2024 Bastyr University Student and Faculty Research Symposium

"From Case to Evidence: A Secondary Analysis Approach to Acupuncture Reports" outlines a mixed-method plan to integrate qualitative and quantitative analyses from real-world data sets. Ten case reports adhering to the CARE guidelines and data from ~130 clinical registry cases will be analyzed to yield meaningful and accurate results, which may help guide clinical understanding and future trials. Rooted as they are in actual practice, the findings may be particularly valuable due to their capacity to preserve the ecological validity of a complex medical system.

Methods: The quantitative analysis will look at the distribution and common characteristics of patient demographics, health complaints, and treatment details within the dataset in which pain was reported. Following this, Spearman's r will assess the correlation between pain scores and patient-reported outcomes. To evaluate the effectiveness of treatments over time, pairwise T-tests will compare pain scores at different follow-up points against baseline measures. In the qualitative portion of the study, thematic analysis will focus on examining changes in diagnostic reasoning and patient-reported self-care behaviors over time, utilizing rich time-series information from electronic health records (EHRs). Additionally, purposive coding is applied to identify specific qualitative fields such as diagnostic reasoning shifts and patient behavior changes. This part of the analysis is particularly focused on detecting complex patterns, such as non-linear symptom changes, consistent with the TCM model.

Data Integrity and Bias Mitigation: Adherence to CARE and CARE-CHM guidelines is necessary to ensure high-quality data collection and reporting. A comparative approach between case reports and clinical registry data enhances validity and addresses potential biases in reporting and outcomes selection. This comprehensive analytical framework aims to reveal not just the efficacy of acupuncture treatments but also the underlying reasoning and patient behaviors associated with successful outcomes. The study's timeline projects that findings will be ready for presentation in 2025, promising significant contributions to academic knowledge and practical acupuncture applications, and may serve as a model for larger dataset analyses of holistic practice outcomes.

Kathleen Lumiere

Bastyr University

In the realm of immune-related research on natural products to protect against growing public health concerns, the influence of market-available vitamin D supplements, with lipid additives, on cellular behavior, remains a mystery. This project looks at the relationship between supplemental vitamin D serum with lipid ingredients, and cell death of human cancerous T cells (type of white blood cell). This study aims to unveil potential modulatory influences of this vitamin D, with lipid additive, on cellular responses, contributing crucial insights to the broader understanding of supplements’ public health implications. The effect of varying concentrations of vitamin D (independent variable) is measured on the stimulated cells by measuring the resulting optical density values (dependent variable), which represents the viability of the cells. Cancerous T cells stimulated with certain compounds (PMA and Ionomycin) secrete a protein, IL-2, representing T cell growth. To further assess cell viability, this protein’s secretion is measured in wells with given optical densities, and interpolation with a standard curve. It is hypothesized that the presence of varying vitamin D, and therefore lipid additive, concentrations would significantly affect the T cells’ viability. Specifically, a potential linear relationship, with higher concentrations resulting in higher cell viability and more IL-2 secretion. Post-experimentation, ANOVA analysis for concentration-specific analysis of cell viability returned a significant p = 0.000000018, concluding that the concentrations of vitamin D used as experimental conditions significantly influenced cellular response. Cell viability analyzed in correlation with IL-2 secretion, a concentration dependent inhibitory effect on PMA+IO induced IL-2 secretion is found.

Pavitra Dave <pia.dave@outlook.com>

Tesla STEM High, Bastyr University

​Effect of Storage Duration on the Phytochemical Potency of Withania somnifera Root as an Herbal Supplement ​ Student: Kyla Little, Olivia Owens, Crescent Pilewski ​ Faculty Advisor: Kaleb Lund, PhD, RH (AHG)​ ​

