The Role of Sunscreen in Mitigating UV-Induced Damage in Saccharomyces cerevisiae Yeast Respiration and Survival, as a Eukaryotic Model Organism

Abstract

Mutagens are agents that cause damage to DNA and have the potential to permanently alter (mutate) its sequence, depending on the organism’s ability to repair the damage. UV radiation is a mutagen in cells. This mutagen relates to both yeast cells and human skin cells, since they have similar reactions. UV radiation can cause cell mutations, but also cell death. This is examined with the absence or presence of sunscreen when in contact with cells. Since yeast cells and human cells have almost identical metabolisms, data results of yeast experiments can be associated with real life. Hypothesis for the presence of sunscreen in yeast solutions includes the exposure of yeast cells with or without sunscreen for different time periods in UV radiation. However, the role of sunscreen in yeast cell mutations, in relation to cancer prevention, may not be directly positive. Here, I show that sunscreen has a positive effect on yeast cells and prevents mutations. I found that the respiration rate differs for yeast cells without or without the presence of sunscreen when exposed to radiation. Yeast cells without sunscreen respired faster than those exposed to UV radiation. However, with sunscreen, the rate of CO2 production was higher, with a higher respiration rate. These results may be connected with skin cancer to some extent, promoting or not the use of sunscreen to protect the skin cells from mutating. This experiment may be the base for further experimentation with different yeast cells, providing clearer and more assuring data about the association of sunscreen, yeast cells, and skin cancer. Such experiments may avoid implications with weather conditions, such as slightly different temperatures, sunlight intensity, and clouds, or with the time between the end of the time period of exposure of the yeast cells to UV radiation, and the measurement of CO2 and density, which my experiment had.

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Kampani, T. (2024) The Role of Sunscreen in Mitigating UV-Induced Damage in Saccharomyces cerevisiae Yeast Respiration and Survival, as a Eukaryotic Model Organism. Advances in Bioscience and Biotechnology, 15, 696-723. doi: 10.4236/abb.2024.1512044.

1. Introduction

1.1. Yeasts

Yeasts, a group of around 1500 single-celled fungi species, are found in sugary media like fruit nectar and flower nectar, and have been traditionally used in the production of bread, beer, and wine [1]. They are eukaryotic organisms with a diameter of around 0.075 mm and can be spherical, egg-shaped, or filamentous [1]. Most yeasts reproduce asexually through budding, while some split into two equal cells through fission [1].

Yeasts are used in food production, where they produce carbon dioxide and ethanol through fermentation. These byproducts are used in bakery products, beer, and wine making. Yeast cells can ferment about their own weight of glucose in an hour [1]. Yeast for baking is available in two forms: compacted cakes with starch or dry grains mixed with cornmeal [1]. Commercial yeast is rich in niacin, folic acid, vitamins B1, B2, and B2, and has a 50% protein content. Deactivated brewer’s yeast and nutritional yeast can be taken as vitamin supplements [1].

Figure 1. Diversity of outlets involving yeast biotechnology roles [3].

Yeast identification involves physiological and morphological assays, including auxanography for determining carbon and nitrogen sources. Auxanography is a study that determines the growth of mutants which require specific substances to develop [2]. Systems like BCCM/Allev 2.00 and API strips analyze sugar absorption and fermentation. Yeasts inhabit diverse environments such as plant tissues, air, water, and land, with some thriving in solute-rich, extreme conditions. Species like S. cerevisiae colonize wine, while others are found in hospitals or cause food spoilage [3] (Figure 1).

Saccharomyces cerevisiae relies on fermentation for energy, even in the presence of oxygen. When glucose is scarce, it switches to using ethanol as a carbon source, triggering a shift in gene expression to favor gluconeogenesis (metabolic reactions that maintain blood glucose levels constant after digestion) [4] and the glyoxylate cycle (a variant of the tricarboxylic cycle found in plants, fungi and protists, permitting the use of two carbon compounds, when glucose is not present) [5], while reducing fermentation-related genes. Zinc cluster proteins like Cat8, Sip4, Rds2, and Adr1 drive this gene reprogramming.

NAD+ + 2e + H+ ◊ Reduced NAD (NADH + H+) (Figure 2)

Figure 2. Cellular respiration and fermentation overview [6].

1.2. Sunscreens

The standard erythema dose (SED) measures the biological efficiency of UV radiation, specifically its ability to cause erythema (skin reddening). Sunscreens, although often applied less thoroughly than in SPF testing, can reduce sunburn cells, DNA damage, and risks of skin cancer such as actinic keratoses and squamous cell carcinomas. Chronic UV exposure is the leading cause of malignant melanoma and contributes to both photoaging and photo carcinogenesis [7]. Inorganic sunscreens like zinc oxide and titanium dioxide are effective and increasingly popular due to improved formulations [7]. SPF remains a key measure of sunscreen efficacy, and recent advancements have combined UV filters with DNA repair agents to enhance skin protection [7].

Extraterrestrial sunlight at sea level includes electromagnetic radiation from 290 to 3000 nm, with UV radiation categorized into UVA (320 - 400 nm), UVB (290 - 320 nm), and UVC (200 - 290 nm). While UVA predominates at the Earth’s surface, UVC is fully absorbed by the ozone layer. Variations in UV radiation depend on latitude, time of day, and season. The depletion of the ozone layer due to pollutants like nitric oxides and chlorofluorocarbons could increase UVB and UVC exposure, raising risks of skin cancer, photo immunosuppression, premature aging, and photosensitive diseases [8].

