Active Compounds of Panax ginseng in the Improvement of Alzheimer’s Disease and Application of Spatial Metabolomics

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The authors have done a very thorough reviewed on the P. ginseng in AD. It included with comprehensive writeup of different active compounds in P. ginseng and how it can help in the progression of AD. 

The title can be revised to reflect the title more appropriately. The title state the mechanism of Panax ginseng but it should be pharmacological action of active compound of P. ginseng. The title mentioned treatment but the explanation is more to improvement of AD. 

The study included the factors and the underlying mechanism of AD, however, there is a lack of signs & symptoms and current diagnosis of AD. 

The review highlighted the potential of P. ginseng in AD, however, the selection of P. ginseng is not clearly elucidated and is that due to the cost or easily available.

The review highlighted mostly ginsenosides, there is not much on toerh compounds such as gintonin, polysaccharides, peptides .. in AD.

All the figure & table should be included into the text or to the manuscript. 

Table 1to 5 , the effect only show target protein, it will be more complete to inlcude whether suppress, improve, enhance or etc, in more detailed

Figure 3-7 : signalling pathways

The consistency of scientific written for scientific name in italic  (including P. ginseng, in vitro, in vivo)

Paragraph 2.5- consistency of using glycoconjugates or glycoconjugated compounds 

subtitles of 3. Mechanism ... is actully more to pharmalogical effect

There are many wrong in text citation in the text (Error! ... ) 

e.g. Line 106-107; 124; line 281-282; line 432-433; Line 474; Line 547; Line 551; Line 604 ; line 647-648; Line 763

Line 18-22 Some redundancy in abstract, can be improvised. 

conclusion state about volatile oil and total phenols and flavanoids which never mentioned in the text. please check on conclusion that conclude the review correctly

Line 44- 46 some redundancy in writeup, can be improvised. 

Line 174 revision of sentence needed 

Line 255-257 was found repeated as in page 2 line 60-65

Line 285 revision of sentence (cleave what into AB) , spacing between secretase cleaved and app should be APP 

Line 328-330 tenses need to be revised and bit redundant 

Line 358 sentence was found a bit redundant 

Line 399-408 is more to decoction which is not reflected in the title. decoction might not reflect ginseng alone as synergetics or antagonistic. (suggest remove from manuscript)

Line437-438 redundant as repeated in Line 444 

Paragraph in 3.3 , the consistency of writeup for ACHE or AChE; BCHE or BChE; or Ach or ACh

Line 491-492; line 506-508 sentences can be rephrased 

Line 536 -538 is on combination which is not reflecting ginseng , suggest remove from manuscript

repeated statement between 562-567 with 358-376

Line 581 , proper citation for ZHAO 

Line 664 binding to it ,unclear what is this

Line 705 elusive or inconclusive 

Line 716-718 proper writen for Lac 

Line 746-750 please counter check whether DuShen tang is really P. ginseng or it can be other type of ginseng 

 

 

Author Response

Dear Editor and Reviewers:

Thank you for your works and the reviewer’s comments on our manuscript entitled “The research progress of the mechanism of Panax ginseng C. A.Meyer in the treatment of Alzheimer's disease and Application of Spatial Metabolomics” (2743371). Those comments are valuable and helpful for revising and improving our paper. We have adjusted the article to reduce the repetition rate and carefully revised the manuscript according to the comments. The details of revisions are listed below point-by-point and the main corrections in the manuscript were marked in red.

Thanks for your time and consideration.

 

Yours Sincerely,

Wei Wu

 

Reviewer 1:

The authors have done a very thorough reviewed on the P. ginseng in AD. It included with comprehensive writeup of different active compounds in P. ginseng and how it can help in the progression of AD. 

1. The title can be revised to reflect the title more appropriately. The title state the mechanism of Panax ginseng but it should be pharmacological action of active compound of  ginseng. The title mentioned treatment but the explanation is more to improvement of AD. 

Reply: For the title of the article, we have made changes according to your suggestions and marked those changes in red in the manuscript.

2. The study included the factors and the underlying mechanism of AD, however, there is a lack of signs & symptoms and current diagnosis of AD. 

Reply: We have made the following changes to your suggestion:We added Paragraph 4 to introduce signs & symptoms and the current diagnosis of AD in paragraphs 4.1 and 4.2.

