Clinical trial NCT01996943
Conditions: Atrial Fibrillation Alcohol
Keywords: Atrial Fibrillation Alcohol Electrophysiologic properties of the atria Randomized Clinical Study
Sponsor: University of California, San Francisco
Start date: 2012-09-01; Completion date: 2020-05-01
Enrollment: 100 participants; Allocation: RANDOMIZED; Intervention model: PARALLEL

Brief summary:
Atrial fibrillation (AF) is the most common sustained arrhythmia in the United States and it has been associated with ethanol use. Understanding how ethanol affects the electrical properties of the heart and induces AF has important public health implications. The objective of this research is to investigate the mechanistic relationship between ethanol and atrial fibrillation in humans by performing a placebo controlled study looking at the electrical properties of the heart in patients receiving intravenous ethanol or placebo. The investigators hypothesize that ethanol increases the susceptibility of human myocardium to atrial fibrillation through electrophysiologic changes in the atrial myocardium in the acute setting.

Detailed description:
The purpose of this study is to look for changes in the electrical properties of heart that may be caused by ethanol (commonly referred to as alcohol) and specifically how ethanol may trigger episodes of the most common abnormal heart rhythm, atrial fibrillation (AF). This study will demonstrate the mechanism of ethanol induced atrial fibrillation and clarify the health effects of one of the worlds' most popular drugs (ethanol). With this understanding, physicians may be able to better identify those patients most at risk for ethanol induced AF and target public health campaigns towards this vulnerable population.

Patients in this study will undergo an electrophysiologic study both prior to and after receiving either an ethanol or placebo infusion. This electrophysiology study will measure AF inducibility (the primary outcome), left and right atrial conduction times, and the atrial effective refractory period in multiple locations (AERP). The changes in the conduction times and AERPs (before and after study drug infusion) will be recorded as secondary outcomes.

About 100 people will participate in this study. 50 people will be randomized to receive intravenous ethanol, and 50 people will be randomized to receive an intravenous placebo. The placebo will be in the form of 0.45% saline solution ("half normal saline") and the alcohol will be in the form of 6% volume/volume ethanol in 0.45% saline solution.

Arms: arm 1: The placebo will be administered to participants after the baseline electrophysiologic measurements are recorded. arm 2: Ethanol (alcohol) will be administered to participants after the baseline electrophysiologic measurements are recorded.
Interventions: intervention 1: Ethanol intervention 2: Placebo
Intervention descriptions: intervention 1: 6% volume/volume ethanol in 0.45% saline solution. intervention 2: The placebo with be 0.45% saline solution ("half normal saline").

Protocol documents:
Prot_000.pdf: 
 
The HOLIDAY (HOw ALcohol InDuces Atrial TachYarrhythmias) Study 
 
 
 
NCT Number: NCT01996943 
Document Type: Study Protocol 
Document Date: 03/30/2017 
 


HOLIDAY Protocol 
 
 
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The HOLIDAY Study 
(HOw ALcohol InDuces Atrial TachYarrhythmias) 
Single-Center Randomized Controlled Trial 
 
 
 
Author(s): 
Gregory M. Marcus, MD, MAS 
Principal Investigator, UCSF Division of Cardiology  
 
Trisha F. Hue, PhD, MPH 
Project Director, UCSF Coordinating Center 
 
Emily Lee 
Clinical Research Coordinator, UCSF Division of Cardiology 
 
Shannon Fan 
Clinical Research Coordinator, UCSF Division of Cardiology 
 
Document type: 
Protocol  
 
Version: 
1.1 
 
Document status: 
Final 
 
Release date: 
30 March 2017 
 
Clinical Site 
University of California, San Francisco 
505 Parnassus Ave, San Francisco, CA 94143 
 
 
 
 


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Signature Page for Principal Investigators 
 
 
Protocol name:  The HOLIDAY Study 
 
 
I have read this protocol and agree to conduct this study in accordance with all stipulations of 
the protocol and in accordance with the World Medical Association Declaration of Helsinki 
Ethical Principles for Medical Research Involving Human Subjects. 
 
 
Gregory M. Marcus, MD, MAS 
(Principal Investigator) 
 
Signature  
 
Date 
 
 
 


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LIST OF ABBREVIATIONS  
 
AUDIT 
The Alcohol Use Disorders Identification Test 
AERP 
atrial effective refractory period 
AF 
atrial fibrillation 
BAC 
blood alcohol concentration 
BrAC 
breath alcohol concentration 
CHADS2 
CRF 
stroke risk index: Cardiac Failure, Hypertension, Age, Diabetes, Stroke [Doubled] 
case report form 
CS 
coronary sinus 
CT 
computed tomography 
EC 
ECG 
Ethics Committee 
electrocardiogram 
PAC 
premature atrial contraction 
PV 
pulmonary vein 
TEE 
transesophageal echocardiogram 
TIA 
transient ischemic attack 
TTE 
transthoracic echocardiogram 
SAE 
serious adverse event 
UCSF  
University of California, San Francisco  
 
 
 