UV radiation, prevalent in the environment, contributes to skin diseases like inflammation, aging, and cancer. Personal exposure depends on sunlight intensity, time outdoors, and protective measures like clothing and sunscreen [9].

UVB and UVA radiation damage skin biomolecules, with UVB directly causing DNA lesions that can lead to mutations and skin cancer, particularly involving p53 mutations. The majority of p53 mutations are missense mutations, which produce full-length mutant p53 proteins. In addition to losing their ability to inhibit malignancies in a way dependent on wild-type p53, mutant p53 (Mutp53) proteins frequently acquire oncogenic gain-of-functions (GOF) that promote tumor growth [10]. UVA contributes to photoaging and suppresses the immune system. Photoprotection, including sun avoidance, protective clothing, and sunscreen, is central for preventing skin damage, photo immunosuppression, and skin cancers, and has become a major public health approach [10] [11].

Octinoxate, a common UVB absorber, is well-tolerated but degrades under sunlight, reducing its effectiveness. Encapsulation in nanoparticles can enhance their photostability. Avobenzone (Parsol 1789) is a strong UVA filter, but stabilizers may be needed to prevent degradation. Benzophenone-3, widely used and highly bioavailable, has a higher incidence of photodermatitis. Diethylamino hydroxybenzyl hexyl benzoate, more photostable than avobenzone, offers similar protection [12].

Photostability and Water Resistance

Photostability is essential for sunscreen effectiveness, ensuring it maintains its protective properties under sunlight. Some chemical filters, like octyl dimethyl PABA and avobenzone, can be photoreactive, reducing stability. Other filters, such as zinc oxide (ZnO), titanium dioxide (TiO2), salicylates, and methyl benzylidene camphor, enhance photostability, helping sunscreens better absorb, reflect, and scatter UV rays while remaining stable [13].

Sunscreen effectiveness in water is assessed by its ability to maintain SPF after immersion. In Europe, sunscreens are classified as “water-resistant” or “extra water-resistant” if post-immersion SPF remains at least 50% of the initial value after 40 or 80 minutes in water. The SPF label in the US reflects the pre-water exposure value [14].

Sunscreens and Melanoma

The link between sunscreen use and melanoma risk is debated, with studies showing conflicting results. Some suggest lower melanoma incidence with sunscreen use [15], while others indicate no significant impact [16]. A meta-analysis found little correlation, likely due to varying study methods and early sunscreens providing only UVB protection [17]. Modern sunscreens with broad-spectrum protection are still essential for preventing sunburn and mutations linked to melanoma, though their effectiveness against melanoma needs further study. Recent trials hint that sunscreens might reduce the risk of developing melanocytic naevi, a melanoma precursor [15] [16].

Cutaneous Responses to UV

UV radiation impacts skin physiology both immediately and over time [18]. Acute effects include inflammation, or “sunburn,” triggered by cytokines, which are small proteins that control the development and operation of blood cells and other immune system components [19], and other mediators, leading to keratinocyte apoptosis [19]. UV exposure also causes hyperkeratosis (thickening of the epidermis) and activates damage responses, such as p53-mediated cell cycle arrest and DNA repair [18]. The organism used in this investigation, yeast, has a similar mechanism. Tanning, an adaptive response, increases melanin production to protect against further UV damage, though defects in this process can raise cancer risk. UV light also affects immune function and converts 7-dehydrocholesterol into vitamin D3 [9]. UVA mainly causes oxidative damage, while UVB directly damages DNA, with ongoing research into their effects on the skin [9].

Oxidative Injury

Figure 3. UV photons generate oxidative free radicals, causing structural and functional changes in macromolecules like DNA, RNA, protein, and lipids. Enzymes like glutathione peroxidase, catalase, and superoxide dismutase detoxify these species. Adapted from: https://www.researchgate.net/figure/UV-generates-oxidative-free-radicals-UV-photons-interact-with-atomic-oxygen-to-promote_fig5_237095045

UV light generates reactive oxygen species (ROS), leading to mutations [20]. ROS causes nucleotide damage, resulting in mispairing and mutagenesis, such as the guanine-to-thymine mutation via 8-hydroxy-2’-deoxyguanine (8-OHdG), linked to skin cancer [21]. The base excision repair (BER) pathway repairs DNA damage, with glycosylases identifying and removing altered bases [20]. Antioxidant systems, including glutathione, superoxide dismutase (SODs), and catalase, detoxify ROS, protecting DNA and other macromolecules from UV-induced damage. These processes are crucial in managing the skin’s response to UV radiation [20] (Figure 3).

2. Research Question

How does exposure time to ambient ultraviolet (UV) radiation (0, 15, 30, 60 minutes) affect the rate of respiration, indicating the death toll of the yeast cells, and survival of yeast (Saccharomyces cerevisiae) by measuring carbon dioxide production (in ppm, ±10) and cell viability through the density in spectrophotometer, in the presence and absence of sunscreen?

3. Hypothesis

It is expected that without the presence of sunscreen, as exposure to ultraviolet radiation increases the rate of respiration of yeast and the number of yeast cells surviving will decrease. This is because exposure to UV radiation, damages the yeast cells. This is expected to be more evident at the high exposure time (60 minutes). On the other hand, in the presence of sunscreen it is expected that most yeast cells will survive, and the rate of respiration will also increase. Hence, there will be an increase in the amount of carbon dioxide produced.

