Alessandra Aragon '22

Archer School for Girls

The Chick Chorioallantoic Membrane as an In Vivo Model to Study the Quantitative Effect of Varying Doses of Cynomorium songaricum in Cancer Angiogenesis

Cancer is the second leading cause of death and a total of 609,360 cancer-related deaths are expected to occur in 2022 in the United States. There are several biological capabilities impacted by cancer development, such as angiogenesis, which is the formation of new blood vessels. Vascular endothelial growth factor (VEGF) is a signaling protein that regulates angiogenesis. The overexpression of VEGF correlates with the angiogenic and metastatic characteristics of cancer. Angiogenesis inhibitors have been significant in advancing cancer drug therapy, however, drug resistance is still a leading obstacle. Therefore, it is essential that VEGF and angiogenesis inhibitors are further researched to develop a wider range of individualized treatment options for cancer patients. Cynomorium songaricum (CS) is a traditional Chinese medicine that has demonstrated PI3K/AKT pathway suppression, and thus decreases in levels of VEGF. In this study, the angiogenic response to varying doses of CS using the chick chorioallantoic membrane was quantified through macroscopic imaging. This research contributes to the body of knowledge used to develop more refined therapies for treating cancer and it provides further insight into the use of traditional Chinese medicines as natural angiogenesis inhibitors.

Carolyn Awad '22, Maya Ismail '22, and Riley Peterson '22

Flintridge Sacred Heart

Preventing Bacterial Infections: One Plant Extract at a Time

More than 80% of bacterial infections are caused by the formation of biofilms in the body. These infections are becoming increasingly more difficult to treat or eradicate because of bacteria’s growing antibiotic resistance. Biofilm formation is mediated by quorum sensing, a mechanism of bacterial communication. A new approach for combating biofilm formation, which could be a source for supplementary therapy through vitamins and minerals, targets the steps involved in quorum sensing. One way to inhibit biofilm formation is through the use of plant extracts and naturally occurring peptides. Specifically, our research was aimed at combating canine periodontal disease which is caused by both Gram-negative and Gram-positive bacteria. Current approaches for treating this disease include adding thymoquinone, a seed extract, and nisin, a lantibiotic, to various bacterial strains. In our study, we proposed to research the effects of these naturally derived compounds on the biofilm formation of a model bacterial system called Vibrio harveyi. We hoped to understand the inhibiting mechanisms of these compounds and to uncover any synergistic effects when they are added together. We measured bacterial growth, bioluminescence and biofilm formation of Vibrio Harveyi in the presence of various concentrations of thymoquinone and nisin.

Faith Blea '22 and Emilie Zhang '22

Flintridge Sacred Heart

Another Reason to Like Red Wine -- It Might Make Medical Implants Safer

Biofilm, a complex collection of bacterial colonies, presents many challenges to current drugs targeting bacterial infections (Barzegari et al., 2020). Resveratrol, a compound found in grape plants, has reduced biofilm formation and interfered with quorum sensing mediated behaviors in several bacterial species (Vestergaard & Ingmer, 2019). In Vibrio harveyi, biofilm formation is regulated by quorum sensing, which is a bacterial communication system (Frederix & Downie, 2011). Thus, we propose to study the inhibitory effects of resveratrol on the mechanisms of quorum sensing in Vibrio harveyi by observing biofilm formation and bioluminescence. Conclusions were made regarding BB120 (wild-type) and BB721 (constitutively glowing mutant). Our result suggests that resveratrol is a promising quorum sensing inhibitor within a specific concentration range, as it didn’t kill the bacteria and yet disabled quorum-sensing-associated phenotypes.

Evan Bowman '22

Archer School for Girls

The Effects of Curcumin and Paeonol on Oxidative Stress induced by Dibutyl phthalate (DBP) and Bis(2-ethylhexyl) phthalate in the Chicken Chorioallantoic Membrane Assay.