3. The review highlighted the potential of  ginseng inAD, however, the selection of P. ginseng is not clearly elucidated and is that due to the cost or easily available.

Reply: We have made the following changes to your suggestion:We added the reason for choosing P. ginseng in lines 36-38.:  P. ginseng has a long history of use and is characterized by many pharmacological effects and few side effects. And marked it in red in the text.

4. The review highlighted mostly ginsenosides, there is not much on toerh compounds such as gintonin, polysaccharides, peptides in AD.

Reply: We have made the following changes to your suggestion:We have added Gintonin in AD treatment in lines 157-158 and 373-374, and polysaccharides in AD treatment in lines 274-276, and are represented in the corresponding graphs. All relevant changes have been marked in red in the article.Although compounds such as ginsenosides, polysaccharides and peptides have been found to have ameliorative effects on attention deficit disorder, there are far fewer studies related to them than to ginsenosides, and therefore fewer descriptions of these three components in this paper.

5. All the figure & table should be included into the text or to the manuscript. 

Reply: We have made the following changes to your suggestion:All the figure & table covered in the text have been inserted into the manuscript, including the Graphical Abstract.In addition, we cite and describe them in the text.

6. Table 1to 5, the effect only show target protein, it will be more complete to inlcude whether suppress, improve, enhance or etc, in more detailed

Reply: We have made the following changes to your suggestion. We have enriched the content of Table 1 to 5 according to the reviewer's request by adding the description of support, improve and enhance of target proteins.

7. Figure 3-7: signalling pathways.

Reply: We have made the following changes to your suggestion. We've made changes to the title of Figure and marked it in red in the text.

8. The consistency of scientific written for scientific name in italic (including  ginseng, in vitro, in vivo)

Reply: We have made the following changes to your suggestions: we have changed all P. ginseng to italics, removed all italics from “in vitroand in vivo” to “in vitro and in vivo”, and reddened them in the text.

9. Paragraph 2.5- consistency of using glycoconjugates or glycoconjugated compounds 

Reply: We have made the following changes to your suggestion:We have consistent glycoconjugates and glycoconjugated compounds in the text as glycoconjugates. And marked it in red.

10. subtitles of 3. Mechanism ... is actully more to pharmalogical effect

Reply: We have made the following changes to your suggestion:For subtitles of 3., we have amended the section title as requested. And marked it in red.

11. There are many wrong in text citation in the text (Error! ...) g. Line 106-107; 124; line 281-282; line 432-433; Line 474; Line 547; Line 551; Line 604 ; line 647-648; Line 763

Reply: We have made the following changes to your suggestion:For the portion of the citation that contained errors, we corrected errors in the formatting of images and table citations in the text in lines 95,109,214,215,317,347,409,414,455,478,546. And marked it in red.

12. Line 18-22 Some redundancy in abstract, can be improvised. 

Reply: We have made the following changes to your suggestion:We have revised this section of the abstract in line 15-17 to make it more concise and understandable. and highlighted in red.

13. conclusion state about volatile oil and total phenols and flavanoids which never mentioned in the text. please check on conclusion that conclude the review correctly

Reply： We thank you for this important suggestion, which was significant in the revision of this article. In this section of the original manuscript on P. ginseng constituents, we summarized almost all of the constituents, including some that have little to do with Alzheimer's disease. These now seem somewhat redundant, so we have removed this section.And a change was made to Figure1 and Graphical Abstract.

14. Line 44- 46 some redundancy in writeup, can be improvised. 

Reply：We have revised this section based on your suggestion to make the article neater.

15. Line 174 revision of sentence needed 

Reply：We have made the following changes to your suggestion:We have revised this sentence in line 151-156 and marked it red in the text.

16. Line 255-257 was found repeated as in page 2 line 60-65

Reply：We have made the following changes to your suggestion:For duplicates,We have deleted the sentence in lines 60-65 and retained the sentence in lines 195-198.

17. Line 285 revision of sentence (cleave what into AB), spacing between secretase cleaved and app should be APP 

Reply：We have made the following changes to your suggestion: We have restated the sentence in line 217-219 more clearly and accurately, and revised app to APP.It has been marked in red in the manuscript.

18. Line 328-330 tenses need to be revised and bit redundant

Reply：We have made the following changes to your suggestion:We have modified the tense of the sentence in line 250-251 and made simplifying changes, and marked it red in the text.