4. Variables

4.1. Independent Variables

1) Presence of 9 ml/50ml yeast solution, or absence of Frezyderm Seaside sunscreen UV protection level 50+. The reason that that 9 ml of sunscreen were used per 50 ml of the yeast solution is that it was the minimum amount of sunscreen that created a thin layer on the surface of the solution. The aim was for this surface to be as thin as possible for it to not be embedded in the yeast solution and affect it even more. Also, after having tested different amounts of sunscreen and mixing the sunscreen with the yeast solution, the best outcome was with 9 ml, where only a thin layer was formed. In the other cases, there needed more titrations, leading to a greater error.

2) Time in minutes (0.0 min, 15.0 min, 30.0 min, 60.0 min, ±0.1 min) of exposure to ultra-violet radiation. The solution was placed in natural UV light, so that it could be exposed in the variety of wavelengths and natural conditions, and not only in the UV chamber which has only UVC. Also, these specific time periods were chosen for various of reasons. Firstly, 0.0 was chosen, to have a control variable which would not be exposed to the UV light at all, so that it could be comparable with the rest of the results. Also, this way, there is a better understanding of if the increase or decrease in CO2, or in the number of cells, was caused by contact inhibition of the yeast, mutations, or actually due to the UV radiation. More specifically, in contact inhibition, there is the factor of the toxic products that are attained and finally create a toxic environment which causes the death of the cells. Overpopulation is the main cause of this, when mutations may also be fatal.

In Table A1 (Appendix A), the ingredients of the sunscreen Frezyderm Seaside sunscreen UV protection level 50+ [22] are presented. To further understand its role, the purposes and the specific wavelength of UV radiation they present are listed.

4.2. Dependent Variables

1) Rate of respiration of yeast (Saccharomyces cerevisiae) by measuring the volume of carbon dioxide gas produced (ppm) using a carbon dioxide sensor (±10 ppm). CO2 will be measured so as to understand if the UV radiation causes cells to die or increases their rate of division. If cells die, then CO2 will decrease, since the rate of respiration will decrease, and even stop for some cells (the ones that will die). If UV radiation causes mutations, then CO2 will increase, since more cells will be respiring.

2) Number of yeast cells surviving measured using a spectrophotometer (absorbance). This number will be measured to understand if the UV radiation causes cells to die or mutate and increase in number. To convert absorbance to density, N = 0.125 * r was used, where N is the density, and r is the number of times which the sample is diluted [23]. If UV radiation kills the cells, then the density after exposure will decreases. On the other hand, if it causes mutations, then the cell density will increase.

In Table 1, the controlled variables are stated. Specifically, the reason and the way they were held constant is presented.

4.3. Controlled Variables

Table 1. Controlled variables, reasons for control, and the method of control.

Controlled variable

Reason for control

Method of control

Volume of yeast solution used per trial

So that the same number of yeast organisms are present in each trial

50 ml measured using a volumetric cylinder

Type of yeast (Saccharomyces cerevisae) used per trial

Different yeasts may carry out respiration at different rates and may be affected by UV radiation in different ways

Dry baker’s yeast

Volume of Frezyderm seaside sunscreen used per trial

So that the same number of sunscreen molecules are present for each different time of exposure

9.0 ml measured using a volumetric cylinder

Number of trials per exposure to UV radiation

So that the experiment is fair and so that sufficient data can be collected

5 trials per condition

Temperature of water bath used for the control trials

As all samples which were exposed to UV radiation where at an ambient temperature of 36˚C, the control yeasts were placed in an electronic water bath at the same temperature

Water bath set at 36˚C and monitored using a thermometer

Duration that the yeast was allowed to respire for before and after UV exposure

So that the experiment is fair and so that each yeast is given the same time to respire

10 minutes each measured using a digital timer

Speed of stirring yeast solution

So that the mixing of yeast and water was at the same intensity

Magnetic stirrer set at speed number 9

Speed of stirring while yeast solution was allowed to respire

So that the mixing of yeast and water was at the same intensity

Magnetic stirrer set at speed number 2

Location of experimental setup while exposing yeast to UV radiation

To allow for the same ambient temperature and amount of UV radiation

All trials were carried out outdoors, while also checking daily for the weather, UV index, and ambient temperature (information can be found in Figures A1-A3 in Appendix A)

5. Materials and Apparatus

  • Dry baker’s yeast (Saccharomyces cerevisiae), 7.5 grams

  • Tap water, 2000 ml

  • Frezyderm Seaside sunscreen UV protection level 50+, 54 ml

  • Electronic weighing scale (±0.0002 g)

  • Magnetic stirrer

  • Carbon dioxide sensor

  • PASCO Capstone program

  • Conical flask 200 ml (±25 ml)

  • Volumetric cylinder 50 ml (±2.5 ml)

  • Volumetric cylinder 250 ml (±10 ml)

  • Beaker 50 ml (±5 ml)

  • Beaker 500 ml (±50 ml)

  • Spatula

  • Hot plate

  • Electronic water bath set at 36˚C

  • Thermometer

  • Digital timer (±0.1 s)

  • Plastic pipette

  • Glass stirring rod

  • Permanent marker

  • Tweezer

6. Method

6.1. Part A: Measuring Carbon Dioxide

1) 500 ml of tap water was measured using a volumetric cylinder and added to a beaker.

