Effects of Ketone Supplement and Alcohol on Brain Metabolism

Screening Visit: Participants will complete a screening visit at the CSA (Center for Studies of Addiction). At the visit participants will be asked to show legal photo identification and undergo a breath alcohol test to ensure a breath alcohol concentration (BrAC) of <0.000%, to ensure that the participant is not intoxicated, which would make it difficult to provide informed consent. Participants will be given the informed consent form to read, which a study staff member will then review, as well as provide an explanation of the study protocol, risks, potential benefits, and alternative treatments. Following resolution of any questions, participants will be asked to sign the consent form and an entire copy of it will be given to the participants. The participant will be reminded that signing the consent form represents their willingness to participate in the study but that the subsequent screening process will determine their eligibility to do so.

Next, the participant will meet with a study clinician who will obtain a medical history and conduct a physical examination. Trained staff will measure subjects' weight, height, and vital signs (blood pressure and heart rate). Study staff trained in phlebotomy will draw approximately 3 tablespoons (44 mL) of blood for baseline laboratory measure. Baseline laboratory samples will include:

• Complete blood count (CBC)
• Blood chemistries (i.e., standard blood work and lipids)
• Urinalysis
• Urine toxicology (presence of psychoactive drugs)
• Urine pregnancy test for women In cases where blood samples are drawn and potential lab errors or abnormal findings occur, subjects will be asked to provide an additional blood sample to repeat the test(s).

Study staff will complete the following assessments with participants during Consent/Screening:

• Sociodemographic/general participant information: An assessment of medical history, personal and family history of alcohol use disorder, marital status, educational and occupational information, and substance abuse treatment history will be obtained, along with the individual's self-identified ancestry.
• A structured diagnostic psychiatric interview for DSM-5, which will be used to identify current substance use- and psychiatric disorders.
• Timeline Follow-Back (TLFB) is a 90-day assessment of use of alcohol, opioids, caffeine, nicotine, marijuana, cocaine, amphetamine, methamphetamine, and benzodiazepines.
• MRI safety form: This standard assessment form, created by the Department of Radiology at the Hospital of the University of Pennsylvania, assesses the history of specific prostheses, surgical implants, and other MRI contraindications.
• Oral HIV test: participants will be administered an oral HIV test. If the HIV test positive, study staff will inform the participants of the results, counsel the participants regarding the need to confirm the finding with a blood test and to avoid potential exposure of others to HIV, and draw an additional 1/2 tablespoon of blood to conduct a confirmatory HIV blood test. If the blood test is positive for HIV, the participants will be referred for medical care and given a copy of the test to provide to the doctor for the follow-up visit.

After the screening visit is complete, staff will call participants within 5 business days to let them know whether or not the study is a good fit. If it is a good fit for staff will schedule the 3 study lab visits.

Study lab visits: Staff will contact the participant the day before the lab visit to take the breakfast order, snack order, and tell subjects if they need to take a Lyft into the study visit located at Stellar Chance Laboratories (440 Curie Blvd.). On the lab visit that participants receive alcohol beverages, they will need to take a Lyft to the visit and back home after the visit.

Each visit will start at approximately 8:30 am, and participants will meet staff at Stellar chance Lab. They will be asked to not eat or drink anything except water after midnight the day before and on the day of each of the study lab visits. Staff will provide a light breakfast,

Table 2. Timeline of events on Lab days Approximate time Study procedure 08:30 AM Participant arrives at the study site and breathalyzer (BrAC must be 0.000) 8:35 AM Vital signs, finger stick for blood glucose, and ketone levels 8:40 AM Urine sample for UDS and urine pregnancy test if you are a woman who is able to become pregnant. 8:45 AM Light breakfast 9:00 AM Staff will review with you any recently used drugs, alcohol, medications, or vitamins since your last visit. 9:45 AM Administration of KS (20g), or alcohol, or no intervention 10:00AM MRI 11:30 AM Breathalyzer, finger stick for blood glucose, and ketone levels 11:45 AM Snack 12:20 AM Breathalyzer (repeated until below 0.020), finger stick for blood glucose (repeated if glucose is below 70 mg/dl, until it returns to normal). 12:30 AM Once BrACH is below 0.020 and blood glucose levels return to normal levels a study clinician will meet with you to ask you some questions and complete a brief sobriety assessment to make sure it is safe for you to go home.

Follow-up (~5 minutes): On the first business day following the study visits, participants will receive a follow-up phone call from study staff to ask about any changes to their health and to ensure that they did not experience any side effects from the MRI scan or other laboratory procedures. If the next business day falls on a holiday, then the follow-up call may occur on the next business day.