Reviewer’ s report
Title: Elevated plasma levels of lysophosphatidic acid and aberrant expression of lysopho-
sphatidic acid receptors in adenomyosis
Version: 0Date: 09 Aug 2017
Reviewer: Jemma Evans
Reviewer's report:
This manuscript presents some moderately interesting findings regarding LPA and adenomyosis.
Their investigation of adenomyosis is commendable as this disease is common but not wellunderstood or investigated. However, there are a number of issues which preclude publication ofthe manuscript in its current form. Corrections and suggestions for further experiments areprovided below1) Abstract: LPA should be defined at first mention rather than third2) Abstract:rationale for IL-8/VEGF studies should be summarized in background/objectives section. Indeed,the reason for these studies is essentially a mystery until the very end of the discussion andtherefore rationale for examination of these factors should also be provided in the introductionsection3) Abstract: results need to also elaborate on the localization of the LPA receptors (i.e.stromal or epithelial or both) to provide the rationale for treating endometrial stromal cells only4)Methods: provide information on the fertility status of the adenomyosis subjects5) Methods:reference to ectopic endometrial tissue - more explanation is required here, ectopic usually refersto distant sites as in the case of ectopic endometrium in endometriosis, presumably herein ectopicrefers to endometrium within myometrium? This should be elaborated and the method if samplingto gain myometrium expanded6) Methods: it is stated that control subjects had normal hormoneprofiles, did the adenomyosis subjects also have normal hormone profiles?7) Methods: I doubt the
sections for immunohistochemisty were 4mm thick8) Methods: elaborate on details within
immunohistochemistry section, e.g. antigen retrieval, secondary antibody species and9) Methods:PBS is not an appropriate control for immunohistochemistry, non-immune IgG should be used.Negative controls are not presented therefore it is difficult to assess the specificity of theimmunostaining10) Methods: brief explanation on quantification of immunohistochemistryrequired11) Methods: more detail needed in cell isolation section to clarify how stromal versusepithelial cells were separated12)Methods: more detail needed for cell proliferation assay e.g. cellseeding density, culture conditions, time points examined, LPA concentrations used13) Methods:ELISA's - details needed as above for cell proliferation assay14) Results: Impossible to see nuclearLPA receptor staining from images presented - also need to refer to specific parts of image whendescribing results (panel A,B,C etc)15) Results: Rationale for using stromal cells only forexperimental purposes when LPA receptors clearly present on/in epithelial cells - results forepithelial cells also required 16) Discussion: Unfocused - needs to be more tightly honed in onadenomyosis17) Discussion: section on infertility/miscarriage in women with adenomyosis - notclear from the wording if this section refers to the specific women in the study or generalpopulation with adenomyosis18) Discussion: references to cell invasion, uncontrolled growth andneovasculariazation in adenomyosis and faciliatation of these cellular functions by LPA - whywere these functions not investigated in the current manuscript - these experiments would greatly
enhance the quality of the manuscript19) Figure legends and results section requires n-numbers20)
Not clear if data presented as mean +/- SD or SEM
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
NoDoes the work include the necessary controls?
If not, please specify which controls are required in your comments to the authors.
NoAre the conclusions drawn adequately supported by the data shown?
If not, please explain in your comments to the authors.
YesAre you able to assess any statistics in the manuscript or would you recommend an
additional statistical review?If an additional statistical review is recommended, please specify what aspects require furtherassessment in your comments to the editors.
I am able to assess the statisticsQuality of written English
Please indicate the quality of language in the manuscript:
Needs some language corrections before being published
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