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Table of Contents 
ADMINISTRATIVE INFORMATION SUMMARY ............................................................................................ 6 
1.0 
BACKGROUND ............................................................................................................................................ 7 
1.1 PREVIOUS STUDY DATA .................................................................................................................................................... 7 
2.0  
INVESTIGATION PLAN ............................................................................................................................ 8 
2.1 STUDY DESIGN .................................................................................................................................................................... 8 
2.2 RANDOMIZATION AND TREATMENT GROUPS ............................................................................................................... 8 
2.4 STUDY VISITS AND WINDOWS ......................................................................................................................................... 9 
2.4 STUDY OBJECTIVES AND AIMS ......................................................................................................................................... 9 
2.6  INCLUSION/EXCLUSION CRITERIA .............................................................................................................................. 11 
2.6.1 Inclusion Criteria ....................................................................................................................................................... 11 
2.6.2 Exclusion Criteria ...................................................................................................................................................... 11 
3.0  
STUDY TREATMENT .............................................................................................................................. 11 
3.1 ANESTHESIA PROTOCOL ................................................................................................................................................. 12 
3.2 CATHETER PLACEMENT ................................................................................................................................................. 12 
3.3 PRE-INFUSION EP STUDY .............................................................................................................................................. 12 
3.4 INFUSION ........................................................................................................................................................................... 13 
3.5 POST-INFUSION EP STUDY ............................................................................................................................................ 13 
3.6  ASSESSMENT OF STUDY TREATMENT COMPLETION ................................................................................................ 14 
4.0 
PARTICIPANT EARLY TERMINATION ............................................................................................... 14 
4.1 UNTREATED RANDOMIZED PARTICIPANTS ................................................................................................................ 14 
4.2  UNSUCCESSFULLY TREATED PARTICIPANTS ............................................................................................................. 14 
4.3  PARTICIPANT WITHDRAWALS ..................................................................................................................................... 15 
4.4  OTHER EARLY TERMINATIONS .................................................................................................................................... 15 
5.0  
STUDY PROCEDURES ............................................................................................................................ 15 
5.1  SCREENING FOR INCLUSION/EXCLUSION CRITERIA & RANDOMIZATION ............................................................ 16 
5.2  FOLLOW-UP VISITS (MONTHS 3, 6, AND 12) ........................................................................................................... 16 
5.3  TARGETED CONCOMITANT MEDICATIONS ................................................................................................................. 16 
5.4  TARGETED ADVERSE EVENT (AE) AND SERIOUS UNEXPECTED ADVERSE EVENT (SAE) EVALUATION AND 
REPORTING ............................................................................................................................................................................... 17 
6.0 
RISKS AND BENEFITS ............................................................................................................................. 17 
6.1  POTENTIAL RISKS ........................................................................................................................................................... 17 
6.2  POTENTIAL BENEFITS .................................................................................................................................................... 18 
7.0 
SAFETY MONITORING ............................................................................................................................ 19 
7.1  DATA AND SAFETY MONITORING BOARD (DSMB) ................................................................................................. 19 
7.2  SAFETY MONITORING FOR TARGETED ADVERSE EVENTS ........................................................................................ 19 
7.2.1 Intra procedure monitoring ................................................................................................................................. 19 
7.2.2 Post-Procedure Monitoring .................................................................................................................................. 20 
8.0  
PROTOCOL DEVIATIONS ...................................................................................................................... 20 
9.0 
QUALITY ASSURANCE AND DATA MANAGEMENT ........................................................................ 20 
9.1 CLINICAL SITE INVESTIGATOR AND COORDINATOR TRAINING .............................................................................. 20 
9.2 DATA HANDLING AND CONFIDENTIALITY .................................................................................................................. 20 
9.3  SOURCE DOCUMENTS ..................................................................................................................................................... 21 
9.4  DATA COLLECTION AND MANAGEMENT .................................................................................................................... 21 
10.0 
STATISTICAL METHODS ..................................................................................................................... 21 


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11.0 
ETHICAL CONSIDERATIONS .............................................................................................................. 21 
11.1  INSTITUTIONAL REVIEW BOARD (IRB) APPROVAL .............................................................................................. 22 
11.2  INFORMED CONSENT ................................................................................................................................................... 22 
11.3  DECLARATION OF HELSINKI ...................................................................................................................................... 22 
BIBLIOGRAPHY ................................................................................................................................................... 23 
 
 
 
 
 


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ADMINISTRATIVE INFORMATION SUMMARY 
 
 Project Title: 
The HOLIDAY (HOw ALcohol InDuces Atrial TachYarrhythmias) 
Study 
 
 
Principal Investigator: 
Gregory M. Marcus, MD, MAS, FACC, FAHA, FHRS 
 
 
Mailing Address: 
University of California, San Francisco 
505 Parnassus Avenue, M-1180B 
San Francisco, CA 94143-0124 
USA 
 
 
Telephone: 
1 (415) 476-5706 
 
 
E-Mail: 
marcusg@medicine.ucsf.edu 
 
 
 
 
 


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1.0 
BACKGROUND 
 
Atrial Fibrillation (AF), the most common sustained arrhythmia in the United States, has 
profound public health implications: the lifetime risk of AF is 1 in 4, the disease affects several 
million Americans, and it is the most common cause of embolic stroke.1-3 Unfortunately, the 
pathogenesis of AF is still poorly understood and has multiple influences, with ethanol likely 
being one of particular importance. Ethanol is the most commonly consumed drug in the United 
States. In the 2001 US National Household Survey on Drug Abuse, 84% of people surveyed 
above the age of 12 admitted to having consumed ethanol in the past, with a quarter of men, 
and nearly a fifth of women, admitting to at least one episode of binge ethanol drinking (>5 
drinks in a single sitting) during the previous month.4 The mechanism(s) by which ethanol 
increases the risk for AF, or “triggers” AF in certain individuals are unknown. 
  
The widest body of literature is epidemiological, and those studies have suggested an 
association between ethanol use and the risk for AF since the initial cases of “holiday heart” 
were described in the 1970s5. The patients in the initial case reports were generally male, 
middle-aged or younger chronic alcoholics who were without structural heart disease. The 
temporal association of acute onset paroxysmal AF with ethanol binges, in patients with 
otherwise normal hearts, led to the premise that AF paroxysms could be related to ethanol use. 
This premise was brought into question when large epidemiologic studies, such as the 
Framingham Heart Study, the Manitoba Follow-up Study, and the Renfrew/Paisley study, found 
no association between chronic ethanol use and AF6-8. Later studies, such as the Copenhagen 
Heart Study and a reanalysis of the Framingham cohort, demonstrated that when controlled for 
the quantity of ethanol consumed and sex, an association with chronic and paroxysmal AF was 
seen with moderate to heavy ethanol use (>3 drinks per day) in men9-12. From these studies it is 
apparent that the effect of ethanol is both multifactorial and complex. The case series and 
anecdotal evidence suggest that acute ethanol binging can induce temporary paroxysms of AF 
in younger patients without structural heart changes (consistent with acute electrophysiologic 
changes)5, 13, 14, while the epidemiological evidence suggests that chronic moderate to heavy 
ethanol use in men is associated with more chronic AF (consistent with a structural effect on the 
atria)9-12. It is unclear if these two conditions are part of continuum of risk or if they represent the 
diverse effects that ethanol may have on different patient populations. In order to investigate the 
mechanistic relationship between acute ethanol use and AF, we will perform a study of the 
acute electrical effects of ethanol in a non-alcoholic patient population with baseline paroxysmal 
AF. 
 
1.1 
Previous Study Data 
 
 
The prevailing theory is that a PAC triggers AF, representing multiple, partially-reentrant 
wavelets. As experiments have been limited to animal models, a few small comparative 
observational (not experimental) studies in humans, and mechanistic concepts inferred by the 
success of certain ablation procedures, there is no human experimental model for in vivo atrial 
EP changes in AF. The data suggest that three readily measurable EP properties are important 
in the initiation and perpetuation of AF: automaticity, the effective refractory period (ERP), also 
reflected in the action potential duration (APD), and conduction velocity. 
 
The changing nature of ERP (or APD) relative to the previous diastolic interval, a property called 
restitution, may be critical to the initiation of fibrillation. There is some evidence that when the 
restitution curve becomes steep (>1), meaning that the ERP (or APD) shortens disproportionate 
to the diastolic interval, susceptibility to fibrillation is particularly enhanced. No human study has 
demonstrated a causal relationship wherein a change in restitution leads to a greater propensity 
to AF. In isolated animal myocardial tissue preparations, ethanol consistently reduces APD. 


