Talk:WikiJournal of Medicine/Impact of xenogenic mesenchymal stem cells secretome on a humoral component of the immune system

From Wikiversity
Jump to navigation Jump to search

WikiJournal of Medicine
Open access • Publication charge free • Public peer review • Wikipedia-integrated

WikiJournal of Medicine is an open-access, free-to-publish, Wikipedia-integrated academic journal for Medical and Biomedical topics. <seo title=" WJM, WikiJMed, Wiki.J.Med., WikiJMed, Wikiversity Journal of Medicine, WikiJournal Medicine, Wikipedia Medicine, Wikipedia medical journal, WikiMed, Wikimedicine, Wikimedical, Medicine, Biomedicine, Free to publish, Open access, Open-access, Non-profit, online journal, Public peer review "/>

<meta name='citation_doi' value='10.15347/WJM/2023.004'>

Article information

Authors: Vitalii Moskalov[b][i] , Olena Koshova[a] , Sabina Ali[c] , Nataliia Filimonova[a] , Irina Tishchenko[a] 

See author information ▼
  1. 1.0 1.1 1.2 Ministry of Health of Ukraine, National University of Pharmacy, Kharkiv, Ukraine
  2. V. N. Karazin Kharkiv National University, Kharkiv, Ukraine
  3. National Academy of Sciences of Ukraine, Institute for Problems of Cryobiology and Cryomedicine, Kharkiv, Ukraine
  1. mosrus93@gmail.com

 

Review invitations sent

Dear authors,

My name is Athikhun Suwannakhan, I am the Editorial Board member of WikiJournal of Medicine who will be responsible for handling the Peer Review of your submission.

We strive to make the review process as transparent and useful for our authors as possible. At the moment, your manuscript has already been sent to a panel of experts, and we will let you know as they return their comments. The final decision for publication lies with the Editor in Chief and the rest of the Editorial Board.

Thank you for choosing WikiJournal of Medicine. I hope you value our peer review process, and please do not hesitate to contact me should you have any further comments or questions.

Kindest regards,

Athikhun Suwannakhan, PhD.

--Athikhun.suw (discusscontribs) 00:26, 2 January 2023 (UTC)[reply]

Dear colleagues!
We finalized the manuscript of the article in accordance with the recommendations of the reviewers. All recommendations were taken into account (perhaps the reviewers will consider that we have not fully taken into account them, but we have carefully worked out all the issues). I am attaching a list of improvements made:
Refinement of the manuscript taking into account the review 1
Material and methods
1) We have attached a photo of cells differentiated in three directions. We added information about the identification of mesenchymal stem cells.
2) We have added information about experimental animals (including species, sex, age and body weight).
3) We have clarified the methodology for modeling secondary immunodeficiency in mice, and corrected technical errors.
4) We have clarified the methodology for calculating AFC.
Refinement of the manuscript taking into account the review 2
Introduction
1)               We have added literature data on the immunobiological properties of the MSCs.
Material and methods
2)               We have specified the FBS origin used in this study.
3)               We have included speed and time for centrifugation.
4)               We`ve attached a photo of cells differentiated in three directions. We added information about the identification of mesenchymal stem cells.
5)               We have added information about cell density of MSCs seeded for secretome preparation.
6)               We have clarified secretome preparation methods.
7)               We have described a method for measuring the proteins in this study.
8)               We have clarified the methodology for modeling secondary immunodeficiency in mice, and corrected technical errors.
Results
9)               We have explained the results linking with figures
10)             The sentence “As for the hemagglutinin (HA) titers, the control and reference drug were on the same level by this indicator.” was corrected.
11)            The sentence "the intramuscular route of administration also turned out to be more effective than the subcutaneous one.” has been clarified with p-value; we have added a link on figure.
12)            We have clarified dose dependence of the observed effects on the humoral arm of immunity with statistical analysis data.
13)            We have explained why we had decided to test the survival rate, FCA count, and HA titer in mice with secondary immunodeficiency in the minimal concentration.
15)            We have added revised illustrations.
Discussion
14)            We have explained and discussed more about the recusing effect of the secretome in secondary immunodeficiency mice.
16)            We have marked the most important parameters of humoral immunity in our opinion with justification.
17)            We have discussed more references to support the effect of xenogeneic MSCs on the humoral immune response, but we have not found literature sources about such effects of secretome.
We ask you to forward the revised manuscript for final consideration
Sincerely, Vitalii Moskalov Vitalii moskalov (discusscontribs) 04:18, 15 May 2023 (UTC)[reply]


Peer review 1


Review by anonymous peer reviewer , Published papers on stem cell differentiation
These assessment comments were submitted on , and refer to this previous version of the article

