Green Chemistry in Medical Applications: Preliminary Assessment of Kuzu Starch Films with Plant-Based Antiseptics
Round 1
Reviewer 1 Report
Comments and Suggestions for Authors1-at page 3 in materials and methods section you must wright the materials and chemicals used in your experiments with their source and not mentioned the methods of preparation in this section at all. The preparation writes under tittle preparation of composite films.
2-silver nanoparticles which you used are prepared? or you bring from company ? if you prepare it please write the method of preparation and if you bring its ready write the name of company which you being it,
3-at page 5 line 168 you write method no this section consider "Tests and Characterization"
4-at page 8 line 276 it is not table 1 it is table 2.
5-at page 6 line 201 write the tittle"Thermal analysis of starch films"
6-at table 3 what is TM please clear it under table .
7-why you are not made morphology test by SEM (scan electron microscope) its necessary to show the change of surface.
8-conclusions need to be rewrite again and show the ideal condition .
9-why you are not applied it on rats?
Author Response
Thank you very much to the reviewer for such an insightful review. Comments and suggestions will undoubtedly help us improve our manuscript and are also a valuable guide for future research. We have marked all changes in the manuscript in yellow. We hope that the new version of the manuscript meets the reviewer's expectations.
1-at page 3 in materials and methods section you must wright the materials and chemicals used in your experiments with their source and not mentioned the methods of preparation in this section at all. The preparation writes under tittle preparation of composite films.
Answer: Thank you very much for this comment. In fact, the materials and methods section was written chaotically. We corrected it in new version of manuscript.
2-silver nanoparticles which you used are prepared? or you bring from company ? if you prepare it please write the method of preparation and if you bring its ready write the name of company which you being it,
Answer: Commercial Nonionic colloidal nanosilver solution, was produced by Vitacolloids. This information was actually missing in the content. We have added it to the materials subsection.
3-at page 5 line 168 you write method no this section consider "Tests and Characterization"
Answer: We agree with the reviewer that Tests and Characterization sounds better. This has been changed in the text.
4-at page 8 line 276 it is not table 1 it is table 2.
Answer: Thank you for noticing this error. Of course, it should be Table 2. It has been corrected in the text.
5-at page 6 line 201 write the tittle "Thermal analysis of starch films"
Answer: The subtitle "Thermal analysis" has been added to the new version of the manuscript.
6-at table 3 what is TM please clear it under table .
Answer: Thank you for this comment. The explanation for Tm was given in the original version of the manuscript in the content (l. 393), but we absolutely agree that it should be placed under the table. The abbreviations are explained under the table.
7-why you are not made morphology test by SEM (scan electron microscope) its necessary to show the change of surface,
and 9-why you are not applied it on rats?
Answer: Of course, we agree with the reviewer that it would be worth adding the results of SEM analysis and the impact on cells or living organisms, such as rats. The research material shown in this manuscript is partial material. Our idea is to obtain a series of different starch films, modified with various antibacterial additives (not only those shown in this manuscript). Unfortunately, for financial reasons, we will be able to perform a full analysis (included the surface analysis) only for the best foils among all received. At the same time, we believe that it is worth showing these partial results, also so that other research groups can use them as a direction of research in searching for the best dressing materials.
8-conclusions need to be rewrite again and show the ideal condition .
Answer: Thank you very much for this comment. The content of the conclusions, but also the abstract, has been corrected in the new version of the manuscript.
Reviewer 2 Report
Comments and Suggestions for AuthorsThe manuscript entitled “Preparation of Kuzu Starch-Based Films with Antibacterial Additives for Wound Dressings” was submitted to the journal Sustainability for possible consideration as a research article. After reading the main text, I have some concerns on the results mainly antibacterial experiments, and questions are as follows,
1) Title of the manuscript was somehow misleading and incorrect to certain degree. So far, the effectiveness and functions of Kuzu films as wound dressings have not been tested and validated in the real circumstance. Only the antibacterial tests were conducted separately on a single Gram+ and a single Gram- strains. Anyway, the title of this manuscript must be revised properly, unless the wound dressing tests were done.
2) Figure 1 was belonging to the background and showed two typical structures of starch (or Kuzu starch). But what’s the connection to the topic of the manuscript? I have no ideas why it was here. Any reasons, or necessities? Maybe we want to dig out more and deeper information on Starch or doing starch modification on its fine structures? Or any of the three bioactive additives may re-define and modify the Kuzu starch?
