Effects of Hydrolysates from Silkworms Fed Cudrania tricuspidata Leaves on Improvement of Memory in Rats with Impaired Memory Induced by Scopolamine

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

1. The resolution of Figures 2-7 is not high, it is recommended to replace the high-resolution images

2.Is the hydrolysis product of Cudrania tricuspidata leaves a pure substance? If not, would the author consider obtaining a pure substance for subsequent activity experiments, which would make the activity experiment more convincing.

3.In the SDS experiment, the author only provided a molecular weight marker with a minimum of 5Kpa. Should we consider using a marker with a smaller molecular weight to make the comparison more convincing.

Comments on the Quality of English Language

Minor editing of English language required

Author Response

We are sincerely grateful for your thorough consideration and scrutiny of our manuscript, “Effects of hydrolysates from silkworm fed Cudrania tricuspidata leaves on improvement of memory in rats with impaired memory induced by scopolamine”, manuscript ID applsci-2621735. Through the accurate comments made by the reviewer, we better understand the critical issues in this paper. We have revised the manuscript according to the reviewer’s suggestions. We acknowledge that the scientific quality of our manuscript was improved by the scrutinizing efforts of the reviewers. We did best to make modifications based on the reviewers' suggestions and point-by-point responses to the reviewer’ comments are provided below.

Point-by-point response to Comments and Suggestions for Authors

Comment 1: The resolution of Figures 2-7 is not high, it is recommended to replace the high-resolution images.

Response 1: Thank you for pointing this out and we agree with this comment. Therefore, we replaced the resolution in figure 2-7 with the highest possible.

Comment 2: Is the hydrolysis product of Cudrania tricuspidata leaves a pure substance? If not, would the author consider obtaining a pure substance for subsequent activity experiments, which would make the activity experiment more convincing.

Response 2: Yes, it is a pure substance. It is a product that has been dried and powdered without any additives. 

Comment 3: In the SDS experiment, the author only provided a molecular weight marker with a minimum of 5Kpa. Should we consider using a marker with a smaller molecular weight to make the comparison more convincing.

Response 3: Thank you for pointing this out and we agree with this comment. It would have been appropriate to use a marker with a smaller molecular weight. Please understand that the major protein size in silkworm fed Cudrania tricuspidata leaves is 17~48 kDa, and our research was focused on effectively hydrolyzing proteins larger than 17 kDa through enzymatic treatment.

I would appreciate your understanding if there is anything insufficient about the author's response and please give us more detailed comments. 

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The paper by Gyu-Mi An et al is a great work designed and presented.

 

doubts and suggestions

 

1. I suggest putting the Organ weight data

2. SDS-PAGE determinations are part of the studies. The relevant thing would be to carry out studies with Western Blot. Are they able to carry them out? in different experimental conditions?

Author Response

We are sincerely grateful for your thorough consideration and scrutiny of our manuscript, “Effects of hydrolysates from silkworm fed Cudrania tricuspidata leaves on improvement of memory in rats with impaired memory induced by scopolamine”, manuscript ID applsci-2621735. Through the accurate comments made by the reviewer, we better understand the critical issues in this paper. We have revised the manuscript according to the reviewer’s suggestions. We acknowledge that the scientific quality of our manuscript was improved by the scrutinizing efforts of the reviewers. We did best to make modifications based on the reviewers' suggestions and point-by-point responses to the reviewer’ comments are provided below.

Point-by-point response to Comments and Suggestions for Authors

Comment 1: I suggest putting the Organ weight data.

Response 1: Thank you for pointing this out and we agree with this comment. Therefore, we added organ weight data to the manuscript as a table (line 213~222)

Comment 2: SDS-PAGE determinations are part of the studies. The relevant thing would be to carry out studies with Western Blot. Are they able to carry them out? in different experimental conditions?

Response 2: Please understand that we are unable to perform western blots at this time. The major protein size in silkworm fed Cudrania tricuspidata leaves is 17~48 kDa and we focused on using SDS-PAGE to determine whether these proteins were effectively hydrolyzed. In addition, experiments measuring non-protein nitrogen, which we did not write about in the manuscript, showed that enzymatic treatment effectively hydrolyzed proteins in silkworm fed Cudrania tricuspidata leaves.

