Next Article in Journal
Dietary Oncopharmacognosy as a Crosswalk between Precision Oncology and Precision Nutrition
Previous Article in Journal
Beta Cell Dysfunction in Youth- and Adult-Onset Type 2 Diabetes: An Extensive Narrative Review with a Special Focus on the Role of Nutrients
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Nutrients
Volume 15
Issue 9
10.3390/nu15092218
nutrients-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Brief Report

Evaluation of Primary DNA Damage in Young Healthy Females Based on Their Dietary Preferences

by
Goran Gajski
*,
Katarina Matković
,
Luka Delić
and
Marko Gerić
Mutagenesis Unit, Institute for Medical Research and Occupational Health, 10000 Zagreb, Croatia
*
Author to whom correspondence should be addressed.
Nutrients 2023, 15(9), 2218; https://doi.org/10.3390/nu15092218
Submission received: 13 April 2023 / Revised: 4 May 2023 / Accepted: 5 May 2023 / Published: 8 May 2023
(This article belongs to the Section Nutrition and Public Health)
Browse Figures
Versions Notes

Abstract

:
DNA damage is known to be associated with many adverse health outcomes, including cancer and chronic diseases, but also with the process of aging. Empirical evidence has shown that environmental exposures, such as certain lifestyle factors, can affect a variety of health-related biomarkers and also impact the stability of DNA through the upregulation of the antioxidant defense system and alteration of its repair capacity. In addition to exercising, diet is an important lifestyle factor that can affect the development of a variety of chronic diseases and growing evidence suggests that plant-based diets, including vegetarianism, may promote health, longevity, and well-being. Therefore, we aimed to assess the primary DNA damage in 32 young healthy females from Zagreb, Croatia, based on their dietary preferences. The participants were divided into two groups: vegetarians and non-vegetarians, where the non-vegetarian group was further divided into omnivores (traditional mixed diet) and pescatarians (consumption of fish and seafood). According to statistical analysis, the DNA damage measured in whole blood cells expressed as the % tail DNA was significantly (p < 0.05) higher in vegetarians (3.6 ± 1.1%) compared to non-vegetarians (2.8 ± 1.0%). When further dividing the participants into specific sub-groups, lower DNA damage was observed amongst omnivorous subjects (3.2 ± 0.8%) compared to vegetarians, with the lowest DNA damage found in females practicing a pescatarian diet (2.4 ± 1.1%). Although a vegetarian diet can lead to a higher intake of specific vitamins and micronutrients, it can also lead to a deficiency of iron, calcium, and total proteins, which may affect genome stability and induce oxidative stress. Even though our results have shown that the pescatarian diet would be more beneficial in terms of maintaining DNA integrity, further research should be carried out to assess how specific dietary preferences affect DNA integrity on a larger scale.

1. Introduction

Damage to DNA molecules is recognized as one of the initial steps in the development of chronic diseases, cancer, and the aging process [1,2,3]. Increasing evidence indicates that dietary factors and exercise may impact various health-related biomarkers and can also affect the stability of DNA molecules by upregulating the antioxidant defense system and modulating repair capacity [4,5]. Numerous bioactive compounds detected in plant foods have the potential to reduce oxidative stress and inflammation as well as protecting against DNA damage through different modes of action, such as induction of DNA repair or by increasing resistance to oxidative damage [6,7]. In many clinical trials, foods and beverages rich in antioxidants have shown the ability to reduce DNA damage and increase DNA repair or antioxidant capacity [8,9,10,11]. This indicates that diet itself can be used as a modulator of different cell processes, especially in individuals with molecular signatures that can create excess DNA damage, oxidative stress, as well as inflammation.
Previously, we reported variances in different health-related biomarkers among people of both sexes practicing either traditional mixed or vegetarian diets. Our results showed that vegetarians have a lower nutritional status of some nutrients such as vitamins B12 and D as well as Zn, Ca, and Cu. This was accompanied by decreased antioxidant defenses in terms of glutathione as well as increased homocysteine levels. Moreover, vegetarians displayed increased DNA damage in the form of micronuclei and strand breaks as well as shorter telomeres. These findings suggest that the supplementation of animal-derived nutrients to this specific dietary group could lead to beneficial effects in terms of certain health-related biomarkers. On the contrary, our results also revealed that non-vegetarians displayed a higher level of some toxic metals, such as As and Hg [12]. Moreover, we also revealed that vegetarianism resulted in improved pancreatic β-cell function in women. The favorable adiponectin and insulin sensitivity responses in females reveal a distinct outcome of diet-to-metabolic homeostasis, demonstrating an interesting pattern of sexual dimorphism concerning the positive metabolic effect of vegetarianism [13].
Therefore, in this study, we wanted to evaluate primary DNA damage in young healthy female volunteers based on their dietary preferences. The subjects were divided into vegetarians (lacto–ovo vegetarians) and non-vegetarians, with the non-vegetarian group being further divided into omnivores (traditional mixed diet) and pescatarians (consumption of fish and seafood). The comet assay was used to evaluate the level of primary DNA damage in the female subjects. The assay itself is a well-known and accepted biomonitoring instrument used for the assessment of the effects of different lifestyle, environmental, occupational, and even dietary exposures on the levels of DNA damage in various human cells [14,15,16]. The comet assay is capable of recognizing DNA damage resulting from single- and double-strand breaks, single-strand breaks related to incomplete excision repair sites, DNA cross-links, as well as alkali-labile sites [17,18]. Furthermore, the measurement of DNA damage with this method could serve as a tool to predict the likelihood of developing any form of cancer and the associated risk of mortality. Such epidemiological evidence can emphasize the importance of incorporating the comet assay as a preventative measure for various non-communicable diseases [19,20].

