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On Sep 2018




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"Journal of Clinical and Diagnostic Research is at present a well-known Indian originated scientific journal which started with a humble beginning. I have been associated with this journal since many years. I appreciate the Editor, Dr. Hemant Jain, for his constant effort in bringing up this journal to the present status right from the scratch. The journal is multidisciplinary. It encourages in publishing the scientific articles from postgraduates and also the beginners who start their career. At the same time the journal also caters for the high quality articles from specialty and super-specialty researchers. Hence it provides a platform for the scientist and researchers to publish. The other aspect of it is, the readers get the information regarding the most recent developments in science which can be used for teaching, research, treating patients and to some extent take preventive measures against certain diseases. The journal is contributing immensely to the society at national and international level."



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Saraswati Dental College
Lucknow
On Sep 2018




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On Aug 2018




Dr. Arundhathi. S
"Journal of Clinical and Diagnostic Research (JCDR) is a reputed peer reviewed journal and is constantly involved in publishing high quality research articles related to medicine. Its been a great pleasure to be associated with this esteemed journal as a reviewer and as an author for a couple of years. The editorial board consists of many dedicated and reputed experts as its members and they are doing an appreciable work in guiding budding researchers. JCDR is doing a commendable job in scientific research by promoting excellent quality research & review articles and case reports & series. The reviewers provide appropriate suggestions that improve the quality of articles. I strongly recommend my fraternity to encourage JCDR by contributing their valuable research work in this widely accepted, user friendly journal. I hope my collaboration with JCDR will continue for a long time".



Dr. Arundhathi. S
MBBS, MD (Pathology),
Sanjay Gandhi institute of trauma and orthopedics,
Bengaluru.
On Aug 2018




Dr. Mamta Gupta,
"It gives me great pleasure to be associated with JCDR, since last 2-3 years. Since then I have authored, co-authored and reviewed about 25 articles in JCDR. I thank JCDR for giving me an opportunity to improve my own skills as an author and a reviewer.
It 's a multispecialty journal, publishing high quality articles. It gives a platform to the authors to publish their research work which can be available for everyone across the globe to read. The best thing about JCDR is that the full articles of all medical specialties are available as pdf/html for reading free of cost or without institutional subscription, which is not there for other journals. For those who have problem in writing manuscript or do statistical work, JCDR comes for their rescue.
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Reviewing articles is no less a pain staking process and requires in depth perception, knowledge about the topic for review. It requires time and concentration, yet I enjoy doing it. The JCDR website especially for the reviewers is quite user friendly. My suggestions for improving the journal is, more strict review process, so that only high quality articles are published. I find a a good number of articles in Obst. Gynae, hence, a new journal for this specialty titled JCDR-OG can be started. May be a bimonthly or quarterly publication to begin with. Only selected articles should find a place in it.
An yearly reward for the best article authored can also incentivize the authors. Though the process of finding the best article will be not be very easy. I do not know how reviewing process can be improved. If an article is being reviewed by two reviewers, then opinion of one can be communicated to the other or the final opinion of the editor can be communicated to the reviewer if requested for. This will help one’s reviewing skills.
My best wishes to Dr. Hemant Jain and all the editorial staff of JCDR for their untiring efforts to bring out this journal. I strongly recommend medical fraternity to publish their valuable research work in this esteemed journal, JCDR".



Dr. Mamta Gupta
Consultant
(Ex HOD Obs &Gynae, Hindu Rao Hospital and associated NDMC Medical College, Delhi)
Aug 2018




Dr. Rajendra Kumar Ghritlaharey

"I wish to thank Dr. Hemant Jain, Editor-in-Chief Journal of Clinical and Diagnostic Research (JCDR), for asking me to write up few words.
Writing is the representation of language in a textual medium i e; into the words and sentences on paper. Quality medical manuscript writing in particular, demands not only a high-quality research, but also requires accurate and concise communication of findings and conclusions, with adherence to particular journal guidelines. In medical field whether working in teaching, private, or in corporate institution, everyone wants to excel in his / her own field and get recognised by making manuscripts publication.


Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journals–No manuscripts–No authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
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In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."



Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011




Dr. Shankar P.R.

"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."



Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011
Anuradha

Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.


Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020

Important Notice

Original article / research
Year : 2023 | Month : January | Volume : 17 | Issue : 1 | Page : BC11 - BC14 Full Version

Evaluation of Oxidative DNA Damage among Type 2 Diabetes Mellitus Patients and Healthy Individuals in Duhok, Iraq: A Case-control Study


Published: January 1, 2023 | DOI: https://doi.org/10.7860/JCDR/2023/53508.17344
Hivi Mohammed Mahmoud, Ardawan Fathi Ali, Wahid Mohammed Hassan, Idris Haji Ahmed, Dhia Jaafer Altimimi

1. PhD Lecturer, Department of Medical Chemistry, College of Medicine/University of Duhok, Duhok, Kurdistan Region, Iraq. 2. PhD Lecturer, Department of Medical Lab. Technology, Shekhan Technical College of Health/Duhok Polytechnic University, Duhok, Kurdistan Region, Iraq. 3. Assistant Professor, Department of Orthopaedic Surgery, College of Medicine/University of Duhok, Duhok, Kurdistan Region, Iraq. 4. Specialist Diabetologist, Department of Duhok Diabetes Center, Azadi General Teaching Hospital, Duhok, Kurdistan Region, Iraq. 5. Professor, Department of Medical Chemistry, College of Medicine/University of Duhok, Duhok, Kurdistan Region, Iraq.

Correspondence Address :
Hivi Mohammed Mahmoud,
KRO Street, Duhok, Kurdistan Region, Iraq.
E-mail: hivi.mahmoud@uod.ac

Abstract

Introduction: The association between oxidative Deoxy Ribonucleic Acid (DNA) damage and diabetes is well established. Increased glucose levels can stimulate free radical production. However, data regarding DNA damage in type 2 Diabetes Mellitus (DM) patients and healthy individuals are controversial and scarce in Iraq.

Aim: To assess the DNA damage among patients with type 2 Diabetes Mellitus (DM) and healthy individuals and to analyse its relationship with oxidative stress biomarkers.

Materials and Methods: This case-control study was conducted at the Duhok Diabetes Center, Duhok, Kurdistan Region, Iraq, from September 2016 to March 2018. In this study, biomarkers of both oxidative stress and DNA damage including Total Antioxidant Capacity (TAC), Malondialdehyde (MDA) and serum 8-Hydroxy-2-deoxiguanosine (8-OHdG) were measured in 297 patients with type 2 diabetes and 188 healthy individuals. Selection of cases and healthy individuals was done using random sampling technique. Statistical analysis was done using Statistical Package for Social Sciences (SPSS) version 18.0 and a p-value <0.05 was set as a cut off value of statistical significance.

Results: The mean age, sex and Body Mass Index (BMI) were similar between patients and healthy individuals. Significantly higher 8-OHdG and MDA levels (p<0.001 and p<0.010, respectively) together with lower TAC levels (p=0.010) were found in diabetics compared to healthy individuals. In diabetic patients, a positive correlation of 8-OHdG was observed with MDA (r=0.220), and a negative correlation was observed with TAC (r= -0.47). Based on the estimated cut-off point of DNA damage (8-OHdG of 4.0 ng/mL), 84.51% of patients had high levels of DNA damage compared with healthy individuals (28.7%).

Conclusion: Oxidative DNA damage increased in diabetic patients, and was associated with lower antioxidant capacity. Antioxidant supplementation may be an effective public health intervention to reduce DNA damage and oxidative stress.

