Calamintha incana (Sm.) Helder: A New Phytoextract with In Vitro Antioxidant and Antidiabetic Action

Round 1

Reviewer 1 Report

The manuscript of Arwa Althaher and colleagues extracted some natural products from Calamintha incana (Sm.) Helder leaves. The antioxidant, antidiabetic and cytotoxicity properties of ethanolic phytoextracts were systematically evaluated. Overall, this manuscript maybe a meaningful work that helps to discover some new active skeleton with unique pharmacological activity. However, despite of these interestingly results, many problems need to be addressed. Thus, I think this manuscript needs major revisions before possible publication in Applied Sciences. The comments are provided as below:

 

1.      In Abstract section, “50” of “IC50” should be written in subscript.

2.      2. Materials and Methods. “3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT)” should be written as “3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H(italic)-tetrazolium bromide (MTT)”.

3.      2.5 Cell culture. “5% CO2” should be written as “5% CO₂(subscript).”.

4.      2.6.2 Reducing Power Ability. What is meaning of D. H2O?

5.      3.2. Cytotoxicity. “C. incana” should be written in italic. “IC50 value” should be written as “IC₅₀ (subscript) value ”. In addition, the positive control should be added.

6.      For these pharmacology assay including cytotoxicity, and antioxidant activity, why not choose all chemical composition of C. incana ethanolic extract to test?

7.      4. Discussion. “S and R” of “(2S,4R)-p-Mentha-6,8-diene 2-hydroperoxide” should be written in italic.

 

Author Response

REVIEWERS#1

The manuscript of Arwa Althaher and colleagues extracted some natural products from Calamintha incana (Sm.) Helder leaves. The antioxidant, antidiabetic and cytotoxicity properties of ethanolic phytoextracts were systematically evaluated. Overall, this manuscript maybe a meaningful work that helps to discover some new active skeleton with unique pharmacological activity. However, despite of these interestingly results, many problems need to be addressed. Thus, I think this manuscript needs major revisions before possible publication in Applied Sciences. The comments are provided as below:

 

1. In Abstract section, “50” of “IC50” should be written in subscript.

Reply: The correction was done (highlighted).

2. Materials and Methods. “3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT)” should be written as “3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H(italic)-tetrazolium bromide (MTT)”.

Reply: The correction was done (highlighted)

3. 5 Cell culture. “5% CO2” should be written as “5% CO₂ (subscript).”

Reply: The correction was done (highlighted).

4. 6.2 Reducing Power Ability. What is meaning of D. H2O?

Reply: The correction was done (highlighted). D.H2O: Distilled water

5. 2. Cytotoxicity. “C. incana” should be written in italic. “IC50 value” should be written as “IC₅₀(subscript) value ”. In addition, the positive control should be added.

Reply: The correction was done (highlighted). Doxorubicin was used as positive control.

6. For these pharmacology assay including cytotoxicity, and antioxidant activity, why not choose all chemical composition of  incanaethanolic extract to test?

Reply: In this study, we used the leaves of the mentioned plant then form ethanolic extract and tested this extract as it is for all assays

7. 4. Discussion. “S and R” of “(2S,4R)-p-Mentha-6,8-diene 2-hydroperoxide” should be written in italic.

Reply: The correction was done (highlighted).

 

Reviewer 2 Report

The manuscript by Althaher and Mastinu describes the identification of chemical compounds in an ethanolic extract of Calamintha incana (Sm.) Helder leaves and its in vitro antioxidative and antidiabetic activities. There are a lot of points as indicated below that need to be addressed by the authors before publication in the journal.

1. What is “(Sm.”)? Most readers don’t know this term.

2. “alpha-” and “α-” are mixing throughout the text. Please be consistent with the terminology.

3. Line 42: “Diabetes mellitus (DM)” should be “DM.”

4. Line 59: “α” is missing.

5. Line 65: “diabetes mellitus” should be “DM.”

6. Line 282: Remove “-” and “-hydrate” to be “2,2-diphenyl-1-picrylhydrazyl.”

7. Lines 282-284: The mechanism of the DPPH test is wrong. Antioxidants donate a hydrogen atom to the DPPH to produce the hydrogenated DPPH (DPPH-H).

8. “3+” in line 292 and “2+” in line 293 are in superscript.

