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Effects of Hemp Extract on Markers of Wellness, Stress Resilience, Recovery and Clinical Biomarkers of Safety in Overweight, But Otherwise Healthy Subjects

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  • AxoGen, Inc.
  • Center for Applied Health Sciences (CAHS)

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We determined the effects of a commercially available, GRAS (Generally Recognized As Safe) by independent conclusion, CBD-containing hemp oil extract on stress resilience, perceived recovery, mood, affect, body composition, and clinical safety markers in healthy human subjects. Methods: Using a randomized, placebo-controlled, double-blind design, 65 overweight, but otherwise healthy men and women (35.2 ± 11.4 years, 28.5 ± 3.3 kg/m²) ingested either Hemp Oil Extract [Hemp, 60 mg/d PlusCBDTM Extra Strength Hemp Extract Oil (15 mg hemp-derived CBD)] or a placebo (PLA) every day for six weeks while continuing to follow their normal diet and physical activity patterns. Outcome variables included changes in stress resilience, a 14-item panel of various psychometric parameters, heart-rate variability, plasma chromogranin A, body composition, and general markers of health. Data were analyzed using mixed factorial ANOVA, t-tests with 95% confidence intervals, and effect sizes (ES). Results: HDL cholesterol significantly improved in the Hemp group (p = 0.004; ES = 0.75). No other statistically significant group x time interaction effects were observed. Statistical tendencies for between-group differences were found for ‘I Get Pleasure From Life’ (p = 0.06, ES = 0.48) and ‘Ability to Cope with Stress’ (p = 0.07, ES = 0.46). Sleep quality (Hemp, p = 0.005, ES = 0.54) and sleep quantity (Hemp, p = 0.01, ES = 0.58) exhibited significant within-group changes. All values for hepato-renal function, cardiovascular health, fasting blood lipids, and whole blood cell counts remained within normal clinical limits with no between-group differences over time being identified. Conclusions: Hemp supplementation improved HDL cholesterol, tended to support psychometric measures of perceived sleep, stress response, and perceived life pleasure and was well tolerated with no clinically relevant safety concerns. Registered at clinicaltrials.gov: NCT04294706.
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Effects of Hemp Extract on Markers of Wellness,
Stress Resilience, Recovery and Clinical
Biomarkers of Safety in Overweight, But
Otherwise Healthy Subjects
Hector L. Lopez, Kyle R. Cesareo, Betsy Raub, A. William Kedia, Jennifer E.
Sandrock, Chad M. Kerksick & Tim N. Ziegenfuss
To cite this article: Hector L. Lopez, Kyle R. Cesareo, Betsy Raub, A. William Kedia,
Jennifer E. Sandrock, Chad M. Kerksick & Tim N. Ziegenfuss (2020): Effects of Hemp
Extract on Markers of Wellness, Stress Resilience, Recovery and Clinical Biomarkers of
Safety in Overweight, But Otherwise Healthy Subjects, Journal of Dietary Supplements, DOI:
10.1080/19390211.2020.1765941
To link to this article: https://doi.org/10.1080/19390211.2020.1765941
Published online: 27 May 2020.
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Effects of Hemp Extract on Markers of Wellness, Stress
Resilience, Recovery and Clinical Biomarkers of Safety
in Overweight, But Otherwise Healthy Subjects
Hector L. Lopez, MD, MS
a
, Kyle R. Cesareo, MS, CCRC
a
, Betsy Raub, RN, BSN
a
,
A. William Kedia, MD
a
, Jennifer E. Sandrock, MS, RD, CCRC
a
, Chad M. Kerksick,
PhD
b
, and Tim N. Ziegenfuss, PhD
a
a
The Center for Applied Health Sciences, Stow, OH, USA;
b
Exercise and Performance Nutrition
Laboratory, School of Health Sciences, Lindenwood University, St. Charles, MO, USA
ABSTRACT
We determined the effects of a commercially available, GRAS
(Generally Recognized As Safe) by independent conclusion, CBD-con-
taining hemp oil extract on stress resilience, perceived recovery,
mood, affect, body composition, and clinical safety markers in
healthy human subjects.
Methods: Using a randomized, placebo-controlled, double-blind
design, 65 overweight, but otherwise healthy men and women
(35.2 ± 11.4 years, 28.5 ± 3.3 kg/m
2
) ingested either Hemp Oil Extract
[Hemp, 60 mg/d PlusCBD
TM
Extra Strength Hemp Extract Oil (15 mg
hemp-derived CBD)] or a placebo (PLA) every day for six weeks while
continuing to follow their normal diet and physical activity patterns.
Outcome variables included changes in stress resilience, a 14-item
panel of various psychometric parameters, heart-rate variability,
plasma chromogranin A, body composition, and general markers of
health. Data were analyzed using mixed factorial ANOVA, t-tests with
95% confidence intervals, and effect sizes (ES).
Results: HDL cholesterol significantly improved in the Hemp group
(p¼0.004; ES ¼0.75). No other statistically significant group x time
interaction effects were observed. Statistical tendencies for between-
group differences were found for I Get Pleasure From Life(p¼0.06,
ES ¼0.48) and Ability to Cope with Stress(p¼0.07, ES ¼0.46).
Sleep quality (Hemp, p¼0.005, ES ¼0.54) and sleep quantity
(Hemp, p¼0.01, ES ¼0.58) exhibited significant within-group
changes. All values for hepato-renal function, cardiovascular health,
fasting blood lipids, and whole blood cell counts remained within
normal clinical limits with no between-group differences over time
being identified.
Conclusions: Hemp supplementation improved HDL cholesterol,
tended to support psychometric measures of perceived sleep, stress
response, and perceived life pleasure and was well tolerated with no
clinically relevant safety concerns. Registered at clinicaltrials.gov:
NCT04294706.
KEYWORDS
affect; body composition;
cannabis; CBD;
endocannabinoid; health;
marijuana; mood;
phytocannabinoids;
supplementation
CONTACT Hector L. Lopez hl@appliedhealthsciences.org The Center for Applied Health Sciences, 4302 Allen
Road, Suite 120, Stow, OH, USA.
ß2020 Taylor & Francis Group, LLC
JOURNAL OF DIETARY SUPPLEMENTS
https://doi.org/10.1080/19390211.2020.1765941
Introduction
Recent interest in the potential wide-ranging health benefits of non-intoxicating or
minimally psychoactive extracts and constituents of the Cannabis plant has spurred add-
itional research leading to a greater articulation of the endocannabinoid system, the
broader endocannabinoidome and several potential mechanisms of action. The endocan-
nabinoid system (ECS) consists of membrane G-protein coupled cannabinoid (CB1 and
CB2 and others that interact via the expanded endocannabinoidome) receptors, their
lipid-based endocannabinoid ligands, and the enzymes responsible for synthesis and
metabolism is found ubiquitously throughout the central and peripheral nervous sys-
tems and most other tissues throughout the body (Pertwee 1997).