2. Materials and Methods

2.1. Study Sample and Participant Selection

This study was conducted on a group of young female subjects (n = 32). All recruited subjects were from the general Zagreb (Croatia) population. At the time of the interview and blood sampling, all subjects were in a healthy condition and were free from any acute illnesses. Prior to donating blood for the study, all participants were occupationally not exposed to any hazards, including diagnostic ionizing radiation, and had not taken any medication such as antibiotics and/or steroid therapy for at least one year. All participants provided their written informed consent and completed a questionnaire intended to obtain demographic data, health status, and lifestyle factors such as smoking habits and alcohol consumption. Moreover, they gave information regarding their family history of cancer as well as data on previous or existing exposure to medication and diagnostic radiation that could interfere with the comet assay results. The study was approved by the Ethics Committee of the Institute for Medical Research and Occupational Health, Zagreb (Croatia) with ensured data privacy.

2.2. Blood Sampling

A trained medical professional collected venous blood into sterile heparinized tubes (Becton Dickinson, Franklin Lakes, NJ, USA) in the morning hours (between 7 and 10 a.m.). Subsequently, we handled their blood samples uniformly, by assigning random codes, storing them at 4 °C in light-protected conditions, and processing them as soon as possible (within 3 h after the sampling).

2.3. Alkaline Comet Assay

2.3.1. Procedure

The alkaline comet assay was performed based on the protocol by Collins et al. [17] following MIRCA guidelines [21]. Whole-blood samples (5 µL) were embedded in an agarose sandwich gel in the 0.5% low melting point (LMP) agarose layer (Sigma, St. Louis, MO, USA) on fully frosted slides. Afterward, they were placed in freshly made cold lysis solution (1% Triton X-100 (Sigma, St. Louis, MO, USA), 1% sodium N-lauroyl sarcosinate (Sigma, St. Louis, MO, USA), 2.5 M NaCl (Kemika, Zagreb, Croatia), 10% dimethyl sulfoxide (Kemika, Zagreb, Croatia), 10 mM Tris−HCl (Sigma, St. Louis, MO, USA; Kemika, Zagreb, Croatia), and 100 mM disodium EDTA (Sigma, St. Louis, MO, USA), pH 10), and left in the refrigerator (at 4 °C) during the night. After the cells were lysed and the DNA was unfolded, the slides were placed in electrophoresis solution (1 mM disodium EDTA (Sigma, St. Louis, MO, USA), 300 mM NaOH (Kemika, Zagreb, Croatia), pH 13) for 20 min (at 4 °C) and then they were electrophoresed for another 20 min (at 1 V/cm). Subsequently, the slides were washed with 0.4 M Tris−HCl buffer (pH 7.5) and dyed with ethidium bromide (10 μg/mL; Sigma, St. Louis, MO, USA) before the inspection.

2.3.2. Image Analysis

We created two slides for each participant and randomly evaluated a total of 100 comets per slide (200 comets in total). Selected comets were analyzed using an epifluorescence microscope (Olympus BX-51, Tokyo, Japan) armed with suitable filters as well as the Comet Assay IV (Perceptive Instruments Ltd., Haverhill, Suffolk, UK) software intended to analyze them. Tail intensity (i.e., % tail DNA) was used as the preferred comet assay descriptor which is expressed as % of total DNA fluorescence in the tail of the comet.

2.4. Statistical Analysis

Statistical analysis was performed using R statistical programming language version 4.1.2 and TIBCO Statistica™ (Palo Alto, CA, USA). Elementary statistical parameters were obtained using descriptive statistics. We tested the normality of the data with Shapiro–Wilk’s W test. The data were then log-transformed. The homogeneity of variance for each variable was tested with Bartlett’s Test. The differences in % tail DNA between the three tested groups (omnivorous, vegetarians, and pescatarians) were tested using ANOVA (type III) and the Scheffe test. The differences in % tail DNA between vegetarians and non-vegetarians were tested using a t-test with Bonferroni correction for p-value. The significance level for all tests was set at p < 0.05.

