Ameliorating Effect of Combined Cinnamon and Ginger Oils against the Neurotoxicity of Nicotine Administration on the Prefrontal Cortex of Adult Albino Rats: Immunohistochemical and Ultrastructural Study
, Mohie Mahmoud Ibrahim 1, Mamdouh Eldesoqui 1, Mohamed A. M. Iesa 3, Tourki A. S. Baokbah 4 and Hany M. A. Sonpol 1,5
Round 1
Reviewer 1 Report
The manuscript Ameliorating Effect of Combined Cinnamon and Ginger Oils against the Neurotoxicity of Nicotine Administration on the Prefrontal Cortex of Adult Albino Rats: Immnuohistochemical and Ultrastructural Study fits the journal's scope. The authors present the influence of two volatile oils and their mixture on the antioxidant status and on the morphology of prefrontal cortex. The study design is simple and correct, and the results are presented in sufficient detail. Before publication, the authors should make the following minor corrections:
2.3. Biochemical analysis, Histopathological Examination please add the references
If it is possible, please add the quality specification from the supplier for the volatile oils as supplementary materials.
2.1. Please specify if the protocol was carried out in accordance with the guidelines of the Declaration of Helsinki regarding animal experimentation.
Lines 242, 263, 274 please correct the typing errors
316-318 please rephrase. Also, please formulate the conclusions in accordance with your results.
Lines 67, 95 please correct the error
Author Response
Dear Ms. Milica Stankovic
Thank you for giving me the opportunity to submit a revised draft of my manuscript titled (Ameliorating Effect of Combined Cinnamon and Ginger Oils against the Neurotoxicity of Nicotine Administration on the Prefrontal Cortex of Adult Albino Rats: Immnuohistochemical and Ultrastructural Study.) to (Scientia pharmaceutica). We appreciate the time and effort that you and the reviewers have dedicated to providing your valuable feedback on my manuscript. We are grateful to the reviewers for their insightful comments on my paper. We have been able to incorporate changes to reflect most of the suggestions provided by the reviewers.
Here is a point-by-point response to the reviewers’ comments and concerns.
Point 1: 2.3. Biochemical analysis, Histopathological Examination please adds the references
if it is possible, please add the quality specification from the supplier for the volatile oils as supplementary materials.
Response 1: regarding to references of
- Biochemical analysis reference: Esterbauer H, Cheeseman KH (1990) Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Methods Enzymol 186: 407–421.
- Histopathological examination reference: Drury, R., Wallnigton, E., 1976. Carleton’s Histological Technique. Oxford University Press, London.
- Quality specification of volatile oils: Cinnamon and Ginger oils were purchased from the local market of Agricultural Herbs, Spices and Medicinal plants (Cairo, Egypt).
The commercial ginger essential oil was characterized by warm, spicy, and woody notes, with slight lemony notes. The oils were pale yellow, low viscosity liquids,
Essential oils extracted from commercial cinnamon barks were yellow, with warm-spicy notes characterized simply as “cinnamon notes
Point 2: 2.1. Please specify if the protocol was carried out in accordance with the guidelines of the Declaration of Helsinki regarding animal experimentation.
Lines 242, 263, 274 please correct the typing errors
316-318 please rephrase. Also, please formulate the conclusions in accordance with your results.
Lines 67, 95 please correct the error
Response 2:
All protocols of the present study were approved by the institutional review board (IRB) of
Mansoura Faculty of Medicine which follow the guidelines of the Declaration of Helsinki regarding animal experimentation.
Conclusion: Cigarette smoking have many health hazard one of this its neurotoxicity, which is characterized by apoptosis of neurocytes of prefrontal cortex might be due to enhanced oxidative stress, upregulation of inflammatory cytokines (TNF-α) and neuronal gliosis (GFAP).The coadminstration of ginger and cinnamon oils protected the Prefrontal cortex neurocytes from apoptosis better than single oil used. The neuroprotective effects of both agents might be due to inhibition of oxidative stress, the inflammatory process, neuronal gliosis.
Line 242: B) Treatment with combined ginger and cinnamon oils significantly improves the previous markers through its antioxidative
Line 263: in agreement with (6) who founded that chronic exposure to nicotine stimulates brain lipid peroxidation
Line 274: The results of the present study confirms the destructive effect of nicotine on the morphology
Line 67: In our study, we tried to examine the neurotoxicity induce by chronic nicotine administration in rats and ameliorative effect of combination of cinnamon and ginger oils through its antioxidant and inflammatory properties.
Line 95: Group V, Nicotine+ Cinnamon + Ginger (10 rats): rats of this group treated with nicotine in same dose as group II and cinnamon and ginger oils as same as group II and group IV, respectively.
Author Response File: 
Author Response.docx
Reviewer 2 Report
The manuscript presented the ameliorating effect of combined cinnamon and ginger oils against the neurotoxicity of nicotine administration on the prefrontal cortex of adult albino rats. Some specific comments need to be addressed:
Line 31: marked improvement was not the correct word to use
In the introduction, so many paragraphs are unnecessary.
Materials and methods: toxicity study was missing. How did the authors select doses for the study?
The activities seem to be additive effects of the compounds, largely dependent on their antioxidant and anti-inflammatory activities. What was the novelty of the study?
Line 101: ''other parts were used'' unclear
Line 116-118: I could not find quantification
Line 271-272: It would be interesting if the author discusses ferroptosis
Line 266 & 314: How was it synergistic?
Why did the authors not include behavioural data?
Line 281: ''greatly improved'' is not the correct word to use
Line 317: ''improve'' wrong. It should ameliorate/attenuate
The writing styles need to be corrected by an English editor. The current form of the manuscript deserves substantial language editing.