2) 2.5 grams of dry yeast were measured using an electronic weighing scale and added to the beaker containing the water.

3) The beaker was placed on a hot plate and a magnetic stirrer was added.

4) The stirrer was switched on at speed 9 and was allowed to stir until all the yeast had dissolved.

5) 50 ml of yeast solution was measured using a volumetric cylinder and added to a 250 ml conical flask

6) 9 ml of sunscreen was measured using a volumetric cylinder and added into the conical flask containing the 50 ml yeast forming a protective layer.

7) A small magnetic stirrer was added and switched on at speed 2.

8) The carbon dioxide sensor was placed in position at the opening of the conical flask.

9) RUN was pressed on the PASCO Capstone program and the levels of carbon dioxide (in ppm) were measured for 10 minutes.

10) The flask was then taken outdoors and sat under UV radiation for 60 minutes.

11) Steps 8 and 9 were repeated.

12) Steps 5 - 10 were repeated four more times.

13) Steps 5 - 12 were repeated for the remaining exposure times (15 min, 30 min)

14) Steps 1 - 13 were repeated in the absence of sunscreen (without step 6).

15) For the control (0 min exposure) steps 1 - 6 were repeated.

16) The conical flask was placed in an electronic water bath set at 36˚C until the mixture reached the desired temperature.

17) Steps 7 - 9 were repeated.

18) Steps 15 - 17 were repeated in the absence of sunscreen.

6.2. Part B: Measuring Number of Yeast Cells

1) 500 ml of tap water was measured using a volumetric cylinder and added to a beaker.

2) 2.5 grams of dry yeast were measured using an electronic weighing scale and added to the beaker containing the water.

3) The beaker was placed on a hot plate and a magnetic stirrer was added.

4) The stirrer was switched on at speed 9 and was allowed to stir until all the yeast had dissolved.

5) 3 ml of distilled water were measured using a plastic pipette and poured in 1 glass cuvette as the control measurement.

6) 3 ml of the yeast solution were measured using a plastic pipette and poured in 1 glass cuvette.

7) The two glass cuvettes were placed in the 2nd and 3rd slots of the spectrophotometer respectively.

8) Calibration at the distilled water took place, until it reached exactly 100%.

9) The drawer with the two cuvettes was moved and the cuvette with the solution was now in front of the light source.

10) The % mode was changed to the absorption setting.

11) Time passed until the absorbance was stable.

12) Dilutions were done until the absorbance was equal to 0.125.

13) Calculations were made to convert the absorbance into density (N = 0.125 * r).

14) 50 ml of yeast solution was measured using a volumetric cylinder and added to a 250 ml conical flask.

15) 9 ml of sunscreen was measured using a volumetric cylinder and added into the conical flask containing the 50 ml yeast forming a protective layer.

16) 4 conical flasks were prepared without sunscreen and 4 with sunscreen, with each the 50 ml of yeast solution.

17) One of each category of the conical flaks were the control measurements which were not exposed to the sun at all.

18) 1 of each flask was then exposed to the sun for 15 min, 30 min, 60 min.

19) After each period steps 5 - 13 were repeated.

7. Data Collection

The following Table 2 is an example of the data collected from the concentration of carbon dioxide released from the yeast cells with no exposure to UV. The remaining data can be found in the Appendix B (Tables B1-B15).

Table 2. Concentration of carbon dioxide released with no exposure to UV (control) without sunscreen.

Time of recording (min)

Concentration of carbon dioxide/ppm

Duration of culture growth

15 min

30 min

45 min

60 min

0

900

404

702

676

1.0

1620

1206

1024

1022

2.0

2118

1640

1474

1322

3.0

2970

2006

1786

1620

4.0

3574

2466

2096

1892

5.0

4114

2880

2464

2192

6.0

4624

3266

2794

2512

7.0

5084

3620

3126

2796

8.0

5604

3930

3412

3062

9.0

5994

4192

3644

3328

10.0

6520

4510

3946

3568

8. Processing of Data

In each table the rate of the change of CO2 is calculated, per time, and then the average rate is estimated together with the standard deviation values.

1) The rate is calculated with the following formula (sample calculation, Table B1, Trial 1):

rateofcarbondioxideconcentrationchange= ( finalvalueinitialvalue )ppm 10minutes = ( 7526528 )ppm 10min =700 ppm min 1

2) The average rate from the five trials is calculated with the following formula (sample calculation, Table 1, Trials 1 - 5):

Averagerateofcarbondioxideconcentrationchange=( Ratetrial1++Ratetrial5 5 )ppm min 1 =741ppm min 1

3) With an SD value of 94 ppm·min1.

These results are summed up and presented in the following Table 3.

The following figure illustrates the changes in CO2 concentration with both the presence or absence of sunscreen over the yeast cells. The results presented in these tables are introduced to Excel to construct relevant graphs (Figures 4-7).

Table 3. Rate of CO2 concentration change over time in different UV exposure duration and in the presence or absence of sunscreen.