Within hemp and other Cannabis sativa cultivars, two phytocannabinoids, cannabi-
diol (CBD) and delta-9-tetrahydrocannabinol (THC), have undergone the most investi-
gation. Cannabinoid research accelerated in the late 1960s after THC was isolated,
synthesized, and identified as a primary psychoactive constituent of Cannabis
(Mechoulam et al. 1967; Mechoulam et al. 1963; Schultes 1969). Another phytocannabi-
noid discovered in 1940, cannabidiol (CBD), accounts for up to 40% of the plants
extract making it a major constituent of the cannabis plant (Schultes 1969; Schultes
1973; Maroon and Bost 2018). Psychoactive effects and impairment are commonly
reported with THC ingestion. Interestingly, CBD use does not elicit the same psycho-
active or impairing effects that are observed with THC (Maroon and Bost 2018). This
difference is thought to be primarily due to THCs affinity as an agonist of CB1 recep-
tors, which are located throughout various portions of the brain (Morales et al. 2017).
Alternatively, interaction with the CB2 receptor (more widely distributed across
immune tissues) is thought to explain the link between THC and CBD use and immune
function (Morales et al. 2017).
Both endogenous cannabinoids and phytocannabinoids as well as synthetic agents
that act on the endocannabinoid system have the potential to impact the function of a
wide range of organ systems and clinical conditions, including: anxiety and stress disor-
ders, immune function, chronic pain and musculoskeletal conditions, mood regulation
or depression, insomnia or sleep quality, and appetite signaling/food intake disorders.
In this regard, research involving CBD has demonstrated various benefits such as mood
regulating, neuroprotective, anti-inflammatory, anxiolytic, immunomodulatory, anti-
seizure and vascular effects (Schultes 1969; Campos et al. 2016; Hillard 2000).
In the past decade, use and interest in hemp extracts, CBD-containing products, and
Cannabis has increased markedly. In response to this interest and diffuse regulatory
guidance by governments, the number of commercially available products providing
CBD has increased several-fold. Currently, very limited amounts of research are avail-
able that outline the safety of CBD-containing hemp extract supplementation (in any
form or dosing). More importantly, there is a dearth of human clinical data on its
potential ability to impact various outcomes of interest such as changes in stress
response, affect and mood, sleep hygiene, body composition, and clinical markers of
health and safety. Searching the biomedical literature database PubMed (www.ncbi.nih.
gov/pubmed/) for key words such as cannabidiol, hemp extract, hemp oil, and hemp
oil extractrevealed no human investigations that have examined these outcomes.
Logical extensions of the well-established pharmacology surrounding the
2 H. L. LOPEZ ET AL.
endocannabinoid system have led to assertions that supplementation with cannabidiol
(CBD) might exert various physical and psychological effects. However, and as men-
tioned previously, the literature base to support or refute many of the claims is largely
unexplored to date. For these reasons, the purpose of this study was to determine the
effects of a commercially available, CBD-containing CO2 supercritical hemp extract of
aerial parts with GRAS by independent conclusion (per criteria as outlined in US FDAs
Final GRAS Rule of 2016) on markers of stress resilience, perceived recovery, mood,
affect, body composition, blood chemistry, and cell counts in healthy human subjects.
Methods
Overview of study design
The study design employed for this protocol was a randomized, double-blind, parallel
groups, placebo-controlled investigation where each study participant supplemented for six
weeks. Each participant completed four study visits (Table 1). The first visit was for screen-
ing purposes and consisted of signing an IRB-approved consent form, completing a med-
ical history, evaluating the presence of inclusion and exclusion criteria, and assessing
routine blood work (comprehensive metabolic panel, complete blood panel, lipid panel)
and resting vitals (heart rate and blood pressure). The next three study visits were identi-
cal, and they took place 0, 3, and 6 weeks after supplementation of the assigned test prod-
uct. Each study visit consisted of measuring vitals (resting heart and blood pressure),
collection of venous blood for assessment of metabolic panel, cell counts, and chromogra-
nin-A, body composition (via DEXA), completion of the Brief Resilience Scale (BRS),
Stroop test, heart rate variability, and visual analog scales for appetite, cravings, relaxation,
mental clarity, positive mood, perceived feelings of stress, sleep quality, well-rested, ability
to handle physical work, readiness to perform physical exercise, readiness to perform job
tasks, and well-being. Once supplementation began, dietary analysis and physical activity
status was assessed before each visit while compliance to the protocol and adverse events
were monitored throughout the supplementation protocol. To facilitate replication for all
measured endpoints, study participants were asked to replicate their diet (including absten-
tion from caffeine and alcohol) for 24 h prior to each study visit, fast for 12 h prior to
each visit, and refrain from exercise for 48 h prior to each study visit. Upon screening and
determination of eligibility, study participants were matched according to gender and body
mass index before randomly being assigned to ingest either a hemp oil extract or placebo
oil (olive oil) for six weeks in a double-blind fashion. After completion of baseline testing
and beginning supplementation, participants were asked to accumulate at least 30 min of
physical activity at least five days per week before returning back to the laboratory for their
subsequent study visits. Table 1 provides a tabular view of the study protocol.
Study participants
Overweight and obese men and women were recruited as participants in this study pri-
marily from a local suburban community in Ohio. All participants were randomly
assigned to one of two supplementation groups and all participant demographics can be
found in Table 2. Prior to commencing the study, participants read and signed an IRB-
JOURNAL OF DIETARY SUPPLEMENTS 3
approved informed consent form (Integreview, Austin, TX, Protocol # CVSI-001-2018,
Approval date: August 27, 2018) and this study was registered with www.clinicaltrials.
gov as NCT04294706. All study participants were required to be in good health as
determined by review of their medical history and routine blood chemistries by the
study physician. Inclusion criteria indicated that all male and female participants
were between the ages of 18 55 years, had body mass index levels between 25
34.99 kg/m
2
, were normotensive (systolic pressure between 100 139 mm Hg and
Table 1. Overview of study design.
Study Activity Screening
Baseline
(Day 0)
Mid-Study
(3 weeks)
Post
(6 weeks)
Visit 1 2 3 4
Informed Consent X
Inclusion/Exclusion Criteria X
Medical History X
Height, weight, heart rate, blood pressure X X X X
Safety screen (CMP, CBC, lipids, uric acid) X X X
Plasma Chromogranin-A X X X
BRS Brief Resilience Scale X X X
Stroop Test X X X
Heart Rate Variability X X X
Body composition (DEXA) X X X
Visual analog scales (appetite, cravings, relaxation,
mental clarity, positive mood, perceived feelings of
stress, sleep quality, sleep quantity, well-rested,
ability to handle physical work, readiness to perform
physical exercise, readiness to perform job tasks, and
well-being
XX X
Dietary Analysis X X X X
Framingham Physical Activity Index X X X
Dispense Test Product X X X
Protocol Compliance X X X X
Collection of Unused Test Product X X
Adverse Event Monitoring X X
Product Side Effect Questionnaire X X X
Table 2. Study participant demographics.
