Malus baccata var. gracilis and Malus toringoides Bark Polyphenol Studies and Antioxidant, Antimicrobial and Anticancer Activities
, Agnieszka Szopa 4,*, Paweł Kubica 4, Diaa O. El-Ansary 5, Halina Ekiert 4 and Fahed A. Al-Mana 1Round 1
Reviewer 1 Report
It is an excellent presentation of an important research. The results give further informations about natural cmpounds. It would be useful to show the structures of the more important compounds and compile a list of abbrevations What possibilities can be applicable to isolate these compounds in greater scale from this barks, wether they could be used at all
Author Response
Reviewer 1
It is an excellent presentation of an important research. The results give further information about natural cmpounds. It would be useful to show the structures of the more important compounds and compile a list of abbrevations What possibilities can be applicable to isolate these compounds in greater scale from this barks, wether they could be used at all.
Comment.
Thank you for your high evaluation of our study. We appreciate it very much. As you requested we added the chemical structures of detected compounds on a new Figure 1.
Author Response File: 
Author Response.docx
Reviewer 2 Report
Materials and Methods
4.1. Plant Material and Preparation
In which season were the bark samples collected? (Line 181)
How long were the extracts left to lose all methanol at room temperature approximately? (Line 186)
The temperature of sample evaporation is missing. (Line 188)
4.2. Chemicals
A lot of standards (acids and flavonoids) are listed: “…for qualification and quantification of phenolics…” and “The flavonoids were quantified…”. However, no data on quantities of these compounds in bark samples are showed in the chapter 2. Results of the manuscript, except for protocatechuic acid, gallic acid, and catechin (in addition, the last one isn’t mentioned among the used standards).
4.4. Anticancer activities
Experiment conditions are missing: culture media and supplements for cells, density of cells used in experments, time of addition of the tested samples after subculturing of cells, exposure time of cells to the tested samples, etc.
4.6. Antibacterial activity
Details of the procedure are missing: incubation time with tested samples, temperature, and culture medium, etc.
Any information on testing of antifungal activity isn’t provided.
References
Some citations, e.g., 22, 42, and 43 aren’t correct.
According to “Instructions“, all Journal Names should be abbreviated.
Latin names of organisms in titles of the articles should be in italics.
Formal errors should be corrected
Line 62 cinere
Line 101, Table 2 mL-1
Lines 106 and 111 Table 2 or Table 3?
Lines 111 and 112 μg mL-1 or mg mL-1?
Line 112 gentisic acid?
Lines 120 and 123 Table 3 or Table 4?
Lines 127 and 131 Table 4 or Table 5?
Line 164 ureas
Line 166 and
Line 172 M. baccata and M. toringoides
Line 185 xtracts
Author Response
Reviewer 2
Materials and Methods
4.1. Plant Material and Preparation
In which season were the bark samples collected? (Line 181)
How long were the extracts left to lose all methanol at room temperature approximately? (Line 186)
The temperature of sample evaporation is missing. (Line 188)
Comment.
We added requested information.
4.2. Chemicals
A lot of standards (acids and flavonoids) are listed: “…for qualification and quantification of phenolics…” and “The flavonoids were quantified…”. However, no data on quantities of these compounds in bark samples are showed in the chapter 2. Results of the manuscript, except for protocatechuic acid, gallic acid, and catechin (in addition, the last one isn’t mentioned among the used standards).
Comment
We applied the analysis of DAD-HPLC. We searched for compounds from our data base, within these listed compounds. Compound identification was accomplished by comparing UV spectra and retention times of reference substances or using co-chromatography. The quantification was determined using calibration curves. We added missing information about lacking catechin derivatives.
4.4. Anticancer activities
Experiment conditions are missing: culture media and supplements for cells, density of cells used in experments, time of addition of the tested samples after subculturing of cells, exposure time of cells to the tested samples, etc.
Comment
We added the required information.
4.6. Antibacterial activity
Details of the procedure are missing: incubation time with tested samples, temperature, and culture medium, etc. Any information on testing of antifungal activity isn’t provided.
Comment
We added all required information. We didn’t investigate antifungal activities.
References
Some citations, e.g., 22, 42, and 43 aren’t correct.
According to “Instructions“, all Journal Names should be abbreviated.
Latin names of organisms in titles of the articles should be in italics.
Comment
Done.
Formal errors should be corrected
Line 62 cinere
Line 101, Table 2 mL-1
Lines 106 and 111 Table 2 or Table 3?
Lines 111 and 112 μg mL-1 or mg mL-1?
Line 112 gentisic acid?