3) The selection of rooibos extract and chili extract looks like maybe reasonable or almost fine, but rather weak, seeming to be lack of scientific thinking and technical consideration. Better to show some overwhelming results that both are economic and accessible, standing out from many daily natural materials and antiseptic vegetables.
4) According to the authors, Kuzu-Ag was made by using nonionic colloidal nanosilver solution (Vitacolloids, 50 ppm). My question is why the authors chose this kind material not others, and how did they know their single choice correct, would this commercial product perfectly fit and function well just for one shot? There are many various nanosilver colloid products, why this not that, no testing, and no comparison at all. Generally, in the studies, people must try several materials several times. Most of the time the results would not be predicted, so in the beginning, they would never ascertain the results and therefore need to optimize the materials and methods until further confirm the effects.
5) If simple preparations are made, or commercial nano products are purchased, the only left task is to use but not to create. Only simple anti-microorganism tests were made. The innovation and significance are limited. I did not see in this study that any existed drug or effective compound was selected for comparison even just in the anti-bacteria test conditions. No creation and no cell-based data. Even the combination of two of current three approaches has not been tried or shown up. Then, they hardly have the mechanism discussion.
6) Line 489-line 497, including Table 4. The description of inhibition area by Kuzu rooibos on Gram- E. coli ATCC 25922 was incorrect. The inhibitory effect was calculated by the Diameter of Area, was 4 times larger than effect of rooiks on Gram+ 25923. So, the words between line 493 and line 494 should be properly revised.
7) By the way, the original images to Table 4 were not seen but should be provided to double-check the resulted information.
8) There was overlapping description between the “Abstract” and the “Conclusions”.
9) Antibacterial tests were truly important results in this manuscript. Before the antibacterial tests, all chem-physical analysis were necessary but of low significance. I expect to see more results by using not only single composite with Kuzu starch film, but also the combined composites with Kuzu starch film.
10) If oil chili extract or chili oil extract or nanosilver colloid was used for real wound curing, would the cells present cytotoxic effects or acute inflammatory effects? At least cultured cell-based experiments should be performed, to show cellular effectiveness or significance.
11) If possible, as authors announced in the original title, animal experiments or the pre-clinical should be tested.
A substantial revision must be carried out on the current manuscript.
Comments on the Quality of English Languageminor revision could be made to improve the English language.
Author Response
We would like to thank the reviewer for his insightful analysis. We hope that our corrections and replies in the cover letter will dispel all his doubts, and the new manuscript will meet the reviewer's expectations. We have marked all changes in the manuscript in yellow.
1) Title of the manuscript was somehow misleading and incorrect to certain degree. So far, the effectiveness and functions of Kuzu films as wound dressings have not been tested and validated in the real circumstance. Only the antibacterial tests were conducted separately on a single Gram+ and a single Gram- strains. Anyway, the title of this manuscript must be revised properly, unless the wound dressing tests were done.
Answer: We would like to thank the reviewer for this comment. We agree that we worded the title of the manuscript somewhat exaggeratedly and imprudently, suggesting a full analysis of the tested materials in terms of their use as wound dressings. We have changed this in the new version of the manuscript.
2) Figure 1 was belonging to the background and showed two typical structures of starch (or Kuzu starch). But what’s the connection to the topic of the manuscript? I have no ideas why it was here. Any reasons, or necessities? Maybe we want to dig out more and deeper information on Starch or doing starch modification on its fine structures? Or any of the three bioactive additives may re-define and modify the Kuzu starch?
Answer: Of course, we agree with the reviewer that such simple and well-known patterns are unnecessary here. They have been removed from the manuscript.
3) The selection of rooibos extract and chili extract looks like maybe reasonable or almost fine, but rather weak, seeming to be lack of scientific thinking and technical consideration. Better to show some overwhelming results that both are economic and accessible, standing out from many daily natural materials and antiseptic vegetables.
Answer: The research material shown in this manuscript is partial material. Our idea is to obtain a series of different starch films, modified with various, natural, antibacterial additives (not only those shown in this manuscript). We believe it is prudent to find such natural substitutes for most of the products used by humanity in order to be prepared when sources of oil and gas are depleted. From an ecological point of view, sustainable agriculture has less impact on the environment, so the search for appropriate compounds from plants is very desirable.