Conditions

Rate of CO2 concentration change over time/ppm·min1

Trials

1

2

3

4

5

Average

SD

0 exposure, no sunscreen

700

776

873

770

616

741

94

0 exposure, with sunscreen

1117

1105

1239

1118

1222

1160

65

Before 15 min exposure no sunscreen

531

520

476

455

485

493

32

After 15 min exposure no sunscreen

154

156

141

129

142

145

11

Before 15 min exposure with sunscreen

1212

1135

1166

1103

1272

1178

66

After 15 min exposure with sunscreen

1158

956

1035

1008

1005

1032

76

Before 30 min exposure no sunscreen

1065

906

928

952

851

904

79

After 30 min exposure no sunscreen

765

492

572

691

634

631

94

Before 30 min exposure with sunscreen

1343

1363

1475

1588

1477

1449

99

After 30 min exposure with sunscreen

1337

1342

1377

1564

1288

1381

107

Before 60 min exposure no sunscreen

702

637

599

495

473

581

97

After 60 min exposure no sunscreen

483

471

415

398

549

462

60

Before 60 min exposure with sunscreen

1146

1237

1311

1351

1273

1284

93

After 60 min exposure with sunscreen

1328

1209

1476

1227

1288

1326

92

Figure 4. Concentration of carbon dioxide released with no exposure to UV (control) without sunscreen.

Figure 5. Average rate of carbon dioxide concentration change before and after exposure of yeast to UV for increasing times, with and without sunscreen (Error bars show ±1sd).

Figure 6. Average rate of carbon dioxide concentration change after exposure to UV for increasing times, with and without sunscreen (Error bars show ±1sd).

Figure 7. Cell density assessed spectrophotometrically with and without sunscreen versus times of UV exposure.

9. Statistical Analysis

1) Rate of CO2 Concentration Change:

The t-test comparing the rates of CO2 concentration change between the trials with sunscreen and without sunscreen shows that:

  • t-statistic: 6.577

  • p-value: 2.62 × 105

This is a very low p-value that indicates a statistically significant difference between the rates of CO2 concentration change with and without sunscreen across all exposure times.

2) Cell Density:

The t-test comparing the cell densities with sunscreen and without sunscreen shows that:

  • t-statistic: 0.055

  • p-value: 0.958

This high p-value suggests there is no statistically significant difference in cell density between the groups with and without sunscreen.

The important finding in this case is that the sunscreen appears to have a significant effect on the rate of CO2 production, but not on cell density.

3) To analyze whether there are significant differences before and after UV exposure both with and without sunscreen, separate t-tests were run for the before and after groups.

The paired t-test results comparing the rate of CO2 concentration change before and after UV exposure show the following:

  • Without Sunscreen:

  • t-statistic = 7.76

  • p-value = 1.94 × 106

This p-value is much smaller than the significance level of 0.05, indicating a statistically significant difference between the rates before and after UV exposure without sunscreen.

  • With sunscreen:

  • t-statistic = 1.81

  • p-value = 0.091

This p-value is larger than the significance level of 0.05, indicating no statistically significant difference between the rates before and after UV exposure with sunscreen.

In conclusion:

1) There is a significant change in the rate of CO2 concentration without sunscreen after UV exposure.

2) There is no significant change in the rate with sunscreen after UV exposure, indicating sunscreen helps decrease the adverse effect of UV on respiration rates in yeast.

10. Discussion

The observed phenomena that the cell density differences are not statistically significant, while the respiration rates are, can be explained by taking into account the effects of UV radiation on cellular metabolic processes apart from cell survival. UV radiation, mostly UVB and UVC, can cause significant damage to cellular molecules like DNA, proteins, and membranes, which would detrimentally affect the cell’s ability to function normally, even in the case that the cell survives.

1) Metabolic Machinery Damage: Although the overall number of yeast cells shown by the cell density remains intact between groups with and without sunscreen, UV radiation can damage cellular molecules essential for respiration. UV radiation causes damage to DNA, proteins, and enzymes, especially those involved in respiration like cytochrome c oxidase or enzymes in the glycolytic pathway [24]. The damage of such proteins, disrupts the electron transport chain. Thus, the cells may remain alive (thus the density remains the same), but their ability to produce ATP is compromised. Moreover, damage to mitochondria or other key organelles involved in energy production can lead to reduced metabolic efficiency [25] [26]. These damages in DNA impair the yeast cells’ ability to carry out oxidative phosphorylation, which happens in human cells, too.

2) Sublethal Damage: UV exposure may cause sublethal damage, where the yeast cells are not instantly killed but experience dysfunction. In such cases the cells may divide or survive but with impaired metabolic functions [27]. This may explain why the respiration rates are significantly lower without sunscreen, as UV-damaged cells struggle to perform effectively their cellular respiration [27] [28].

3) Heterogeneity of Damage: Not all cells may be equally influenced by UV radiation. Some cells in the population may be more resistant to UV, while others may be significantly damaged. This in turn could create a population where some cells continue to respire normally, whereas others would show severely decreased respiration, keeping the density constant but decreasing the overall CO2 production [24] [29].

4) Apoptosis or Programmed Cell Death: UV radiation may also induce apoptosis or programmed cell death mechanisms in yeast, which could result in cells that remain intact, contributing to the cell density, but being metabolically inactive, therefore contributing less to respiration. In this case, cells damaged by UV can still be detected by the spectrophotometer measuring density, but they are not functioning normally in terms of metabolism [30] [31].