3. Results

3.1. Population Characteristics

This study encompassed a group of young female non-smoking volunteers (n = 32) aged between 25 and 35 years (the average age was 31.2 years and their median age was 30 years) with comparable socioeconomic statuses. They were divided into several dietary groups: vegetarian (n = 16) and non-vegetarian (n = 16) dietary groups. Vegetarian participants were practicing a lacto–ovo vegetarian diet which includes eggs and dairy as products of animal origin but excludes meat, poultry, fish, and seafood. The mean number of years practicing a vegetarian diet was 10.7 years. Moreover, the non-vegetarian group was divided into two subgroups, namely, omnivores (n = 8) and pescatarians (n = 8). Detailed population and lifestyle characteristics and their comet assay values are presented in Table 1.
All participants were selected from the general Croatian population and lived in the same city of Zagreb (the capital of Croatia). Female volunteers displayed comparable patterns of physical activity (regular exercise at least two times per week for at least half an hour) and comparable education level (high school and university). The participants in our study had no history of smoking, exposure to ionizing radiation, steroid therapy, or antibiotics for at least one year prior to blood donation. In addition, they were free of any occupational exposures that might have hindered the results of our study and had no known chronic illnesses or inherited genetic disorders. They were all in the normal weight range with their body mass index (BMI) above 18.5 and below 25 kg/m2.

3.2. Dietary Preferences and Genome Damage

According to our results (presented as box plots), damage to DNA molecules measured in whole blood cells, as evaluated by the alkaline comet assay and expressed as the % of tail DNA, was significantly (p < 0.05) higher in vegetarian subjects (3.6 ± 1.1%) compared to non-vegetarian ones (2.8 ± 1.0%; Figure 1). Moreover, when divided into specific sub-groups, we observed lower DNA damage in omnivorous subjects (3.2 ± 0.8%) compared to vegetarians with the lowest DNA damage observed in females practicing a pescatarian diet (2.4 ± 1.1%). It has to be pointed out that the only statistically significant (p < 0.05) difference was detected between vegetarians and pescatarians (Figure 2).

4. Discussion

Diet is regarded as one of the most important environmental contributors that can influence the occurrence of a variety of chronic illnesses [22,23] and there is increasing evidence that plant-based diets including vegetarianism may promote health, longevity, and well-being. Moreover, it is postulated that vegetarian and vegan diets and their specific components can lower the rate of different chronic disorders including diabetes, cancer, and cardiovascular disease [4,24,25,26]. Today we have many different types of vegetarian diets that are based on the level to which one is avoiding the consumption of animal products [27]. This can subsequently influence micronutrient intake; however, there are also differences in omnivorous diets regarding the quantity and the type of meat that is preferentially consumed. Although a vegetarian diet may result in a higher intake of specific micronutrients, as well as vitamins that may provide antioxidant defenses, there is also a possibility of a deficiency in some other micronutrients that are involved in DNA metabolism and its stability [4,12]. In line with the above-mentioned studies, this particular study aimed to investigate whether there is a possible difference in the level of DNA damage among young women practicing different diets, namely vegetarian and non-vegetarian ones.
To date, several studies have evaluated the impact of the vegetarian diet on genome integrity with non-conclusive results (reviewed in Gajski et al. [4]). Vegetarian Seventh-Day Adventists displayed lower chromosomal damage, expressed as the frequency of sister chromatid exchanges, in comparison with the general public. Still, inside the group of Seventh-Day Adventists, the authors did not observe differences among vegetarians and omnivores, suggesting that the overall lifestyle could be the cause for such differences [28]. Studies from Slovakia showed a higher amount of oxidative DNA damage, particularly concerning elderly omnivores but failed to observe differences in total DNA strand breaks, chromosome aberrations, or micronuclei frequency (an indication of chromosome breakage [29,30,31,32,33]) levels between different dietary groups [34,35,36]. In the papers by Kotova et al. [37] and Dhavan et al. [38] from Swedish and Indian populations, respectively, vegetarians had a lower baseline cytogenetic status and a reduced amount of DNA strand breaks compared to omnivore subjects.
It has to be pointed out that there are studies that failed to find differences between eating preferences and genome damage, such as another study from India showing no differences in DNA damage between vegetarian versus omnivore groups [39]. No differences were found regarding DNA damage and micronuclei frequency among subjects practicing uncooked veganism and omnivores [40] as well as the difference in micronucleated cells among vegetarians and meat eaters in research done by Davies et al. [41]. Concerning DNA damage and age, a study done by Fenech and Rinaldi [42] showed that young males practicing a vegetarian diet had a higher micronuclei frequency and middle-aged males practicing vegetarianism had a lower micronuclei frequency whereas, in older male and female subjects practicing a vegetarian diet, there was no difference in micronuclei compared to omnivores. Our previous results [12] on both males and females showed significantly higher DNA strand breaks and micronuclei frequencies in vegetarians compared to matched non-vegetarians, together with the several other blood and plasma biomarkers tested.
Generally speaking, vegetarians could have a higher intake of certain specific micronutrients as well as vitamins that are necessary for the proper functioning of the antioxidant defense system. On the other hand, they could be experiencing a deficiency in other micronutrients with a role in DNA metabolism and stability [4]. For instance, dietary antioxidants are involved in the cellular antioxidant defense system, which could strengthen the protection of DNA molecules by modulating several signaling pathways and gene expression, protecting and repairing DNA damage, and increasing the free radical scavenging capacity that arises through metabolic reactions [43,44]. In addition, it has been shown that higher consumption of antioxidant-rich vegetables and/or fruits can lower levels of oxidatively damaged DNA [45,46,47]. Since DNA damage is regarded as the initiating event in carcinogenesis, eating larger quantities of vegetables and fruits could be helpful in cancer protection in line with the prevention of free radical attacks on macromolecules [48,49], although there are carnivorous diets rich in fruits and vegetables as well as different plant-based types of diets which do not necessarily completely avoid meat and other animal products, such as the Mediterranean type of diet [50]. Moreover, antioxidants found in plants, such as flavonoids, carotenoids, and certain vitamins (C and E), can help protect from against various diseases by lowering the level of oxidative damage [51]. A certain downfall of avoiding meat consumption is that meats and dairy products are sources of vitamins belonging to the B group that are vital for cell division and erythropoiesis [52,53] as well as for DNA metabolism and stability [48]. Moreover, vitamin D that exists mostly in animal food sources is also beneficial in maintaining DNA integrity. Its role is primarily connected with DNA damage prevention but it also regulates the growth rate of cells [54]. Additionally, vegetarians tend to lack iron, calcium, and total proteins which may lead to genome instability as well as oxidative stress [55,56]. Hence, minimizing or eliminating animal foods from one’s diet may lead to the accumulation of unrepaired DNA damage that can initiate genome instability and possibly induction of cancer [57] and its related costs [58].
We also have to acknowledge several limitations of the study. This study was cross-sectional in its design; hence, causation could not be established. As in many other studies, the number of participants was limited; however, such studies are valuable to use in future meta-analyses where the statistical power of the results would be higher. The volunteers self-reported their dietary preferences, which leaves room for the potential of underestimating or overestimating the intake of certain food items, as well as confusion about the classification of certain dietary categories.