Phthalates such as Bis(2-Ethylhexyl) phthalate (DEHP) have been reported to increase oxidative stress (OS) levels in-vivo, putting those exposed at a higher risk for tumors such as fibroids, or Uterine Leiomyomas (ULs)). However, it has not been investigated whether Dibutyl phthalate (DBP), one of the most common environmental exposures, increases OS, and whether traditional remedies for fibroids, that have been historically believed to prevent and reduce symptoms, impact the OS biomarkers. This study tested the MDA and SOD concentrations in the sera and cardiac tissue of chicken embryos that were previously exposed to varying levels of DEHP and DBP and treated with either curcumin or paeonol. Preliminary results are available, however, this study is in the process of interpreting this data. These results will indicate how DBP and DEHP impacted the OS of the organisms and whether the curcumin and paeonol mitigated these effects. This information will provide critical guidance for those at risk for and experiencing ULs by indicating the efficacy of the two most well-known homeopathic remedies against OS, as well as guidance for the general public as to how this common environmental exposure may be adversely affecting their health.

Kate Chiang '23 & Anjali Mani '23

Gretchen Whitney High School

Protective effects of omega-3 fatty acids against oxidative stress-induced collagen degradation of Tilapia fish scales

Background:

Aging-associated oxidative stress and free radical generation leads to collagen degradation and subsequent reduced skin elasticity, bone degeneration and organ dysfunction. Omega-3 fatty acids are potent free radical scavengers and exhibit anti-inflammatory properties in numerous pathologies. However, the protective role of omega-3 fatty acids against oxidative stress-induced collagen degradation during aging remains to be established.

Methods:

Collagen samples were isolated from tilapia fish scales sourced from a local 99 Ranch Market, Artesia, CA, USA. Samples were treated with hydroxyl free radicals generated with Fenton reaction and were subjected to eicosapentaenoic (EPA) acid, docosahexaenoic acid (DHA), EPA:DHA (1:4) or oil tablets (320 mg EPA: 200 mg DHA). SDS-PAGE electrophoresis (Bio-Rad Laboratories, Inc., Hercules, CA) was carried out with or without dithiothreitol or chloroacetamide at Gretchen Whitney High School, Cerritos, CA.

Results:

Hydroxyl-treated collagen samples exhibited increased electrophoretic mobility compared to non-treated controls in both the chloroacetamide and non-chloroacetamide groups. Control samples run with or without DTT did not noticeably differ. In the non-chloroacetamide group, electrophoretic mobility of collagen samples pretreated with EPA or DHA was comparable to hydroxyl-treated samples, but was noticeably reduced in EPA:DHA- and oil-treated samples similar to controls. In samples run in chloroacetamide, hydroxyl-induced increase in electrophoretic mobility was further promoted by pretreatment with EPA, DHA, EPA:DHA or oil.

Conclusion:

A combination of DHA and EPA protects against hydroxyl-induced collagen damage, suggesting the potential benefit of prophylactic fatty acid supplementation to preserve collagen integrity during aging.

Anna Entin '24

Archer School For Girls

Reducing Symptoms of Parkinson's Disease in Fruit Flies

Objectives

My objective was to determine whether Kaempferol or Apigenin is more successful at reducing the symptoms of Parkinson’s disease (PD) in fruit flies and at what dosage. I observed and recorded how these medications affected the activity of the flies.

Materials and Methods

I used fruit flies with the PARK8 gene, which is the fly equivalent to the LRRK2 gene in humans, that causes PD. PD often presents itself as loss of mobility. I put 28 flies, with different medication and dosages mixed into their culture, in the Trikinetic Activity Monitor and recorded their activity over a 100 hour period. Higher activity means the medication is more successful. I had 7 groups: a non-medicated group, 3 groups for Kaempferol (560µM, 840µM, and 1120µM), and 3 groups for Apigenin (1µL, 2µL, and 3µL).

Results

The results were based on the average activity of the flies for every 5 hours. The flies that received the 840µM of Kaempferol had the highest and longest activity out of both Kaempferol and Apigenin. Throughout the entire experiment the control that received no medication had the lowest activity.