Objective: Hydroethanolic tinctures and capsules containing the dried powdered root are common uses for administering Withania somnifera as an herbal supplement. However, there is little research indicating the best practice for processing and storage of the root to maximize potency and therapeutic potential. The change in potency of phytochemical constituents over time in dried Withania somnifera root as potentially indicated by the results of this study will inform best storage and preparation methods for ashwagandha producers and consumers.​ ​ Methods: Tinctures of dried Withania somnifera were prepared after intervals of storage as powder and cut and sifted root. After maceration, the tinctures were pressed out into amber glass dropper bottles. Withanolide A and Withaferin A contents were analyzed using HPLC. ​ ​ Results: Withaferin A (WA) and withanolide A (WDA) concentrations varied in both sample groups, with a trend indicating decrease in constituent levels over five months of storage. WA and WDA content ranged between 0.092–0.179 mg/mL and 0.008–0.015 mg/mL respectively.​ At many intervals of storage, pre-powdered root retained higher levels of WA and WDA. ​ Conclusions: The results suggest that pre-powdered Withania somnifera root maintains comparable potency over time. However, both WA and WDA content of dried root samples decreased within 5 months of storage, suggesting products are best prepared soon after harvest to maximize potency. Further studies should be conducted expanding storage duration and exploring other constituents. ​ ​ Funding: This project was funded by Bastyr University and a donation of Withania somnifera root from River Raven Herb Farm.​

Kyla Little

Bastyr University

Analysis of Plant Growth and Constituent Levels in Medicago sativa Following the Application of Human Saliva to Seeds Prior to Planting Students: Hannah Jackson, Carolyn Soley, Adrianne Smith, Keith Jacobsen, Alessandra Villegas, Mattaya Miller Faculty Advisor: Kaleb Lund, PhD, RH (AHG)

Objective: Many cultures have ideas about how saliva applied to seeds before planting will activate individualized constituent profiles based on a person’s unique health needs. While this idea may seem esoteric, this project’s purpose is to investigate the possibility of these interactions and measure them.

Methods: Four group members applied their saliva to three samples of pre-weighed seeds which were planted in three separate pots and cultivated for four weeks, as well as three control pots without saliva. All plants were grown under identical conditions in the Bastyr greenhouse. At the end of the growth period, each sample was measured for height and weight, harvested, and tinctured in high-proof alcohol at a 1:5 ratio. We ran each sample through Thin Layer Chromatography (TLC) and High-Pressure Liquid Chromatography (HPLC) analysis to find any differences in the resulting levels of diadzen, genistein, and other constituents.

Results: While subtle, differences in our samples are evident when compared to each other, as well as the control, on both the HPLC and TLC analyses. Analysis of Variance (ANOVA) of the data demonstrated a statistically significant difference between samples for one of the HPLC peaks. A two-tailed independent T-test revealed a statistically significant difference in HPLC peak area of ~9.7 minutes between Hannah and Adrianne's plants, Hannah and Mattaya's plants, and Hannah and Keith's plants.

Conclusion: While there are potential variables and/or sources of error that could explain the differences between our samples, it is possible that exposure to subject’s saliva resulted in significant differences in the growth and chemical composition. Further research into the specific constituents that changed and correlated to the subject’s health conditions is warranted. Additionally, studying this effect on other plants could demonstrate other specific interactions not observed with alfalfa.

Hannah Jackson <Hannah.Jackson@bastyr.edu>

Bastyr University

Investigating the efficacy of psilocybin in psychological disorders

Student: LeRaih Beard-Appling

Faculty Advisor: Kaleb Lund, PhD, RH (AHG)

Objective: According to the World Health Organization (WHO), approximately 280 million people in the world have depression. SSRI’s are first line therapy to major depressive disorders (MDD), unfortunately they only have a 60% response rate with a 30-45% chance of remission. Researchers are thus exploring alternative treatments, including therapeutics from the fungi kingdom. Psilocybin has a well-documented historical use within indigenous culture, and is currently being heavily researched. This project aims to investigate the current published literature exploring the use of psilocybin in major depressive disorders.

Methods: The literatures highlighted in this presentation were chosen based on relevancy, and methods of research. Databases, PubMed, NIH, and Google Scholar were searched using the key phrases; ‘psilocybin mechanism of action’, ‘Psilocybin as treatment in major depression disorder’, psilocybin and cognitive function, 'SSRI pharmacokinetics'.