19. Line 358 sentence was found a bit redundant 

Reply：We removed this redundant sentence.

20. Line 399-408 is more to decoction which is not reflected in the title. decoction might not reflect ginseng alone as synergetics or antagonistic. (suggest remove from manuscript)

Reply：We have made the following changes to your suggestion:We have removed this section, including changes to Table 4, Table6, and Figure 6.

21. Line437-438 redundant as repeated in Line 444 

Reply：We have made the following changes to your suggestion:We have removed line 444and made changes to this part of lines 323-327

22. Paragraph in 3.3, the consistency of writeup for ACHE or AChE; BCHE or BChE; or Ach or Ach

Reply：We have made the following changes to your suggestion:In this section, we have chosen AChE,BChE,and ACh as the uniform writing style. And marked it in red.

23. Line 491-492; line 506-508 sentences can be rephrased 

Reply：Based on your suggestions, we have revised these two sections in line 364-365 and 377-378. And marked in red in the text.

24. Line 536 -538 is on combination which is not reflecting ginseng, suggest remove from manuscript

Reply： We have made the following changes to your suggestion:We have removed relevant content in this area, including Table 3 and Table 6.

25. repeated statement between 562-567 with 358-376

Reply： We have made the following changes to your suggestion:We have simplified both sections in line 423-424 and 270-271, and removed some repetitive descriptions and marked them red in the text.

26. Line 581, proper citation for ZHAO 

Reply： We have made the following changes to your suggestion:We have corrected the error in the citation in line 435.

27. Line 664 binding to it,unclear what is this

Reply： We have made the following changes to your suggestion:We have modified the expression of the language in line 507 to make the meaning of the sentence clearer. And marked them in red in the text.

28. Line 705 elusive or inconclusive 

Reply： We have made the following changes to your suggestion:We have restated this sectioninline 540-541 more clearly.And marked them in red in the text.

29. Line 716-718 proper writen for Lac 

Reply：We have made the following changes to your suggestion:We have added Lac.in line 577-578 to its full nameto make the article more accurate. And we also marked the changes as red in the article.

30. Line 746-750 please counter check whether DuShen tang is really  ginseng or it can be other type of ginseng 

Reply： In response to your suggestion, we conducted a literature search and found that although there is a paucity of literature on DuShen tang, the literature we do have suggests that DuShen tang is indeed P. ginsengand not some other ginseng, and the literature we refer in line553-556 also clearly states that Panax ginseng C.A. Meyer was used.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The review describes properly the aspects of P. ginseng in Alzheimer disease and shows important data to encourage researchers to see P. ginseng as a remarkable source of active ingredient to development of medicines or delivery systems. The work was weel presented and has quality enough to be published. My only advice to authors is improve the data discussions critically in order to present different point of views to readers. Additionally, the text cited references need to be improved.

Comments on the Quality of English Language

A few of gramatical erros need to be revised

Author Response

Dear Editor and Reviewers:

Thank you for your works and the reviewer’s comments on our manuscript entitled “The research progress of the mechanism of Panax ginseng C. A.Meyer in the treatment of Alzheimer's disease and Application of Spatial Metabolomics” (2743371). Those comments are valuable and helpful for revising and improving our paper. We have adjusted the article to reduce the repetition rate and carefully revised the manuscript according to the comments. The details of revisions are listed below point-by-point and the main corrections in the manuscript were marked in red.

Thanks for your time and consideration.

 

Yours Sincerely,

Wei Wu

 

 Reviewer 2:

The review describes properly the aspects of P. ginseng in Alzheimer disease and shows important data to encourage researchers to see P. ginseng as a remarkable source of active ingredient to development of medicines or delivery systems. The work was weel presented and has quality enough to be published. My only advice to authors is improve the data discussions critically in order to present different point of views to readers. Additionally, the text cited references need to be improved.

