined the effect of tVNS on the inflammatory response in a VPA mouse model. The findings are of interest but there are some issues in the communication of the manuscript.

First – there is no ‘autistic’ mouse. would change ‘autistic’ to ‘VPA’- as that is the actual model- in the Abstract and elsewhere in the paper. Also in the abstract add ‘were’ before ‘significantly higher’, change ‘indifferent’ to ‘not different’ and fix spelling on ‘autistism’.

In the Introduction, ‘multiple factors cause autism’- might acknowledge that there are rare high penetrance variance that are known causes alone, and acknowledge that there is a high degree of heterogeneity in the effects on neurotransmitter profiles. Change ‘Biomarker is’ to Biomarkers are’. Most studies are NOT based on the VPA model. The last sentence in paragraph 4 is poorly phrased. Change ‘a complete cure has’ to ‘treatment for core features have’- as a complete cure is just not a reasonable thing. Change ‘path ways’ to ‘pathways’. Might note the heterogeneity of autonomic dysregulation, as the decrease in vagal tone is not universal. Need reference for association between vagal activity and language and behavior. Change ‘it shows’ to ‘all show’, and don’t capitalize ‘Fields’. Change ‘This research aim’ to ‘This research aims’, and change ‘have more precise’ to ‘provide’- not really a precision issue. Remove ‘In addition, it contributes to research in the field of biomarkers by’ and just say ‘while’, and adjoin to the previous sentence. This data doesn’t really help with autism biomarkers clinically.

Methods- first line insert ‘the’ before ‘experiments’. Clarify the wording on the first 2 sentences after ‘2.2 Drugs’. Change ‘In PND28’ to ‘On PND28’. Again change ‘Autistic’ to ‘VPA’ throughout. Were there one male and one female from each litter to avoid litter effects? Fix spelling ‘anestesia’. First few sentences after ‘2.6.ELISA’ the sentence structure is poor- please fix. What is meant by ‘In the incubations’?

Results- again change ‘Autistic’ to ‘VPA’. Fix ‘grups’. On figures 2, 3, and 4, p values are above each figure image- be clear which finding these p values represent among the significant findings in the image. Also- for each of those figures, fix ‘Comperison’ in the figure legend. Change ‘levels significantly’ to ‘levels were significantly’.

Discussion- change ‘single VPA’ to ‘a single dose of VPA’ and add ‘the’ before ‘increased brain NLRP3’. Need reference for association between cytokines and cognition and behavior. Change ‘aberrations’ to ‘alterations’, again change ‘Autistic’ to ‘VPA’ throughout. Change ‘the different effect of gender’ to ‘a difference in effect between genders’, change ‘suggested’ to ‘suggests’ here. Change the ‘It also increased slightly in females, but the difference was not significant’ to just ‘The difference in females was not significant’- if it is not significant there is not a difference. Fix the sentence structure at the beginning of the subsequent paragraph starting with ‘Various studies have described…’. Insert ‘and’ before ‘is primarily found in immune…’ Please fix ‘Zhao et al’- seems stuck in randomly. Is reference 55 with VPA? Please specify. Last paragraph, change ‘on autism’ to ‘on one model of autism’- this doesn’t necessarily speak to clinical autism in a broad manner. Remove ‘so it think it will shed light on future studies in the field’- does not add meaning. Similarly, the sentence starting ‘However, it has contributed…’ does not add meaning. Not sure this study points to ‘suggesting the need for further research on cytokine modulation as a therapeutic approach in autism’. One additional weakness is the lack of assessments of behavior in this model or other aspects such as neurotransmitters.

Conclusions- change ‘augmented’ to mitigated’- right? Change ‘crucial tasks’ to ‘a crucial role’. Remove ‘to provide its early diagnosis’- this study has no relation to that aspect.

Re: FEBSOPEN-24-0067R1

The authors have examined the effect of tVNS on the inflammatory response in a VPA mouse model. The findings are of interest but a few issues remain in the communication of the manuscript.

In the Introduction, the last sentence in paragraph 4 is still poorly phrased.

Discussion- The first sentence of the second paragraph (new text in the revision) is also poorly phrased. Change ‘autism fields’ to just ‘autism’.

my comments are now addressed