Results: Existing research demonstrates the potential of psilocybin in MDD and suggests it may provide additional benefits by improving cognitive flexibility.   Conclusion: Psilocybin represents a safe and efficacious treatment for MDD and warrants continued research. It would be advantageous for the Naturopathic Doctoral and Herbal Science communities to extend their knowledge and training to this field of study, as they can provide a perspective unique to what has been published.

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LeRaih Beard-Appling

University of Bastyr, Department of Botanical Medicine

INTRODUCTION Ramadan is a month-long Islamic ritual of fasting, prayer, giving and self-valuation. Ramadan seemed to be a naturalistic way to study intermittent fasting. Intermittent fasting during the Islamic month of Ramadan was studied by Bastani, et. al. in 2017. It involved 22 females and 7 males and four neurochemicals in their blood were analyzed. Four blood factors were: Brain-derived Neurotropic Factor (BDNF), Nerve Growth Factor (NGF), dopamine, and serotonin. From their published data, the effect sizes were calculated for the pre-post changes before and after Ramadan.

In 2021, the first Ramadan research protocol included blood drawing and psychometric measurements was written. It was necessary to obtain a cooperative agreement from religious communities in order to be compliant with the Institutional Review Board (IRB) for the safety of vulnerable human research subjects.

In 2022, another research question was addressed about blood glucose profile and one volunteer subject wore the continuous glucose monitor during Ramadan. In 2023, with Jay Little Research Funding, a control group was added and data were obtained.

This poster presents the learning experience through the progression of studying intermittent fasting during Ramadan.

METHODS: This narrative report was not protocoled based on a formal quantitative research methodology. Also, this is not a compendium of outcome data from each year. Data and stories shared on this poster represent how the processes of conducting studies on a particular research question evolved by generating data every year, reviewing the literature, and designing the best possible study for investigating the health effects of intermittent fasting with a realistic budget.

RESULTS Planning for a research study and thinking through logistics required dedication and commitment. Diagrams convey ideas easily and can speak 1000 words instantly. Presenters just need to narrate what is depicted and clarify questions. Researchers always look for data consistency and how to deal with outliers became a debate. Outliner deletion must be consistent and justified. Another consistency was searched between 2021 and 2023 data on blood neurochemical measurement. BDNF and NGF were declined during Ramadan for both year 2021 and 2023 data. A single group provided data in 2021 while three groups were found in 2023 data.

CONCLUSIONS Researchers must critically evaluate their own logical decisions and keep improving the research protocols. Re-analyzing existing data and generating new datasets sometimes take years of trials. However, despite many challenges and frustration, sharing the findings and looking back at the accomplishment is satisfying when a project is completed. Searching for new knowledge is a humbling and rewarding experience. We hope you can partake in challenging tasks if your endeavors lead to knowledge that is useful for other people.

Masa Sasagawa

Bastyr University

INTRODUCTION We investigated the health effects of intermittent fasting (IF) by a non-interventional and observational method. IF means abstaining from food and/or water intake and resuming the intake in a scheduled cycle.

OBJECTIVES: Enzyme linked immunosorbent assay (ELISA) measured blood BDNF, NGF, dopamine, and serotonin concentration. Urinalysis pictures are used to monitor hydration status. REDCap online survey on mood, stress, religiosity, empathy, and sense of coherence were assessed. The interview generated some qualitative data

METHODS The protocol was approved by the Institutional Review Board as described in the diagram below. Two local organizations, IMAN Center of Kirkland and Muslim Association of Puget Sound (MAPS) were contacted for recruitment efforts. With 20 Muslim participants, two subscales (dopamine and mood scale) were supposed to be powered for pre-post measurements. Because Ramadan restricted water intake during fasting, urinalysis was added to monitor hydration status. Self-administered urine strips were provided to participants. A handful of control subjects provided normal fluctuations of all variables without IF. The control subject also provided data for between-group statistics.

Because Ramadan restricted water intake during fasting, urinalysis was added to monitor hydration status. Self-administered urine strips were provided to participants. A handful of control subjects provided normal fluctuations of all variables without IF. The control subject also provided data for between-group statistics.

RESULTS A total of 24 participants were recruited. The study asked, “Are you going to observe Ramadan this year?” The answer to this question was meant to categorize groups into Ramadan IF and control groups.