Reply：We have made the following changes to your suggestion:We have revised the data discussions of the article. In terms of references, we have corrected errors in literature citation and added citations to more valuable literature. Example:

1. Grabrucker, S.; Marizzoni, M.; Silajdžić, E.; Lopizzo, N.; Mombelli, E.; Nicolas, S.; Dohm-Hansen, S.; Scassellati, C.; Moretti, D.V.; Rosa, M.; et al. Microbiota from Alzheimer’s Patients Induce Deficits in Cognition and Hippocampal Neurogenesis. Brain 2023, 146, 4916–4934, doi:10.1093/brain/awad303.
2. Vandereyken, K.; Sifrim, A.; Thienpont, B.; Voet, T. Methods and Applications for Single-Cell and Spatial Multi-Omics. Nat Rev Genet 2023, 1–22, doi:10.1038/s41576-023-00580-2.
3. Scheltens, P.; Strooper, B.D.; Kivipelto, M.; Holstege, H.; Chételat, G.; Teunissen, C.E.; Cummings, J.; van der Flier, W.M. Alzheimer’s Disease. Lancet 2021, 397, 1577–1590, doi:10.1016/S0140-6736(20)32205-4.
4. Lee, C.H.; Ko, M.S.; Kim, Y.S.; Ham, J.E.; Choi, J.Y.; Hwang, K.W.; Park, S.-Y. Neuroprotective Effects of Davallia Mariesii Roots and Its Active Constituents on Scopolamine-Induced Memory Impairment in In Vivo and In Vitro Studies. Pharmaceuticals 2023, 16, 1606, doi:10.3390/ph16111606.

Comments on the Quality of English Language

A few of gramaticalerros need to be revised

Reply: English language of the manuscript has been carefully checked and polished by the editing service to improve readability of the manuscript.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

This review paper covers a wide range of topics related to the effects of Panax ginseng on Alzheimer's disease (AD), discussing mitochondrial damage, regulation of gut microbiota, and the application of spatial metabolomics. However, there are some areas that could be refined for clarity and precision. Here are a few suggestions:

1)      The title is a bit lengthy and could be more concise. Consider simplifying it while retaining the essential focus of the paper.

2)      Display Panax ginseng in italics in certain areas while leaving it in regular font elsewhere. Apply italics to "Panax ginseng" throughout the entire document.

3)      Some sentences are long and complex (also in abstract). Breaking them into smaller, more digestible segments can improve readability.

4)      There are a few instances of grammatical errors and awkward phrasing that could be revised for better readability. Proofread for grammar and sentence structure, ensuring clarity and coherence of ideas throughout the paper.

5)      Maintain consistency in the terminology used throughout the paper. For example, some sections refer to "Alzheimer's disease" while others mention "AD." Choose one and use it consistently.

6)      Break down the sections into more manageable parts with clear subheadings for each mechanism of action (Aβ aggregation, tau hyperphosphorylation, neurotransmitter regulation, mitochondrial damage, gut microbiota regulation, and spatial metabolomics etc.).

7)      Include a brief introductory paragraph summarizing the key aspects of Alzheimer's disease pathogenesis before delving into specific mechanisms and the role of P. ginseng.

8)      Provide more context and explanations where necessary, especially for less-known abbreviations, technical terms, or complex pathways, to aid readers in understanding.

9)      Emphasize the most significant or recent findings in the field related to P. ginseng's mechanisms in treating Alzheimer's disease.

10)  The figures need to be appropriately referenced. Each table/figure should be introduced and discussed in the main text where relevant.

11)  The conclusion can be further strengthened by summarizing key findings in a more concise and organized manner. It should reiterate the main points discussed in the paper and emphasize the significance of the findings.

Comments on the Quality of English Language

Extensive editing of English language required

Author Response

Dear Editor and Reviewers:

Thank you for your works and the reviewer’s comments on our manuscript entitled “The research progress of the mechanism of Panax ginseng C. A.Meyer in the treatment of Alzheimer's disease and Application of Spatial Metabolomics” (2743371). Those comments are valuable and helpful for revising and improving our paper. We have adjusted the article to reduce the repetition rate and carefully revised the manuscript according to the comments. The details of revisions are listed below point-by-point and the main corrections in the manuscript were marked in red.

Thanks for your time and consideration.

 

Yours Sincerely,

Wei Wu

 

 Reviewer 3:

This review paper covers a wide range of topics related to the effects of Panax ginseng on Alzheimer's disease (AD), discussing mitochondrial damage, regulation of gut microbiota, and the application of spatial metabolomics. However, there are some areas that could be refined for clarity and precision. Here are a few suggestions:

• The title is a bit lengthy and could be more concise. Consider simplifying it while retaining the essential focus of the paper.