Group Both Sexes Men Women
Age
(years)
Hemp 33.8 ± 11.8 34.6± 10.0 33.0 ± 13.4
PLA 36.6 ± 10.9 31.8 ± 9.9 41.4 ± 10.0
Height
(centimeters)
Hemp 171.1 ± 10.7 179.5 ± 6.5 163.2 ± 7.2
PLA 169.6 ± 10.0 177.3 ± 6.6 161.9 ± 6.2
Body Mass
(kilograms)
Hemp 83.3 ± 14.2 91.3± 12.3 75.9 ± 11.9
PLA 82.5 ± 14.0 90.0 ± 12.5 75.1 ± 11.4
Body Mass Index
(kg/m
2
)
Hemp 28.3 ± 3.1 28.3 ± 3.0 28.4 ± 3.2
PLA 28.6 ± 3.6 28.6 ± 3.2 28.6 ± 4.1
DEXA % Fat Hemp 32.6 ± 8.0 27.1 ± 6.0 37.8 ± 6.0
PLA 33.7 ± 10.5 25.8 ± 7.0 41.6 ± 6.7
Systolic Blood Pressure
(mm Hg)
Hemp 120.5 ± 10.8 126.1 ± 10.4 115.2 ± 8.4
PLA 121.5 ± 10.4 124.3 ± 9.4 118.7 ± 11.0
Diastolic Blood Pressure
(mm Hg)
Hemp 75.2 ± 8.8 75.5 ± 8.6 74.9 ± 9.3
PLA 76.6 ± 8.9 74.4 ± 8.9 78.8 ± 8.6
Resting HR
(beats/min)
Hemp 66.2 ± 10.0 62.6± 7.0 69.7 ± 11.3
PLA 66.9 ± 8.3 65.7 ± 8.5 68.1 ± 8.2
HEMP ¼Hemp oil extract (n¼33; Men ¼16; Women ¼17); PLA ¼Placebo (n¼32; Men ¼16; Women ¼16); Data
presented as means ± SD;
¼PLA >Hemp, p¼0.05.
4 H. L. LOPEZ ET AL.
diastolic pressure between 65 89 mm Hg) with a normal resting heart rate (90 beats/
min), and agreed to abide by all requirements of the study protocol.
Alternatively, participants were excluded if they reported regular exercise of more
than three days per week, had reported using any medication indicated for weight loss
for three months prior to commencing the study protocol or had gained or lost more
than five pounds within the past 30 days. Additionally, any participant regularly taking
a prescribed thyroid medication at a non-stable dose for 90 days prior to beginning the
study protocol and any individual with a metabolic disorder (electrolyte abnormalities,
diabetes (type I or II), thyroid disease or hypogonadism), a history of hepato-renal, can-
cer, chronic inflammatory conditions, autoimmune, dyssomnia or other diagnosed sleep
disorder, neurologic, gastrointestinal, or musculoskeletal disease were excluded.
Participants with known allergies, currently using tobacco or nicotine within 12 previ-
ous months, or females who were nursing, lactating, postpartum (<120 days), or cur-
rently pregnant were excluded. Participants using anti-coagulants, anti-platelets, or fish
oil supplements were excluded due to any concern with clotting complications and par-
ticipants who had a history of psychiatric or neurological disorders and the use of any
medications that impacted central nervous system function within two weeks of starting
the study protocol were excluded. Lastly, participants with syndromes or medications
that may influence body or cardiovascular disease and any orthopedic limitations were
excluded from participating. A CONSORT diagram is provided in Figure 1 for all study
participants through the study protocol.
Height, weight, heart rate, blood pressure
Standing height was determined using a wall-mounted stadiometer with each study par-
ticipant in their socks with heels together. Body weight was measured using a Seca
767
TM
Medical Scale (Hamburg, Deutschland). Resting heart rate and blood pressure
was measured in duplicate using an automated blood pressure cuff (Omron HEM-780).
Venous blood collection and processing
Whole blood and serum samples were collected using standard phlebotomy techniques at
all study visits. Whole blood samples were collected into K
2
-EDTA treated Vacutainer
tubes. Upon collection, each sample was slowly inverted ten consecutive times prior to
immediate refrigeration. Serum samples were collected in serum separation tubes and
allowed to clot for 30 min at room temperature prior to being centrifuged (Horizon mini
E Centrifuge, Drucker Diagnostics, Port Matilda, PA) for 15 min at 3,200 rpm.
Biochemical analysis
For screening purposes, blood collected at visit one was analyzed for a comprehensive
metabolic panel, complete blood count with platelet differentials, and lipid panel.
Components of the comprehensive metabolic panels consists of glucose, blood urea
nitrogen [BUN], creatinine, aspartate aminotransaminase [AST], alanine aminotransa-
minase [ALT], creatine kinase, lactate dehydrogenase, total bilirubin, alkaline
JOURNAL OF DIETARY SUPPLEMENTS 5
phosphatase [ALP], uric acid, sodium, potassium, total protein, albumin, globulin, and
iron. Complete blood counts were analyzed for absolute and percentage of contribution
for neutrophils, eosinophils, basophils, lymphocytes, and monocytes in addition to over-
all white blood cell and red blood cell count, hemoglobin, hematocrit, mean corpuscle
Figure 1. Consolidated standards of reporting trials (CONSORT) diagram.
6 H. L. LOPEZ ET AL.
volume, mean corpuscle hemoglobin, red cell dimension width, and mean corpuscle
hemoglobin content. Lipid panel components consist of triglycerides [TG], total choles-
terol [TC], LDL cholesterol, HDL cholesterol. All analyses were completed using auto-
mated clinical chemistry analyzers (LabCorp, Dublin, OH branch). Additionally, serum
levels of chromogranin-A were assessed by LabCorp using the ThermoFisher/BRAHMS
KRYPTORV
Rtime resolved amplified cryptate emission (TRACE) between energy donor
and acceptor in a sandwich immunofluorescent assay using two mouse monoclonal
antibodies (van der Knaap et al. 2015). All samples from the same day were batch ana-
lyzed with test-retest reliabilities commonly reported using internal quality control data
from clinical laboratories and associated automated analyzers within a range of 3 5%
(Cuka et al. 2001).
Brief resilience scale
Adhering to the procedures of Smith et al. (Smith et al. 2008) the Brief Resilience Scale (BRS)
was administered to all study participants after 0, 3, and 6 weeks of supplementation. The BRS is
a six-item questionnaire each consisting of a five-item Likert scale ranging from 1¼Strongly
Disagreeto 5¼Strongly Agree. Each question is scored 1 five providing a range of total
scores from 6 30. The average score from all five scales was used for data analysis.
The stroop test
The Stroop Color and Word Test (SCWT), a neuropsychological test, was administered 0,
3, and 6 six weeks after supplementation to assess performance during mental stress
(Stroop 1935). Briefly, this test was administered for five continuous minutes via an online
app. One of four words (red, yellow, green, blue) appeared at random, until a selection was
made, then a new word appeared at random. Subjects selected the response which matched
the color of the ink of the randomly selected word (not the color that the word described).
The speed at which each test was completed (time per score) and the number of errors (%
of correct scores) allowed for the computation of a total Stroop score.