Conclusion/Discussions

I rejected my hypothesis which was that Apigenin and Kaempferol would be equally successful in reducing the symptoms of PD and the highest dosage would be most successful. The data I collected demonstrated that Kaempferol is more successful than Apigenin at reducing PD symptoms in fruit flies and is most successful at the second highest dosage (840µM). This suggests that taking Kaempferol can help people with PD improve their condition.

Zoe Epps '23

Archer School For Girls

The effects of phytoestrogen on mortality, fertility, and oxidative stress in female Drosophila melanogaster in comparison to the xenoestrogen progesterone

Women produce many hormones, most notably estrogen and progesterone, which play an important role in reproduction. The body naturally regulates their production and will sometimes elevate levels of these hormones to prevent oocytes from maturing; however, by introducing additional xenoestrogens, like progesterone, birth control mimics the same process. Birth control uses xenoestrogens that mirror natural hormones produced within one’s body; however, it subjects it’s consumers to the risk of experiencing any number of the birth control’s side effects and is very costly. Given that some plants produce phytoestrogens, or natural estrogens, that have in vivo estrogenic effects, when introduced into the diet of a female Drosophila melanogaster the genistin could potentially act as birth control. As genistin is a natural alternative, it is less expensive and could potentially could result in less side effects. Thus, this research aims to find an economical and safer alternative progesterone by comparing fertility, mortality, and oxidative stress in D. melanogaster after ingesting the isoflavone genistin and progesterone. Data is yet to be analyzed.

Avery Fox '22

Archer School For Girls

The Effect of a CRISPR/Cas9 System to Edit the TCS1 Gene in Camellia sinensis var. assamica on the Lifespan of Caenorhabditis elegans

Camellia sinensis, the tea plant, naturally contains the drug caffeine (1,3,7-Trimethylpurine) because of its tea caffeine synthase (TCS) genes, particularly TCS1. Researchers are using clustered regularly interspaced short palindromic repeats (CRISPR) to remove the TCS1 gene from C. sinensis, eliminating caffeine and thus its side effects that are linked to caffeinated and decaffeinated beverages. To do so, the researchers wanted a sequence with a difference of two base pair sequences or less from TCS1. One of these potential editing targets is the theacrine synthase gene, CkTcS. Without CkTcS, there would be no theacrine but more caffeine since TCS1 wasn’t edited out. Additionally, there would be more theobromine and theophylline (two other substances in C. sinensis). The purpose of this study was to measure the effect of an unintended CRISPR editing of the CkTcS gene by measuring the lifespan of C. elegans while the worms were exposed to the amounts of substances present after the CRISPR edit. Worms exposed to caffeine and worms not exposed to any chemicals had slightly lower mortality rates than worms exposed to caffeine, theobromine, and theophylline. However, the study was not ompleted due to unforeseen circumstances. Only one days worth of data was collected, making these results inconclusive. Nevertheless, more research on this topic is critical because CRISPR’s recentness and the long breeding time of C. sinensis has prohibited research. Results should be clearly understood before tea from CRISPR-edited, naturally decaf C. sinensis plants is made available for consumption.

Mohuli Ganguly '24

La Salle High School

Examining the effects of heart supplements on the differentiation process of human-induced pluripotent stem cells to cardiomyocytes

My research tests to see if any of the three heart supplements added to human induced pluripotent would have an advantageous effect on cell growth and/or the differentiation process. The pluripotent stem cells are derived from genetically reprogrammed skin cells which created cells with similar properties to those of the early embryo. The already developed strategy for differentiating hIPSCs into heart cells involves treating confluent cells with CHIR 99021 for 24 hours and IWP2 for 48 hours in RPMI with B27 lacking insulin. The hIPSCs are plated, treated with treatments that allow the cells to transform to cardiomyocytes, and then immunolabeled to visualize the differentiation yield. However, this approach is not fully optimal since it causes a lot of cell death leading to low yields of cardiomyocytes. The goal of this experiment is to determine if the supplements had an effect on cell yield, growth, and differentiation. Differentiating hIPSCs will be treated with the supplements L-Carnitine, Coenzyme Q10, and Omega 3 Fatty acid 24 hours before and throughout the differentiation process. Differentiation efficiency will be assayed using immunocytochemistry for NKX2.5, a cardiac differentiation marker. My results showed that the cell culture that had the omega 3 fatty acid added had the most amount of differentiation compared to the other supplements and the control. It also shows Omega 3 had the most cell death. Thus, the results did confirm that one of the supplements does affect the differentiation process and cell yield.