9. Line 312: Please explain why acarbose was used at this stage, because most readers don’t know acarbose.

10. Table 1 is confusing and should be improved.

11. Line 418-419: “In addition to…” is nonsense. Please rewrite it.

12. Lines 450-454: “DPP-IV” and “DPPIV” are mixing.

13. More discussion about the extract is needed in the “4.4 Antidiabetic activity” section, because most sentences are for the explanations for the enzymes. Furthermore, the proposed mechanisms should be provided. Why do the components of the extract act as an antioxidant and as an inhibitor of the enzymes? Are there any similarities in the molecular shapes between the positive controls and the components of the extract?

14. Line 468: This reviewer disagrees with the authors’ comment that the toxicity of the extract was low. The authors need to rephrase this statement.

15. “C. incana” should be in italic throughout the text.

Author Response

REVIEWER#2

The manuscript by Althaher and Mastinu describes the identification of chemical compounds in an ethanolic extract of Calamintha incana (Sm.) Helder leaves and its in vitro antioxidative and antidiabetic activities. There are a lot of points as indicated below that need to be addressed by the authors before publication in the journal.

 

1. What is “(Sm.”)? Most readers don’t know this term.

Reply: "Sm.", author abbreviation for "Smith". The surname Smith, as a taxonomic authority

2. “alpha-” and “α-” are mixing throughout the text. Please be consistent with the terminology.

Reply: The correction was done (highlighted).

3. Line 42: “Diabetes mellitus (DM)” should be “DM.”

Reply: The correction was done (highlighted).

4. Line 59: “α” is missing.

Reply: The correction was done (highlighted).

5. Line 65: “diabetes mellitus” should be “DM.”

Reply: The correction was done (highlighted).

6. Line 282: Remove “-” and “-hydrate” to be “2,2-diphenyl-1-picrylhydrazyl.”

Reply: The correction was done (highlighted).

7. Lines 282-284: The mechanism of the DPPH test is wrong. Antioxidants donate a hydrogen atom to the DPPH to produce the hydrogenated DPPH (DPPH-H).

Reply: The correction was done (highlighted).

8. “3+” in line 292 and “2+” in line 293 are in superscript.

Reply: The correction was done (highlighted).

9. Line 312: Please explain why acarbose was used at this stage, because most readers don’t know acarbose.

Reply: Acarbose s an anti-diabetic drug used to treat diabetes mellitus type 2 and, in some countries. Acarbose lowers blood sugar by preventing the breakdown of starch into sugar.

10. Table 1 is confusing and should be improved.

Reply: Reply: The correction was done (highlighted).

11. Line 418-419: “In addition to…” is nonsense. Please rewrite it.

Reply: The sentence was removed

12. Lines 450-454: “DPP-IV” and “DPPIV” are mixing.

Reply: The correction was done (highlighted).

13. More discussion about the extract is needed in the “4.4 Antidiabetic activity” section, because most sentences are for the explanations for the enzymes. Furthermore, the proposed mechanisms should be provided. Why do the components of the extract act as an antioxidant and as an inhibitor of the enzymes? Are there any similarities in the molecular shapes between the positive controls and the components of the extract?

Reply: The exact mechanism need more studies to be confirmed. But we add the suggested mechanism of action.

medicinal plants improve the body's antioxidant system and insulin control, making them a safer choice for treating obesity and diabetes [19]. The antioxidant activity of this plant extract was confirmed in this investigation. Moreover, the extract contains several phytochemicals such as phenols, terpenes, fatty acids, and others that may be responsible for its antiradical and antidiabetic properties.

14. Line 468: This reviewer disagrees with the authors’ comment that the toxicity of the extract was low. The authors need to rephrase this statement.

Reply: The correction was done (highlighted).

15. “C. incana” should be in italic throughout the text.

Reply: The correction was done (highlighted).

 

Reviewer 3 Report

I have evaluated the manuscript entitled " Calamintha incana (Sm.) Helder: a new phytoextract with invitro antioxidant and antidiabetic action". the study aimed to evaluate the antioxidant and antidiabetic properties of ethanolic phytoextracts derived from Calamintha incana (Sm.). Idea of study is interesting and falls within the scope of journal; however the manuscript cannot be accepted for publication in its current form. Moreover the manuscript can be considered for publication after drastic revisions.

General comments:

1.     Currently manuscript look like a thesis rather than a research article. The manuscript should be rewrite.

2.     Hypothesis and novelty of study should be elaborated in more detail.

3.     Use scientific language and significant editing of language is also recommended.

*Introduction : must be improve with more information

*Materials and methods: clear statement and good written

*Result : must rewrite using scientific language

* discussion: must be more depth contain mor information

*References : it have recently Ref. so that it is good statement

 

   

 

 

 

Author Response

REVIEWER#3

 

I have evaluated the manuscript entitled " Calamintha incana (Sm.) Helder: a new phytoextract with invitro antioxidant and antidiabetic action". the study aimed to evaluate the antioxidant and antidiabetic properties of ethanolic phytoextracts derived from Calamintha incana (Sm.). Idea of study is interesting and falls within the scope of journal; however the manuscript cannot be accepted for publication in its current form. Moreover the manuscript can be considered for publication after drastic revisions.