Heart rate variability
Heart rate variability (low frequency [LF], high frequency [HF], LF/HF ratio, and root
mean square) were captured using a Polar H10 heart rate sensor (Polar, Finland) placed
over the xiphoid process. While being worn, the monitor automatically calculated the
R-R interval. This data was record and analyzed by Elite HRV (2014-2019) (Usui and
Nishida 2017). All HRV measurements were completed at three separate times. HRV
was first assessed while resting quietly in the laboratory for ten minutes where the final
five minutes were recorded. HRV was assessed a second time for five minutes while
completing the Stroop test. HRV was assessed a third and final time immediately fol-
lowing completion (typically within 30 s) of the Stroop and was assessed for approxi-
mately five minutes after completing the Stroop test.
JOURNAL OF DIETARY SUPPLEMENTS 7
Body composition
Lean mass, fat mass, % fat, and android/gynoid ratio were determined by dual-energy
x-ray absorptiometry (DEXA; General Electric Lunar DPX Pro) after 0, 3, and 6 weeks
of supplementation. All DEXA scans were performed by the same technician and ana-
lyzed by the manufacturers software (enCORE version 13.31). Briefly, subjects were posi-
tioned in the scanner according to standard procedures and remained motionless for
approximately 15 min during scanning. DEXA segments for the upper and lower limbs
and trunk were directed using standard anatomical landmarks. Percent fat was calculated
by dividing fat mass by total scanned mass. Lean to fat mass ratio was computed using a
simple ratio between the two values. Quality control calibration procedures were
performed prior to all scans using a calibration block and procedures provided by the
manufacturer. Prior to this study, we determined testretest reliability for repeated meas-
urements of lean mass, bone mineral content, and fat mass using this DEXA using intra-
class correlation coefficients; all values were 0.98 (Ziegenfuss et al. 2006).
Visual analog scales
Visual analog scales (VAS) were completed by each study participant after 0, 3, and
6 weeks of supplementation. All visual analog scales were similarly constructed using a
100-mm line anchored by Lowest Possibleand Highest Possibleto assess subjective
ratings of appetite, cravings, relaxation, mental clarity, positive mood, perceived feelings
of stress, sleep quality, well-rested, ability to handle physical work, readiness to perform
physical exercise, readiness to perform job tasks, and well-being. The validity and reli-
ability of VAS to assess fatigue and energy have been previously established (Lee et al.
1991) and our methods have been published elsewhere (Ziegenfuss et al. 2017;
Ziegenfuss et al. 2018; Ziegenfuss et al. 2017).
Dietary intake and physical activity monitoring
All subjects were instructed to maintain their normal dietary habits throughout the entire
study. During baseline screening and after six weeks of supplementation, participants
were asked to complete a 3-day dietary record. Dietary records were analyzed for average
daily energy and macronutrient intake by trained study investigators and NutriBase IX
(Clinical Edition) software (CyberSoft, Inc. Phoenix, AZ). In addition, study participants
were asked to complete a 24-hour dietary recall after arriving for baseline screening. The
recalled log of food and fluid intake was copied and provided back to the study partici-
pant. Study participants were then instructed to duplicate their food and fluid intake for
the 24 h prior to each subsequent study visit (i.e. at week 3 and week 6).
As part of the study protocol, participants were asked to adapt their daily physical
activity to 30 min of walking at least five days per week throughout the study.
Compliance was monitored by having each subject document their exercise and physical
activity in a training log. Each completed log was returned to study investigators at
each study visit. Additionally, physical activity was assessed using the Framingham
physical activity questionnaire after 0 and six weeks of supplementation. Briefly, phys-
ical activity scores were computed first by having study participants record how much
8 H. L. LOPEZ ET AL.
time was spent completing various types of activity (e.g. rest, job/occupation, and extra-
curricular activities). Within each category, participants then indicated how much of
that time was classified as sedentary, slight, moderate, and heavy activity. Next, factors
of 1.1, 1.5, 2.4, and 5.0 were multiplied by the indicated number of hours spent to rep-
resent sedentary, slight, moderate, and heavy activity, respectively. All computed scores
are then summed to achieve the final physical activity score which was subsequently
used in statistical analysis.
Supplementation
As mentioned previously, all participants were matched according to sex and body mass
index prior to being randomly assigned in a double-blind fashion to one of two supple-
mentation groups: placebo (olive oil) or hemp oil extract. Participants were randomized
and assigned to ingest either hemp oil extract or an olive oil placebo. Both groups were
subsequently instructed to ingest one soft gel each day with breakfast for six weeks. Each
soft gel delivered 60mg/day of CBD-containing CO2 supercritical extract of aerial parts
of hemp oil extract (PlusCBD
TM
Extra Strength Hemp Extract) which delivered 15 mg of
hemp-derived CBD. The investigational product has been designated as an independent
GRAS (Generally Recognized As Safe) conclusion CBD-containing hemp oil extract, per
the US Food & Drug Administration (FDA) Final GRAS Rule guidelines (https://www.
fda.gov/food/generally-recognized-safe-gras/about-gras-notification-program). All study
materials were prepared following current good manufacturing practices (cGMP) accord-
ing to Code of Federal Regulations of US Food and Drug Administration Title 21 CFR
part 111 in blinded gel caps and packaged in coded generic containers for double-blind
administration. Compliance to the supplementation regimen was monitored by daily logs,
communication with study participants at each study visit, and counting all returned soft
gels at each subsequent study visit. Certificate of analysis for multiple lots of production
are available online (www.pluscbdoil.com).
Adverse events
During weekly phone calls, the frequency and intensity of local and systemic non-serious
and serious adverse events (AEs) were recorded by study team members. All reported
events were coded using the Medical Dictionary for Regulatory Activities (MedDRA)
while the intensity of recorded adverse events were graded using standardized criteria.
Statistical analysis
All data were entered into two separate Microsoft Excel spreadsheets (i.e. manual dou-
ble-key data entry) and compared to assure data quality prior to analysis. SPSS 23
(Armonk, NY USA) was used for all analyses. Normality assumptions were checked on
all variables using a one-sample Shapiro-Wilk test. Non-normal distributions were
transformed using natural logarithms, cubed, and square root transformations. Outliers
were checked via visual inspection of studentized calculations on the residuals (thresh-
old value of ± 3 SD) of each dependent variable. Independent t-tests were used to assess
JOURNAL OF DIETARY SUPPLEMENTS 9
baseline differences. Separate 2 x two mixed factorial ANOVA with repeated measures
on time were assessed for all outcomes after three and six weeks of supplementation.
When the sphericity assumption was not met, the Huynh-Feldt correction was applied
when epsilon was greater than 0.75 and the Greenhouse-Geiser correction was applied
when epsilon was less than 0.75. In addition, delta values were computed and independ-
ent t-tests were completed to assess group differences after three and six weeks of sup-
plementation. Mean differences of the change scores and 95% confidence intervals were
calculated on the difference between groups. Within-group effects were compared using
paired samples t-test. Effect sizes (ES) were also used to assess the magnitude of change,
and values of 0.2, 0.5 and 0.8 were considered small, medium and large effects, respect-
ively. All data are presented as means ± standard deviations. Results were considered
statistically significant at p0.05 and trends were declared at 0.051 p0.10.