Lines 120 and 123 Table 3 or Table 4?
Lines 127 and 131 Table 4 or Table 5?
Line 164 ureas
Line 166 and
Line 172 M. baccata and M. toringoides
Line 185 xtracts
Comment
Thank you. All corrections were applied.
Thanks for accepting the publishing of this paper.
Author Response File: Author Response.docx
Reviewer 3 Report
This study by Elansary et al., evaluates potential antimicrobial and anticancer activity of M. baccata and M. toringoides bark extracts. While the results seem significant, its presentation lacks some details as mentioned below:
For antiproliferative activity, what were the concentrations and its range used to determine IC50 values in Table 2. Also, the extracts were solubilized in 1% DMSO, what was the effect of DMSO on cancer and normal cells. Basically, 1% DMSO can have an impact on its own and makes it important to have a negative control in these experiments. Figure 3 legend needs much more elaboration. There is no mention of concentration of extracts used, number of replicates, etc. In Table 3 & 4, what are units for MIC and MBC or MFC. How were these values calculated from absorbance measurements. In general, n for each experimental study is missing. This needs to be added to every table and figure. In methods, there is no description of what media was used for various cell types and what solution was used to solubilize the extracts except use of 1% DMSO.
Author Response
Reviewer 3
This study by Elansary et al., evaluates potential antimicrobial and anticancer activity of M. baccata and M. toringoides bark extracts. While the results seem significant, its presentation lacks some details as mentioned below:
For antiproliferative activity, what were the concentrations and its range used to determine IC50 values in Table 2.
Also, the extracts were solubilized in 1% DMSO, what was the effect of DMSO on cancer and normal cells. Basically, 1% DMSO can have an impact on its own and makes it important to have a negative control in these experiments. Figure 3 legend needs much more elaboration. There is no mention of concentration of extracts used, number of replicates, etc. In Table 3 & 4, what are units for MIC and MBC or MFC. How were these values calculated from absorbance measurements. In general, n for each experimental study is missing. This needs to be added to every table and figure. In methods, there is no description of what media was used for various cell types and what solution was used to solubilize the extracts except use of 1% DMSO.
Comment
Done.
A negative control was used and added to the material and methods. All concentrations and media were added. The n was added as you requested.
Thanks for accepting the publishing of this paper.
Author Response File: Author Response.docx
Round 2
Reviewer 2 Report
Please see the attachment. My notes to authors’ comments (to my original suggestions) are in yellow.
Comments for author File: 
Comments.pdf
Author Response
Ref.: Processes-716341
Title: Malus baccata var. gracilis and Malus toringoides bark polyphenol studies and antioxidant, antimicrobial and anticancer activities
Dear Eva Huang
I would like to thank for the Reviewer 2 comments. With reference to your decision I'm sending the responses below. I hope that you find the results satisfying.
Sincerely yours,
Agnieszka Szopa
Responses For Reviewer No. 2:
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Reviewe 2 comment |
Authors response |
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The authors list 40 compounds in 4.2. Chemicals. They write that 21 were screened in Results, paragraph 2.1. What is true? Data on quantities of 3 compounds in bark samples are in Table 1. If the rest of screened compounds (of 21 or 40?) is not present in the bark, they cannot be quantified, and the authors cannot write that the listed compounds were quantified. If some of them were present, except for 3 in Table 1, and quantification of them was really done, the result can be shown in Results.
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Thank you for your valuable comment. We apologize for our oversight. We check one again compounds which were search for in the Malus extracts. That was 37 compounds it total. We corrected this information in the Materials and Methods section, and in the whole manuscript. We also corrected wrong statement that we quantified all 37 compounds. We qualified and quantified only 3 confirmed substances. |
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Incubation time of cells with the tested substances, among others, is very important information. The experiment cannot be reproduced according to authors’ description.
Was the density of cells for experiments 4 x 10-4 per well?
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The incubation time was added which is 2 days or 48 hours
Corrected, thanks.. (per micro) |
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In case that the authors didn’t investigate antifungal activities, the results in paragraph 2.4. and Table 5 are false. |
We investigated the antifungal activity and this was added to the material and methods part as you requested. Thanks again |
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It wasn’t done. Journal Names are and aren’t abbreviated, i.e. they aren’t arranged according to “Instructions“. In addition, some citations aren’t still correct , e.g., Journal Names are missing in (18, 52, and 55). |
We checked and corrected the Reference list carefully.
Thank you for acceptance. |
Author Response File: Author Response.docx
Round 3
Reviewer 2 Report
no comment