5. Conclusions

In summary, a number of studies failed to associate specific dietary preferences with DNA damage. Nevertheless, conflicting reports mentioned above indicate that there could be some differences regarding dietary preferences and measured genome instability biomarkers that could also be dependent on the country of origin and/or urban and rural differences in dietary intake. Based on our results, it seems that a pescatarian diet, which could be defined as plant-based if it contains an adequate intake of fish and seafood, could be a more beneficial diet in terms of maintaining DNA integrity. Although toxic metals such as mercury, increasing microplastics, as well as other persistent toxins found in fish could increase our risk for DNA damage and other side effects, a considerable body of evidence including our study still suggests that the benefits of fish consumption may outweigh the risks. Moreover, our results indicate the necessity of conducting further research on a larger scale to investigate how specific dietary preferences may impact DNA integrity and consequently our health to contribute to the everlasting discussion that started more than 130 years ago [59,60].

Author Contributions

Conceptualization, G.G. and M.G.; methodology, G.G. and M.G.; formal analysis, G.G. and K.M.; investigation, G.G. and M.G.; resources, G.G.; writing—original draft preparation, G.G. and M.G.; writing—review and editing, G.G., M.G., K.M. and L.D.; visualization, K.M.; supervision, G.G. All authors have read and agreed to the published version of the manuscript.

Funding

This study was funded by the Institute for Medical Research and Occupational Health, Zagreb, Croatia.

Institutional Review Board Statement

The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Ethics committee of the Institute for Medical Research and Occupational Health, Croatia (approval code: 01-74/1-11, date of approval: 16 March 2011).

Informed Consent Statement

Informed consent was obtained from all subjects involved in the study.

Data Availability Statement

The original contributions generated for this study are included in the article. Further inquiries can be directed to the corresponding author upon reasonable request.

Acknowledgments

The authors would like to thank all of the volunteers who donated their blood samples. We would also like to thank Maja Nikolić for her excellent technical assistance. All authors would declare themselves as omnivores. This study was funded by the Institute for Medical Research and Occupational Health and was supported by the European Cooperation in Science and Technology (CA COST Action CA15132–hCOMET).