Results
As seen in Table 2, no baseline differences (p>0.05) were noted between groups any of
the demographic variables (age, height, body mass, body mass index, percent body fat,
blood pressure (systolic & diastolic), resting heart rate, and physical activity score.
Additionally, weekly compliance checks by the research study team revealed >95% com-
pliance to the supplementation (data not shown) regimen. A brief summary table of
adverse events (AEs) is provided (See Table 3).
Anthropometrics and hemodynamic
As seen in Table 4, main and group x time interaction effects for body mass, body mass
index, and hemodynamic variables were assessed by mixed factorial ANOVA with
repeated measures on time. No statistically significant within-group changes or differen-
ces between groups were identified for body mass (p¼0.13; 95% CI: 0.38, 3.03; ES ¼
0.39) and body mass index (p¼0.15; 95% CI: 0.20, 1.24; ES ¼0.36). Similarly, no
within-group or between-group differences were identified for heart rate (p¼0.85; 95%
CI: 4.15, 3.43; ES ¼0.05) and diastolic blood pressure (p¼0.15; 95% CI: 1.24,
Table 3. Summary of adverse events (AEs).
Active (n¼33) Placebo (n¼32)
Severity
Mild 1 2
Moderate ——
Severe ——
Relationship to Test Article
Not Related ——
Possible 1 2
Definite ——
Body System and AEs
Nervous System
Headache 2
Syncope ——
Respiratory
Allergic rhinitis 1
Total Number of Adverse Events Experienced During Study 1 2
Total Number of Subjects Experiencing Adverse Events: n (%) 1/33 (3%) 2/32 (6%)
10 H. L. LOPEZ ET AL.
7.98, 1.24; ES ¼0.36). Between-group changes in systolic blood pressure tended to be
different between groups (p¼0.06; 95% CI: 0.29, 10.48; ES ¼0.48).
Body composition
As seen in Table 5, main and group x time interaction effects of all body composition
variables were assessed using mixed factorial ANOVA with repeated measures on time.
No statistically significant group x time interaction effects were observed for any of the
body composition outcomes. Notably, DEXA lean mass (p¼0.09; 95% CI: 0.09, 1.37;
ES ¼0.27), DEXA fat-free mass (p¼0.08; 95% CI: 0.08, 1.38; ES ¼0.31), and DEXA
total scanned mass (p¼0.06; 95% CI: 0.02, 1.74; ES ¼0.48) tended to be different
between groups. Within-group changes for DEXA lean mass (p¼0.90, ES ¼0.00),
DEXA fat-free mass (p¼0.92, ES ¼0.00), and DEXA total scanned mass (p¼0.71, ES
¼0.01) revealed no change in the Hemp group while significant decreases were
observed in the PLA group for DEXA lean mass (p¼0.04, ES ¼0.06), DEXA fat-free
mass (p¼0.04, ES ¼0.05), and DEXA total scanned mass (p¼0.04, ES ¼0.06).
Additionally, between-group changes in the android: gynoid ratio tended to be different
(p¼0.09; 95% CI: 0.01, 0.11; ES ¼0.43). Observed changes within each group for
this variable revealed a tendency for Hemp to increase (p¼0.08) while no change was
observed in the PLA (p¼0.56).
Visual analog and brief resilience scales
Main and group x time interaction effects of all visual analog and average Brief
Resilience Scales were assessed using mixed factorial ANOVA with repeated measures
on time. No statistically significant (p<0.05) group x time interaction effects were
Table 4. Anthropometrics, hemodynamic, and physical activity data.
Within-Group Between-Group
Variables N
Week 0
(Visit 2)
Week 6
(Visit 4) p-value ES 95% CI
p-value
(ES)
Body Mass (kg)
Hemp 33 83.3 ± 14.1 84.2 ± 12.9 0.27 0.06 (0.38, 3.03) 0.13
PLA 32 82.8 ± 13.9 82.3 ± 13.4 0.15 0.03 0.39
Body Mass Index (kg/m
2
)
Hemp 33 28.3 ± 3.0 28.7 ± 3.3 0.26 0.13 (0.20, 1.24) 0.15
PLA 32 28.6 ± 3.6 28.5 ± 3.5 0.20 0.03 0.36
Heart Rate (beats/min)
Hemp 33 64.9 ± 9.6 65.5 ± 8.8 0.71 0.06 (4.15, 3.43) 0.85
PLA 32 65.6 ± 9.8 66.5 ± 10.5 0.41 0.09 0.05
Systolic Blood Pressure (mm Hg)
Hemp 33 117.7 ± 13.2 119.6 ± 15.7 0.41 0.13 (0.29, 10.48) 0.06
PLA 32 122.8 ± 11.3 119.6 ± 12.0 0.03 0.27 0.48
Diastolic Blood Pressure (mm Hg)
Hemp 33 73.1 ± 9.2 75.2 ± 11.0 0.23 0.21 (1.24, 7.98) 0.15
PLA 32 76.8 ± 7.9 75.5 ± 8.7 0.42 0.15 0.36
Framingham Physical Activity
Hemp 33 37.4 ± 7.1 37.6 ± 6.3 0.84 0.02 (1.33, 1.95) 0.71
PLA 32 37.1 ± 6.8 37.0 ± 6.3 0.73 0.03 0.05
Raw data is presented as mean ± standard deviation. ES ¼Effect size; 95% CI ¼95% Confidence intervals computed
using the differences between groups for each within-group change.
JOURNAL OF DIETARY SUPPLEMENTS 11
observed for any of the scales measured. I Get Pleasure from LifeVAS (p¼0.06; 95%
CI: 0.26, 14.8; ES ¼0.48) and Ability to Cope with StressVAS (p¼0.07; 95% CI:
0.73, 19.1; ES ¼0.46) did exhibit a statistical tendency to be different between groups.
Additionally, ratings in the Hemp group for I Get Pleasure from Lifeillustrated a stat-
istically significant increase after supplementation (p¼0.007) while the PLA group did
not change (p¼0.44). Also, ratings in the Hemp group for Ability to Cope with Stress
did not change across the supplementation protocol (p¼0.27) while PLA exhibited a
statistically significant decrease (p¼0.001) in this scale. Within-group changes in sleep
quality for Hemp (p¼0.005, ES ¼0.54) and PLA (p¼0.06, ES ¼0.47) as well as sleep
quantity for Hemp (p¼0.01, ES ¼0.58) and PLA (p¼0.09, ES ¼0.37) exhibited sig-
nificant (for Hemp) and marginally significant (for PLA) within-group changes.
Increases from baseline in PLA were observed for I Have a Clear Mind(PLA, p¼0.05,
ES ¼0.35), whereas Hemp approached within-group statistically significant increases
(Hemp, p¼0.07, ES ¼0.40). Other variables exhibited similar within-group changes
between the two groups, see Table 6.
Table 5. Body composition.
