Comparative Study of Three Raspberry Cultivar (Rubus idaeus L.) Leaves Metabolites: Metabolome Profiling and Antioxidant Activities
Round 1
Reviewer 1 Report
General comments:
The manuscript titled “Comparative study of three Raspberry Cultivars (Rubus idaeus L.) leaves: Metabolome profiling and antioxidant activities” is a study focused on discriminating three different raspberry varieties. To achieve this goal, the authors performed several analyses of the leaves of each variety, including target metabolites by LC-MS/MS and the antioxidant activity using mouse fibroblast L929 cells.
There is a lack of caring when spelling Latin names and/or expressions. Please revise the manuscript regarding missing italic forms.
Also, it would be great if the authors make some efforts to present their manuscript for peer review considering the journal standards and quality.
Abstract:
English spelling and grammar are confusing. Furthermore, this section must be improved regarding the way authors describe their work (in a brief).
Introduction:
The same as abstract, English must be revised and written improved.
Materials and Methods
Q1: the described methodology was developed/optimized by the authors? Was inspired by other authors? Refs are missing along this section.
Q2: How does the authors identified the studied varieties? Can the authors provide some info in this regard, please?
Q3: Why the authors do not express their results as IC50 values?
Line 86: “(…) solid-liquid ratio of 1:40 (v/v)”; do the authors means w/v?
Line 117, 126: CO2 (please revise through the manuscript)
Line 122; 131: 2*104/well (please revise through the manuscript)
Sections 2.3.1 and 2.3.2: Do the authors used a positive control? How to compare the achieved results without a positive control?
Line 166 to 179: why this info is presented?
Tables: Be careful when presenting data. It must be coherent. Values expressed as (ex.) “2.0E-03” is obsolete. Please, restructure this table.
Figures 1 and 3: Please improve the quality of the images.
Line 220: T-test; it should be t-test.
Figure 4: Positive controls should be presented as well as, it could be great if higher concentrations were tested to verify at which concentration 50% of the population was inhibited.
Lines 486-488: this section is not mandatory.
References
Please revise according to the journal format.
Author Response
Please see the attachment.
Author Response File: 
Author Response.docx
Reviewer 2 Report
Manuscript ID: applsci-1485742
After careful review of the ms entitled “Comparative study of three Raspberry Cultivars (Rubus idaeus L.) leaves: Metabolome profiling and antioxidant activities” (Manuscript ID: applsci-1485742), this reviewer recommends it for publication after the suggested revisions.
The fruits and leaves of raspberry (Rubus idaeus L.) are an excellent source of secondary metabolites with antioxidant functions. This study performed a metabolome analysis using LC-MS/MS to investigate the components of leaves from the three known cultivars of raspberry 'Autumn Britten', 'Autumn Bliss', and 'Red Autumn', and tested their antioxidant activity on mouse fibroblast L929 cells. The cytotoxicity of raspberry leaves ethanol extracts (RLEE) from the three cultivars in study had no hazardous effects on L929 fibroblasts in the concentration range of 0-50 µg/mL. Moreover, RLEE at 50 µg/mL induced an increase of the levels of superoxide dismutase (SOD), glutathione (GSH), and catalase (CAT), after UV-B exposition of L929 fibroblasts. RLEE is confirmed to be rich in bioactive metabolites with important antioxidant effects in vitro.
This reviewer suggests important revisions of the text, also by improving English language, and by removing inaccuracies and repetitions.
Further suggestions are listed below:
TITLE
L2: insert italics type for "Rubus idaeus"
ABS
This reviewer suggests rewriting the abstract all over again. It has to well summarize and reflect the main findings of work.
L16-18: rewrite this sentence, why “which has caused some researchers to be concerned”?
L16: “a vital role”? Are you sure about that?
L16: here and wherever it occurs, insert italics type for "Rubus idaeus"
L17: explain better what you mean for “multi-dimensional functions”, here it is not clear
L20: change “types” to “cultivars”
L21: change “in-vivo” to “in-vitro”
24-25: explain better what you mean for “and a certain research basis was provided for the subsequent functional development of raspberry leaves”
L27: change “g” to “µg”
L27: remove “and 50 g/mL”
L30-31: explain better what you mean for “This study provides theoretical support for the variety screening of active metabolites 30 three raspberry varieties leaves and the utilization of leaf resources,”; why “theoretical”?
L31: insert “from” before “three”
L31: remove “while” and insert full stop
INTRO
L37: here and wherever it occurs, insert italics type for "Rubus idaeus", use the Latin name in M&M section, check for “Ideaus”, change it to “idaeus” in lowercase letters
L37: insert italics type for "Rubus"
L44: here, insert citations for “several studies”
L46: garbage or waste?
L49: insert italics type for bacteria
L51: explain better these effects
L55: insert year and number for reference
L56: remove repetition “for anti-oxidation”
L57: insert year and number for reference; check for this reference: Aleksandra is the name, use surname Pavlovi
L62: insert “extracellular” before “matrix”
L66-67: the first time cited, explain acronyms SOD, CAT, GSH
L68: change “Antioxidant” to “These”, insert “their” before “antioxidant”
L69: remove “enzyme”
M&M
L81-82: remove brackets and “, all”
L84: check for English in “Preparing…”
L87: please, indicate the Hz
L88: indicate the RCF instead of “rpm”
L114: remove brackets
L116: remove comma in “USA,)”
L117,126: use subscript in CO2: CO2
L120: remove double brackets, use only a pair
L122,131,136: use superscript in 2x104: 2x104, and cm2: cm2
L123-125: check for number of treatments, 9 groups of 6 replicates, describe them more clearly
128: insert “and” after “450 nm”
L129: insert “CCK-8” before “kit”
L141: is lysate for lysis buffer?
