Case Report: Molecular Analyses of Cell-Cycle-Related Genes in Cortical Brain Tissue of a Patient with Rasmussen Encephalitis
, Eliseu Paglioli Neto 3, André Palmini 3 and Daniel Rodrigo Marinowic 1,2,*
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsCase Report: Molecular analyses of cell cycle-related genes in 2 cortical brain tissue of a Rasmussen Encephalitis patient
Dear authors, your study is interesting because the pathogenesis and the aetiology of Rasmussen´encephalitis remains elusive.
I just have minor comments about the case report that I hope will improve its quality.
1. Abstract and methods
In the results section you said that the tissue obtained from the functional hemispherotomy was compared to control samples. Please describe in the methods section the control samples characteristics (number of control patients, tissue from epileptic patients or from other diseases?).
2. Case study description
Patients MRI is described as progressive right hemisphere and caudate atrophy but only one MRI is shown , the same with the EEG. Please consider to include two or more MRIs showing these progression because progression is included in the RE criteria.
Consider to include more information about differential diagnosis studies (viral serologies, immunological results…) if it is possible.
3. Discussion
I guess that patient underwent surgery in the RE residual phase. Could you comment which differences could be seen between the inflammatory phase and the residual phase?
4. Minor points.
-“Here, we report a RE patient that underwent hemispherectomy and remains seizure free after more than 6 months after seizure, with progressive motor improvement” Substitute hemispherectomy to hemispherotomy and after seizure to after surgery.
-“functional hemispherectomy” the same change hemispherectomy to hemispherotomy.
-Please add the meaning of the acronyms of all the genes the first time they appear in the text and always use the same nomenclature (for exemple CCDN gene I guess is the same as CCND3 gene).
Author Response
Reviewer 1
- Abstract and methods
In the results section you said that the tissue obtained from the functional hemispherotomy was compared to control samples. Please describe in the methods section the control samples characteristics (number of control patients, tissue from epileptic patients or from other diseases?)
Answer: We added the control patient’s information. The cortex sample of control group were obtained of residual resection of TLE-HS.
- Case study description
Patients MRI is described as progressive right hemisphere and caudate atrophy but only one MRI is shown , the same with the EEG. Please consider to include two or more MRIs showing these progression because progression is included in the RE criteria.
Consider to include more information about differential diagnosis studies (viral serologies, immunological results…) if it is possible.
Answer: We added a few tests for viral serologies and autoimmunity. Since our main hypothesis when he arrived was Rasmussen’s encephalitis, he did not undergo a lumbar puncture, as our intent was to take him to surgery as soon as possible. His clinical history, EEG and Brain MRI all pointed to this diagnosis. Indeed, we have added the figures of his subsequent MRI, showing the rapid progression of atrophy in the right hemisphere (Figure 1A-F), which significantly improved the image. Thank you for the suggestion.
- Discussion
I guess that patient underwent surgery in the RE residual phase. Could you comment which differences could be seen between the inflammatory phase and the residual phase?
Answer: Regarding the stage of RE, we believe that our patient was still in the Acute stage, since he was still developing progressive weakness in the right, as well as continuous status epilepticus, with preserved cognitive function. Nonetheless, the expression of genes may change throughout the illness, and samples from other cases may highlight this difference, especially in the residual phase, which shows almost complete loss of cortical neurons with fibrillary astrogliosis. At this stage, it is possible that our finding of increased BDNF would be reversed, perhaps showing a severe decrease in expression.
- Minor points.
-“Here, we report a RE patient that underwent hemispherectomy and remains seizure free after more than 6 months after seizure, with progressive motor improvement” Substitute hemispherectomy to hemispherotomy and after seizure to after surgery.
-“functional hemispherectomy” the same change hemispherectomy to hemispherotomy.
Answer: We changed the word hemispherectomy to hemispherotomy in all text long and seizure for surgery in the indicated place.
-Please add the meaning of the acronyms of all the genes the first time they appear in the text and always use the same nomenclature (for exemple CCDN gene I guess is the same as CCND3 gene).
Answer: We added the meaning of the genes and inflammatory cytokines acronyms and standardize the gene CCND3.
All changes were highlighted to facilitate your revision.
I hope you will find my response satisfactory and consider the manuscript of interest to the readership of International Journal of Molecular Sciences.
Reviewer 2 Report
Comments and Suggestions for AuthorsSpecific Comments to Authors:
· In the Results section: Authors have not mentioned anything about the characteristics of control patients. If control patients are clinically not patients then they should be mentioned as control individuals rather than control patients in results.
· The methodology regarding the PCR array and measurement of cytokines are NOT mentioned.
· In the discussion: Authors have mentioned that “In this work, we found a general decreased expression of cell cycle-related genes in brain tissue from the RE patient, where the genes BRCA1, BIRC5, CASP3, E2F4 and CCDN were more than 5-fold downregulated”. However, according to figure 2C (results) it is CCND3, not CCDN. Please clarify.
· According to the heat map MCM3 gene also showed changes which is involved in the “genome replication, cell proliferation, and neuronal cell cycle activation”. Please discuss about it.
Author Response
Reviewer 2
In the Results section: Authors have not mentioned anything about the characteristics of control patients. If control patients are clinically not patients then they should be mentioned as control individuals rather than control patients in results.
Answer: We added the control patient’s information. The cortex sample of control group were obtained of residual resection of TLE-HS.
- The methodology regarding the PCR array and measurement of cytokines are NOT mentioned.
Answer: We apologize for the oversight in not detailing the methodology for the PCR array and cytokine measurements. We have now included a comprehensive description in the text, prior to the description of the results encountered on the experiments
- In the discussion: Authors have mentioned that “In this work, we found a general decreased expression of cell cycle-related genes in brain tissue from the RE patient, where the genes BRCA1, BIRC5, CASP3, E2F4 and CCDN were more than 5-fold downregulated”. However, according to figure 2C (results) it is CCND3, not CCDN. Please clarify.
Answer: We apologize for the typographical error in the Discussion section. The correct gene mentioned should indeed be CCND3, not CCDN. We have revised the text accordingly. Thank you for bringing this to our attention
- According to the heat map MCM3 gene also showed changes which is involved in the “genome replication, cell proliferation, and neuronal cell cycle activation”. Please discuss about it.
Answer: We appreciate your observation regarding the MCM3 gene, which is involved in genome replication, cell proliferation, and neuronal cell cycle activation. In the revised manuscript, we have expanded our discussion to include the significant downregulation of MCM3 observed in the heat map. This gene's reduction in expression may indicate a broader dysregulation of the cell cycle machinery in RE, potentially contributing to the observed pathological features.
All changes were highlighted to facilitate your revision.
I hope you will find my response satisfactory and consider the manuscript of interest to the readership of International Journal of Molecular Sciences.