  • English language editing is required throughout the manuscript including the title.
  • Concerning the isolation of mesenchymal stem cells (MSCs) from bovine’s bone marrow, characterization of MSCs is lacking such as by observing cell morphology, cell surface antigen molecule profiling, and multi-lineages differentiation. How do verify that these isolated cells are MSCs? Are there pictures of the isolated cells?
  • Information on the animals are required including species, sex, age, and body weight.
  • The following sentence is unclear: “Secondary immunodeficiency was modeled by daily subcutaneously administration in the nuchal region of 2.5 mg/kg of hydrocortisone acetate (HCA).” Please rephrase.
  • How long was the daily subcutaneous administration of HCA?
  • How do you verify the AFC?
  • Is there any other evidence of AFC characterization other than macroscopically visible zones of hemolysis counting?

Peer review 2


Review by anonymous peer reviewer , Published papers on human induced pluripotent stem cells
These assessment comments were submitted on , and refer to this previous version of the article

  1. Literature review about xenogenic mesenchymal stem cells (MSCs)-derived secretome is lacking.
  2. In the methods, please specify the FBS origin used in this study.
  3. Authors should include speed and time for centrifugation.
  4. What are the criteria used to test the characteristics of MSCs? Have authors characterized the isolated bovine MSCs? The authors may include the results of MSC characteristic tests as supplement data.
  5. What is the cell density of MSCs seeded for secretome preparation?
  6. Secretome preparation methods must be clearly explained.
  7. How did the authors measure the proteins in this study?
  8. To create a secondary immunodeficiency mice model, the authors injected HCA daily. How long?
  9. The results were not explained linking with figures. Readers must guest with themselves. Please read and correct throughout the result section.
  10. The sentence “As for the hemagglutinin (HA) titers, the control and reference drug were on the same level by this indicator.”, please explain more in terms of statistics.
  11. The sentence” the intramuscular route of administration also turned out to be more effective than the subcutaneous one.”, what figure indicates this sentence? How about p-value?
  12. Could the authors clarify about dose dependence of the observed effects on the humoral arm of immunity? What statistics are used for analyzing this effect?
  13. Why did the authors choose to test the survival rate, FCA count, and HA titer in mice with secondary immunodeficiency? How about higher concentrations?
  14. Please explain and discuss more about the recusing effect of the secretome in secondary immunodeficiency mice.
  15. There are no axis lines and unit labeling in Figures 1 and 2. Moreover, the symbol legends are too small to identify. Wrong label of figure 2B at 10 uL s/c. Figure legends are not clearly explained and difficult to understand the statistical analysis. What is the meaning of each number above the box plot? What is the difference between the immunized control in figure 2A and the control in figure 2B-D? Redundant data is used in each figure. The authors should not separate graphs if use the same control group. I recommended using a scatter plot with trend line addition for a correlation plot.
  16. Based on the objective of this study, please discuss in detail of parameters of the humoral arm of immunity under the action of the secretome. What parameter is most important and why?
  17. The authors must discuss and find more references to support the effect of xenogeneic secretome on the activation of humoral immune response regarding B cells such as this reference: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9396016/

Tech Editor Comment

I noticed that the spelling of "mesenchimal", while consistent with many published papers, is not consistent with the Wikipedia spelling, which is with a Y, mesenchymal. Should this be kept as the author stated it, or use the more common/Wikipedia standard spelling?

Additionally, consider adding redirects for both spellings. Treenxorr (discusscontribs) 11:46, 8 February 2023 (UTC)[reply]

The authors do not object to writing the term "mesenchimal" through "Y" (as "mesenchymal"), but we don't know how to fix the title of the article in the "Wiki" system. Vitalii moskalov (discusscontribs) 17:51, 27 April 2023 (UTC)[reply]

Peer review 3


Review by anonymous peer reviewer , Published papers on human induced pluripotent stem cells
These assessment comments were submitted on , and refer to this previous version of the article

  1. What strain of mice were used (not specified)
  2. What do the panels in Figure 1 indicate (no legend).
  3. There is mention of randomization without a randomization procedure (classic mistake in these kinds of studies)
  4. What number of animals were used in each experiment? (not specified).
  5. Why was a Mann-Whitney test used as some kind of post-test on multigroup data and not ANOVA?
  6. Why are there no statistics on the survival data, and at which time point was this gauged?