Variables N
Week 0
(Visit 2)
Week 6
(Visit 4)
Within-Group Between-Group
p-value ES 95% CI
p-value
(ES)
Lean Mass-Arms (kg)
Hemp 33 6.3 ± 2.4 6.4 ± 2.4 0.70 0.02 (0.24, 0.47) 0.51
PLA 32 6.3 ± 2.5 6.2 ± 2.3 0.58 0.03 0.16
Lean Mass-Legs (kg)
Hemp 33 18.9 ± 4.2 18.9 ± 4.5 0.95 0.00 (0.48, 0.91) 0.54
PLA 32 17.8 ± 4.2 17.6 ± 3.8 0.50 0.05 0.15
Lean Mass (kg)
Hemp 33 54.0 ± 11.1 54.0± 11.1 0.90 0.00 (0.09, 1.37) 0.09
PLA 32 53.1 ± 12.4 52.5 ± 11.5 0.04 0.06 0.27
Fat-Free Mass (kg)
Hemp 33 56.9 ± 11.6 56.8± 11.6 0.92 0.00 (0.08, 1.38) 0.08
PLA 32 56.1 ± 13.0 55.4 ± 12.1 0.04 0.05 0.31
Total Scanned Mass (kg)
Hemp 33 83.0 ± 14.0 83.1± 14.2 0.71 0.01 (0.02, 1.74) 0.06
PLA 32 83.0 ± 13.8 82.3 ± 13.3 0.04 0.06 0.48
Fat Mass (kg)
Hemp 33 26.2 ± 8.1 26.3 ± 8.4 0.50 0.02 (0.41, 0.83) 0.49
PLA 32 26.9 ± 9.7 26.9 ± 9.2 0.73 0.01 0.17
Percent Fat (%)
Hemp 33 32.6 ± 8.0 32.7 ± 8.2 0.67 0.01 (0.72, 0.51) 0.73
PLA 32 33.7 ± 10.5 33.9 ± 9.9 0.45 0.02 0.08
Lean: Fat Mass Ratio
Hemp 33 2.28 ± 0.95 2.29± 1.00 0.78 0.01 (0.02, 0.18) 0.12
PLA 32 2.32 ± 1.25 2.25 ± 1.11 0.11 0.06 0.39
Android: Gynoid Ratio
Hemp 33 1.01 ± 0.25 1.06± 0.29 0.08 0.15 (0.01, 0.11) 0.09
PLA 32 1.12 ± 0.18 1.11 ± 0.21 0.56 0.06 0.43
Visceral Adipose Tissue (grams)
Hemp 33 660 ± 679 701 ± 741 0.12 0.06 (33.2, 178.2) 0.18
PLA 32 844 ± 623 813 ± 630 0.52 0.05 0.34
Bone Mineral Content (grams)
Hemp 33 2872 ± 601 2876 ± 610 0.63 0.01 (25.0, 41.1) 0.63
PLA 32 2949 ± 881 2944± 860 0.78 0.00 0.12
12 H. L. LOPEZ ET AL.
Heart rate variability
As can be seen in Table 7, main and group x time interaction effects of all heart rate
variability metrics were assessed using mixed factorial ANOVA with repeated measures
on time. No statistically significant (p<0.05) group x time interaction effects were
observed for any of the variables measured. Changes in low frequency power during the
Stroop test did tend to be different between groups (p¼0.07; 95% CI: 25.8, 538; ES ¼
0.46), which appear to be largely mediated by a statistically significant within-group
decrease in this variable in the PLA group (p¼0.05). Two other statistically significant
Table 6. Visual analog and brief resilience scale.
Variables N
Week 0
(Visit 2)
Week 6
(Visit 4)
Within-Group Between-Group
p-value ES 95% CI
p-value
(ES)
Appetite
Hemp 33 63.9± 17.8 59.9 ± 18.7 0.23 0.22 (11.3, 6.6) 0.60
PLA 32 62.7 ± 20.4 61.0 ± 24.3 0.61 0.07 0.13
Cravings for Sweets
Hemp 33 42.8 ± 29.2 44.2 ± 23.2 0.74 0.05 (10.7, 11.3) 0.96
PLA 32 45.9 ± 25.6 47.1 ± 23.7 0.76 0.05 0.01
I Feel Relaxed
Hemp 33 61.2 ± 29.3 71.2 ± 20.7 0.05 0.39 (11.1, 11.8) 0.95
PLA 32 64.1 ± 23.0 73.7 ± 15.6 0.004 0.49 0.02
I Have a Clear Mind
Hemp 33 63.4 ± 29.3 71.4 ± 20.5 0.07 0.40 (10.6, 11.8) 0.92
PLA 32 64.2 ± 23.9 71.6 ± 18.0 0.05 0.35 0.02
Ability to Cope with Stress
Hemp 33 38.3 ± 25.8 34.6 ± 21.7 0.27 0.16 (0.73, 19.1) 0.07
PLA 32 41.1 ± 21.2 28.2 ± 17.1 0.001 0.67 0.46
Ability to Perform Physical Work
Hemp 33 19.2 ± 19.6 22.4 ± 15.8 0.25 0.18 (8.2, 10.9) 0.78
PLA 32 20.5 ± 22.8 22.4 ± 20.0 0.64 0.09 0.07
Willingness to Perform Exercise
Hemp 33 66.2 ± 25.6 73.2 ± 17.5 0.08 0.32 (1.73, 17.0) 0.11
PLA 32 78.4 ± 19.8 77.7 ± 15.6 0.80 0.04 0.41
Willingness to Perform Job Tasks
Hemp 33 70.6 ± 23.4 74.5 ± 16.7 0.35 0.19 (5.9, 16.6) 0.34
PLA 32 79.9 ± 18.9 78.3 ± 18.5 0.70 0.08 0.24
Current State of Well-Being
Hemp 33 72.8 ± 22.3 77.2 ± 16.6 0.13 0.23 (6.1, 8.8) 0.71
PLA 32 77.1 ± 12.8 80.2 ± 12.6 0.19 0.24 0.09
Sleep Quality
Hemp 33 53.4 ± 23.8 65.2 ± 19.7 0.005 0.54 (8.1, 15.0) 0.56
PLA 32 60.8 ± 18.9 69.2 ± 16.6 0.06 0.47 0.15
Sleep Quantity
Hemp 33 53.3 ± 20.7 64.7 ± 18.7 0.01 0.58 (5.0, 16.4) 0.29
PLA 32 59.0 ± 15.3 64.7 ± 15.2 0.09 0.37 0.26
I Feel Rested Upon Waking
Hemp 33 50.9 ± 25.3 66.8 ± 19.2 <0.001 0.71 (9.7, 12.6) 0.80
PLA 32 54.8 ± 23.1 69.3 ± 17.3 0.001 0.71 0.06
I Get Pleasure From Life
Hemp 33 71.5 ± 22.6 80.4 ± 15.1 0.007 0.46 (0.26, 14.8) 0.06
PLA 32 82.1 ± 15.7 83.8 ± 13.4 0.44 0.11 0.48
Mood This Past Week
Hemp 33 66.5 ± 21.8 70.1 ± 17.6 0.46 0.18 (10.6, 12.5) 0.87
PLA 32 73.9 ± 17.4 76.5 ± 14.0 0.44 0.16 0.04
Brief Resilience Scale (BRS)
Hemp 33 3.57 ± 0.77 3.85 ± 0.69 0.01 0.39 (0.29, 0.28) 0.98
PLA 32 3.73 ± 0.48 4.02 ± 0.51 0.005 0.59 0.01
JOURNAL OF DIETARY SUPPLEMENTS 13
within-group changes were observed, both within the PLA group, with one variable
(High Frequency Power During the Stroop) increasing (p¼0.04) and the other
Table 7. Heart rate variability.
Variables N
Week 0
(Visit 2)
Week 6
(Visit 4)
Within-Group Between-Group
p-value ES 95% CI
p-value
(ES)
HRV Low Frequency At Rest (m/s
2
)
Hemp 33 1809 ± 1475 1948 ± 1823 0.67 0.08 (466, 1357) 0.33
PLA 32 1983 ± 1878 1677 ± 1249 0.35 0.19 0.24
HRV Low Frequency During Stroop (m/s
2
)
Hemp 33 837 ± 774 904 ± 870 0.53 0.08 (25.8, 538) 0.07
PLA 32 1022 ± 1082 834 ± 931 0.05 0.19 0.46
HRV Low Frequency Recovery from Stroop (m/s
2
)
Hemp 33 1939 ± 1903 2207 ± 2265 0.35 0.13 (484, 1218) 0.39
PLA 32 2101 ± 1752 2002 ± 2124 0.76 0.05 0.21
HRV Low Frequency - Rest (%)
Hemp 33 15.0 ± 5.5 15.7 ± 4.4 0.42 0.14 (2.95, 1.97) 0.69
PLA 32 15.5 ± 5.4 16.7 ± 4.3 0.19 0.25 0.10
HRV Low Frequency During Stroop (%)
Hemp 33 14.8 ± 4.2 15.2 ± 4.3 0.64 0.08 (2.21, 1.62) 0.76
PLA 32 15.1 ± 4.0 15.8 ± 3.9 0.34 0.16 0.08
HRV Low Frequency - Recovery from Stroop (%)
Hemp 33 16.4 ± 4.9 17.3 ± 4.4 0.31 0.19 (1.29, 3.05) 0.42
PLA 32 16.7 ± 4.4 16.7 ± 3.9 0.99 0.00 0.20
HRV High Frequency Power At Rest (m/s
2
)
Hemp 33 1339 ± 1418 1216 ± 1247 0.58 0.09 (781, 1062) 0.76
PLA 32 1347 ± 1634 1084 ± 1652 0.53 0.16 0.08
HRV High Frequency During Stroop (m/s
2
)
Hemp 33 661 ± 688 616 ± 645 0.74 0.07 (165, 465) 0.34
PLA 32 669 ± 712 474 ± 496 0.04 0.35 0.24
HRV High Frequency Recovery from Stroop (m/s
2
)
Hemp 33 1062 ± 1090 997 ± 1197 0.74 0.06 (680, 701) 0.98
PLA 32 1093 ± 1462 1018 ± 1219 0.80 0.06 0.01
HRV High Frequency Rest (%)
Hemp 33 10.0 ± 5.5 9.52 ± 3.99 0.56 0.10 (2.32, 2.78) 0.98
PLA 32 9.50 ± 5.3 8.78 ± 4.75 0.47 0.14 0.05
HRV High Frequency During Stroop (%)
Hemp 33 10.76 ± 4.71 9.73 ± 4.08 0.22 0.23 (2.74, 1.25) 0.46
PLA 32 9.78 ± 4.08 9.50 ± 0.32 0.62 0.08 0.19
HRV High Frequency Recovery from Stroop (%)
Hemp 33 8.85 ± 4.47 8.30 ± 4.30 0.51 0.12 (2.91, 1.57) 0.55
PLA 32 8.25 ± 4.41 8.38 ± 4.23 0.87 0.03 0.15
HRV Low-to-High Frequency Ratio At Rest
Hemp 33 2.66 ± 2.52 2.33 ± 2.35 0.50 0.14 (1.60, 0.92) 0.59
PLA 32 2.69 ± 2.22 2.71 ± 1.89 0.98 0.01 0.13
HRV Low-to-High Frequency Ratio During Stroop
Hemp 33 2.05 ± 1.97 2.05 ± 1.41 0.99 0.00 (0.97, 0.77) 0.82
PLA 32 2.10 ± 1.60 2.19 ± 1.72 0.71 0.06 0.06
HRV Low-to-High Frequency Ratio Recovery from Stroop
Hemp 33 2.88 ± 2.50 3.10 ± 2.74 0.64 0.08 (1.07, 1.49) 0.74
PLA 32 2.97 ± 2.23 2.98 ± 2.40 0.98 0.00 0.08
HRV RMSSD At Rest
Hemp 33 56.1 ± 31.7 53.8 ± 26.9 0.67 0.08 (8.2, 22.7) 0.35
PLA 32 55.0 ± 31.1 45.4 ± 23.7 0.10 0.35 0.23
HRV RMSSD During Stroop
Hemp 33 51.0 ± 57.5 41.2 ± 27.4 0.36 0.22 (27.0, 17.0) 0.65
PLA 32 38.9 ± 19.4 34.1 ± 19.4 0.03 0.25 0.11
HRV RMSSD Recovery from Stroop
Hemp 33 49.8 ± 26.2 46.4 ± 22.9 0.29 0.14 (11.6, 10.4) 0.92
PLA 32 49.5 ± 23.9 46.6 ± 28.0 0.53 0.11 0.03
14 H. L. LOPEZ ET AL.
variable (Root Mean Square of the Standard Deviation During the Stroop) statistically
decreasing (p¼0.03).
Perceived recovery and stroop color task test
Main and group x time interaction effects of the perceived recovery scale and Stroop Color
Task were assessed using mixed factorial ANOVA with repeated measures on time and are
found in Table 8. No statistically significant (p<0.05) group x time interaction effects or
within-group changes over time were observed for any of the variables measured.
Serum and whole blood metabolic and hematological markers
As can be seen in Table 9, main and group x time interaction effects of all serum and
whole blood metabolic and hematological markers were assessed using mixed factorial
ANOVA with repeated measures on time. Statistically significant between-group differ-
ences were identified for changes observed in AST (p¼0.04; 95% CI: 8.10, 0.18; ES
¼0.52) and HDL cholesterol (p¼0.004; 95% CI: 1.66, 8,24, ES ¼0.75). Changes in
AST were largely governed by a tendency for PLA values to increase (p¼0.08) while
changes in HDL cholesterol were mediated by statistically significant increases in the
Hemp group (p¼0.005). Additionally, bilirubin (p¼0.07; 95% CI: 0.01, 0.22; ES ¼
0.46), ALT (p¼0.08; 95% CI: 9.43, 0.52; ES ¼0.44), and total cholesterol (p¼0.09;
95% CI: 1.7, 20.2; ES ¼0.42) all exhibited statistical tendencies for differences
between groups. For bilirubin, a statistical tendency (p¼0.07) for PLA to decrease was
observed while the Hemp group exhibited a tendency to have lower ALT values
(p¼0.08). Moreover, although not a statistically significant between group interaction,
plasma uric acid levels demonstrated within group decrease from baseline only in the
Hemp group (p¼0.02; 95% CI: 0.61, 0.15; ES ¼0.30) with a moderate effect size.