We also believe that it is worth showing these partial results, also so that other research groups can use them as a direction of research in searching for the best dressing materials. Two plant materials with known antimicrobial and anti-inflammatory properties were selected for the research presented in this manuscript, i.e. rooibos tea leaves and chili peppers. Their properties are described in the "Introduction" section. Looking at the characteristics of these plant materials, it can be assumed that their use may be beneficial to obtain medical products, not only pharmacological ones.
4) According to the authors, Kuzu-Ag was made by using nonionic colloidal nanosilver solution (Vitacolloids, 50 ppm). My question is why the authors chose this kind material not others, and how did they know their single choice correct, would this commercial product perfectly fit and function well just for one shot? There are many various nanosilver colloid products, why this not that, no testing, and no comparison at all. Generally, in the studies, people must try several materials several times. Most of the time the results would not be predicted, so in the beginning, they would never ascertain the results and therefore need to optimize the materials and methods until further confirm the effects.
Answer: We actually used this solution as a comparative material for foils with additions of plant extracts to be able to determine whether the tested foils affect bacteria to a greater or lesser extent than foils without additives (negative control) and foils with silver nanoparticles with proven antimicrobial activity (positive control). This information has been added to the text of the manuscript. When choosing this particular Ag solution, we were guided by the results of tests performed by an accredited laboratory, the reports of which are attached in the response. Unfortunately, they are in Polish.
5) If simple preparations are made, or commercial nano products are purchased, the only left task is to use but not to create. Only simple anti-microorganism tests were made. The innovation and significance are limited. I did not see in this study that any existed drug or effective compound was selected for comparison even just in the anti-bacteria test conditions. No creation and no cell-based data. Even the combination of two of current three approaches has not been tried or shown up. Then, they hardly have the mechanism discussion.
Answer: As previously mentioned, the materials presented in the manuscript and their analysis are part of a larger project aimed at examining various plant materials with long-known antimicrobial and often anti-inflammatory properties, in terms of their potential use to modify the properties of starch. These plant materials are selected based on scientifically documented properties. The preliminary research presented in the manuscript is intended to show whether it is possible to introduce such plant material into the starch matrix and what effect it has on its basic properties. The end result will be the selection of the best materials from all received and their full analysis for use as a dressing material. Unfortunately, for financial reasons this is not possible for all foils.
6) Line 489-line 497, including Table 4. The description of inhibition area by Kuzu rooibos on Gram- E. coli ATCC 25922 was incorrect. The inhibitory effect was calculated by the Diameter of Area, was 4 times larger than effect of rooiks on Gram+ 25923. So, the words between line 493 and line 494 should be properly revised.
Answer: We thank the reviewer for this comment. This means that we expressed ourselves imprecisely. What we meant was that in the case of both S. aureus and E. coli, the order of impact of individual films with additives was as follows: starch discs containing silver colloid > chili pepper extract > rooibos extract. We have made a change to the manuscript. We hope it's clearer now.
7) By the way, the original images to Table 4 were not seen but should be provided to double-check the resulted information.
Unfortunately, when taking photos (after measuring the inhibition diameters), we did not check their quality. It turned out that the matrix and aperture in the camera were broken and it recorded very overexposed photos :-( Therefore, we cannot present photos of plates with samples and bacterial colonies. In attachment is an example of such an overexposed photo (and this is the best picture).
8) There was overlapping description between the “Abstract” and the “Conclusions”.
Answer: Thank you very much for this comment. Both fragments have been corrected in the new version of the manuscript.
9) Antibacterial tests were truly important results in this manuscript. Before the antibacterial tests, all chem-physical analysis were necessary but of low significance. I expect to see more results by using not only single composite with Kuzu starch film, but also the combined composites with Kuzu starch film.
and 10) If oil chili extract or chili oil extract or nanosilver colloid was used for real wound curing, would the cells present cytotoxic effects or acute inflammatory effects? At least cultured cell-based experiments should be performed, to show cellular effectiveness or significance.
and 11) If possible, as authors announced in the original title, animal experiments or the pre-clinical should be tested.