L147: remove double brackets, use only a pair
L154: the first time cited, explain acronym VIP
RESULTS
L182: remove “Metabolites were collected and analyzed using”, and insert “was used” after “technique”
L185: change “Table 1” to “Table S1”
L186: acronym QC in full: quality control (QC)
L200-202: move Table 1 to L215 where it is the first time mentioned; in the title, add “ion mode” after “negative”
Tables 1-3: there are reported the nine (9) most important metabolites, not 10 as indicated in table titles
L212-213: check for English language and meaning
L219-221: check for English language and meaning, use t-test instead T-test
L221: move Figure 2 here
L232: on what basis was the comparison conducted? describe better
L237: move Figure 3 here
L244: move Figure 4 here
L252: remove “and”
L253: lines are not red, change “red” to “vertical”
L254: change “black” to “horizontal”
L268: check for English language and rewrite
L269: move RLEE before “different”, insert “from” after RLEE
L272: remove “different varieties”, change Figure 4A to Figure 5A
L276: remove brackets in excess, change Figure 4B to Figure 5B
L278: consider removing “, while the activity of SOD in the UVB group was 79.67±0.35 U/ mgprot.”
L279: remove repetition of “significantly”
L283: remove “control and”
L284: remove “could”, and change “increase” to “increased”
L289-90: consider removing the repetition “CAT significantly decreased in UVB and UVB + RLEE groups but significantly increase in the RLEE pre-protection groups compared to the control group.”
L291: change “at” to “of”
L295: insert “this” before “indicated”
L295: use acronym ROS
L298-301: remove editorial indications
L311-315: on the Y-axis of Figures 5B, 5C, 5D, as in L317, use “activity” with the enzymes SOD, GSH, and CAT
L317: use “Cell viability” instead of “Cell activity”, as on the Y-axis of Figures 5A
DISCUSSION
L325: remove Discussion, these are results
L326-331: move this part of Discussion before L318
L332: insert new paragraph title for these findings in Results section
L332: insert “molecular” before “species”
L333: insert colon instead of comma, after “varieties”
L333-335: the total number of metabolites is not 399, but 258, please explain better
L333: insert the kind of molecule for (17)
L342-348: this part is confusing, please rewrite; have you compared each variety to itself?
L347: amino acids are not up-regulated
L397: insert year of reference
L398: remove the repetition of “abalone”
L413: change “can” to “does”; remove the repetition “the body” before “from”
L414: remove the repetition “of the body”
L423-424: change “product” to “products”
L430: to avoid missense, change “improve” to “reduce”
L438,453: use superscript in cm2: cm2
L440: change “species” to “types”
L442: change “interspecies” to “varietal”
L444: remove “Excessive”, use only ROS
L445-446: remove the repetition “of the body”
L449: use subscript in O2 and H2O: O2 and H2O
L467-469: check for English language and meaning, then rewrite better
L479: use “varieties” instead of “types”
L486-488: remove
L489-490: insert Supplementary
L494: choose the option
REFERENCES
Remove double list numbering
Use italics for species names
L533: remove “?”
Author Response
Please see the attachment
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
In general, this manuscript need to be revised regarding its Enghish quality of redaction. The exception for the Discussion, in which, in my oppinion, is well redacted and with sufficient quality.
Abstract
This section was reduced but now it lacks of English grammar and spelling correction.
Introduction
Line 89:please add the year after the REF mentioned (parentheses).
Line 90: Why do the authors are reporting the F. rubra features instead of R. idaeus?
Lines 106-116: it would be great if some context were added to this paragraph. It contains important facts, however, poorly linked with the remaining text.
Lines 117-124: please add some intro to this paragraph (ex: This work aims to ... To achieve those goals, several assays were performed including... Therefore, this study represents ...).
Line 241: Table S1. For some reason, SI is not available. However, do the authors perfomed the quantification of the main molecules? There is no info in this regard in the Material and Methods section. If you don't, how do you perform the statistical analysis to have those "significant differences"?
Line 256: "VIP values". This is the first time which "VIP" is presented. It would be better to explain what does VIP means as well as, clarifying at M&M section which parameters will be evaluated for each assay.
Figure 5: it could be more interesting to present this data in a table instead of a image.
Line 498: "U/ mgprot"; please explain what those unit means.
Discussion:
This section is really well done. Good job!
Author Response
Please see the attachment
Author Response File: Author Response.docx
Round 3
Reviewer 1 Report
The authors made efforts to address the previous suggestions.
However, there are a few details that I would like to mention, aiming to contribute to its improvement.
I suggest reviewing punctuations through the manuscript (spaces, dots…).
Furthermore, despite being perceptible, I suggest the revision of this manuscript by a colleague fluent in English at it present some deficiencies in this regard.
Line 18: please use “activity” instead of “property”.
Lines 19-20: “However, different cultivars might processe different phytochemisty and bioactivities and could be used for divers purposes”. I suggest deleting this phrase as the former and next ones perfectly explain the context and aim of this work.
Line 50: the year is missing after “Pradeepa et al.”, please correct references properly. If the same author has two publications, therefore you can redact something like “ Pradeepa et al. (2014) synthesized (…) REF. Later (2019), the same working group showed the delayed effect on plasma (…) by means of an ethanolic extract with XYZ molecules).
Line 59: In vitro should be in italic form. Please revise the manuscript.
Line 72: Please correct the phrase “In order to make more efficient the use of raspberry leaves”.
Line 110: “in a cold bath”
Line 182: “was lower than”
Table 4: it would be great if authors use scientific numbering as the values have a lot of digits.
Author Response
Please see the attachment.
Author Response File: Author Response.docx
This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.