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Intracoronary ethanol prolongs ventricular conduction times in canine models.  
 
 
Small (n<15) case series describing invasive EP studies before and after oral ethanol in 
humans were performed decades ago. The following was observed: in 10 habitual drinkers, oral 
whiskey resulted in a decrease in the atrial ERP that did not reach statistical significance; atrial 
arrhythmias could be induced with atrial extra stimulus pacing in 5 after, but not before, drinking 
whiskey. In 14 patients, whiskey resulted in an increase in atrial conduction time and a decrease 
in the ventricular ERP. And in 14 patients, whiskey resulted in the His-Purkinje system delay, 
and sustained atrial arrhythmias could be induced in 4 of them. Our prior study of 48 
consecutive patients undergoing a standard questionnaire and right atrial ERP measurements 
during invasive EP studies found that greater average ethanol intake was associated with a 
shorter right atrial ERP. 
 
Utilizing the protocol detailed here, we have successfully obtained pilot data in 3 patients during 
AF ablation procedures. For the first patient, the PK model achieved a breath alcohol 
concentration (BrAC) of between 0.079-0.082% within 20 minutes, with successful clamping 
within that range for the duration of the experiments. No pacing maneuvers were performed in 
the first patient in order to focus on safety and feasibility of the ethanol infusion. With EP testing, 
patient 2 exhibited a 45 ms decrease in the proximal CS ERP; while no AF was observed with 
pacing prior to ethanol, AF was reproducibly observed during pacing from both upper pulmonary 
veins after ethanol. Patient 3 exhibited an 85 ms drop in the left upper pulmonary vein ERP after 
ethanol, with repeatedly induced AF after (but not before) the ethanol was administered. All 3 
patients exhibited normal wound healing and recovery without complication. Thus, our 
experience with 3 patients receiving intravenous ethanol undergoing AF ablation procedures 
revealed no complications and normal recovery and wound healing. 
 
This important preliminary data demonstrates: 1) The feasibility of delivering IV ethanol and 
serial EP assessments in patients undergoing invasive AF ablation procedures; 2) Our ability to 
utilize the BrAC method; 3) Dramatic changes in ERPs and AF inducibility in the two patients 
undergoing pacing maneuvers; 4) The safety of these experiments in patients undergoing AF 
ablation procedures. 
 
2.0  
INVESTIGATION PLAN 
 
2.1 
Study Design 
 
This study is a randomized single-center single-blinded placebo controlled trial of patients age 
21-90 with paroxysmal atrial fibrillation. This study will be conducted at University of California, 
San Francisco (UCSF) Medical Center. One hundred patients will be enrolled and will receive 
the study infusion (alcohol or placebo infusion). 
 
2.2 
Randomization and Treatment Groups 
 
Participants will be allocated (1:1) to a study assignment, using alternating blocked 
randomization (in blocks of five). Participants will be assigned to either the Ethanol Infusion 
group or to the Placebo Infusion group.  
 
2.3   
Blinding 
 
To preserve blinding to infusion assignment, as much as possible, the following individuals will 
be blinded, during the trial: 
• 
Study participants 


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• 
UCSF Electrophysiology team, including the electrophysiologist, electrophysiology 
fellow, and nursing staff during the procedure* (not including the individuals noted 
below) 
It will be necessary for the following individuals to be unblinded to individual participant 
randomization assignments only, during the trial: 
• 
UCSF Study Coordinator(s) that administers the infusion, records the infusion 
assignment in the database, and oversees the infusion process during the procedure 
• 
UCSF Anesthesia team  
• 
UCSF Pharmacy team that prepares the study infusion 
 
Only the following individuals will be unblinded to both individual and grouped participant 
randomization assignments, during the trial: 
• 
UCSF Coordinating Center Epidemiologist that is the DSMB liasion 
• 
UCSF Coordinating Center Analyst and Statistician that performs analyses for the DSMB 
reports and interim analysis 
* NOTE:  Breath alcohol readings will be taken during the study procedure for both the alcohol 
and placebo group participants, to help maintain blinding. Treatment group assignment will not 
be indicated on the infusion bags. 
 
2.4 
Study Visits and Windows 
 
After screening and eligibility has been determined, interested participants will be consented at 
either their pre-procedure, standard of care CT scan, or another pre-procedure appointment at 
UCSF Medical Center. At this point, questionnaires addressing alcohol consumption, triggers of 
AF, and medical history will be administered by the research team member performing the 
consent process.  See Figure 1 for study flow.   
 
Safety follow-up will be performed by chart review at 3 months, 6 months, and 12 months post-
procedure, when AF ablation patients normally return to clinic. If participants have no scheduled 
clinic appointment within the visit window (+/- 30 days for 3 and 6 months, + 6 months and - 30 
days for 12 month), a member of the research team will attempt to contact the participant by 
phone to gather follow-up information. The clinical site will make every possible effort to ensure 
that each study visit is conducted within the specified window. 
 
Table 1. Visit Windows 
Study Visit 
Scheduled Timepoint 
Visit Window  
Screening and Enrollment 
 
 
V0 
Randomization and Procedure 
 
V1 
Month 3* 
+/- 30 days 
V2 
Month 6* 
+/- 30 days 
V3 
Month 12* 
+6 months/- 30 days 
*Follow-up visits will be calculated from the date of ablation procedure (i.e., baseline).  
 
 


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Figure 1. Study Flow Diagram 
 
  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
* The 
pharmacy that prepares the ethanol/placebo requires notification of the treatment assignment 1 
business day prior to administration, therefore, participants must be consented and randomized at least 
1 day prior to the scheduled procedure. 


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2.5  
Study Objective and Aims 
 
The objective of this trial is to seek to reveal the mechanisms underlying the relationship 
between ethanol, the most commonly consumed drug in the world, and atrial fibrillation (AF), the 
most common arrhythmia. Specifically, we seek to determine the electrophysiological effects of 
acute ethanol exposure on atrial myocardium via a randomized trial of intravenous ethanol 
versus placebo in paroxysmal AF patients. 
 
Study Aims:  
1.  To assess the acute effects of intravenous ethanol administration on the atrial effective 
refractory period (AERP) and atrial conduction velocity of in vivo human atrial myocardium.  
 
Hypothesis: Intravenous administration of ethanol decreases AERP and atrial conduction 
velocity.  
 
2.  To assess the acute effects of intravenous ethanol administration on the inducibility of AF of 
in vivo human atria.  
 
Hypothesis: Intravenous administration of ethanol increases in vivo susceptibility to AF.   
 