Urinalysis strips were provided to participants to self-test and pictured images of the result were sent from a phone; however, due to the variation of lighting, peculiarity of color temperature, and difference in orientation of the image, analysis became impossible. Therefore, the urinalysis data were not presented.

Statistically significant decreases were found in the pre-post measurement of the NEGATIVE affect scores of the Ramadan group from the baseline to the mid-month and baseline to the end of the Ramadan period. Unfortunately, the dopamine ELISA failed at the lab, and no data were provided.

Fewer neurochemicals were detected as the month progressed for both Ramadan and control groups. Serotonin varied over the course of Ramadan.

Informal interviews during Ramadan and at the end of Ramadan indicated mostly positive comments from Muslim participants. They enjoyed the experience of participating in the scientific study of the Ramadan ritual.

CONCLUSIONS Health Effect of Ramadan Study (HERS) 2023 was successfully and safely concluded. The study recruited 24 participants, 17 in the Ramadan group, 5 in the control group, and 2 in the unexpected no IF Muslim group, with a mean age of 38.3 years old and a standard deviation of 16.8 years for all participants.

Two notable trends in early data analysis include a greater decrease in Negative Affect section of the PANAS in the Ramadan group as compared to the control group.

Urinalysis procedures were unsuccessful. Photographs of urinalysis dipsticks posed difficulties in manually assessing each image and in storing and labeling the images correctly. If the hydration data is needed, this procedure requires modification.

Nerve Growth Factor (biological measurement) and Negative Affect Scores (mood or psychological measurement) can be justifiable for a follow-up study if funding is limited to recruiting about 100 participants.

Masa Sasagawa

Bastyr University

OBJECTIVES The aims of this prospective observational case series are to:

1. Describe both the quality of life (QoL) changes over time associated with complementary and integrative medicine treatments provided by naturopathic physicians.

2. Provide students with a real-world clinical research experience prior to graduation.

METHODS Patients ≥18 years of age presenting to the Bastyr Center for Natural Health (BCNH) for care who meet the following inclusion criteria are eligible to participate; speak English well enough to understand the consent form and able to read and understand the questionnaires.

Patients will be consented and PROMIS Global Health-10 (PGH-10), Global Anxiety Disorder-7 (GAD-7), and the Patient Health Questionnaire-9 (PHQ-9) will be administered at each visit. Questionnaires, ICD-10 codes, CPT codes and recommended treatments will be abstracted from the participants chart at the time of each visit and entered into the Research and Electronic Data Capture (REDCap) database.

RESULTS Results are currently ongoing for our participants enrolled in the study. A total of 33 participants have been enrolled. A total of 72 visits have been completed.

CONCLUSION The purpose of the research shifts is to provide students with a real-world clinical research experience prior to graduation. This is intended to provide medical students with invaluable opportunities to develop essential skills, gain practical exposure to patient care, and contribute to the advancement of medical knowledge, ultimately preparing them for successful careers as evidence-based practitioners.

Collecting this data will help to inform physicians and patients what conditions naturopathic physicians commonly treat, with what therapies, frequency of use, the cost of such treatments, and how these treatments are associated with QoL. This research is disseminated annually at various conferences, which addresses the need to conduct naturopathic clinical outcomes research and the goal of providing students with a research experience during their education and training. In addition to the current shift offered at the Bastyr Center for Natural Health (BCNH) (Hsuing), this project was expanded to include a second research-focused teaching shift at Bastyr University San Diego (BUSD) (Roth) in Fall 2023.

ACKNOWLEGEMENT Funding for these studies has been provided by Bastyr University Research Institute. Contact: Erin Sweet esweet@bastyr.edu

Erin Sweet, ND, MPH, FABNO

Bastyr University

BACKGROUND Despite advances in screening and treatment, breast cancer remains a leading cause of cancer death. Several complementary and integrative medicine (CIM) therapies have been studied for their effect on cancer outcomes; however, few studies have evaluated these therapies delivered collectively by naturopathic doctors (ND). This prospective observational study has the overarching goal of evaluating naturopathic oncology in a real-world setting.