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Helleday, T.; Eshtad, S.; Nik-Zainal, S. Mechanisms Underlying Mutational Signatures in Human Cancers. Nat. Rev. Genet. 2014, 15, 585–598. [Google Scholar] [CrossRef] [PubMed]
  2. Nikfarjam, S.; Singh, K.K. DNA Damage Response Signaling: A Common Link between Cancer and Cardiovascular Diseases. Cancer Med. 2023, 12, 4380–4404. [Google Scholar] [CrossRef] [PubMed]
  3. Ribezzo, F.; Shiloh, Y.; Schumacher, B. Systemic DNA Damage Responses in Aging and Diseases. Semin. Cancer Biol. 2016, 37–38, 26–35. [Google Scholar] [CrossRef] [PubMed]
  4. Gajski, G.; Gerić, M.; Jakaša, I.; Peremin, I.; Domijan, A.-M.; Vučić Lovrenčić, M.; Kežić, S.; Bituh, M.; Moraes de Andrade, V. Inflammatory, Oxidative and DNA Damage Status in Vegetarians: Is the Future of Human Diet Green? Crit. Rev. Food Sci. Nutr. 2021. [Google Scholar] [CrossRef]
  5. Vidaček Škrobot, N.; Nanić, L.; Ravlić, S.; Sopta, M.; Gerić, M.; Gajski, G.; Garaj-Vrhovac, V.; Rubelj, I. Telomeres, Nutrition, and Longevity: Can We Really Navigate Our Aging? J. Gerontol. Ser. A 2018, 73, 39–47. [Google Scholar] [CrossRef]
  6. Mattioli, R.; Mosca, L.; Sánchez-Lamar, A.; Tempera, I.; Hausmann, R. Natural Bioactive Compounds Acting against Oxidative Stress in Chronic, Degenerative, and Infectious Diseases. Oxid. Med. Cell. Longev. 2018, 2018, 3894381. [Google Scholar] [CrossRef]
  7. Teodoro, A.J. Bioactive Compounds of Food: Their Role in the Prevention and Treatment of Diseases. Oxid. Med. Cell. Longev. 2019, 2019, 3765986. [Google Scholar] [CrossRef]
  8. Riso, P.; Del Bo’, C.; Vendrame, S.; Brusamolino, A.; Martini, D.; Bonacina, G.; Porrini, M. Modulation of Plasma Antioxidant Levels, Glutathione S-Transferase Activity and DNA Damage in Smokers Following a Single Portion of Broccoli: A Pilot Study. J. Sci. Food Agric. 2014, 94, 522–528. [Google Scholar] [CrossRef]
  9. Moser, B.; Szekeres, T.; Bieglmayer, C.; Wagner, K.-H.; Mišík, M.; Kundi, M.; Zakerska, O.; Nersesyan, A.; Kager, N.; Zahrl, J.; et al. Impact of Spinach Consumption on DNA Stability in Peripheral Lymphocytes and on Biochemical Blood Parameters: Results of a Human Intervention Trial. Eur. J. Nutr. 2011, 50, 587–594. [Google Scholar] [CrossRef]
  10. Fogarty, M.C.; Hughes, C.M.; Burke, G.; Brown, J.C.; Davison, G.W. Acute and Chronic Watercress Supplementation Attenuates Exercise-Induced Peripheral Mononuclear Cell DNA Damage and Lipid Peroxidation. Br. J. Nutr. 2013, 109, 293–301. [Google Scholar] [CrossRef]
  11. Bakuradze, T.; Lang, R.; Hofmann, T.; Schipp, D.; Galan, J.; Eisenbrand, G.; Richling, E. Coffee Consumption Rapidly Reduces Background DNA Strand Breaks in Healthy Humans: Results of a Short-Term Repeated Uptake Intervention Study. Mol. Nutr. Food Res. 2016, 60, 682–686. [Google Scholar] [CrossRef] [PubMed]
  12. Gajski, G.; Gerić, M.; Vučić Lovrenčić, M.; Božičević, S.; Rubelj, I.; Nanić, L.; Škrobot Vidaček, N.; Bendix, L.; Peraica, M.; Rašić, D.; et al. Analysis of Health-Related Biomarkers between Vegetarians and Non-Vegetarians: A Multi-Biomarker Approach. J. Funct. Foods 2018, 48, 643–653. [Google Scholar] [CrossRef]
  13. Vučić Lovrenčić, M.; Gerić, M.; Košuta, I.; Dragičević, M.; Garaj-Vrhovac, V.; Gajski, G. Sex-Specific Effect of Vegetarian Diet on Adiponectin Levels and Insulin Sensitivity in Healthy Non-Obese Individuals. Nutrition 2020, 79–80, 110862. [Google Scholar] [CrossRef] [PubMed]
  14. Azqueta, A.; Ladeira, C.; Giovannelli, L.; Boutet-Robinet, E.; Bonassi, S.; Neri, M.; Gajski, G.; Duthie, S.; Del Bo’, C.; Riso, P.; et al. Application of the Comet Assay in Human Biomonitoring: An HCOMET Perspective. Mutat. Res. Mutat. Res. 2020, 783, 108288. [Google Scholar] [CrossRef] [PubMed]
  15. Gerić, M.; Gajski, G.; Oreščanin, V.; Garaj-Vrhovac, V. Seasonal Variations as Predictive Factors of the Comet Assay Parameters: A Retrospective Study. Mutagenesis 2018, 33, 53–60. [Google Scholar] [CrossRef]
  16. Gajski, G.; Gerić, M.; Živković Semren, T.; Tariba Lovaković, B.; Oreščanin, V.; Pizent, A. Application of the Comet Assay for the Evaluation of DNA Damage from Frozen Human Whole Blood Samples: Implications for Human Biomonitoring. Toxicol. Lett. 2020, 319, 58–65. [Google Scholar] [CrossRef] [PubMed]