2.6  
Inclusion/Exclusion Criteria 
 
2.6.1  
Inclusion Criteria 
 
1. Aged 21-90 
2. History of paroxysmal or persistent atrial fibrillation (AF)  
3. Planned procedure pulmonary vein isolation (PVI) or electrophysiology study (EPS) 
4. Presenting in sinus rhythm or atrial fibrillation of recent onset (the latter confirmed by 
sinus rhythm documented no more than one week prior) 
 
2.6.2  
Exclusion Criteria 
 
1. History of substance abuse or alcoholism (determined by history, Alcohol Use Disorders 
Identification Test (AUDIT) questionnaire, or chart review) 
2. LVEF <50% 
3. Liver dysfunction (AST, ALT, total bilirubin, or alkaline phosphatase level greater than 
two times the normal range) 
4. Clinical evidence of liver disease (enlarged liver, caput medusa, spider angiomas, or 
other signs of liver disease on exam) 
5. Pregnancy 
6. Inability to give informed consent 
7. Taking or have taken amiodarone within one month prior to procedure 
8. Taking or have taken dofetilide within 24 hours prior to the procedure 
9. General severe intolerance to alcohol 
 
3.0  
STUDY TREATMENT 
 
Demographic and medical history information will be obtained by chart review and by 
questionnaire administered by a member of the research team.  Prior to the procedure, patients 
will be instructed to abstain from alcohol for 48 hours and to stop antiarrhythmic drugs for at 
least 5 half-lives.  Per the standard AF ablation procedure with a double transseptal puncture, 
catheters will be positioned.  Study coordinators measure conduction times using electronic 


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calipers and custom software in regular use in the UCSF EP laboratory, and EP measurements 
are then reviewed offline by the PI blinded to treatment assignment.  
 
 
3.1 
Anesthesia Protocol 
 
To minimize the effects of anesthestic agents on AERP and conduction time (see Operations 
Manual), the anesthesia team will follow a specific anesthesia protocol for HOLIDAY, devised in 
collaboration with the Anesthesia Department at UCSF, from induction to termination of the 
study protocol.  Propofol, remifentanil and nitrous oxide may be used at induction, with cis-
atracurium for paralysis.  Prior to transseptal access, nitrous oxide and cis-atracurium should be 
turned off, though propofol and remifentanil infusions can continue. From transseptal access 
through the end of the experimental portion of the procedure, only a propofol infusion may be 
used.  If remifentanil is necessary after transseptal access, the anesthesia team may also use 
remifentinal so long as remifentinal is also infused for the reminder of the experimental portion 
of the procedure to ensure consistency.  Phenylephrine may be titrated throughout the 
procedure to maintain a mean arterial pressure above 70 mmHg.  
 
The following medications may not be used until after completion of the experimental portion: 
benzodiazepines, fentanyl, rocuronium, succinylcholine, ketamine, scopolamine, and inhaled 
anesthetics other than nitrous oxide.  Only in the case of an emergency, may anticholinergics be 
used. 
 
3.2 
Catheter Placement 
 
Following the initiation of sedation, intra-cardiac multipolar electrocatheters will be placed in the 
following positions under fluoroscopic guidance: decapolar catheter in the coronary sinus (CS), 
decapolar catheter along the crista terminalis, quadripolar catheter along the medial tricuspid 
annulus at the position of the His bundle, duodecapolar circular catheter in the left upper 
pulmonary vein, and quadripolar ablation catheter in the right upper pulmonary vein. If the 
ablation procedure were a cryoballoon PVI, four, instead of five, catheters will be used: decapolar 
catheter in the coronary sinus (CS), decapolar catheter along the crista terminalis, quadripolar catheter 
along the medial tricuspid annulus at the position of the His bundle, and cryoballoon catheter in the left 
upper pulmonary vein. (See Operations Manual for details) 
 
Catheter repositioning should be avoided to prevent disrupting post-procedure conduction time 
measurements.  Prior to transseptal access, the alcohol or placebo solution will be connected to 
minimize left atrial time.  
 
3.3 
Pre-Infusion EP Study 
 
Immediately following catheter placement, a one-minute premature atrial contraction (PAC) 
count will be conducted. AERP measurements will then be taken at the following bipoles: CS 
proximal, CS distal, high right atrium (crista terminalis catheter), left upper pulmonary vein, and 
right upper pulmonary vein. The AERP will be obtained by an incremental technique using a 600 
ms drive cycle length for 8 beats (S1) at twice pacing threshold with incremental steps for the 
atrial extrastimuli (S2). The first drive train at each location should begin with an atrial 
extrastimuli (S2) of 140 ms, which is not expected to result in atrial capture. Once the first atrial 
capture is achieved, the S2 will be decreased by 5 ms. AERP will be defined as the longest S1-
S2 interval that fails to result in atrial capture. (See Operations Manual for details) 
 


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3.4 
Infusion 
 
After the first EP study, the intravenous ethanol or placebo infusion will be initiated while the left 
atrial anatomic map is being constructed to reduce time added to the ablation procedure and 
minimize risk.  The infusion is run by an iMED Gemini PC-2TX pump connected to and 
controlled by the CAIS 2.0 computer software.  The ethanol infusion will consist of a 6% 
volume/volume ethanol solution in 0.45% saline titrated by the BrAC quick clamp method until a 
steady state is achieved at a blood alcohol concentration of 0.08%. The mean time to achieving 
this steady state is regulated by the CAIS 2.0 software to be approximately 20 minutes.  Infusion 
rates are determined by the patient’s height, weight, age, and sex, but are automatically 
adjusted depending on the patient’s breath alcohol measurements, which are taken every three 
to four minutes until steady state is reached.  
 
Patients assigned to the placebo group will receive 5% dextrose (D5) in 0.45% saline.  BrAC 
measurements will be obtained at identical intervals for all participants; BrAC measurements will 
be randomly generated within two standard deviations of the predicted values calculated by the 
CAIS 2.0 software based on height, weight, age, and sex.  The randomly generated BrAC 
values will be input into the CAIS 2.0 software, as for patients receiving an alcohol infusion, and 
the infusion rates will be automatically adjusted accordingly.  
 