OBJECTIVES Primary Aim: 1. Five-year survival compared to a matched cohort from the Surveillance Epidemiology and End Results (SEER) registry.

Secondary Aims: 1. Survival rates among participants based on the type and frequency of naturopathic treatments received (“internal survival”) 2. Description of the type and frequency of naturopathic recommendations provided to participants 3. Incidence of conventional treatments received

METHODS Eligibility Criteria: Adults (≥ 18 years) with confirmed stage IV breast cancer receiving care from a participating naturopathic clinic.

RESULTS 65 participants had MBC with 149 included in the final analysis. From SEER, there were 35,551 patients with stage IV breast cancer diagnosed in 2010 or later. 13,456 (38%) matched with at least one study participant.

Five-Year Survival: 1. No significant differences when compared to SEER (median HR 0.85, 95% CI 0.66 – 1.07, HR > 1.00 favors CUSIOS). 2. No significant differences when including only participants who saw their ND within 6 months of diagnosis (HR 1.15, 95% CI 0.80 – 1.60)

Internal survival comparison: Significantly improved survival odds were seen when participants received more intravenous vitamin C (IVC) treatments (HR = 0.65, 95% CI 0.47 – 0.91, p = 0.01, median treatments = 13) and more ND visits (HR = 0.65 95% CI 0.47 – 0.90, p = 0.01, median visits = 5). No benefits were seen with increasing mistletoe or hyperthermia treatments.

Conventional treatments: 99% of participants received some form of conventional therapy either prior to or during the study.

CONCLUSION No survival benefits were seen for North American patients with MBC who received naturopathic care compared to US registry data. Survival benefits within the cohort were seen in participants who saw their ND more frequently and who received more IVC treatments. Recommendations made by NDs were highly individualized with only 10 items recommended to at least 50% of participants. Results should be interpreted with caution given methodological limitations and biases, which include comparing a small cohort to a large population, lag time between diagnosis and enrolment, and lack of data on adherence to ND recommendations.

ACKNOWLEGEMENT Funding for these studies has been provided by the John & Lotte Hecht Memorial Foundation.

Contact: Erin Sweet esweet@bastyr.edu

Erin Sweet, ND, MPH, FABNO

Bastyr University and the Center for Health Innovation

The fungal species of polyporus shelf mushroom, Fomitopsis officinalis, commonly called Agarikon, is reported to have tumoricidal actions as well as antiviral, antimicrobial, anti-inflammatory and immunomodulatory actions. However, mainly fruiting body extracts have been studied. This study aims to determine the cytotoxic and immune-modulating responses of Jurkat E6.1 T-cell ALL cancer cells treated with mycelial extracts of Fomoptosis officinalis. To establish the optimal experimental conditions, two 96-well assays were performed, including an XTT assay on Jurkat cells and an IL-2 ELISA on supernatants. Cells in 96-well assay plates were either untreated or pre-treated with MG132, a positive control for cytotoxicity and cytokine inhibition, and then treated with PMA and ionomycin alone or together (to stimulate IL-2 production) for 16-20 hr prior to XTT assay and supernatant collection for ELISA. Results of these two assays confirmed that pre-treatment with the positive control MG132 was cytotoxic, and that PMA + Ionomycin stimulation resulted in detectable IL-2 secretion without significant cytotoxic effect. However, the IL-2 concentration of the PMI+ionomycin treated cells was lower than optimal, suggesting that increasing the cell concentration per well or incubating the cells longer will allow better detection of modulating effects of the mushroom extract test agents. With these working assay systems, experiments can proceed to test the effects of differing concentrations of F. officinalis extracts on Jurkat cell viability and IL-2 secretion. These future experiments will determine if mycelial extracts of F. officinalis have potential cytotoxicity and immunomodulatory effects on human Jurkat cancer cells.

Emma Arnold and Luke Markis

Bastyr University

Analysis was conducted on multiple studies aiming to look for successful treatments for healing the mind and body simultaneously to manage Post Traumatic Stress Disorder (PTSD) symptoms and pain responses that are comorbid with trauma. In the present study, 10 peer-reviewed articles examining treatments for PTSD and pain were examined. The research literature was identified in March 2024 through detailed searches in databases such as PsycINFO and PubMed using the following search terms: PTSD and chronic pain, Chronic illness and Trauma, PTSD, and comorbid pain.