Answer: From a technical point of view, for use as wound dressings, the determined physico-chemical properties are very important. It is important whether the introduction of additives will cause, for example, the polymer to have a too low melting or softening point (due to the increased temperature in the wound it may start to stick to it), or to be too weak and will tear in the hands. Water affinity must also be taken into account when selecting a dressing for the type of wound. The preliminary antibacterial tests of these materials, along with their physico-chemical properties, are shown to indicate whether these starch films may be suitable for further testing as wound dressings.
The concentrations of modifying additives used result from previous attempts to obtain starch film. Other starch/additive ratios resulted in films that were either sticky, too brittle, or cracked or with unmixed starch lumps. Hence, the amounts of added modifying solutions with the best physicochemical and organoleptic properties were selected. We do not discuss them in the publication because it would often be difficult to estimate individual properties.
As we mentioned earlier, unfortunately, financial limitations allow us to expand our research to include analysis of the impact on cells (in vitro) and on living organisms for each material. After developing and determining the basic properties of all starch materials planned to be obtained, we will select the best ones for which we will be able to expand research, especially those related to biological determinations.
Author Response File: 
Author Response.pdf
Reviewer 3 Report
Comments and Suggestions for AuthorsThe research is of good quality, contains useful and promising results and has potential to be further improved by providing some additional details.
The justification of nano silver use has not been duly provided taking into consideration its potential toxicity reported e.g. in Zhang J, Wang F, Yalamarty SSK, Filipczak N, Jin Y, Li X. Nano Silver-Induced Toxicity and Associated Mechanisms. Int J Nanomedicine. 2022 Apr 26;17:1851-1864. doi: 10.2147/IJN.S355131. or in Shati AA, Elsaid FG. Biosynthesized silver nanoparticles and their genotoxicity. J Biochem Mol Toxicol. 2020;34(1):e22418. or in Pang S, Gao Y, Wang F, Wang Y, Cao M, Zhang W, Liang Y, Song M, Jiang G. Toxicity of silver nanoparticles on wound healing: A case study of zebrafish fin regeneration model. Sci Total Environ. 2020 May 15;717:137178. doi: 10.1016/j.scitotenv.2020.137178. and many other
The evident weak point is that in vitro tests and animal tests of developed wound dressings have not been provided. So the conclusion in lines 526-528 was not duly justified.
Please make corrections in text in line 84.
It would be beneficial at the end of Introduction more clearly and more specifically state the aim of the research - at present one can see only the description of the work which has been performed.
The "accelerating wound healing process" has been claimed in line 119 but has not been experimentally demonstrated and confirmed in the following main text.
Author Response
We would like to thank the reviewer for his favourable attitude and, above all, for his constructive comments. We hope that the new version of the manuscript meets the reviewer's expectations. Any changes in the text are marked in yellow.
The justification of nano silver use has not been duly provided taking into consideration its potential toxicity reported e.g. in Zhang J, Wang F, Yalamarty SSK, Filipczak N, Jin Y, Li X. Nano Silver-Induced Toxicity and Associated Mechanisms. Int J Nanomedicine. 2022 Apr 26;17:1851-1864. doi: 10.2147/IJN.S355131. or in Shati AA, Elsaid FG. Biosynthesized silver nanoparticles and their genotoxicity. J Biochem Mol Toxicol. 2020;34(1):e22418. or in Pang S, Gao Y, Wang F, Wang Y, Cao M, Zhang W, Liang Y, Song M, Jiang G. Toxicity of silver nanoparticles on wound healing: A case study of zebrafish fin regeneration model. Sci Total Environ. 2020 May 15;717:137178. doi: 10.1016/j.scitotenv.2020.137178. and many other
Answer: We would like to thank the reviewer for this comment. We agree that nanoparticles should be used with caution and avoided whenever possible. Of course, there are situations when the benefits of their use outweigh the associated risks, e.g. in the case of cancer. This certainly does not apply to wound dressings. However, in our work, the foil with nanoparticles was used as a positive control test to determine whether samples with plant additives also have an antibacterial effect. We stressed this too little in the manuscript, but we have now corrected it.
The evident weak point is that in vitro tests and animal tests of developed wound dressings have not been provided. So the conclusion in lines 526-528 was not duly justified.
Answer: We agree with the reviewer that this conclusion was formulated prematurely. The research material shown in this manuscript is partial material. Our idea is to obtain a series of different starch films, modified with various natural antibacterial additives (not only those shown in this manuscript). Unfortunately, for financial reasons, we will be able to perform a full analysis (including research on cells and living organisms) only for the best foils among all received. At the same time, we believe that it is worth showing these partial results, also so that other research groups can use them as a direction of research in searching for the best dressing materials.