  17. Collins, A.; Møller, P.; Gajski, G.; Vodenková, S.; Abdulwahed, A.; Anderson, D.; Bankoglu, E.E.; Bonassi, S.; Boutet-Robinet, E.; Brunborg, G.; et al. Measuring DNA Modifications with the Comet Assay: A Compendium of Protocols. Nat. Protoc. 2023, 18, 929–989. [Google Scholar] [CrossRef]
  18. Azqueta, A.; Collins, A.R. The Essential Comet Assay: A Comprehensive Guide to Measuring DNA Damage and Repair. Arch. Toxicol. 2013, 87, 949–968. [Google Scholar] [CrossRef]
  19. Bonassi, S.; Ceppi, M.; Møller, P.; Azqueta, A.; Milić, M.; Monica, N.; Brunborg, G.; Godschalk, R.; Koppen, G.; Langie, S.A.S.; et al. DNA Damage in Circulating Leukocytes Measured with the Comet Assay May Predict the Risk of Death. Sci. Rep. 2021, 11, 16793. [Google Scholar] [CrossRef]
  20. Milić, M.; Ceppi, M.; Bruzzone, M.; Azqueta, A.; Brunborg, G.; Godschalk, R.; Koppen, G.; Langie, S.; Møller, P.; Teixeira, J.P.; et al. The HCOMET Project: International Database Comparison of Results with the Comet Assay in Human Biomonitoring. Baseline Frequency of DNA Damage and Effect of Main Confounders. Mutat. Res. Mutat. Res. 2021, 787, 108371. [Google Scholar] [CrossRef]
  21. Møller, P.; Azqueta, A.; Boutet-Robinet, E.; Koppen, G.; Bonassi, S.; Milić, M.; Gajski, G.; Costa, S.; Teixeira, J.P.; Costa Pereira, C.; et al. Minimum Information for Reporting on the Comet Assay (MIRCA): Recommendations for Describing Comet Assay Procedures and Results. Nat. Protoc. 2020, 15, 3817–3826. [Google Scholar] [CrossRef] [PubMed]
  22. Willett, W.C.; Koplan, J.P.; Nugent, R.; Dusenbury, C.; Puska, P.; Gaziano, T.A. Prevention of Chronic Disease by Means of Diet and Lifestyle Changes; The International Bank for Reconstruction and Development/The World Bank: Washington, DC, USA, 2006; ISBN 0821361791. [Google Scholar]
  23. Kimokoti, R.W.; Millen, B.E. Nutrition for the Prevention of Chronic Diseases. Med. Clin. N. Am. 2016, 100, 1185–1198. [Google Scholar] [CrossRef] [PubMed]
  24. Craig, W.J. Nutrition Concerns and Health Effects of Vegetarian Diets. Nutr. Clin. Pract. 2010, 25, 613–620. [Google Scholar] [CrossRef] [PubMed]
  25. Craig, W.J. Health Effects of Vegan Diets. Am. J. Clin. Nutr. 2009, 89, 1627S–1633S. [Google Scholar] [CrossRef] [PubMed]
  26. Melina, V.; Craig, W.; Levin, S. Position of the Academy of Nutrition and Dietetics: Vegetarian Diets. J. Acad. Nutr. Diet. 2016, 116, 1970–1980. [Google Scholar] [CrossRef]
  27. Hargreaves, S.M.; Rosenfeld, D.L.; Moreira, A.V.B.; Zandonadi, R.P. Plant-Based and Vegetarian Diets: An Overview and Definition of These Dietary Patterns. Eur. J. Nutr. 2023, 62, 1109–1121. [Google Scholar] [CrossRef]
  28. Wulf, H.C.; Iversen, A.S.; Husum, B.; Neibuhr, E. Very Low Sister-Chromatid Exchange Rate in Seventh-Day Adventists. Mutat. Res. Fundam. Mol. Mech. Mutagen. 1986, 162, 131–135. [Google Scholar] [CrossRef]
  29. Fenech, M. Cytokinesis-Block Micronucleus Cytome Assay. Nat. Protoc. 2007, 2, 1084–1104. [Google Scholar] [CrossRef]
  30. Gajski, G.; Gerić, M.; Oreščanin, V.; Garaj-Vrhovac, V. Cytokinesis-Block Micronucleus Cytome Assay Parameters in Peripheral Blood Lymphocytes of the General Population: Contribution of Age, Sex, Seasonal Variations and Lifestyle Factors. Ecotoxicol. Environ. Saf. 2018, 148, 561–570. [Google Scholar] [CrossRef]
  31. Kopjar, N.; Kašuba, V.; Milić, M.; Rozgaj, R.; Želježić, D.; Gajski, G.; Mladinić, M.; Garaj-Vrhovac, V. Normal and Cut-off Values of the Cytokinesis-Block Micronucleus Assay on Peripheral Blood Lymphocytes in the Croatian General Population. Arh. Hig. Rada Toksikol. 2010, 61, 219–234. [Google Scholar] [CrossRef]
  32. Fenech, M.; Kirsch-Volders, M.; Natarajan, A.T.; Surralles, J.; Crott, J.W.; Parry, J.; Norppa, H.; Eastmond, D.A.; Tucker, J.D.; Thomas, P. Molecular Mechanisms of Micronucleus, Nucleoplasmic Bridge and Nuclear Bud Formation in Mammalian and Human Cells. Mutagenesis 2011, 26, 125–132. [Google Scholar] [CrossRef] [PubMed]