5) Sunscreen Protection: Sunscreen creates protective barriers that absorb, scatter, or reflect UV radiation. The ingredients that do so may be Zinc Oxide, Titanium Dioxide, and other UV filters. This way, the UV cannot penetrate the cell membrane. This membrane can be the membrane of a yeast cell, or the cell of human skin [32].

6) Relation with Human Skin Cells: It is true that yeast cells have similarities with human skin cells. Most similarities are found in the metabolic processes that both types of cells follow. Except for metabolic processes, they follow similar division and growth processes. The similarities are the reasons that yeast cells can be a model to study human cells. But, there are also differences that cause limitations. Such differences are the process of respiration without oxygen present. Yeast cells follow alcoholic fermentation, while human cells undergo anaerobic respiration.

7) General Information: Since the role of sunscreen is so major it must be spread. Schools must implement specific sessions, not only for students, but parents as well. Another way to increase awareness, and cancer prevention, is to directly provide people with sunscreens. On the other hand, there might be a disadvantage concerning the environment. All the plastic bottles and caps, can easily be carried by wind during summer, polluting beaches and nature. This might also kill animals, distracting ecosystems [33].

By looking at the data collected we can make the following observations:

For yeast without sunscreen, the average rate of CO2 concentration did not follow a clear trend. At 0 minutes without UV exposure, the rate was 741 ppm per minute, decreasing after 15 minutes and nearly doubling at 30 minutes, indicating peak respiration. After UV exposure, respiration began later and increased more slowly, showing yeast respired faster without UV exposure.

With sunscreen, CO2 production was generally much higher, both with and without UV exposure. UV radiation slowed the onset of respiration, but CO2 production at 15, 30, and 60 minutes was consistently higher. At 60 minutes, yeast exposed to UV with sunscreen respired faster than those unexposed. The small difference in average CO2 concentration before and after exposure suggests effective protection by sunscreen. The statistical analysis supports this, with a t-statistic (6.577) and a p-value (2.62 × 105) confirming sunscreen’s protective role against UV-induced metabolic damage and a t-statistic (7.76) with a p-value (1.94 × 106) highlighting the harmful effects of UV on yeast respiration. This aligned with the hypothesis, since without sunscreen, the yeast cells’ survival decreased. This was more evident at the high exposure time (60 minutes). On the other hand, in the presence of sunscreen most yeast cells survived, with the rate of respiration increasing.

In conclusion, this study highlights the significant protective role of sunscreen in mitigating the adverse effects of UV radiation on Saccharomyces cerevisiae, a eukaryotic model organism. By preserving metabolic processes and shielding cells from UV-induced damage, sunscreen proves critical in protecting cellular components essential for survival. Furthermore, the findings establish a clear link between UV exposure and metabolic dysfunction, reinforcing the parallels between yeast and human skin cells and emphasizing sunscreen’s importance in promoting cellular health and preventing damage.

11. Evaluation

In Table 4, the limitations of the experiment and ways to improve such problems are suggested.

Table 4. Evaluation of data collected.

Limitations

Suggested improvements

In the control runs, the water bath was used to match the yeast solution’s temperature to the ambient temperature of the other samples. However, some flasks remained in the water bath longer, causing temperature variations that could have affected the cells differently.

To improve this, each trial should be carried out on its own, placing only one flask within the water bath and removing it at the appropriate time.

The effects of UV radiation were examined on only one type of yeast, dry yeast.

To improve this, repeat the procedure using wet yeast, to collect comparable data.

The location where the flasks were placed was not kept constant, due to the absence or presence of sunlight on each particular day.

A solution of this would be to place them in the UV chamber, so that they receive the same amount of radiation, however it would only be UVC.

The UV intensity varied from day to day due to the presence of clouds. (Figure A3, Appendix A)

Yeasts should be placed in the UV chamber, so that they receive the same amount of radiation, however it would only be UVC.

The magnetic stirrer was not on operation while the data was being collected.

To improve this, leave the magnetic stirrer on during the entire duration of data collection.

The flasks were left outdoors for a longer period of time than others, thus receiving more UV radiation.

To improve this, each trial should be carried out on its own, placing only one flask outdoors at each time.

Some specimens of yeast were not measured directly after the appropriate time of exposure to UV radiation, and were left indoors for some period of time.

To improve this, set up more than one stations, or carry out a larger number of trials, for example 10 trials, so as to collect more sufficient data.

Appendix A

Table A1. Chemical composition of Frezyderm seaside sunscreen UV protection level 50+ [22].

Ingredient name

Purpose

Wavelength protection

C13-15 alkane

Solvent, emollient

-

Ethylhexyl Methoxycinnamate

sunscreen

UVB [34]

Dibutyl adipate

Emollient, solvent

Diethylamino hydroxybenzyl Hexyl benzoate

sunscreen

UVA [35]

Octocrylene

sunscreen

UVA, UVB [36]

Ethylhexyl salicylate

sunscreen

UVB [37]

Butyl methoxydibenzoylmethane

sunscreen

UVA [38]

Perfume

Perfuming

-

Benzyl alcohol

Preservation, perfuming, solvent, viscosity, controlling

-

Figure A1. Relative humidity during 2 June-30 June 2024, when I carried out the experiment.

Figure A2. Maximum temperature measurements during 2 June-30 June 2024, when I carried out my experiment.

Figure A3. UV index during 2 June-30 June 2024, when I carried out my experiment.