What would be the follow-up study?
The authors should address the limitations of their study
Author Response
Response to Reviewer 2 Comments
Dear Ms. Milica Stankovic
Thank you for giving me the opportunity to submit a revised draft of my manuscript titled (Ameliorating Effect of Combined Cinnamon and Ginger Oils against the Neurotoxicity of Nicotine Administration on the Prefrontal Cortex of Adult Albino Rats: Immnuohistochemical and Ultrastructural Study.) to (Scientia pharmaceutica). We appreciate the time and effort that you and the reviewers have dedicated to providing your valuable feedback on my manuscript. We are grateful to the reviewers for their insightful comments on my paper. We have been able to incorporate changes to reflect most of the suggestions provided by the reviewers.
Here is a point-by-point response to the reviewers’ comments and concerns.
Point 1: Line 31: marked improvement was not the correct word to use.
Response 1: These changes were observed to a less degree in rats' group (III) and group (IV) while there was a (clear) marked improvement achieved by the combined usage of cinnamon and ginger oils together compared to the nicotine group.
Point 2: In the introduction, so many paragraphs are unnecessary.
Response 2:
Background: Nicotine is the active alkaloid in cigarettes. It was reported that tobacco smoking has many hazards one of these hazards is the affection of cognitive function of prefrontal cortex. The aim of our study was to investigate the antioxidant effects of ginger, cinnamon oils and their combination on morphological changes in prefrontal cortex that were induced by nicotine. Fifty adult male albino rats were divided into five groups: group I (control group), group II (nicotine), group III (nicotine + cinnamon), group IV (nicotine + ginger) and group V (nicotine + cinnamon + ginger). Coronal sections from prefrontal cortex were examined by light microscope for (H&E and immunohistochemichal staining with TNF-α and GFAP) and transmission electron microscopy. Markers of oxidative stress (MDA, GSH) were being assessed. Compared to the control group, the rats those were treated with nicotine (group II) showed a significant oxidative stress in the form of marked elevation of MDA and decrease in GSH, apoptotic changes in e pyramidal cells, together with elevation in the inflammatory marker TNF-α and GFAP expressions. These changes were observed to a less degree in rats' group (III) and group (IV) while there was a marked improvement achieved by the combined usage of cinnamon and ginger oils together.
Point 3: Materials and methods: toxicity study was missing. How did the authors select doses for the study?
According to previous study done by Benowitz NL. Pharmacology of nicotine: addiction, smoking-induced disease, and therapeutics Annu Rev Pharmacol Toxicol. 2009; 49:57-71, doi. 10.1146/annurev.pharmtox.48.113006.094742, indexed in Pubmed: 18834313.
Point 4: The activities seem to be additive effects of the compounds, largely dependent on their antioxidant and anti-inflammatory activities. What was the novelty of the study?
The novelty of our study as we are the first how examine the protective effect of this addition on prefrontal cortex of nicotine exposed rats, the use of this addition examined only on Spermatogenesis Dys-function of Diabetes Rats which done by Khaki A et al 2014.
Point 5: Line 101: ''other parts were used'' unclear
I mean that some rat’s brain at prefrontal cortex was separated and used for the assessment of the MDA and GSH levels, and another rat’s brain at prefrontal cortex were used for histopathological studies as we take 10 rats in each group.
Point 6: Line 116-118: I could not find quantification
The denisty of immunstaining in prefrontal cortex area (region of interest) was quantified as the percent of brain tissue occupied by positive staining (calculated by averaging the values from ten fields at 10x magnification) for each brain area by using image J software.
Point 7: Line 271-272: It would be interesting if the author discusses ferroptosis
Ferroptosis is a newly identified iron-dependent cell death that is different from other cell death forms, including apoptosis and necrosis. The program involves three primary metabolisms involving thiol, lipid, and iron, leading to an iron-dependent generation of lipid peroxidation and, ultimately, Ghio, A. J. et al. (2008) founded that smokers’ lung can affect oxidative stress and inflammation by altering iron homeostasis. This shift our attention for further examination for both combined oils on ferropotosis induced by nicotine exposure
Point 8: Line 266 & 314: How was it synergistic?
I am so sorry I mean that combined antioxidant and anti-inflammatory activities of both oils.
Point 9: Why did the authors not include behavioural data?
Because I focused in my study only on histopathological aspect and this may open door for another research idea about behavioural study.
Point 10: Line 281: ''greatly improved'' is not the correct word to use
In our study the treatment of nicotine exposed rats with combination of cinnamon and ginger oils (improving) greatly improved the brain morphology than treatment with cinnamon and ginger oils separately
Point 11: Line 317: ''improve'' wrong. It should ameliorate/attenuate
the writing styles need to be corrected by an English editor. The current form of the manuscript deserves substantial language editing.
We suggest that usage of ginger and cinnamon with other polyphenol containing herbs such as onion to attenuate nicotine induced oxidative stress to maximize the antioxidant defense.
Point 12: What would be the follow-up study?
I can examine the effect of combined oils in the future on hippocampus of nicotine exposed rats
Point 13: The authors should address the limitations of their study
The limitations of the current study are numerous, starting with some more obvious ones, like how well a nicotine-induced neurotoxicity in animal model reflects the human behavioural changes due to smoking, and some less obvious ones related to difference in brain anatomy between albino rats and humans
Author Response File: Author Response.docx
Round 2
Reviewer 2 Report
The revised version of the manuscript could be considered for publication; however, the manuscript still needs an English proofreading.
This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.