  33. Fenech, M. Cytokinesis-Block Micronucleus Cytome Assay Evolution into a More Comprehensive Method to Measure Chromosomal Instability. Genes 2020, 11, 1203. [Google Scholar] [CrossRef] [PubMed]
  34. Kazimírová, A.; Barancoková, M.; Volkovová, K.; Staruchová, M.; Krajcovicová-Kudlácková, M.; Wsólová, L.; Collins, A.R.; Dusinská, M. Does a Vegetarian Diet Influence Genomic Stability? Eur. J. Nutr. 2004, 43, 32–38. [Google Scholar] [CrossRef] [PubMed]
  35. Krajcovicova-Kudlackova, M.; Valachovicová, M.; Pauková, V.; Dusinská, M. Effects of Diet and Age on Oxidative Damage Products in Healthy Subjects. Physiol. Res. 2008, 57, 647–651. [Google Scholar] [CrossRef]
  36. Krajcovicova-Kudlackova, M.; Dusinská, M. Oxidative DNA Damage in Relation to Nutrition. Neoplasma 2004, 51, 30–33. [Google Scholar]
  37. Kotova, N.; Frostne, C.; Abramsson-Zetterberg, L.; Tareke, E.; Bergman, R.; Haghdoost, S.; Paulsson, B.; Törnqvist, M.; Segerbäck, D.; Jenssen, D.; et al. Differences in Micronucleus Frequency and Acrylamide Adduct Levels with Hemoglobin between Vegetarians and Non-Vegetarians. Eur. J. Nutr. 2015, 54, 1181–1190. [Google Scholar] [CrossRef]
  38. Dhawan, A.; Mathur, N.; Seth, P.K. The Effect of Smoking and Eating Habits on DNA Damage in Indian Population as Measured in the Comet Assay. Mutat. Res. Fundam. Mol. Mech. Mutagen. 2001, 474, 121–128. [Google Scholar] [CrossRef]
  39. Bajpayee, M.; Dhawan, A.; Parmar, D.; Pandey, A.K.; Mathur, N.; Seth, P.K. Gender-Related Differences in Basal DNA Damage in Lymphocytes of a Healthy Indian Population Using the Alkaline Comet Assay. Mutat. Res. 2002, 520, 83–91. [Google Scholar] [CrossRef]
  40. Verhagen, H.; Rauma, A.L.; Törrönen, R.; de Vogel, N.; Bruijntjes-Rozier, G.C.; Drevo, M.A.; Bogaards, J.J.; Mykkänen, H.; Dreve, M.; Bogaards, J.J.; et al. Effect of a Vegan Diet on Biomarkers of Chemoprevention in Females. Hum. Exp. Toxicol. 1996, 15, 821–825. [Google Scholar] [CrossRef]
  41. Davies, H.W.; Kennedy, S.M.; Teschke, K.; Quintana, P.J.E. Cytogenetic Analysis of South Asian Berry Pickers in British Columbia Using the Micronucleus Assay in Peripheral Lymphocytes. Mutat. Res. Toxicol. Environ. Mutagen. 1998, 416, 101–113. [Google Scholar] [CrossRef]
  42. Fenech, M.; Rinaldi, J. A Comparison of Lymphocyte Micronuclei and Plasma Micronutrients in Vegetarians and Non-Vegetarians. Carcinogenesis 1995, 16, 223–230. [Google Scholar] [CrossRef] [PubMed]
  43. Khuda-Bukhsh, A.R.; Das, S.; Saha, S.K. Molecular Approaches toward Targeted Cancer Prevention with Some Food Plants and Their Products: Inflammatory and Other Signal Pathways. Nutr. Cancer 2014, 66, 194–205. [Google Scholar] [CrossRef] [PubMed]
  44. Nosrati, N.; Bakovic, M.; Paliyath, G. Molecular Mechanisms and Pathways as Targets for Cancer Prevention and Progression with Dietary Compounds. Int. J. Mol. Sci. 2017, 18, 2050. [Google Scholar] [CrossRef] [PubMed]
  45. Duthie, S.J.; Ma, A.; Ross, M.A.; Collins, A.R. Antioxidant Supplementation Decreases Oxidative DNA Damage in Human Lymphocytes. Cancer Res. 1996, 56, 1291–1295. [Google Scholar]
  46. Dean, R.T.; Fu, S.; Stocker, R.; Davies, M.J. Biochemistry and Pathology of Radical-Mediated Protein Oxidation. Biochem. J. 1997, 324 Pt 1, 1–18. [Google Scholar] [CrossRef]
  47. Pool-Zobel, B.L.; Bub, A.; Müller, H.; Wollowski, I.; Rechkemmer, G. Consumption of Vegetables Reduces Genetic Damage in Humans: First Results of a Human Intervention Trial with Carotenoid-Rich Foods. Carcinogenesis 1997, 18, 1847–1850. [Google Scholar] [CrossRef]
  48. Kazimírová, A.; Barancoková, M.; Krajcovicová-Kudlácková, M.; Volkovová, K.; Staruchová, M.; Valachovicová, M.; Pauková, V.; Blazícek, P.; Wsólová, L.; Dusinská, M. The Relationship between Micronuclei in Human Lymphocytes and Selected Micronutrients in Vegetarians and Non-Vegetarians. Mutat. Res. 2006, 611, 64–70. [Google Scholar] [CrossRef]
  49. Wasson, G.R.; McKelvey-Martin, V.J.; Downes, C.S. The Use of the Comet Assay in the Study of Human Nutrition and Cancer. Mutagenesis 2008, 23, 153–162. [Google Scholar] [CrossRef]