Keywords

Deoxyribonucliec acid, 8-hydroxy-2-deoxiguanosine, Diabetes mellitus, Malondialdehyde, Oxidative stress, Total antioxidant capacity

Cellular dysfunction occurs as a result of proteins and lipids oxidation as well as DNA modifications (1). When DNA repair system and endogenous antioxidant network are overwhelmed, oxidative DNA damage occurs (2). Association between oxidative stress and DNA damage has been reported (3). Some studies have shown that oxidative stress plays an important role in cellular injury through persistent increased blood sugar (4),(5). The data regarding association of increased glucose and amount of DNA damage are controversial (6),(7). Numerous studies showed an increase in DNA damage in patients with type 2 Diabetes Mellitus (DM) when compared with apparently healthy subjects (8),(9).

Persistent hyperglycemia can produce large amount of free radicals (10). Because the body's defenses are inadequate to deal with the increased production of Reactive Oxygen Species (ROS), an imbalance between ROS production and their defenses develops. This results in oxidative stress (10). These substances that are produced can cause breaks in the strands of DNA molecule as well as cause base modification, including guanine residues oxidation which is most potential base in DNA molecule that is oxidized more than other bases because of imidazole ring in its structure and leads to 8-OHdG production that is an oxidized nucleoside of DNA. The DNA damage by oxidation has been demonstrated by measuring the 8-OHdG levels, a known marker of oxidant induced DNA damage, in diabetic subjects (3),(11). The most frequent studied and detected biomarker of oxidative DNA damage is 8-OHdG (8),(12).

Recently, numerous clinical studies have studied levels of 8-OHdG in human organs, leukocyte DNA and urine concerning oxidative stress and diabetes mellitus (8),(13). Since 8-OHdG is an established biomarker of oxidative DNA damage (14), it is pivotal to study the association of oxidative DNA with other related factors such as lipid oxidation by-product (such as MDA) and total antioxidant capacity in patients with type 2 DM and healthy individuals.

To the author's knowledge little data are available in this area investigating oxidative stress and DNA damage in patients with diabetes (8). This study is a part of previously published study (8). Thus, this study was conducted to assess oxidative DNA damage, as well as antioxidant capacity in patients with type 2 DM and healthy individuals in Duhok, Kurdistan Region (Iraq).

Material and Methods

This case-control study was conducted at the Duhok Diabetes Center, Duhok, Kurdistan Region, Iraq, from September 2016 to March 2018. Medical Ethics Committee approval (Reference No: 07062016-4) and informed written consent was obtained for all participants.

Inclusion criteria: For patients it was fasting glucose (>125 mg/dL and HbA1c >7.0%). For healthy individuals inclusion criteria were: no history of chronic diseases or history of diabetes mellitus among first-degree relatives , they were included as controls.

Exclusion criteria: Exclusion criteria for cases were: cardiovascular, respiratory, rheumatoid, renal and hepatic diseases, history of malignancy, recent infections, pregnancy, smoking, and alcoholics. Individuals who had CRP>6 mg/dL, HbA1c>5.5% and glucose levels >100mg/dL were excluded from the study.

Sample size calculation: The sample size for this study was calculated with 95% confidence interval with 80% power at a 5% level of statistical significance (15). Four hundred-eighty five subjects, 297 patients with type 2 DM and 188 apparently healthy individuals (aged 35-65 years) were enrolled in the study. During the sampling period of this study all patients (n=5783) who attended Duhok diabetes center were interviewed and asked to participated in the study. At the beginning, a total of 337 participate in the study. After exclusion of 40 responders who were unfit according to the inclusion criteria, the remaining participants were enrolled in this study.

A pre tested proforma was designed to obtain information on gender, age, weight, and height. The BMI was measured for each subject. Participants were instructed to avoid strenuous physical activity for more than two hours before the examinations.