Once the BrAC reading is ±0.002% from 0.080% (steady state), BrAC measurements will be 
taken every 10 minutes and input into the CAIS 2.0 software. Breath alcohol measurements will 
be taken using the Intoximeters Alco Sensor IV fitted with a one-way valve adaptor for intubated 
patients. (See Operations Manual for details) 
 
3.5 
Post-Infusion EP Study 
 
As soon as the blood alcohol content reaches a steady state of 0.080% ±0.002%, a one-minute 
premature atrial contraction (PAC) count will be conducted.  AERP measurements will then be 
taken at the following bipoles: CS proximal, CS distal, high right atrium (crista terminalis 
catheter), left upper pulmonary vein, and right upper pulmonary vein.  The AERP will be 
obtained by an incremental technique using a 600 ms drive cycle length for 8 beats (S1) at twice 
pacing threshold with incremental steps for the atrial extrastimuli (S2).  The first drive train at 
each location should begin with an atrial extrastimuli (S2) of 140 ms, which is not expected to 
result in atrial capture.  Once the first atrial capture is achieved, the S2 will be decreased by 5 
ms.  AERP will be defined as the longest S1-S2 interval that fails to result in atrial capture. (See 
Operations Manual for details) 
 
Following the PAC count and AERP measurements, a phenylephrine infusion will be started 
prior to the initiation of the isoproterenol infusion to maintain a mean arterial pressure of >60 
mmHg.  Isoproterenol hydrochloride will then be infused at the following rates for 2 minutes 
each: 3 mcg/min, 6 mcg/min, 12 mcg/min, and 20 mcg/min.  If AF or another arrhythmia initiates 
at any point during the isoproterenol infusion and is sustained over 30 seconds, the 
isoproterenol infusion will be stopped, the rhythm documented, and the study terminated.  
 
If neither AF nor another arrhythmia lasting over 30 seconds occurs after 2 minutes of 20 
mcg/min isoproterenol infusion, the isoproterenol infusion will be stopped.  Once the heart rate 
falls below 100 bpm, up to 3 bursts of rapid atrial pacing will be performed from the CS proximal 
bipole.  If AF or another arrhythmia initiates at any point during the burst pacing and is sustained 
over 30 seconds, the study will be terminated and the rhythm documented.  


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3.6  
Assessment of Study Treatment Completion 
 
At the Baseline Visit, upon completion of the study treatment, the Investigator will complete the 
Study Treatment CRF. The Investigator will record details from the study treatment procedure 
(e.g., pre-procedure medications, details regarding pericardial and transseptal access, and 
fluoroscopy time) and will indicate whether the study procedure was completed as intended.  
 
4.0 
PARTICIPANT EARLY TERMINATION  
 
Full details regarding procedures for early termination will be outlined in the Operations Manual. 
 
4.1 
Untreated Randomized Participants 
 
After randomization, if an exclusionary criterion is identified prior to or at the start of the study 
procedure, the participant will not undergo the study infusion procedure.  Randomized 
participants may be early terminated, prior to commencing the study procedure, and will not 
receive the study infusion if any of the following study exclusions are identified:   
• 
Participants who present in AF sustained >1 week  
• 
If a participant does not present in sinus rhythm at the procedure, but he or she has 
proven to be in sinus rhythm at the enrollment visit, it is up to the attending whether or 
not to cardiovert the patient and proceed with study protocol.  If the participant is to be 
cardioverted, the cardioversion should take place prior to transseptal puncture and 
following right catheter placement to minimize potential effects on AERP and conduction 
times.  If a second AF episode initiates and is sustained over 2 minutes prior to initiation 
of study procedure, the study will be aborted. 
 
These participants will be considered as “Untreated Participants”.  Unrandomized participants 
who do not commence/receive the study infusion will not continue in follow-up and will not be 
included in analyses.  
 
4.2  
Unsuccessfully Treated Participants 
 
After enrollment, if the study infusion commenced and was stopped and/or determined by the 
study Principal Investigator to be unsuccessful (i.e., immediately after the procedure, the 
investigator determines that the protocol was not completely executed) for any reason, the 
participant will continue in follow-up.  
 
4.2.1 Early Stopping of Study Infusion Protocol ONLY 
 
If the study infusion commenced and the participant develops AF that is sustained over 2 
minutes during study procedure (excluding the AF susceptibility testing section), the patient 
should be cardioverted to sinus rhythm, at the discretion of his or her attending physician.  If a 
second AF episode initiates and is sustained over 2 minutes, the study will be stopped.  If a 
participant develops a non-AF tachyarrhythmia that persists over 2 minutes, it will be up to the 
attending physician to map and/or ablate the rhythms.  
 
If the study infusion (ethanol or placebo) protocol commenced prior to AF development, the 
participant will not be early terminated and will continue in follow-up.  These participants will be 
considered as “Unsuccessfully Treated Participants”.   
 


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4.3  
Participant Withdrawals 
 
Each participant will be informed that participation in the study is voluntary and that he or she 
may withdraw from the study at any time without effect on subsequent medical treatment or 
relationship with the treating physician. Participants who discontinue follow-up at any time after 
study treatment will be included in analyses. 
 
4.4  
Other Early Terminations 
 
Participants who discontinue follow-up at any time after study treatment will be included in 
analyses. 
 
Lost to Follow-Up: If a participant fails to return for at least one follow up visit (3, 6, and 12 
months post-procedure) and cannot be reached by phone within the visit windows, the 
participant will be designated an early termination. The study personnel will show "due 
diligence" by documenting in the source documents, all steps taken to contact the participant, 
e.g., dates of telephone calls, registered letters, etc. Vital status for participants lost to follow-up 
may be later obtained using outside records/death registries.  
 
Principal Investigator Determination: Participants should also be early terminated at any time if 
the Principal Investigator concludes that it would be in the participant’s best interest for any 
reason. Protocol deviations should not lead to early termination unless they indicate a significant 
risk to the participant’s safety.  
 
5.0  
STUDY PROCEDURES 
 
Table 2 provides an overview of the evaluations to be performed at each of the study visits. Full 
details regarding the all study measurements and timing of data collection will be outlined in the 
Operations Manual. 
 
Table 2. Schedule of Evaluations and Visits  
 
Screening 
Baseline 
Follow-Up Visits 
 
 
Study 
Infusion  
Month 3 
 
Month 6 
 
Month 12/ 
Close-Out 
Informed consent 
x 
 
 
 
 
Inclusion/ Exclusion 
Evaluation 
x 
 
 
 
 
Other laboratory 
evaluations 
x 
 
 
 
 
Medical History 
x 
 
 
 
 
Echo (TTE)* 
x1 
 
 
 
 
Height 
x 
 
 
 
 
Weight 
x 
 
 
 
 
ECG 
x 
x 
 
 
 
Vital signs 
 
x 
 
 
 
Concomitant Medications 
 
x 
x 
x 
x 
Targeted Adverse 
Events  
 
x 
x 
x 
x 
Serious Unexpected 
Adverse Events 
 
x 
x 
x 
x 


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Collection of materials 
for  Targeted AEs/SAEs 
(if applicable) 
 
x 
x 
x 
x 
Targeted medical care 
utilization (e.g., cardiac 
care during follow-up) 
 
 
x 
x 
x 
1 Must be performed > 1 year prior to study procedure 
 
5.1  
Screening for Inclusion/Exclusion Criteria & Randomization 
 
Each potential participant’s medical record will be reviewed for available inclusion and exclusion 
criteria data.  After the participant provides informed consent, his or her medical history and 
additional demographic data will be obtained.  Initial screening will occur at least 1 day prior to 
the patient’s scheduled ablation procedure.   
 
The pharmacy that prepares the ethanol/placebo infusion bags requires notification of the 
treatment assignment 1 business day prior to administration in order to prepare the infusion 
bag.  Therefore, participants must be consented and randomized at least 1 day prior to the 
scheduled procedure.  
 
Baseline measures will include:  
• 
Inclusion/Exclusion criteria assessment 
• 
Demographics and medical history 
• 
World Health Organization AUDIT questionnaire 
• 
CARDIA alcohol use questionnaire (shortened version) 
• 
Vital signs 
• 
Infusion, procedure, BrAC, and EP study measurements 
• 
Arrhythmia episodes pre/during/post-procedure 
• 
Adverse event assessment 
 
5.2  
Follow-up Visits (Months 3, 6, and 12)  
 
Following an ablation, patients usually return to clinic at 3, 6, and 12 months post-procedure as 
per standard of care.  HOLIDAY safety follow-up data will be collected by chart review.  We will 
collect data on targeted AEs, serious, unexpected AEs, use of antiarrhythmic drugs, use of 
anticoagulation, incidence and burden of AF or atrial flutter, incidence of ablation or 
cardioversion, and emergency room visits since the last visit. Participants who do not return to 
clinic at 3, 6, and 12 months will be contacted by the Study Coordinator to conduct the visit by 
phone.   
 
Follow-Up Visit measures will include: 
• 
Medication use 
• 
Arrhythmia episodes  
• 
Cardioversions/ablations  
• 
Adverse event assessment 
 
5.3  
Targeted Concomitant Medications 
 
Current/regular use (since their last study visit) of a specific list of concomitant 
medications/therapies must be documented at Baseline (pre-study treatment) and at all follow-
up visits. Medication data will be collected by chart review and will include type of medication, 


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frequency, and dosage. The list of targeted medications/therapies includes cardiovascular-
related medications (e.g., anticoagulants and antiarrhythmics). 
 
5.4  
Targeted Adverse Event (AE) and Serious Unexpected Adverse Event (SAE) 
Evaluation and Reporting 
 
An adverse event is any undesirable sign, symptom or medical condition occurring after 
initiation of the study treatment, even if the event is not considered to be related to study 
treatment.  Medical conditions/diseases present before starting study treatment are only 
considered AEs if they worsen after starting study treatment (or any procedures specified in the 
study protocol).  
 
A targeted adverse event (AE) is an event that may be reasonably anticipated to occur as a 
result of the study procedures or study participation or is part of the normal disease process or 
progression. Specific targeted AEs are documented in detail in the Operations Manual.  
 
A serious unexpected adverse event (SAE) is defined as being unexpected if the event exceeds 
the nature, severity or frequency described in the current IRB Application, protocol, and consent 
form.  An unexpected AE also includes any AE that meets any of the following criteria: 
§ 
Results in subject withdrawal from study participation, or  
§ 
Due to a deviation from the IRB approved study protocol 
 
After completion of the study treatment (at the Baseline Visit) and at all follow-up visits, safety 
evaluations will include monitoring for targeted adverse events and recording all serious 
adverse events for the assessment of the secondary exploratory study objective.  
 
Information about targeted adverse events, whether volunteered by the subject, discovered by 
Principal Investigator questioning, or other means, will be collected and recorded and followed 
as appropriate.  
 
As possible, each targeted adverse event will be described by: 
● duration (start and end dates), 
● severity grade (mild, moderate, severe),  
● relationship to the study treatment and/or procedure (suspected/not suspected), 
● action(s) taken and, as relevant, the outcome. 
 
Even though planned/anticipated pregnancy is a study exclusion, pregnancy during follow-up 
may occur. If a female study participant should report pregnancy, this should be reported on a 
Protocol Deviation CRF. The pregnancy should be followed up to determine details of birth, 
outcome, including spontaneous or voluntary abortion, the presence or absence of any birth 
defects or congenital abnormalities, or any maternal/newborn hazards.  
 
6.0 
RISKS AND BENEFITS 
 
6.1  
Potential Risks 
 
The primary risk to participants involves the receipt of IV ethanol. The procedure for IV ethanol 
infusion is well described and has previously been used for studies in the intoxication 
literature.86-88  The target blood alcohol level of 0.08% that will be achieved is the legal limit for 
driving in California, and is equivalent to approximately 4 drinks of ethanol in a 180 pound man 
(equivalent to moderately heavy social drinking in one sitting).  The use of IV ethanol in clinical 
settings is well established, with a long history of use in the treatment of such conditions as 


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methanol and ethylene glycol poisoning. The blood alcohol levels that have been used in such 
situations (0.1 to 0.2%) are higher than the level that we will use in our study (0.08%).156, 157  
Side effects from the ethanol infusion at our targeted dose may include mild headache, but it is 
unlikely to cause nausea and vomiting.  Early studies with high dose IV ethanol infusions used 
for surgical anesthesia in the 1960s targeted to blood ethanol concentrations of 0.2 to 0.4% 
resulted in dizziness, nausea and/or vomiting.158-160  Most of these symptoms resolved by 24 
hours. In more recent experiments using the moderate levels of ethanol that we will target 
(0.08% blood alcohol concentration), only mild headache, with no nausea or vomiting, has been 
reported.86-88  There have been no reports of serious adverse events with IV ethanol titrated by 
the BrAC quick clamp method. In addition, previous studies in which oral whiskey was 
administered to patients undergoing invasive electrophysiology (EP) studies (discussed above) 
did not result in any serious adverse events.  While the total volume infused in either the ethanol 
group or the placebo (D5 half normal saline) group is not expected to exceed 500 cc, there is 
the theoretical possibility of causing volume overload; of note, patients with a history of heart 
failure or LVEF < 50% will be excluded.  It is possible that ethanol may potentiate the side 
effects of other sedatives in patients undergoing AF ablation procedures, but this is unlikely.  All 
of these cases are staffed by board certified anesthesiologists (with continuous 12 lead 
electrocardiograms, pulse oximetry, CO2 monitoring, and blood pressure monitoring per an 
arterial line per standard of care) as well as a cardiac EP attending, a cardiac EP fellow (board 
certified or board eligible in cardiology) and two nurses. All patients have several large sheaths 
in place for venous access, defibrillation pads already in place, and the intracardiac catheters 
can provide pacing if necessary.  An echocardiography machine is immediately available in the 
EP laboratory and at least one on call interventional cardiologist and interventional cardiology 
fellow are available in the cardiac catheterization laboratory in the same suite  
 
We observed no complications and no difficulties with recovery or wound healing in the 3 AF 
ablation patients receiving IV ethanol titrated to a blood ethanol concentration of 0.08% using 
the BrAC quick clamp method.  We are not aware of any evidence to suggest that ethanol 
should adversely affect the efficacy of the AF ablation or the recovery or wound healing 
processes.  
 