Karen Garcia '23

Archer School For Girls

The Effect of Piperlongumine as an Inhibitor for the BRAF V600E Mutation from the Ras/MAPK Signaling Pathway in Caenorhabditis elegans

The Ras/MAPK signal transduction pathway plays a vital role in the cell cycle, thus, mutations in this pathway lead to cancers and tumor cell proliferation. The BRAF V600E mutation in the BRAF gene creates a constitutively active B-Raf protein, occurring in cancers such as melanoma and colon cancer. Piperlongumine, an active alkaloid in long pepper, has been shown to suppress tumor cells in colon cancer and has helped BRAF inhibitor resistance. In this study, Piperlongumine was used to treat Caenorhabditis elegans with the lin-45 mutation, the mutation being a homolog of the BRAF mutation in humans. Since the lin-45 mutation of the worm’s Ras/MAPK pathway homolog decreases its olfaction, a chemotaxis assay was performed on wild-type N2 worms and lin-45 mutants (both worm-types exposed or not exposed to Piperlongumine) to determine if Piperlongumine can suppress the BRAF homolog protein in lin-45. If a treated lin-45 mutant strain exhibited a higher olfactory ability than a non-treated mutant, Piperlongumine would show to be an efficient inhibitor of the BRAF V600E protein in the Ras/MAPK pathway. However, all worms were presumed dead one day after liquid medium culturing, therefore, no worms reacted to the chemotaxis procedure. The goal of this study would determine if Piperlongumine could inhibit the protein products of the BRAF V600E mutation in the Ras/MAPK pathway, furthering the knowledge of this subject because Piperlongumine was solely used as an inhibitor for the faulty BRAF V600E protein, in contrast to combining Piperlongumine with predetermined treatments.

Siena Guralnick '23

De Toledo High School

What makes e-cigarette taste so good and at what cost: finding the effects of flavoring agents on cellular respiration using Aliivibrio fischeri bacteria.

Objective: Study the potentially harmful effects of flavoring ingredients in e-cigarette products.

Methods: Aliivibrio Fischeri is a bioluminescent bacteria which emits light as a marker of healthy cellular respiration. In order to establish control parameters, this bacteria was grown in a photobacterium agar plate at 21 degrees celsius. The light transmittance of this healthy control culture was measured with a spectrophotometer and found to be maximal at a wavelength of 380 nm. Subsequently, each week, over the course of three weeks, three cultures were grown in the same agar medium and at the same temperature: one culture being the control, one culture infused with four drops of the flavoring agent acetoin and one culture with four drops of the flavoring agent benzaldehyde. Each week, the bioluminescence of the individual cultures was measured by spectrophotometer at 380 nm. The bioluminescence measures of the three groups were compared as an attempt to determine whether the added flavoring molecules impaired bacterial respiration in the form of bioluminescence.

Results: Those bacterial subcultures infused with acetoin emitted significantly less light compared to healthy control. The bacterial subcultures infused with benzaldehyde showed varied transmittance levels.

Conclusion: Aliivibrio Fischeri respiration as measured by light transmittance is negatively impacted by the addition of Acetoin to culture medium. The addition of benzaldehyde only variably impacted bacterial cellular respiration. This study adds support that the flavoring ingredients in e-cigarettes negatively impact bacterial respiration. This opens up the question of whether these same additives impact human cells when vaporized and inhaled.