Study Procedure

Morning blood samples after overnight fasting for 12-14 hours were collected between 9:00-11:30 AM at the Lab Department of Clinical Biochemistry at Azadi General Teaching Hospital. About 10 mL of blood was withdrawn by venipuncture, using vacutainer from the antecubital vein and collected in Becton Dickinson (BD) Vacutainer System CAT- plain tubes. Total 2 mL of blood was collected in an Ethylenediamine Tetraacetic Acid (EDTA) tube for measurement of HbA1c%. The sera were separated by centrifugation using a HITACHI centrifuge, Model O5P-21 at 5000 rpm for 10 min at room temperature and was collected into two tubes- one was processed immediately for measuring serum Fasting Blood Glucose (FBG), MDA and TAC levels. The MDA and TAC were estimated by colorimetric methods at 532 nm wavelength for MDA and at 570 nm for TAC. The second tube with liquid sera was stored at -800°C for measurement of serum 8-OHdG levels by using Enzyme Linked Immunosorbent Assay (ELISA) kit (Catalog number: E-EL-0028, ELABSCIENCE. USA). Assessment of BMI was done. Subjects with BMI less than 25 Kg/m2 were considered normal, while those with BMI between 25 Kg/m2 and 2 9.9 Kg/m2 were considered overweight and those with BMI ≥30 Kg/m2 were considered obese. (16). Assessment of MDA and TAC was done. A cut-off point of MDA<1.4 nmol/L and of TAC>1.77 mmol/L (as obtained from the laboratory) were used to classify patients as having low oxidative stress (17). Assessment of DNA damage was based on the levels of 8-OHdG. Subjects with 8-OHdG >4.0 ng/mL were considered to have a high level of DNA damage (8).

STATISTICAL ANALYSIS

Analysis of data was done using version 18.0 SPSS computer software. Means±Standard Deviation (SD) adopted to present data. Student’s t-test was used to evaluate the differences between groups. Pearson’s® correlation coefficient was used to estimate the correlation between 8-OHdG levels and other variables. A p-value at ≤ 0.05 was used as statistical significance cut off value.

Results

(Table/Fig 1) illustrates the demographic and laboratory characteristics of the studied subjects. Age, sex distribution were statistically similar between patients and healthy individuals. The mean+SD of serum 8-OHdG of patients and healthy individuals was 6.04±2.7 ng/mL and 3.59±2.9 ng/mL, p<0.001. Significantly higher 8-OHdG and MDA levels (p<0.001 and p<0.01 respectively) together with lower TAC levels (p=0.01) were found in diabetics compared to healthy individuals. The DNA damage quantitated by the 8-OHdG was detected in 251 (84.5%) of patients with type 2 DM as compared to 54 (28.7%) in healthy individuals, p<0.01.

(Table/Fig 2) illustrates the mean±SD of 8-OHdG levels for age, sex, BMI and oxidative stress biomarkers of patients with type 2 DM. No statistically significant differences were found in the mean values of 8-OHdG with respect to age and gender, p=0.250 and p=0.680. A significantly higher mean 8-OHdG level was found in the overweight and obese group compared with the normal weight group (p=0.045), also the mean 8-OHdG level was significantly higher (p=0.020) for patients with low antioxidant capacity as compared to that of high antioxidant capacity patients.

In the patient group, 8-OHdG negatively correlated with TAC (r=-0.47, p<0.001) and it correlated positively with MDA (r=0.22, p=0.015). No significant correlation was found with respect to age and BMI in cases group (Table/Fig 3).

Discussion

The results of this study confirm that DNA damage in type2 DM patients characterised by the low antioxidant levels which may play an important role in the disease progression and disease related complications. A significantly higher serum 8-OHdG levels were seen in overweight and obese diabetic subjects when compared to those with normal weight group. Further, high MDA level was seen in diabetic subjects when compared to healthy individuals. These results are in accordance with a previous study (18). In this study, 84.51% of the patients and 28.7% of the healthy individuals had high degree of DNA damage (8-OHdG>4.0ng/mL). Such a high prevalence of DNA damage is worthy of discussion. It is favorably comparable to values from the developing countries (18),(19), but it is markedly higher than values of diabetic and normal populations in other studies (14),(20),(21). The discrepancy may be related partly to the low antioxidant status. In a study done in Iraq, significantly higher levels of 8-OHdG was found in patients with type I diabetes than in healthy controls. (14) Significantly higher levels of oxidative stress markers (like MDA, glutathione peroxidase) and 8-OHdG were reported in type 2 diabetic patients when compared to healthy individuals in a study in Saudi Arabia (22). Similar findings were also reported from another study from Baghdad (23). In a review from India, authors have stated that hyperglycemia leads to overproduction of reactive oxygen species thus leading to microvasculature damage (20). However, in a study from United Kingdom, authors did not find any association between increased DNA damage markers in diabetic patients (24). The difference in results possibly could be because of the difference in the duration of diabetes and glycemic control (25).