Appendix B

Table B1. Concentration of carbon dioxide released at 0 min exposure without sunscreen.

Control

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

528

922

582

826

956

1.0

1686

2214

1794

1774

1770

2.0

2488

3290

3032

2864

2740

3.0

3332

4232

4118

3814

3532

4.0

4058

4976

4980

4598

4194

5.0

4706

5788

5842

5408

4750

6.0

5334

6478

6648

6200

5300

7.0

5962

7074

7414

6892

5792

8.0

6532

7612

8100

7480

6216

9.0

7058

8092

8744

8000

6610

10.0

7526

8682

9316

8522

7116

Table B2. Concentration of carbon dioxide released at 0 min exposure with sunscreen.

Control

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

1238

1208

4244

1776

4496

1.0

1980

2052

5370

1818

5482

2.0

3442

3642

6026

3106

6462

3.0

4716

4924

7704

4410

8060

4.0

6124

6248

9172

5620

10,064

5.0

7350

7360

11,366

7006

11,920

6.0

8488

8402

13,072

8364

13,354

7.0

9492

9352

14,386

9556

14,502

8.0

10,472

10,196

15,294

10,734

15,516

9.0

11,482

11,136

16,034

11,764

16,214

10.0

12,406

12,260

16,632

12,956

16,718

Table B3. Concentration of carbon dioxide released before 15 min exposure without sunscreen.

Before

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

528

982

664

742

846

1.0

1200

1346

1192

1012

1190

2.0

1608

1748

1532

1354

1576

3.0

2158

2214

2028

1868

1990

4.0

2782

2860

2596

2416

2588

5.0

3364

3492

3096

2934

3182

6.0

3916

4086

3560

3430

3738

7.0

4456

4650

4082

3922

4284

8.0

4940

5188

4560

4398

4786

9.0

5408

5692

5004

4854

5264

10.0

5842

6178

5426

5288

5700

Table B4. Concentration of carbon dioxide released after 15 min exposure without sunscreen.

After

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

500

600

550

480

510

1.0

764

854

792

728

770

2.0

814

902

840

780

818

3.0

844

934

872

812

848

4.0

874

966

904

838

872

5.0

898

992

926

864

894

6.0

926

1012

950

888

918

7.0

1096

1194

1064

994

1040

8.0

1432

1504

1348

1240

1344

9.0

1738

1816

1654

1502

1648

10.0

2044

2156

1964

1774

1934

Table B5. Concentration of carbon dioxide released before 15 min exposure with sunscreen.

Before

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

564

1012

944

550

3538

1.0

912

1532

1572

1200

4372

2.0

1562

2340

3058

2954

5470

3.0

2328

4144

4386

3030

6506

4.0

3194

5282

5654

4240

8230

5.0

4970

6596

6954

5500

9836

6.0

5682

7642

8118

6768

11,550

7.0

6430

8618

9294

7922

13,072

8.0

7212

9526

10,428

8066

14,348

9.0

9970

10,380

11,570

9504

15,388

10.0

12,688

12,366

12,602

11,586

16,260

Table B6. Concentration of carbon dioxide released after 15 min exposure with sunscreen.

After

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

1318

1030

1132

1024

1262

1.0

2416

1832

2156

2394

2250

2.0

4136

2878

3654

3800

3806

3.0

5838

4026

4966

4972

5218

4.0

7326

5206

6326

6196

6572

5.0

8512

6428

7450

7266

7622

6.0

9526

7454

8350

8214

8532

7.0

10,534

8360

9220

9098

9330

8.0

11,430

9160

9966

9784

10,000

9.0

12,240

9828

10,762

10,442

10,712

10.0

12,900

10,590

11,486

11,106

11,308

Table B7. Concentration of carbon dioxide released before 30 min exposure without sunscreen.

Before

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

1740

1694

1756

1588

878

1.0

2922

2890

2744

2954

1652

2.0

5354

4696

4818

4736

3346

3.0

7164

6158

6356

6250

4680

4.0

8394

7216

7472

7412

5734

5.0

9208

8006

8268

8266

6582

6.0

9824

8680

8884

8946

7250

7.0

10,510

9224

9432

9502

7800

8.0

11,234

9728

9916

9986

8346

9.0

11,878

10,246

10,488

10,566

8896

10.0

12,390

10,756

11,036

11,108

9386

Table B8. Concentration of carbon dioxide released after 30 min exposure without sunscreen.

After

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

500

1544

1342

938

916

1.0

2298

1124

1816

1956

2210

2.0

3926

1108

3102

3478

3454

3.0

4910

1780

4096

4474

4342

4.0

5714

2974

4746

5196

4952

5.0

6254

3938

5272

5874

5486

6.0

6558

4620

5766

6384

5908

7.0

6980

5162

6122

6828

6272

8.0

7414

5672

6464

7212

6622

9.0

7798

6092

6792

7540

6968

10.0

8148

6466

7060

7844

7256

Table B9. Concentration of carbon dioxide released before 30 min exposure with sunscreen.

Before

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

1686

1478

1936

5874

4380

1.0

2210

2806

2178

7342

5228

2.0

4642

4328

4864

8668

6516

3.0

6900

5832

7210

9600

8764

4.0

8662

7340

8956

12,518

9714

5.0

10,116

8688

10,296

14,824

12,776

6.0

11,616

9844

11,616

16,546

14,502

7.0

12,770

11,148

13,620

17,876

15,776

8.0

13,682

12,360

14,400

18,930

17,120

9.0

14,440

14,278

15,060

19,814

18,112

10.0

15,120

15,106

16,690

21,750

19,152

Table B10. Concentration of carbon dioxide released after 30 min exposure with sunscreen.

After

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

2350

2662

2420

2440

3400

1.0

3298

3560

3812

3748

4102

2.0

4564

4889

5186

5772

5606

3.0

6090

6410

6930

8250

6898

4.0

7744

8054

8542

10,900

8994

5.0

9442

9762

10,266

12,820

10,854

6.0

11,036

11,290

11,562

14,354

12,466

7.0

12,450

12,778

12,820

15,586

13,640

8.0

13,710

13,050

13,980

16,540

14,672

9.0

14,784

15,043

15,106

17,264

15,510

10.0

15,720

16,080

16,182

18,080

16,284

Table B11. Concentration of carbon dioxide released before 60 min exposure without sunscreen.

Before

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

640

1274

724

696

814

1.0

1476

1894

1144

1066

1066

2.0

2180

2690

1866

1574

1486

3.0

3148

3476

2682

2132

1946

4.0

3940

4202

3412

2782

2534

5.0

4630

4872

4050

3344

3116

6.0

5300

5564

4614

3862

3634

7.0

5992

6184

5160

4352

4138

8.0

6604

6726

5740

4790

4594

9.0

7160

7212

6254

5202

5034

10.0

7660

7648

6710

5644

5542

Table B12. Concentration of carbon dioxide released after 60 min exposure without sunscreen.

After

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

500

438

622

1288

810

1.0

1124

1032

894

1714

972

2.0

1758

1572

1320

2256

1728

3.0

2392

2032

1834

2796

2502

4.0

2984

2604

2376

3276

3256

5.0

3444

3130

2892

3732

3880

6.0

3904

3606

3336

4120

4436

7.0

4310

4050

3758

4476

4948

8.0

4674

4420

4122

4796

5502

9.0

5002

4776

4462

5122

6028

10.0

5332

5146

4770

5268

6302

Table B13. Concentration of carbon dioxide released before 60 min exposure with sunscreen.

Before

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

530

550

1144

1828

2374

1.0

1314

1740

2092

2370

2808

2.0

2118

3660

4198

4464

5472

3.0

3172

5262

5898

6792

7858

4.0

4244

6622

7478

8650

9722

5.0

5256

7746

8892

10,236

11,364

6.0

6334

8824

10,072

11,778

12,892

7.0

7232

9962

11,368

12,884

14,054

8.0

8074

10,886

12,468

13,804

14,926

9.0

9844

11,812

13,336

14,658

15,808

10.0

11,992

12,916

14,250

15,334

16,116

Table B14. Concentration of carbon dioxide released after 60 min exposure with sunscreen.

After

Concentration of carbon dioxide/ppm

Time (min)

Trial 1

Trial 2

Trial 3

Trial 4

Trial 5

0

6582

5422

2572

4608

1856

1.0

7268

6076

4882

6558

3824

2.0

8174

7160

8042

7938

6270

3.0

9784

9622

10,640

10,118

8160

4.0

12,730

11,854

12,666

12,212

9712

5.0

12,750

13,712

14,166

13,766

11,164

6.0

16,206

15,088

15,162

14,774

12,392

7.0

17,444

16,132

15,972

15,464

13,334

8.0

18,432

17,036

16,648

16,118

13,902

9.0

19,204

17,870

16,994

16,468

14,314

10.0

19,869

18,508

17,328

16,882

14,738

Table B15. Cell density with and without sunscreen for each exposure time.

Before exposure: N = 18.92 × 104

Time of exposure in UV (min)

With sunscreen (9.0 ml/50)

Without sunscreen

Control

12.73 × 104

12.5 × 104

15

15.5 × 104

15.86 × 104

30

18.08 × 104

24.24 × 104

60

11.71 × 104

4.46 × 104

Conflicts of Interest

The author declares no conflicts of interest regarding the publication of this paper.

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https://ocean.si.edu/ecosystems/coral-reefs/sunscreen-environment#:~:text=Coral%20reefs%20are%20particularly%20susceptible,ingredients%20like%20oxybenzone%20and%20octinoxate
[34] Ethylhexyl Methoxycinnamate.
https://www.aako.nl/products/sun-care/ethylhexyl-methoxycinnamate/#:~:text=Ethylhexyl%20Methox-ycinnamate%20is%20organic%2C%20oil,a%20peak%20protection%20at%20310nm
[35] Ataman Kimya (n.d.) Diethylamino Hydroxybenzoyl Hexyl Benzoate.
https://www.atamanchemicals.com/diethylamino-hydroxybenzoyl-hexyl-benzoate_u25308/
[36] Octocrylene.
https://www.sciencedirect.com/topics/medicine-and-dentistry/octocrylene
[37] Ataman Kimya (n.d.) Ethylhexyl Salicylate.
https://www.atamanchemicals.com/ethylhexyl-salicylate_u26075/#:~:text=Ethylhexyl%20salicylates%20are%20weak%20UVB,peak%20at%20307%2D310%20nm
[38] COSMILE Europe (2023) Ingredient.
https://cosmileeurope.eu/inci/detail/2000/butyl-methoxydibenzoylmethane/

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