I would appreciate your understanding if there is anything insufficient about the author's response and please give us more detailed comments. 

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The manuscript from An et al describes the effects of hydrolysates of silkworms that have been fed Cudrania tricuspidata on rats with impaired memory. The work appears to have been carefully performed and is clearly described. The conclusions drawn are justified by the results presented.

The authors appear to assume that the effects are due to peptides or small proteins in the hydrolysate. The possibility that a non-peptidic small molecule could be responsible does not seem to have been considered. A more thorough characterization of the hydrolysate would enable a better understanding of the active substance.

It would also be interesting to know if hydrolysates from silkworms fed a different diet showed similar effects. The text suggests that these studies have been carried out but does not discuss them.

Comments on the Quality of English Language

A few suggestions to help improve the English

line 2   replace "silkworm" with "silkworms"

line 56   Perhaps "silkworm" should be replaced with "silkworm extract", as it seems that the silkworms are not used whole but are processed.

line 64   italicize "Morus alba"

line 72   Are "ordinary mulberry leaves" leaves from Morus alba?

line 230   Define "latency", perhaps in the Materials and Methods section.

 

Author Response

We are sincerely grateful for your thorough consideration and scrutiny of our manuscript, “Effects of hydrolysates from silkworm fed Cudrania tricuspidata leaves on improvement of memory in rats with impaired memory induced by scopolamine”, manuscript ID applsci-2621735. Through the accurate comments made by the reviewer, we better understand the critical issues in this paper. We have revised the manuscript according to the reviewer’s suggestions. We acknowledge that the scientific quality of our manuscript was improved by the scrutinizing efforts of the reviewers. We did best to make modifications based on the reviewers' suggestions and point-by-point responses to the reviewer’ comments are provided below.

Point-by-point response to Comments and Suggestions for Authors

Comment 1: The authors appear to assume that the effects are due to peptides or small proteins in the hydrolysate. The possibility that a non-peptidic small molecule could be responsible does not seem to have been considered. A more thorough characterization of the hydrolysate would enable a better understanding of the active substance.

Response 1: Thank you for pointing this out and we agree with this comment. Therefore, we have added to the manuscript that further studies are needed to improve our understanding of the active substance (line 401~403).

Comment 2: It would also be interesting to know if hydrolysates from silkworms fed a different diet showed similar effects. The text suggests that these studies have been carried out but does not discuss them.

Response 2: Thank you for pointing this out and we agree with this comment. Therefore, we wrote as a limitation that we did not compare the functionality of silkworm hydrolysate fed a different diet with that of silkworm hydrolysate fed Cudrania tricuspidata leaves (line 397~399).

Comment 3: A few suggestions to help improve the English
line 2 replace "silkworm" with "silkworms"
line 56 Perhaps "silkworm" should be replaced with "silkworm extract", as it seems that the silkworms are not used whole but are processed
line 64 italicize "Morus alba"
line 72 Are "ordinary mulberry leaves" leaves from Morus alba?
line 230 Define "latency", perhaps in the Materials and Methods section.

Response 3: Thank you for pointing this out and we agree with this comment. Therefore, we have modified the English you suggested (line 2, 56, 64, 72, 142~143, 234)

I would appreciate your understanding if there is anything insufficient about the author's response and please give us more detailed comments. 

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Line 303  mU/m should be  mU/mL;

  line 418 missing a period.

Author Response

We are sincerely grateful for your thorough consideration and scrutiny of our manuscript. We did best to make modifications based on the reviewers' suggestions and point-by-point responses to the reviewer’ comments are provided below.

Point-by-point response to Comments and Suggestions for Authors

Comment 1: Line 303 mU/m should be mU/mL

Response 1: Thank you for pointing this out and we agree with this comment. Therefore, we corrected mU/m to mU/mL (line 303).

Comment 2: line 418 missing a period

Response 2: Thank you for pointing this out and we agree with this comment. Therefore, we wrote a period on line 418.

Author Response File: Author Response.pdf