  50. Gerić, M.; Matković, K.; Gajski, G.; Rumbak, I.; Štancl, P.; Karlić, R.; Bituh, M. Adherence to Mediterranean Diet in Croatia: Lessons Learned Today for a Brighter Tomorrow. Nutrients 2022, 14, 3725. [Google Scholar] [CrossRef]
  51. Smolková, B.; Dusinská, M.; Raslová, K.; Barancoková, M.; Kazimírová, A.; Horská, A.; Spustová, V.; Collins, A. Folate Levels Determine Effect of Antioxidant Supplementation on Micronuclei in Subjects with Cardiovascular Risk. Mutagenesis 2004, 19, 469–476. [Google Scholar] [CrossRef]
  52. Herrmann, W.; Schorr, H.; Purschwitz, K.; Rassoul, F.; Richter, V. Total Homocysteine, Vitamin B(12), and Total Antioxidant Status in Vegetarians. Clin. Chem. 2001, 47, 1094–1101. [Google Scholar] [CrossRef] [PubMed]
  53. Obersby, D.; Chappell, D.C.; Dunnett, A.; Tsiami, A.A. Plasma Total Homocysteine Status of Vegetarians Compared with Omnivores: A Systematic Review and Meta-Analysis. Br. J. Nutr. 2013, 109, 785–794. [Google Scholar] [CrossRef]
  54. Nair-Shalliker, V.; Armstrong, B.K.; Fenech, M. Does Vitamin D Protect against DNA Damage? Mutat. Res. 2012, 733, 50–57. [Google Scholar] [CrossRef] [PubMed]
  55. Ames, B.N. DNA Damage from Micronutrient Deficiencies Is Likely to Be a Major Cause of Cancer. Mutat. Res. 2001, 475, 7–20. [Google Scholar] [CrossRef] [PubMed]
  56. Ames, B.N. Micronutrient Deficiencies: A Major Cause of DNA Damage. Ann. N. Y. Acad. Sci. 1999, 889, 87–106. [Google Scholar] [CrossRef]
  57. Kapiszewska, M. A Vegetable to Meat Consumption Ratio as a Relevant Factor Determining Cancer Preventive Diet. The Mediterranean versus Other European Countries. Forum Nutr. 2006, 59, 130–153. [Google Scholar] [CrossRef]
  58. Viegas, S.; Ladeira, C.; Costa-Veiga, A.; Perelman, J.; Gajski, G. Forgotten Public Health Impacts of Cancer—An Overview. Arh. Hig. Rada Toksikol. 2017, 68, 287–297. [Google Scholar] [CrossRef]
  59. Anonymous. Annotations: Irish Distressed Ladies.-Vegetarianism.-“Sweating”. Hospital 1888, 3, 446. [Google Scholar]
  60. Anonymous. Vegetarianism and Cancer. Hospital 1888, 5, 124. [Google Scholar]
Nutrients 15 02218 g001 550
Figure 1. The difference in % tail DNA among vegetarians and non-vegetarians; * p < 0.05.
Figure 1. The difference in % tail DNA among vegetarians and non-vegetarians; * p < 0.05.
Nutrients 15 02218 g001
Nutrients 15 02218 g002 550
Figure 2. The differences in % tail DNA among three tested groups (omnivorous, vegetarians, and pescatarians); ns—non-significant, * p < 0.05.
Figure 2. The differences in % tail DNA among three tested groups (omnivorous, vegetarians, and pescatarians); ns—non-significant, * p < 0.05.
Nutrients 15 02218 g002
Table
Table 1. Population characteristics in vegetarians versus non-vegetarians (n = 32) and their related sub-groups.
Table 1. Population characteristics in vegetarians versus non-vegetarians (n = 32) and their related sub-groups.
Vegetarians
(n = 16)		Non-Vegetarians
(n = 16)
Omnivores
(n = 8)		Pescatarians
(n = 8)
Age (years)
Mean ± SD31.4 ± 3.131.0 ± 3.531.0 ± 3.931.0 ± 3.27
Range25–3526–3526–3526–35
Vegetarianism (years)
Mean ± SD10.7 ± 3.8---
Range 4–18---
Tail intensity (% tail DNA)
Mean ± SD3.6 ± 1.1 *,#2.8 ± 1.03.2 ± 0.82.4 ± 1.1
Range1.9–6.00.9–4.62.1–4.60.9–3.7
* Statistically significant increase compared to non-vegetarians (p < 0.05); # statistically significant increase compared to pescatarians (p < 0.05).
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Gajski, G.; Matković, K.; Delić, L.; Gerić, M. Evaluation of Primary DNA Damage in Young Healthy Females Based on Their Dietary Preferences. Nutrients 2023, 15, 2218. https://doi.org/10.3390/nu15092218

AMA Style

Gajski G, Matković K, Delić L, Gerić M. Evaluation of Primary DNA Damage in Young Healthy Females Based on Their Dietary Preferences. Nutrients. 2023; 15(9):2218. https://doi.org/10.3390/nu15092218

Chicago/Turabian Style

Gajski, Goran, Katarina Matković, Luka Delić, and Marko Gerić. 2023. "Evaluation of Primary DNA Damage in Young Healthy Females Based on Their Dietary Preferences" Nutrients 15, no. 9: 2218. https://doi.org/10.3390/nu15092218

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop