## Protocol Section
### Identification Module
**NCT ID:** NCT06416228

**Acronym:** NCS

**Brief Title:** Beetroot Juice NO Cold Study

**Official Title:** Beetroot Juice Supplement for Boosting Mucosal Immunity - The NO Cold Study

#### Organization Study ID Info
**ID:** 23-108
#### Organization
**Class:** OTHER
**Full Name:** Southern Methodist University
### Status Module
#### Completion Date
**Date:** 2026-05-15

**Type:** ESTIMATED

#### Expanded Access Info
#### Last Update Post Date
**Date:** 2024-05-16

**Type:** ACTUAL

**Last Update Submit Date:** 2024-05-10

**Overall Status:** RECRUITING

#### Primary Completion Date
**Date:** 2026-05-15

**Type:** ESTIMATED

#### Start Date
**Date:** 2024-04-02

**Type:** ACTUAL

**Status Verified Date:** 2024-05

#### Study First Post Date
**Date:** 2024-05-16
**Type:** ACTUAL
**Study First Submit Date:** 2024-04-29
**Study First Submit QC Date:** 2024-05-10
### Sponsor Collaborators Module

#### Collaborators

**Class:** OTHER

**Name:** Baylor University

#### Lead Sponsor

**Class:** OTHER

**Name:** Southern Methodist University

#### Responsible Party

**Investigator Affiliation:** Southern Methodist University

**Investigator Full Name:** Thomas Ritz

**Investigator Title:** Professor

**Type:** PRINCIPAL_INVESTIGATOR

### Oversight Module

**Is FDA Regulated Device:** False

**Is FDA Regulated Drug:** False

**Oversight Has DMC:** True

### Description Module

**Brief Summary:** Currently, there is a lack of evidence-based prevention strategies for respiratory infection and management of these conditions can be costly to the public. Airway nitric oxide provides a first line of defense against pathogens, and beetroot juice, a source of dietary nitrate, has been shown to elevate nitric oxide. The main objective of this project is to demonstrate that one week of supplementation with beetroot juice elevates airway nitric oxide during stressful periods in young adults and thereby can protect against respiratory viral infections.

**Detailed Description:** Study Title Beetroot Juice Supplement for Boosting Mucosal Immunity: The NO Cold Study

Objectives Aim 1: Demonstrate that a 7-day trial of daily beetroot juice or nitrate depleted placebo beetroot juice is feasible with acceptable retention and adherence during a period of real-life stress, using adherence monitoring of beetroot juice intake by cell phone recorded video; acceptable burdensomeness for participants and success of team coordination and study logistics should also be demonstrated.

Aim 2: Generate initial estimates of effect size for a) elevations in exhaled nitric oxide (FENO) and b) the correlation between changes in FENO and both infection reduction and cold symptom reduction (biological signature).

Aim 3: a) Examine if 2 daily doses of beetroot juice are more potent than 1 daily dose in elevating FENO, and b) investigate whether each beetroot juice dosage (1 dose and 2 doses) increases FENO more than the placebo.

Exploratory aims: Explore sex as a moderator of all the Aims. Also explore effects of competitors/inhibitors of NO (arginase, asymmetric dimethylarginine) in a) reducing FENO under stress, and b) whether beetroot juice buffers any of these potential adverse effects on FENO; c) investigate whether effects of stress cortisol, which negatively impact NO, cold symptoms, and respiratory infections, are also buffered by beetroot juice.

Design and Outcomes This is a double-blind, placebo controlled clinical trial to test the efficacy of beetroot juice as elevating airway NO, which is associated with reduced cold symptoms and respiratory viral infection rates, in undergraduate students aged 18-30.

Interventions and Duration Participants will be receiving 1 active daily dose of beetroot juice and 1 dose of nitrate-depleted placebo beetroot juice, 2 active daily doses of beetroot juice, or 2 daily doses of nitrate-depleted placebo beetroot.

Cold symptoms will be explored by questionnaire at baseline (in a low stress period during the semester) and subsequently twice during the final exam period, once at an early stage of the finals (days 1-3 of the final exam period), and once at a later stage (days 4-6). A follow-up online questionnaire packet will be administered 3 days after the last final day. FENO, sampling for viral PCR, salivary cortisol, and exhaled breath condensate will be undertaken at baseline, early finals, and late finals (in-person assessments are not feasible at follow-up, because students leave campus after finals).

Sample Size and Population The sample size is 150 students (n=66 at SMU site, n=84 at Baylor site). Female and male students 18-30 years old will be recruited from Baylor and SMU. The investigators will make an extra effort to guarantee equal representation of both genders. Participants can be from any ethnic or cultural background, as long as they can understand and read English adequately. The student population of both universities combined is diverse (28.5-38.5% minorities).

The investigators will stratify by sex and by site, to randomize participants to receiving 1 active daily dose of beetroot juice and 1 dose of nitrate-depleted placebo beetroot juice, 2 active daily doses of beetroot juice, or 2 daily doses of nitrate-depleted placebo beetroot juice (n=50 per group).

### Conditions Module

**Conditions:**

- Respiratory Tract Infections

- Cold Symptoms

- Exhaled Nitric Oxide

### Design Module

#### Design Info

**Allocation:** RANDOMIZED

**Intervention Model:** PARALLEL

**Intervention Model Description:** Double blinded, placebo controlled study

##### Masking Info

**Masking:** QUADRUPLE

**Masking Description:** Both participating students and assessors will be blinded to the participants' group assignment. Since students and research staff will not know whether their beetroot juice is depleted or not, treatment conditions are double-blinded (juice "shots" will be pre-labeled by study statistician with a participant number based on the pre-study randomization, so neither the staff nor the participant know the contents). Students will also be assured of full blinding of course instructors, including the PIs and Co-Is, to names of study participants.

**Who Masked:**

- PARTICIPANT

- CARE_PROVIDER

- INVESTIGATOR

- OUTCOMES_ASSESSOR

**Primary Purpose:** PREVENTION

#### Enrollment Info

**Count:** 150

**Type:** ESTIMATED

**Phases:**

- PHASE2

**Study Type:** INTERVENTIONAL

### Arms Interventions Module

#### Arm Group 1

**Description:** 2 active (nitrate containing) doses of Beet-it Pro 400

**Intervention Names:**

- Dietary Supplement: Beetroot juice

**Label:** Two active doses of beetroot juice

**Type:** EXPERIMENTAL

#### Arm Group 2

**Description:** 1 active (nitrate containing) dose of Beet-it Pro 400 and one placebo (nitrate-depleted) dose of beetroot juice

**Intervention Names:**

- Dietary Supplement: Beetroot juice

**Label:** One active dose of beetroot juice and one placebo dose of beetroot juice

**Type:** ACTIVE_COMPARATOR

#### Arm Group 3

**Description:** 2 placebo (nitrate-depleted) doses of beetroot juice

**Intervention Names:**

- Dietary Supplement: Beetroot juice

**Label:** Two placebo doses of beetroot juice

**Type:** PLACEBO_COMPARATOR

### Interventions

#### Intervention 1

**Arm Group Labels:**

- One active dose of beetroot juice and one placebo dose of beetroot juice

- Two active doses of beetroot juice

- Two placebo doses of beetroot juice

**Description:** Participants will take on dose in the morning and one in the afternoon, for 7 days during their final academic examinations

**Name:** Beetroot juice

**Other Names:**

- Beet-it 400

**Type:** DIETARY_SUPPLEMENT

### Outcomes Module

#### Other Outcomes

**Description:** One-day food record via the Automated Self-Administered 24-Hour platform; control variable

**Measure:** One-day Food Record

**Time Frame:** Baseline (non-stress mid point of semester), 1-3 days into active beetroot shots (during final examination period), 4-6 days into active beetroot shots (during final examination period)

**Description:** From Exhaled breath condensate or saliva, competitors/inhibitor of NO; exploratory mediator of FENO

**Measure:** Arginase

**Time Frame:** Baseline (non-stress mid point of semester), 1-3 days into active beetroot shots (during final examination period), 4-6 days into active beetroot shots (during final examination period)

**Description:** From Exhaled breath condensate or saliva, competitors/inhibitor of NO; exploratory mediator of FENO

**Measure:** Asymmetric dimethylarginine

**Time Frame:** Baseline (non-stress mid point of semester), 1-3 days into active beetroot shots (during final examination period), 4-6 days into active beetroot shots (during final examination period)

#### Primary Outcomes

**Description:** The Fraction of NO in Exhaled Breath (FENO, in ppb) will be measured with an electrochemical gas analyzer (NIOX vero).

**Measure:** Exhaled Nitric Oxide

**Time Frame:** Baseline (non-stress mid point of semester), 1-3 days into active beetroot shots (during final examination period), 4-6 days into active beetroot shots (during final examination period)

**Description:** Binary positive/negative test results for a panel of viral and bacterial respiratory pathogens.

**Measure:** Viral Polymerase Chain Reaction

**Time Frame:** Baseline (non-stress mid point of semester), 4-6 days into active beetroot shots (during final examination period)

**Description:** Assess symptoms of acute upper respiratory tract infections (common cold symptoms) by questionnaire

**Measure:** Wisconsin Upper Respiratory Symptom Survey (WURSS)

**Time Frame:** Baseline (non-stress mid point of semester), 1-3 days into active beetroot shots (during final examination period), 4-6 days into active beetroot shots (during final examination period), 1-3 days after finishing beetroot shots (after final examinations)

**Description:** Participants will upload a cell phone video of their intake for that day to a secure cloud folder, time stamp in comparison to assigned intake windows provides measure of adherence

**Measure:** Adherence monitoring (feasibility)

**Time Frame:** Every day during active beetroot shots (Days 1-7)

**Description:** Follow-up questionnaire will be administered with the online follow-up survey. It explores experience with the best mode of reminder receipt (email or text message), whether the participants guessed their condition, and how burdensome 8 aspects of the study (including daily intake, video recording, upload, questionnaires, physiological assessment) were.