--Athikhun.suw (discusscontribs) 11:05, 28 June 2023 (UTC)[reply]

1) What strain of mice were used (not specified).
The study was performed on outbred white adult mice, as indicated in the manuscript. Unfortunately, the researchers did not have the financial and logistical ability to purchase mice of certain strains. We can consider these mice as "wild type" or the supplier's own population of mice.
2) What do the panels in Figure 1 indicate (no legend).
Figure 1 depicts a three-line differentiation of MSCs (in osteogenic, adipogenic, and chondrogenic directions) as a stage in cell culture identification. A detailed description of the figure is given in the corresponding section (Section "File description"):
The capacity of the mesenchymal stem cells (used in the study to obtain the secretome) to differentiate into three lines:
1 - osteogenic differentiation (1a - control, undifferentiated cells (no differentiation inducer added); 1b - osteocytes, calcium deposits stained with Alizarin red dye, x100); 2 - adipogenic differentiation (2a - control, undifferentiated cells (no differentiation inducer added); 2b - adipocytes, fat deposits stained with Oil Red, x100); 3 - chondrogenic differentiation (3a - control, undifferentiated cells (no differentiation inducer added); 3b - chondrocytes, glycosaminoglycans stained with Alcian blue dye, x100)
Perhaps this legend should be placed elsewhere. Please indicate where this needs to be done.
3) There is mention of randomization without a randomization procedure (classic mistake in these kinds of studies).
Randomization was done by urn design taking into account the body weight of the animals (the deviation in the initial weight of animals did not exceed 10%).
We have clarified this information to the "Materials and Methods" section.
4) What number of animals were used in each experiment? (not specified).
·      n = 6 (experiment in mice with normal immune status);
·      n = 10 (experiment in mice with secondary immunodeficiency at the beginning of the experiment);
·      Taking into account the survival rate in the groups, there were such an end-point number of animals: Positive control – 0 animals; Negative control – 10 animals; “Thymalin” – 4 animals; “10 µl i/m” – 5 animals; “10 µl s/c” – 5 animals.
We have added this information to the "Materials and Methods" section.
5) Why was a Mann-Whitney test used as some kind of post-test on multigroup data and not ANOVA?
At the first stage, the Kruskal-Wallis test (ANOVA) was applied. At the significance level p≤0.001, the null hypothesis about the equality of the samples was rejected. However, this statistical method indicates the presence of a statistically significant difference between groups, but does not indicate between which groups this difference is significant. Therefore, at the second stage, the groups were compared in pairs using the Mann-Whitney (Wilcoxon) U test.
We have added this information to the manuscript.
6) Why are there no statistics on the survival data, and at which time point was this gauged?
Samples were administered 5 days prior to the modeling of immunodeficiency and until the indicators were taken into account. Immunization with a 3% suspension of freshly washed ram erythrocytes was carried out for 4 days, starting from the next day after modeling the pathology. Accounting for survival was carried out before determining the amount of AFC and antibody titer on the next day after the completion of immunization (5th day after the administration of HCA) by taking into account the number of animals that had died by this time.
We have added this information to the manuscript.
Statistical analysis of survival was performed, as indicated in the manuscript, using the Fisher test for sample shares. A significant increase in survival compared to the positive control was found. Differences in this indicator between experimental groups and negative control, as well as between different experimental groups, are not significant (as indicated in the manuscript). Vitalii moskalov (discusscontribs) 11:31, 28 June 2023 (UTC)[reply]

Editor comments

One of our board - Eric Youngsrtrom - says:

I have reviewed the author responses in more depth than the paper.

Two thoughts on the responses, both small: (a) it seems that characterizing the mice as “supplier's own population of mice” seems most precise, though I defer to others with more expertise whether “wild type” is appropriate.

(b) The statistical methods are both non-parametric alternatives to the ones asked about in Review 3 (ANOVA, t-test). Presumably there was a reason or advantage to using nonparametric methods (non-normal distribution of dependent variable, or outliers, are the two most likely). Kruskal Wallis and Mann-Whitney are good non-parametric options, so the choice is not a problem, but the *rationale* for the choice is still missing (and without a rationale, ANOVA would usually be preferable because of familiarity and a slight statistical power advantage).

I would recommend acceptance with minor revision that clarifies those two points. Rwatson1955 (discusscontribs) 11:55, 20 July 2023 (UTC)[reply]

Editor comment

Eric A Youngstrom comments:

I commend the authors on a thorough and helpful response! The description of the mice is sufficiently detailed, and the logic for the selection of non-parametric tests is clear and appropriate. In fact, it follows best practices (the other good alternative would have been trying transformations to make the normality assumption tenable). One final point of clarification – I think that the authors used Fisher’s exact test for the proportions. Fisher developed multiple different statistical methods, so adding that one word ensures certainty about the choice of method.

Kudos on a strong piece of work! Rwatson1955 (discusscontribs) 08:54, 28 July 2023 (UTC)[reply]