Please make corrections in text in line 84
Answer: The sentence was corrected into: ”The Eskandarinia and others’ group obtained films based on corn starch with the addition of propolis and hyaluronic acid [30]. The films exhibited antimicrobial activity against Staphylococcus aureus and Escherichia coli.”
It would be beneficial at the end of Introduction more clearly and more specifically state the aim of the research - at present one can see only the description of the work which has been performed.
Answer: The aim of the research was added to the content of the manuscript. We hope it is clear in the current version.
The "accelerating wound healing process" has been claimed in line 119 but has not been experimentally demonstrated and confirmed in the following main text.
Answer: Of course, we agree that the statement "accelerating wound healing process" did not result from the research conducted. It was the result of a review of scientific literature on the properties of rooibos tea and chili extract. We concluded that by introducing these extracts into the starch matrix, we can also obtain materials suitable for wound dressings.
This statement has been removed from the manuscript.
Reviewer 4 Report
Comments and Suggestions for Authors1. p2, l54-59: These are well-known facts, so it should be erased or shortened.
2. p3, l85: The obtained films exhibited...
3. The Introduction section is way too long.
4. Either merge the subparagraphs 2.1.1, 2.1.2 and 2.1.3 into one subparagraph, or isolate p4, l152- onwards into a 2.1.4 subparagraph. In my opinion, option 1 is more appropriate.
5. Table 1: 92.5 g Rooibos extract, 20 g chili extract and 46.5 g nanocolloid Ag: Were you planning to obtain approximately such contents?
6. p 5, l168-171: Delete. It will be explained below, so no need to repeat. Actually, organize paragraphs and subparagraphs in the section "Materials and Methods" in a more proper way.
7. p6, l218-222: P, M, t - should be in italic. p7, l260-1: D also in italic.
8. p7, l263: Figure 2 shows photographs of Kuzu starch films and Kuzu starch films with additives - this not the proper way to start the new paragraph, especially the "Results and discussion" section. Please modify.
9. Table 2: One-way ANOVA should be performed.
10. Figure 3 should be primarily mentioned in the manuscript and then presented, not other way around. The same stands for Table 3.
11. Figure 3A and 3B are missing ANOVA, while Figure 3C is missing standard deviation and ANOVA.
12. p11, l695-onwards: This should be organized into a separate subparagraph.
13. Figure 4 Table 4 are missing ANOVA.
14. p13, l432: It should be Figure 5 instead of Figure 4.
15. Kuzu chili films showed good results, but my concern would be related to the real application of such films, due to the known fact that capsaicin is irritant compounds. Could you comment on this?
Author Response
We express our sincere gratitude for the time and effort that you dedicated to reviewing my publication. Your insightful comments and constructive feedback have been invaluable in enhancing the quality and clarity of our work. We greatly appreciate your thorough analysis and thoughtful suggestions, which have significantly contributed to the improvement of the manuscript.
Thank you once again for your valuable contribution.
- 1. p2, l54-59: These are well-known facts, so it should be erased or shortened.
Answer: Amylose is a linear macrochain where glucose units are connected by a(1→4) glycosidic bonds. Amylopectin is highly branched and has a densely packed structure due to the presence of a(1→6) glycosidic bonds. Amylopectin is a much larger molecule than amylose and is one of the largest known biological molecule
Thank you for this comment. The above sentences have been removed from the manuscript.
- p3, l85: The obtained films exhibited...
Answer: The sentence has been corrected.
- 3. The Introduction section is way too long.
Answer: Thank you for your valuable feedback on our manuscript and for the specific comments regarding the length of the introduction. We appreciate your perspective and understand the importance of conciseness in academic publications.
In the introduction, we aimed to provide a detailed overview of the current state of knowledge in two interconnected areas: starch-based materials modified with antibacterial substances and wound dressing materials that accelerate healing. We believe this comprehensive background is essential to establish the context and relevance of the research, particularly for readers who may not be intimately familiar with these rapidly evolving fields.
However, we acknowledge your concerns and have re-examined the introduction to ensure it is as concise as possible while still providing the necessary background. We hope that the revised Introduction content meets your expectations.