Additional risks include prolongation of the AF ablation procedure. We anticipate that the 
additional procedure time will average 40 minutes.  The left atrial anatomic map will be created 
using the three dimensional mapping system per the standard clinical procedure while the 
ethanol or placebo infusion is started and titrated to steady state, helping to minimize any extra 
procedure time.  This additional procedural time may minimally increase the patient’s risk for 
stroke or sedation complications including pneumonia, deep vein thrombosis, or pulmonary 
embolism.  Although the EP tests that will be performed and attempts to induce AF are 
consistent with standard of care during AF ablation procedures per the most recent expert 
consensus10 (as well as standard of clinical practice in the UCSF cardiac EP laboratory), the 
potential induction of AF includes a risk of cardioversion, which may minimally increase the risk 
of skin burns and stroke.  As per standard of care, all patients may undergo transesophageal 
echocardiograms immediately prior to the procedure to exclude the presence of a left atrial 
appendage thrombus and all will receive IV heparin with serial ACT monitoring to minimize the 
risk of stroke.  
 
6.2  
Potential Benefits 
 
There are no direct benefits for participating in this study.  However, the knowledge gained from 
this study may help to elucidate the mechanism underlying AF, which would be pertinent to the 
~4 million Americans with the disease.35, 54  In addition, understanding these particular cardiac 


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effects may ultimately benefit the greater than hundred million individuals that consume alcohol.  
This may be particularly important as the media often touts alcohol as being “heart healthy.”23-31 
 
7.0 
Safety Monitoring  
 
7.1  
Data and Safety Monitoring Board (DSMB) 
 
HOLIDAY has an independent DSMB that is responsible for safeguarding the interests of study 
participants, assessing the safety and validity of study procedures, and for monitoring the 
overall conduct of the study and outcomes data.  The DSMB members are independent 
consultants to the PI at UCSF, and are required to provide recommendations about starting, 
continuing, and stopping the study.  In addition, the DSMB is asked to make recommendations, 
as appropriate, to the PI about: 
• 
The effects of the HOLIDAY study intervention 
• 
Benefit/risk ratio of procedures and participant burden 
• 
Selection, recruitment, and retention of participants 
• 
Adherence to protocol requirements 
• 
Completeness, quality, and analysis of measurements 
• 
Amendments to the study protocol 
• 
Adequacy of and amendments to consent forms 
• 
Performance of the UCSF centers 
• 
Participant safety 
• 
Notification of and referral for abnormal findings 
7.2  
Safety monitoring for targeted adverse events 
 
7.2.1  
Intra procedure monitoring 
 
During the procedure the patient will be monitored by a comprehensive care team. This will 
include the electrophysiology attending, the electrophysiology fellow (who is a board certified or 
board eligible cardiologist), the cardiac anesthesiologist attending (who will be unblinded), and 2 
electrophysiology lab nurses. They will continually watch the patient for signs of distress, 
changes on the continual 12 lead electrocardiogram, hemodynamic changes on the arterial line 
blood pressure, continuous pulse oximetry and heart rate telemetry. If at any time there is a 
concern that the infusion is causing dangerous effects, it will be stopped immediately. Possible 
adverse events (AEs) include: 
1. Hypotension: If hypotension occurs during the ethanol administration, intravenous fluids 
and presser medications will be administered as appropriate by the cardiac 
anesthesiologist. The anesthesiologist will have the prerogative to stop the infusion if 
he/she feels that the infusion is contributing to the hypotension and if it is not controlled 
with fluids or pressers. 
2. Hypertension: If hypertension beyond a systolic blood pressure of 180 mmHg occurs 
during the ethanol administration, the cardiac anesthesiologist may administer 
intravenous blood pressure medications such as metoprolol, hydralazine, or 
nitroglycerin. The anesthesiologist will have the prerogative to stop the infusion if he/she 
feels that the infusion is contributing to the hypertension and if it is not controlled with 
blood pressure medications. 
3. Atrial Fibrillation: This is to be expected and will be dealt with as appropriate. This may 
include electrical cardioversion or observation depending on what phase of the 
procedure the patient is in. It is important to emphasize that some of these procedures 


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are routinely performed while a patient is in atrial fibrillation for several hours or 
throughout the entire case. 
4. Pulmonary compromise: The patients will be monitored by the anesthesia team 
throughout the procedure and the level of ethanol that we will target is not associated 
with respiratory depression. There is no evidence that ethanol (particularly at modest 
doses) affects oxygenation. The ethanol infusion can be stopped and the patient can be 
intubated if there are signs of pulmonary compromise. 
5. Agitation: The patient will be monitored by the anesthesia team and agitation induced by 
ethanol will be dealt with appropriately with sedatives. 
The PI will also monitor for AEs in all study participants. The research team will discuss the 
progress of the study and all AEs at weekly research and MMM conferences. At those 
meetings, feedback will be obtained from attendees. These meetings will also include the Chief 
of Cardiology, Chief of the EP service, head of quality assurance for electrophysiology, nurses, 
other EP faculty, fellows, and staff.  
 
7.2.2  
Post-Procedure Monitoring 
 
We will monitor patients for up to 12 months via chart review and patient phone calls. Targeted 
adverse events will be recorded if reported/found at each study follow-up visit. 
 
If three unexpected adverse events occur, then we will put the study on hold and discuss the 
circumstances associated with the adverse events with the EP group (including the Chief of 
Cardiology and Chief of Electrophysiology), the anesthesiologists that were present during 
those cases, and the DSMB. At that time, alternatives to the study design and the possibility of 
discontinuing the study will be discussed. 
 
8.0  
PROTOCOL DEVIATIONS 
 
The Principal Investigator is required to adhere to the study protocol, the signed Principal 
Investigator’s Agreement, applicable federal (national) or state/local, laws and regulations, and 
any conditions required by the IRB or applicable regulatory authorities. 
 
A protocol deviation is used to describe situations in which the clinical protocol was not followed.  
All deviations from the study protocol must be reported on the Protocol Deviation CRF to the 
UCSF CC, as soon as possible, but no later than 10 working days of notification of the event.  In 
addition, all deviations must be reported to the local IRB per the IRB’s reporting requirements.  
Protocol deviations will also be reported to the DSMB.   
 