Olivia Jarvie '22

Archer School For Girls

Snailed it! : The Effect of Helix aspersa Snail Mucus and Antimicrobial Bacterial Cellulose Hydrogel on Wound Healing in Caenorhabditis elegans

Chronic wounding is a very serious condition, however due to the lack of designated funding for this topic, further research is necessary to combat this pressing issue. This study aimed at testing the efficacy of antimicrobial bacterial cellulose (BC) in combination with snail slime (Helix aspersa) as an effective method to increase the rate of wound healing. To test this, snail slime was extracted from H.aspersa and combined with antimicrobial BC hydrogel to create a 2:1 ratio of BC to snail slime serum. The original plan was to monitor wound healing in Caenorhabditis elegans by applying saline solution, pure snail slime, or pure antimicrobial BC as controls, and the BC/snail slime serum as the experimental group to wounded C.elegans. To visualize the healing process, each group had to be monitored for 24 hours and a UV light was necessary to track the C.elegans’ fluorescence color fluctuation (which represents different stages of healing). However, during control group experimentation the C.elegans did not survive a sufficient amount of time after wounding to track healing. Consequently, experimentation shifted focus and assessed the factors that led to the worm's death. After 5 experimental days of narrowing down the cause of death, it is hypothesized that wounding and injection was not the cause, but rather the method of visualization, with a UV light, was. While advancing the field of wound healing is crucial, future studies should address the limitations of this visualization method while monitoring healing in C.elegans.

Lacey Thompson '24

Archer School For Girls

Beta-BLOCK Hypertrophic Cardiomyopathy (The Effects of Natural Beta Blockers of Daphnia’s Heart Rate)

Objectives

My objective was to see if natural beta-blockers can aid in the treatment of hypertrophic cardiomyopathy by reducing the heart rate. To do this, I placed daphnia in a garlic, hibiscus, and hawthorn berry solution.

Materials and Methods

My experiment used 3 natural beta-blockers: garlic, hibiscus, and hawthorn berry. In total, I had 4 groups. For my control group, I used 350ml of spring water with no beta-blockers added. For my experimental groups, I had 350 ml of spring water and 0.4 g of each substance. I placed 4 daphnia in each beaker. After 24 hours, I measured the heart rate (bpm) of each daphnia under a microscope. Using a pipette, one at a time, I carefully removed the daphnia from their beaker and placed them on a living organism slide. I then counted each heartbeat for 10 seconds and then multiplied the result by 6.

Results

The results were based on the average beats per minute of each group. Hibiscus achieved the lowest heart rate (bpm) in daphnia. Hawthorn berry also reduced the heart rate of the daphnia, however, the garlic raised the heart rate of the daphnia.

Conclusion/Discussions

In conclusion, two out of the three substances lowered the heart rate of the daphnia. This disproves my hypothesis as I hypothesized that each group would decrease the heart rate. Now I know, hibiscus is the most effective in lowering the heart rate of daphnia. Additionally, I learned that garlic is ineffective.

Alyssa Ponrartana '23

Archer School For Girls

"Yay" or "Nay" for BPA? The Effects of Bisphenol A (BPA) on Fragile X Syndrome (FXS) Phenotypes in Male and Female Wild-type and dFMR1 Mutant Drosophila melanogaster

Fragile X Syndrome (FXS) is one of the leading causes of intellectual disability and autism, caused by a mutation in the Fragile X Mental Retardation-1 (FMR1) gene. People with FXS exhibit various behavioral and neurodevelopmental characteristics, such as hyperactivity and social impairments, and males commonly have more severe symptoms than females. While the genetics of FXS are well-studied, little is known about how environmental factors affect the disorder and whether they impact males and females differently. Specifically, this research examined the impact of Bisphenol A (BPA), a near-ubiquitous chemical found primarily in polycarbonate plastics, on two Fragile X Syndrome phenotypes: hyperactivity and oxidative stress. The effects of BPA were studied in male and female wild-type and dFMR1 mutant Drosophila melanogaster using the Rapid Iterative Negative Geotaxis (RING) and Superoxide Dismutase (SOD) assays to assess hyperactivity and oxidative stress respectively. Initial data showed an average increase in hyperactivity in both wild-type and mutant females exposed to BPA in comparison to their controlled counterparts, but further data analysis is required to determine statistical significance. The SOD assay data is also in the analysis process and is planned to be completed before the symposium.