The reviewed studies indicated that interventions incorporating Cognitive Therapy (CBT), Support groups, Biofeedback, Selective Serotonin Reuptake Inhibitors (SSRIs), Serotonin and Norepinephrine Reuptake Inhibitors (SNRIs), and pain medication have been effective when treating both chronic pain and PTSD.

Researchers noted that precautions are needed to be taken for medications treating these conditions as they may cause side effects and may not be effective in treating symptoms for all individuals, especially benzodiazepines which are also a common treatment for PTSD (Guina, 2014).

Emily Flick

Bastyr University Kenmore

Cognitive Impacts of PTSD in Women Who Have Experienced Intimate Partner Violence: A Literature Review Students: Heather Donohoe & Emmi Armstrong Faculty advisor: Naomi Lester, PhD.

Objective: This study explored the research literature on the cognitive impacts of post-traumatic stress disorder (PTSD) due to intimate partner violence (IPV) in American women. Rates of IPV in American women are as high as 46% (HRSA, 2023), and rates of PTSD are notably higher for women who are IPV survivors than the general population (Inslicht et al., 2012).

Methods: Published research was identified via database searches in PubMed, EBSCO, PsychInfo, Data World, ScienceDirect, and Proquest. Twenty-five research reports were initially identified, with seven meeting full inclusion criteria for review.

Results: Although the research in this area is limited, results indicated that accelerated cognitive decline has been observed among survivors of IPV and that IPV-related PTSD adversely impacts several areas of cognition, including working memory, critical thinking/decision-making, and additional areas such as psychomotor speed, interpretive bias, and self-efficacy. Cognitive impacts of trauma related to IPV may also adversely impact decision-making autonomy, general well-being, and physical health.

Conclusion: While research on the cognitive impacts of PTSD from intimate partner violence (IPV) is limited, existing studies suggest this trauma adversely affects cognitive areas such as working memory and critical thinking. There is substantial evidence that IPV-related trauma is common among American women, many of whom develop PTSD, therefore there is a need for further research on the cognitive impacts of this trauma to aid in the development of effective treatments.

Emmi-Nicole Armstrong & Heather Donohoe <Emmi.Armstrong@bastyr.edu>

Bastyr University

Heat and Health in the Valley: Assessing Climate Change Impact Disparities in the Coachella Valley

Student: Lisa Elizondo

Faculty Advisor: Christina Ciupitu-Plath, MSc, DrPH

Objective: This project assessed the vulnerabilities of Coachella Valley residents to climate change events, specifically rising temperatures and prolonged high heat events, and identified strategies for preparing for and responding to these impending events.

Methods: Using the CDC’s Building Resilience Against Climate Effects (BRACE) Framework, this project examined the literature to determine the most pressing effects projected to impact this region, identify related health outcomes, and identify the communities most vulnerable to these effects. Research literature was identified through searches in Google Scholar and PubMed databases. The California Heat Assessment Tool and the Climate Change and Health Visualization tool were used to project potential future climate scenarios for Coachella Valley and assess health and social vulnerabilities. These tools were used to project disease burden over the next 70 years. Mann-Whitney U tests were performed to compare the prevalence of cardiopulmonary disease, asthma, and low birth weight between East Coachella Valley and West Coachella Valley cities.

Results: The study found statistically significant differences in the prevalence of cardiopulmonary disease and asthma between West Coachella Valley and East Coachella Valley cities (p < 0.001), indicating higher risk and vulnerability in eastern cities. However, there was no significant difference in the low birth rate, suggesting a need for further investigation in this area.

Conclusion: The Coachella Valley faces an increasingly dangerous future with a significant rise in extreme heat days and projected increases in heat health events. Communities in the eastern cities are particularly vulnerable and in need of public health interventions that take a multifaceted approach to not only mitigate the projected health impacts but work to address the vulnerability gap between east and west.

Lisa Elizondo

Bastyr University