A study by Ali AF and Altimim DJ reported a lower vitamin D status may increase DNA damage (26). A recent research by Mahmoud HM et al., also reported that micronutrients including zinc element act as a powerful antioxidant and may reduce DNA damage in diabetic patients (8). Several factors are known to increase DNA damage in diabetic patients, such as glycemic controls, duration of diabetes, repair capacity, diet, lifestyle and environment (5),(6). Recently some studies reported a variety of anti DNA damaging effect by dietary antioxidants, including the effects of antioxidant supplements (8),(27). So it is permissible to speculate that the impact of zinc supplement and/or other antioxidants and supplements may have a protective effect against DNA damage by reducing oxidative stress, a finding supported by the current uses of zinc for protection of healthy subjects from the initiation and progression of Coronavirus Disease-2019 (COVID-2019), although the mechanism of action is still not well understood, particularly in diabetic patients who are at a high risk of COVD-19 disease complications (28). Further studies to elucidate this hypothesis could be of concern.

The results of the present study is in accordance with previously published results and suggests that oxidative DNA damage through oxidative stress process is mediated mainly by hyperglycemia (4),(5), and supported that serum level of 8-OHdG is a potentially a valuable marker in estimating the severity of DNA damage, especially in diabetic patients.

Limitation(s)

The major limitation of this study is the relatively small sample size and data from a single population as the duration for the sampling was limited. Hence, the control were less than the cases. Further, lack of using more than one marker of oxidative stress, DNA damage as well as for antioxidant capacity to detect more association between oxidant-antioxidant balance and DNA damage was a drawback of this study.

Conclusion

Elevated levels of serum 8-OHDG with abnormal oxidative stress biomarkers may be associated with reduced antioxidant status in diabetic patients. In this study, a negative correlation between 8-OHdG and TAC in diabetic patients was seen. More than half of the population of this study, particularly diabetic patients had lower antioxidant capacity and a high degree of oxidative DNA damage. Antioxidant supplementation may be an effective public health intervention to reduce DNA damage and oxidative stress.

References

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Dandone P, Thusu K, Cook S, Snyder B, Makowski J, Armstrong D, et al. Oxidative damage to DNA in diabetes mellitus. Lancet. 1996;347:444-55. [crossref] [PubMed]
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Giacco F , Brownlee M. Oxidative stress and diabetic complications. Circ Res. 2010;107(9):1058-70. Doi: 10.1161/CIRCRESAHA.110.223545 [crossref] [PubMed]
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Fiorentino TV, Prioletta A, Zuo P, Folli F. Hyperglycemia-induced Oxidative Stress and its Role in Diabetes Mellitus Related Cardiovascular Diseases. Current pharmaceutical design. 2013; 19(32):5695-03 Doi: 10.2174/1381612811319320005 [crossref] [PubMed]
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DOI and Others

DOI: 10.7860/JCDR/2023/53508.17344

Date of Submission: Dec 02, 2021
Date of Peer Review: Jan 13, 2022
Date of Acceptance: Oct 19, 2022
Date of Publishing: Jan 01, 2023

AUTHOR DECLARATION:
• Financial or Other Competing Interests: None
• Was Ethics Committee Approval obtained for this study? Yes
• Was informed consent obtained from the subjects involved in the study? Yes
• For any images presented appropriate consent has been obtained from the subjects. NA

PLAGIARISM CHECKING METHODS:
• Plagiarism X-checker: Dec 03, 2022
• Manual Googling: Oct 11, 2022
• iThenticate Software: Oct 17, 2022 (10%)

ETYMOLOGY: Author Origin

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