**Measure:** Burdensomeness

**Time Frame:** 1-3 days after finishing beetroot shots (after final examinations)

#### Secondary Outcomes

**Description:** Ad-hoc acute stress rating

**Measure:** Acute stress

**Time Frame:** Baseline (non-stress mid point of semester), 1-3 days into active beetroot shots (during final examination period), 4-6 days into active beetroot shots (during final examination period), 1-3 days after finishing beetroot shots (after final examinations)

**Description:** Cortisol, from saliva sampled with passive drool test

**Measure:** Cortisol

**Time Frame:** Baseline (non-stress mid point of semester), 1-3 days into active beetroot shots (during final examination period), 4-6 days into active beetroot shots (during final examination period)

**Description:** Perceived Stress Scale (PSS)

**Measure:** Perceived stress

**Time Frame:** Baseline (non-stress mid point of semester), 4-6 days into active beetroot shots (during final examination period)

**Description:** Hospital Anxiety and Depression Scale (HADS)

**Measure:** Mood

**Time Frame:** Baseline (non-stress mid point of semester), 4-6 days into active beetroot shots (during final examination period)

**Description:** Measured after at least 10 min of acclimatization to the laboratory

**Measure:** Blood pressure (systolic and diastolic blood pressure) and heart rate

**Time Frame:** Baseline (non-stress mid point of semester), 1-3 days into active beetroot shots (during final examination period), 4-6 days into active beetroot shots (during final examination period)

### Eligibility Module

**Eligibility Criteria:** Inclusion Criteria:

Individuals will be included:

* SMU or Baylor University students
* Ages of 18-30 years old.

Exclusion Criteria:

* Active smokers
* Smoking cannabis or vape
* Students with clinically significant asthma
* COPD and emphysema
* Allergic rhinitis
* High levels of exhaled nitric oxide (FENO ≥40ppb)
* Developing kidney stone
* Clinically significant heart disease
* Cerebrovascular disease
* Thyroid dysfunction
* Out-of-control diabetes
* Significant current problems with schizophrenia, psychosis, mood disorders, suicidality, and drug or alcohol dependence or abuse

**Healthy Volunteers:** True

**Maximum Age:** 30 Years

**Minimum Age:** 18 Years

**Sex:** ALL

**Standard Ages:**

- ADULT

### Contacts Locations Module

#### Central Contacts

**Contact 1:**

**Email:** sspringfield@mail.smu.edu

**Name:** Savannah Springfield, BA

**Phone:** (214) 768-6441

**Role:** CONTACT

**Contact 2:**

**Email:** Danielle_Young@baylor.edu

**Name:** Danielle Young, Ph.D.

**Phone:** (254) 710-2236

**Role:** CONTACT

#### Locations

**Location 1:**

**City:** Dallas

**Contacts:**

***Contact 1:***

- **Email:** sspringfield@smu.edu

- **Name:** Savannah Springfield, BA

- **Phone:** 214-768-6441

- **Role:** CONTACT

**Country:** United States

**Facility:** Southern Methodist University

**State:** Texas

**Status:** RECRUITING

**Zip:** 75206

**Location 2:**

**City:** Waco

**Contacts:**

***Contact 1:***

- **Email:** Danielle_Young@baylor.edu

- **Name:** Danielle Young, Ph.D.

- **Phone:** 254-710-2236

- **Role:** CONTACT

**Country:** United States

**Facility:** Baylor University

**State:** Texas

**Status:** RECRUITING

**Zip:** 76796

#### Overall Officials

**Official 1:**

**Affiliation:** Southern Methodist University

**Name:** Thomas Ritz, Ph.D.

**Role:** PRINCIPAL_INVESTIGATOR

**Official 2:**

**Affiliation:** Baylor University

**Name:** Annie Ginty, Ph.D.

**Role:** STUDY_DIRECTOR

### IPD Sharing Statement Module

**IPD Sharing:** UNDECIDED

## Derived Section

### Condition Browse Module - Ancestors

- ID: D000007239

- Term: Infections

- ID: D000012140

- Term: Respiratory Tract Diseases

### Condition Browse Module - Browse Branches

- Abbrev: BC01

- Name: Infections

- Abbrev: All

- Name: All Conditions

- Abbrev: BC23

- Name: Symptoms and General Pathology

- Abbrev: BC08

- Name: Respiratory Tract (Lung and Bronchial) Diseases

### Condition Browse Module - Browse Leaves

- ID: M10283

- Name: Infections

- Relevance: LOW

- As Found: Unknown

- ID: M6368

- Name: Communicable Diseases

- Relevance: LOW

- As Found: Unknown

- ID: M14978

- Name: Respiratory Tract Infections

- Relevance: HIGH

- As Found: Respiratory Tract Infections

- ID: M14977

- Name: Respiratory Tract Diseases

- Relevance: LOW

- As Found: Unknown

### Condition Browse Module - Meshes

- ID: D000012141

- Term: Respiratory Tract Infections

### Misc Info Module

- Version Holder: 2024-05-31