Reply: We have made the following changes to your suggestion:

For the title of the article, whilepreserving the focus of the paper, we have simplified the title of the article and marked it in red in the manuscript.

• Display Panax ginsengin italics in certain areas while leaving it in regular font elsewhere. Apply italics to "Panax ginseng" throughout the entire docum

Reply: We have made the following changes to this suggestion you made:We have changed "Panax ginseng" and "P.ginseng" to italics in the text.

• Some sentences are long and complex (also in abstract). Breaking them into smaller, more digestible segments can improve readability.

Reply: In response to this suggestion of yours, we have revised most of the sentences throughout the text. We have removed some complex sentences to make the article more readable.

• There are a few instances of grammatical errors and awkward phrasing that could be revised for better readability. Proofread for grammar and sentence structure, ensuring clarity and coherence of ideas throughout the paper.

Reply: In response to your suggestions, we have revised and standardised the grammar, wording and sentence structure throughout the manuscript.

• Maintain consistency in the terminology used throughout the paper. For example, some sections refer to "Alzheimer's disease" while others mention "AD." Choose one and use it consistently.

Reply: We have made the following changes to your suggestion:We have modified this issue by using "AD" consistently throughout the text, except for the application of "Alzheimer's disease" at the beginning.

• Break down the sections into more manageable parts with clear subheadings for each mechanism of action (Aβ aggregation, tau hyperphosphorylation, neurotransmitter regulation, mitochondrial damage, gut microbiota regulation, and spatial metabolomics etc.).

Reply: According to your comments, we have added subheadings for these sections and revised some related content accordingly.

• Include a brief introductory paragraph summarizing the key aspects of Alzheimer's disease pathogenesis before delving into specific mechanisms and the role of ginseng.

Reply: In response to your comment, we have included a summary paragraph at the beginning of paragraph 3and at the beginning of each mechanism paragraph.

• Provide more context and explanations where necessary, especially for less-known abbreviations, technical terms, or complex pathways, to aid readers in understanding.

Reply: We have added explanations of less-known abbreviations, technical terms, or complex pathways when they are first introduced.

• Emphasize the most significant or recent findings in the field related to ginseng's mechanisms in treating Alzheimer's disease. 

Reply: In response to this suggestion made by you, in lines 80-86, andthe paragraphs “Conclusions”we have highlighted the focus and novelty of this paper.

• The figures need to be appropriately referenced. Each table/figure should be introduced and discussed in the main text where relevant.

Reply: We have made the following adjustments in response to your questions. We have placed the table/figure near the relevant content and discussed himin lines 95,109,214,215,317,347,409,414,455,478,546.

• The conclusion can be further strengthened by summarizing key findings in a more concise and organized manner. It should reiterate the main points discussed in the paper and emphasize the significance of the findings.

Reply: We have revised the conclusion.The main points of the article are summarised in the conclusion, removing some of the less important elements.

Comments on the Quality of English Language

Extensive editing of English language required

Reply: English language of the manuscript has been carefully checked and polished by the editing service to improve readability of the manuscript.

Author Response File: Author Response.pdf

Reviewer 4 Report

Comments and Suggestions for Authors

The manuscript "The research progress of the mechanism of Panax ginseng C. A. Meyer in the treatment of Alzheimer's disease and Application of Spatial Metabolomics" describes the main biologically active components of P. ginseng and their possible application to the Alzheimer's disease (AD) treatment. Several main mechanisms of AD genesis were identified including amyloid plaques generation and aggregation, tau hyperphosphorylation and others, and a thorough work was done to show the possible effect of ginseng components onto them. In general, review seem well-written and useful for those who works in the field of ginseng research trying to apply it for fighting against AD. The schemes are of special interest. However, I feel that some point should be still paid attention to:

1. Usually, a review starts with the discussion of the previous reviews devoted to the same topic to show the gaps in the knowledge. Some relevant reviews are referenced (Refs. 6, 97), but are not discussed in the manner mentioned above. In addition, the following reviews are known [10.3390/molecules28155716, 10.2174/1568026615666150813143753]. What new brings this manuscript when compared to the pieces published already?

2. Many research papers studies only the action of a single component of giseng that was previously isolated. However, CTM used ginseng plant as a whole considering synergetic effect of biologicall active compounds. Is there some information available in the literature about such synergy regarding the AD treatment?