- Either merge the subparagraphs 2.1.1, 2.1.2 and 2.1.3 into one subparagraph, or isolate p4, l152- onwards into a 2.1.4 subparagraph. In my opinion, option 1 is more appropriate.
Answer: Thank you for your insightful feedback on the organization of the manuscript. In response to your suggestion, we have carefully reviewed sections 2.1.1, 2.1.2, and 2.1.3. This restructuring was done in accordance with the guidelines provided by the publishing house, ensuring a more streamlined and coherent presentation of the content.
We hope these revisions meet your expectations and enhance the clarity and effectiveness of the manuscript. Could you kindly review the updated version and share your thoughts? Your feedback is invaluable to us, and we aim to ensure that the final product aligns with both editorial standards.
- 5. Table 1: 92.5 g Rooibos extract, 20 g chili extract and 46.5 g nanocolloid Ag: Were you planning to obtain approximately such contents?
Answer: The proportions of additives were calculated as follows, so that the percentage of starch (2%) is maintained in each of the film-forming solutions. This information was also added in the text.
- 6. p 5, l168-171: Delete. It will be explained below, so no need to repeat. Actually, organize paragraphs and subparagraphs in the section "Materials and Methods" in a more proper way.
Answer: Following your suggestion, the redundant content in the "Materials and Methods" section has been removed, and the section has been reorganized for better clarity. Please let us know if these changes meet your approval.
- p6, l218-222: P, M, t - should be in italic. p7, l260-1: D also in italic.
Answer: The font has been changed to italic type.
- p7, l263: Figure 2 shows photographs of Kuzu starch films and Kuzu starch films with additives - this not the proper way to start the new paragraph, especially the "Results and discussion" section. Please modify.
Answer: We understand your concern about the initial sentence structure and its appropriateness for starting a new paragraph in this section. In line with your suggestion, we have revised the beginning of this paragraph to provide a clearer and more appropriate introduction to the results and discussion of the Kuzu starch films and their variants with additives. This change, we believe, enhances the flow and coherence of the section.
- Table 2: One-way ANOVA should be performed.
Answer: One-way ANOVA was performed. The comment has been posted below the table.
- Figure 3 should be primarily mentioned in the manuscript and then presented, not other way around. The same stands for Table 3.
We have changed the location of the figure in the text.
- Figure 3A and 3B are missing ANOVA, while Figure 3C is missing standard deviation and ANOVA.
Answer: One-way ANOVA was performed. The comment has been posted below the figure. Standard deviations have been added to Figure 3C.
- p11, l695-onwards: This should be organized into a separate subparagraph.
Answer: We have reorganized the content from page 11, line 695 onwards into a separate subparagraph, in line with your suggestion.
- Figure 4 Table 4 are missing ANOVA.
Answer: One-way ANOVA was performed. The comment has been posted below the figure.
- p13, l432: It should be Figure 5 instead of Figure 4.
Answer: The numbering of the figures has been changed
- Kuzu chili films showed good results, but my concern would be related to the real application of such films, due to the known fact that capsaicin is irritant compounds. Could you comment on this?
Answer: When selecting natural, plant materials, we took into account their antimicrobial and anti-inflammatory properties. In the case of capsaicin, an additional effect - an analgesic, can be obtained. There are studies showing the use of capsaicin during surgery (e.g. DOI: 10.1213/ane.0b013e3181753276). This minimizes the need to administer opioid analgesics.
Round 2
Reviewer 1 Report
Comments and Suggestions for Authors1-at figure 2 (b,c)&3 why you made the draw with black color how the reader can differentiate between kuzu 0, kuzu rooibos, kuzu chili and kuzu silver?
Author Response
We do not know what could have happened that the reviewer received a manuscript with figures in black and white. To be sure, we attach a pdf version of the manuscript. Thank you once again for your insightful and constructive review.
Reviewer 2 Report
Comments and Suggestions for AuthorsAll my concerned questions have been properly answered or explained by the authors. Maybe more data are better, as I expected to enhance the quality of the paper, but the authors explained that they have reasons for reporting current results in the present manuscript. So, I have no further questions to ask. In general, I may agree that such revised manuscript could be accepted by the journal.
Author Response
Once again, we would like to thank the reviewer for his insightful and constructive review.
Kind regards,
Authors