9.0 
QUALITY ASSURANCE AND DATA MANAGEMENT 
 
9.1 
Clinical Site Investigator and Coordinator Training 
 
Each investigator and coordinator will be trained on the study protocol and procedures to ensure 
accurate and consistent study methods are used study-wide and throughout the entire study 
duration. Trainings will include review of the protocol, operations manual, CRFs, event 
reporting, and data management procedures. If any protocol amendments occur during the 
study, research team trainings will be scheduled as needed.  
 
9.2 
Data Handling and Confidentiality 
 


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Information about study participants will be kept confidential and managed according to the 
requirements of the Health Insurance Portability and Accountability Act of 1996 (HIPAA). Those 
regulations require a signed patient authorization informing the patient of the following:  
● 
What protected health information (PHI) will be collected from patients in this study 
● 
Who will have access to that information and why 
● 
Who will use or disclose that information 
● 
The rights of a research patient to revoke their authorization for use of their PHI. 
 
In the event that a participant revokes authorization to collect or use PHI, the Principal 
Investigator, by regulation, retains the ability to use all information collected prior to the 
revocation of participant authorization. For participants that have revoked authorization to collect 
or use PHI, attempts should be made to obtain permission to collect at least vital status (i.e., 
that the participant is alive) at the end of their scheduled study follow-up. 
 
9.3  
Source Documents  
 
Study files will be kept in a secure location. Participant files should include archives of 
completed CRFs and source documents. The study case report forms (CRFs) are the primary 
data collection instrument for the study and are considered source documents. Source data also 
include original records of clinical findings, observations, or study measurement results (e.g., 
hospital records, clinical and office charts, laboratory notes, dispensing records, recorded data 
from automated instruments, and patient files).  
 
9.4  
Data Collection and Management 
 
A member of the research team will enter data from each CRF into the study REDCap 
database.  The study data is subjected to checks for completeness, consistency and validity. 
Upon completion of data collection and all data cleaning efforts have been performed, the 
database will be locked and analyses will commence.  
 
10.0 
STATISTICAL METHODS 
 
Demographic and baseline characteristics will be described using mean with standard deviation, 
median and interquartile range, or proportions, where appropriate.  The change in AERP and 
conduction times will be compared within cohorts using paired t-tests (if the outcome is not 
normally distributed, the Wilcoxon sign rank test will be used), and the changes in those values 
will be compared by treatment assignment (ethanol versus placebo) using t-tests or Wilcoxon 
rank sum tests as appropriate. The proportion with inducible AF will be compared by treatment 
assignment using the chi-squared test, and a logistic regression model will be utilized to adjust 
for potential confounders that are not addressed by randomization. Full details will be outlined in 
the Statistical Analysis Plan.   
 
The Primary Analyses will be conducted in the “Analysis Population”, which includes all 
participants who are (1) randomized and (2) have received the study infusion.  Secondary 
analyses will be conducted in the full “Randomized Population”, which includes all participants 
who have been randomized, whether or not they received the study infusion   
 
11.0 
ETHICAL CONSIDERATIONS 
 
This study is to be conducted according to US and international standards of Good Clinical 
Practice (FDA Title 21 part 812 and International Conference on Harmonization guidelines), 
applicable government regulations and Institutional research policies and procedures.  


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11.1  Institutional Review Board (IRB) Approval 
 
This protocol and any amendments will be submitted to a properly constituted IRB.  
 
11.2  Informed Consent 
 
Study personnel must explain to each participant (NOTE: consent by proxy is not allowed for 
this study) the nature of the study, its purpose, the procedures involved, the expected duration, 
the potential risks and benefits involved and any discomfort it may entail.  Each participant must 
be informed that participation in the study is voluntary and that he/she may withdraw from the 
study at any time and that withdrawal of consent will not affect his/her subsequent medical 
treatment or relationship with the treating physician.  Patients will have to demonstrate adequate 
reasoning faculties and will have to demonstrate understanding of the implications, risks, 
benefits and alternatives to participating in the study before consent is obtained.  This will 
include asking potential participants questions to assure they understand the ramifications of 
participation. 
 
This informed consent will be given by means of a standard written statement, written in non-
technical language. The subject should read and consider the statement before signing and 
dating it, and should be given a copy of the signed document.  If the participant cannot read or 
sign the document, oral presentation may be made or signature given by the participant’s legally 
appointed representative, if witnessed by a person not involved in the study, mentioning that the 
participant could not read or sign the documents.  No participant will receive a study infusion 
before his/her informed consent has been obtained. 
 
11.3  Declaration of Helsinki 
 
The investigator must conduct the trial in accordance with the Declaration of Helsinki.  


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SAP_001.pdf: 
The HOLIDAY (HOw ALcohol InDuces Atrial TachYarrhythmias) Study 
 
 
 
NCT Number: NCT01996943 
Document Type: Statistical Analysis Plan 
Document Date: 12/7/2020 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


HOLIDAY Study Statistical Analysis 
 
Normally distributed continuous variables are presented as means ± SD and 
were compared using t-tests, and continuous variables with skewed distributions are 
presented as medians and interquartile ranges (IQR) and were compared using 
Wilcoxon rank sum or signed rank tests as appropriate. Categorical variables were 
compared using chi squared and Fisher’s exact tests. The proportion of atrial effective 
refractory periods that decreased with the infusion, the proportion of conduction times 
that decreased with the infusion, and the continuous change in each individual atrial 
effective refractory period and conduction time were assessed. Linear mixed models 
(LMMs) to estimate average and site-specific effects on pre- to post-infusion changes in 
AERP and conduction times were performed, with site and baseline values modeled as 
fixed effects and participants as random effects; analyses were repeated after adjusting 
for covariates statistically significantly different between alcohol and placebo groups. 
Heterogeneous relationships between pulmonary vein and non-pulmonary vein sites 
were assessed using formal tests for interaction. In addition, given evidence of potential 
effect modification by biological sex (19), tests for interaction by sex on the relationships 
between randomization assignment and outcomes were performed. Based on a review 
of repeat AERP measurements in 30 AF patients at 600 ms drive cycle length prior to 
the trial (yielding an average ERP of 230 ms ± 20 ms), we estimated that 22 patients in 
each group would provide 90% power to detect a statistically significant one standard 
deviation difference in the overall AERP using a single interim analysis for efficacy 
(assessed by an independent Data Safety Monitoring Board) and an overall two-sided α 
= 0.05. We based power calculations for the secondary analysis on 25 paroxysmal AF 
patients in whom AF induction occurred in 20%; we therefore estimated that 50 patients 
in each group would provide 84% power to detect a 30% increase in AF induction (20% 
versus 50%) with ethanol, again using a single interim analysis for efficacy and an 
overall two-sided α = 0.05. Analyses were conducted using Stata version 16 (College 
Station, Texas). Two-sided p values < 0.05 were considered statistically significant.