3. I believe, the manuscript would be even more thorough and useful if some data on the clinical trials (if any exist) would be added to the text. Molecular mechanisms and model animals are good, but what about the real effect on human?

4. There are multiple warnings across the text "Error! Reference source not found." on, e.g. lines 107, 125, 281 etc. Please, make the necessary corrections.

Author Response

Dear Editor and Reviewers:

Thank you for your works and the reviewer’s comments on our manuscript entitled “The research progress of the mechanism of Panax ginseng C. A.Meyer in the treatment of Alzheimer's disease and Application of Spatial Metabolomics” (2743371). Those comments are valuable and helpful for revising and improving our paper. We have adjusted the article to reduce the repetition rate and carefully revised the manuscript according to the comments. The details of revisions are listed below point-by-point and the main corrections in the manuscript were marked in red.

Thanks for your time and consideration.

 

Yours Sincerely,

Wei Wu

 

 Reviewer 4:

The manuscript "The research progress of the mechanism of Panax ginseng C. A. Meyer in the treatment of Alzheimer's disease and Application of Spatial Metabolomics" describes the main biologically active components of P. ginseng and their possible application to the Alzheimer's disease (AD) treatment. Several main mechanisms of AD genesis were identified including amyloid plaques generation and aggregation, tau hyperphosphorylation and others, and a thorough work was done to show the possible effect of ginseng components onto them. In general, review seem well-written and useful for those who works in the field of ginseng research trying to apply it for fighting against AD. The schemes are of special interest. However, I feel that some point should be still paid attention to:

 

1. Usually, a review starts with the discussion of the previous reviews devoted to the same topic to show the gaps in the knowledge. Some relevant reviews are referenced (Refs. 6, 97), but are not discussed in the manner mentioned above. In addition, the following reviews are known [10.3390/molecules28155716, 10.2174/1568026615666150813143753]. What new brings this manuscript when compared to the pieces published already?

Reply: In response to your suggestion, we have described the relevant content in line 80-81, compared it with the review literature cited in the article, and proposed the characteristics and novelty of this article.For the two articles you brought up, we compared them as follows:

• In the article [10.3390/molecules28155716], the herb studied was American ginseng (AG), whereas the herb studied in our review was ginseng, and our review covers more mechanisms of action (including modulation of gut microbes) and signalling pathways than this article.
• In the article [10.2174/1568026615666150813143753], a review of clinical studies was mainly provided, while our review was more of a summary of the results of animal and cellular experiments on the mechanism of action. So there are still some differences between the two.

2. Many research papers studies only the action of a single component of ginseng that was previously isolated. However, CTM used ginseng plant as a whole considering synergetic effect of biologicall active compounds. Is there some information available in the literature about such synergy regarding the AD treatment?

Reply: We have searched the relevant literature, but the literature searched is a combination of P. ginseng monomers as well as some ginseng compounds, and there is no synergistic effect or mechanism of P. ginseng searched. However, we do note that the use of ginseng as a whole has some ameliorative effects on AD, as shown in the study on Doksangtangin line553-556 and the clinical study on P. ginseng in paragraph 4.3.

3. I believe, the manuscript would be even more thorough and useful if some data on the clinical trials (if any exist) would be added to the text. Molecular mechanisms and model animals are good, but what about the real effect on human?

Reply: We have made the following changes to this suggestion you made:We have added clinical studies inparagraph 4.3. However, the clinical studies on P.ginseng in the treatment of AD are not deep enough and comprehensive enough, so there is less content in this section. It is hopeful that these researches would provide possibilities and foundation for the future clinical studies.

4. There are multiple warnings across the text "Error! Reference source not found." on, e.g. lines 107, 125, 281 etc. Please, make the necessary corrections.

Reply:  We have made the following changes to this suggestion you made: For the portion of the citation that contained errors, we corrected errors in the formatting of images and table citations in lines 95, 109, 214, 215, 317, 347, 409, 414, 455, 478, 546.. And marked it in red.

Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

I read the entire manuscript, and the authors gave careful consideration to every comment. Along with the earlier, much information was added. The manuscript can be accepted in the present form.

Comments on the Quality of English Language

Minor corrections needed.

Reviewer 4 Report

Comments and Suggestions for Authors

Authors have made the necessary corrections to the text improving it significantly and gave the answers to the concerns raised in the satisfactory manner. I support publishing the manuscript in the revised form.