REPLY: We thank the Reviewer for taking the time to review the manuscript. We have improved the manuscript as requested.

General comments:

1. Currently manuscript look like a thesis rather than a research article. The manuscript should be rewrite.

REPLY: the observation is pertinent; the entire manuscript has been revised and improved in its parts.

2. Hypothesis and novelty of study should be elaborated in more detail.

REPLY: The study should be considered innovative due to the particularity of the plant species studied. Calamintha incana hydroalcoholic extracts were obtained from plants grown in Jordan, in an ecological area that has influenced the production of specific volatile secondary metabolites. This aspect has been emphasized throughout the manuscript.

3. Use scientific language and significant editing of language is also recommended.

REPLY: We followed the Reviewer's suggestion and tried to improve the language of the manuscript by making it more scientific.

*Introduction : must be improve with more information

REPLY: More details have been added in the introduction emphasizing the specificity and importance of our research.

*Materials and methods: clear statement and good written

*Result : must rewrite using scientific language

* discussion: must be more depth contain mor information

*References : it have recently Ref. so that it is good statement

REPLY: the results and discussion section were revised as required. We remain open to further revisions if necessary.

Reviewer 4 Report

The manuscript entitled "Calamintha incana (Sm.) Helder: a new phytoextract with in-vitro antioxidant and antidiabetic action" summarizes the results of a remarkable research. In the framework of the research, a medicinal herb (C. incana) was examined, and the ethanolic extracts made from its leaves were extensively examined. During the tests, the most important antioxidant and antidiabetic effects of the extracts were determined.

The introductory part of the study, as well as the presentation of the material and method, are sufficiently detailed. The presentation of the results is also sufficiently detailed and well supported. The test results represented: phytochemical analysis of C. incana ethanolic extract, its cytotoxicity, antioxidant activity and antidiabetic activity.

The discussion chapter of the manuscript is sufficiently detailed. The novelty of the study is based on the examination of the effects of leaf extracts, in comparison with the previous results of the natural oil of the examined plant.

In order to improve the quality of the work, I recommend the following corrections and modifications:

- in line 63, I recommend indicating the full scientific name of the examined plant (Calamintha incana).

- in line 125, the spelling of CO2 is correctly CO₂

- in lines 171, 267, 269, 281, 308, 323, 367, 371, 372, 378, 389, 393, 402, 404, 407 I recommend the italic spelling of the scientific name of the examined plant (C. incana).

- in line 476, the spelling of in vivo is also suggested in italics.

After the implementation of the modifications suggested above, I recommend publishing the manuscript in the form of a scientific article.

Author Response

Reviewer#4

 

The manuscript entitled "Calamintha incana (Sm.) Helder: a new phytoextract with in-vitro antioxidant and antidiabetic action" summarizes the results of a remarkable research. In the framework of the research, a medicinal herb (C. incana) was examined, and the ethanolic extracts made from its leaves were extensively examined. During the tests, the most important antioxidant and antidiabetic effects of the extracts were determined.

The introductory part of the study, as well as the presentation of the material and method, are sufficiently detailed. The presentation of the results is also sufficiently detailed and well supported. The test results represented: phytochemical analysis of C. incana ethanolic extract, its cytotoxicity, antioxidant activity and antidiabetic activity.

The discussion chapter of the manuscript is sufficiently detailed. The novelty of the study is based on the examination of the effects of leaf extracts, in comparison with the previous results of the natural oil of the examined plant.

In order to improve the quality of the work, I recommend the following corrections and modifications:

REPLY: We thank the Reviewer for his attention in reviewing our manuscript. We believe that after his indications the manuscript has improved considerably.

- in line 63, I recommend indicating the full scientific name of the examined plant (Calamintha incana).

REPLY: done

- in line 125, the spelling of CO2 is correctly CO_2\

REPLY: done

- in lines 171, 267, 269, 281, 308, 323, 367, 371, 372, 378, 389, 393, 402, 404, 407 I recommend the italic spelling of the scientific name of the examined plant (C. incana).

REPLY: done

- in line 476, the spelling of in vivo is also suggested in italics.

REPLY: done

Round 2

Reviewer 1 Report

The authors have improved the paper and answered all the questions. I suggest the paper is ready for publication.