Dietary intake
Main and group x time interaction effects of all dietary intake data were assessed using
mixed factorial ANOVA with repeated measures on time (Table 10). No statistically
Table 8. Perceived recovery scale and stroop color task test.
Variables N
Week 0
(Visit 2)
Week 6
(Visit 4)
Within-Group Between-Group
p-value ES 95% CI
p-value
(ES)
Perceived Recovery Scale
Hemp 33 6.3 ± 2.4 6.4 ± 2.4 0.70 0.08 (0.52, 1.07) 0.49
PLA 32 6.3 ± 2.5 6.2 ± 2.3 0.58 0.13 0.17
Stroop Total Score
Hemp 33 249 ± 62.5 284 ± 58 <0.001 0.59 (13.1, 18.8) 0.72
PLA 32 234 ± 53.9 267 ± 49.0 <0.001 0.63 0.09
Stroop Accuracy (% Correct)
Hemp 33 98.6 ± 1.38 98.9 ± 0.84 0.08 0.28 (0.79, 0.55) 0.72
PLA 32 98.3 ± 1.84 98.8 ± 1.19 0.13 0.29 0.09
Stroop Time Per Score
Hemp 33 1.28 ± 0.33 1.10 ± 0.23 <0.001 0.66 (0.11, 0.10) 0.90
PLA 32 1.34 ± 0.27 1.16 ± 0.21 <0.001 0.73 0.03
JOURNAL OF DIETARY SUPPLEMENTS 15
Table 9. Serum and whole blood metabolic and hematological markers.
Variables N
Week 0
(Visit 1)
Week 6
(Visit 4)
Within-Group Between-Group
p-value ES 95% CI
p-value
(ES)
White Blood Cell Count
Hemp 33 5.37 ± 1.59 5.65 ± 1.48 0.11 0.19 (0.17, 0.84) 0.19
PLA 32 5.59 ± 1.69 5.54 ± 1.41 0.78 0.03 0.33
Red Blood Cell Count
Hemp 33 4.81 ± 0.33 4.77 ± 0.37 0.21 0.12 (0.10, 0.10) 0.99
PLA 32 4.83 ± 0.37 4.79 ± 0.42 0.27 0.10 0.00
Hemoglobin (g/dL)
Hemp 33 14.3 ± 1.2 14.1 ± 1.3 0.16 0.11 (0.29, 0.30) 0.96
PLA 32 14.4 ± 1.1 14.3 ± 1.3 0.17 0.13 0.01
Hematocrit (%)
Hemp 33 42.1 ± 3.1 42.0 ± 3.2 0.73 0.03 (0.88, 0.91) 0.97
PLA 32 42.4 ± 2.9 42.3 ± 3.3 0.72 0.04 0.01
Glucose (mg/dL)
Hemp 33 85.9 ± 10.0 87.2 ± 9.9 0.56 0.13 (3.04, 6.96) 0.44
PLA 32 88.2 ± 7.8 87.5 ± 7.3 0.57 0.09 0.20
Blood Urea Nitrogen (mg/dL)
Hemp 33 14.15 ± 3.63 13.55 ± 4.73 0.28 0.14 (2.24, 1.09) 0.49
PLA 32 14.22 ± 4.29 14.19 ± 4.84 0.96 0.01 0.17
Creatinine (mg/dL)
Hemp 33 0.92 ± 0.16 0.92 ± 0.19 0.69 0.04 (0.03, 0.06) 0.57
PLA 32 0.89 ± 0.18 0.88 ± 0.19 0.68 0.04 0.14
BUN: Creatinine
Hemp 33 15.7 ± 3.9 14.8 ± 4.6 0.13 0.20 (2.73, 1.09) 0.40
PLA 32 16.3 ± 4.2 16.3 ± 4.8 0.97 0.01 0.21
Sodium (mEq/L)
Hemp 33 140.4 ± 2.0 140.0 ± 1.9 0.28 0.20 (1.20, 0.85) 0.74
PLA 32 140.0 ± 1.6 139.8 ± 1.6 0.56 0.14 0.08
Potassium (mEq/L)
Hemp 33 4.20 ± 0.28 4.23 ± 0.25 0.40 0.14 (0.10, 0.16) 0.65
PLA 32 4.30 ± 0.28 4.30 ± 0.24 0.90 0.02 0.11
Chloride (mEq/L)
Hemp 33 102.4 ± 1.93 102.5 ± 2.55 0.75 0.05 (1.22, 1.15) 0.95
PLA 32 102.0 ± 1.86 102.2 ± .96 0.74 0.08 0.01
Carbon Dioxide (mEq/L)
Hemp 33 22.2 ± 2.06 21.7 ± 2.08 0.15 0.25 (1.45, 0.55) 0.37
PLA 32 22.7 ± 1.47 22.6 ± 1.93 0.86 0.04 0.22
Calcium (mg/dL)
Hemp 33 9.47 ± 0.32 9.45 ± 0.35 0.73 0.05 (0.14, 0.17) 0.87
PLA 32 9.36 ± 0.39 9.33 ± 0.35 0.60 0.09 0.04
Total Protein (g/dL)
Hemp 33 7.14 ± 0.40 7.14 ± 0.38 0.96 0.01 (0.08, 0.29) 0.25
PLA 32 7.10 ± 0.333 6.99 ± 0.37 0.13 0.29 0.29
Albumin (g/dL)
Hemp 33 4.66 ± 0.29 4.64 ± 0.26 0.72 0.06 (0.07, 0.20) 0.34
PLA 32 4.63 ± 0.25 4.54 ± 0.30 0.15 0.29 0.24
Globulin (g/dL)
Hemp 33 2.48 ± 0.31 2.50 ± 0.35 0.71 0.05 (0.08, 0.16) 0.52
PLA 32 2.47 ± 0.26 2.45 ± 0.31 0.58 0.08 0.16
Albumin: Globulin
Hemp 33 1.91 ± 0.27 1.90 ± 0.31 0.77 0.04 (0.13, 0.09) 0.75
PLA 32 1.89 ± 0.24 1.90 ± 0.31 0.87 0.02 0.08
Bilirubin (mg/dL)
Hemp 33 0.50 ± 0.33 0.51 ± 0.31 0.70 0.04 (0.01, 0.22) 0.07
PLA 32 0.57 ± 0.34 0.48 ± 0.29 0.07 0.29 0.46
Alkaline Phosphatase (IU/L)
Hemp 33 60.6 ± 14.8 63.0 ± 14.8 0.02 0.16 (2.63, 4.98) 0.54
PLA 32 66.3 ± 15.1 67.6 ± 19.3 0.47 0.07 0.15
AST (U/L)
Hemp 33 25.2 ± 8.8 23.5 ± 6.1 0.25 0.22 (8.10, 0.18) 0.04
(continued)
16 H. L. LOPEZ ET AL.
significant (p<0.05) group x time interaction effects or within-group changes over time
were observed for any of the variables measured.
Discussion
The present study is one of the first controlled investigations using human participants
to examine the impact of supplementation with a commercially available, CBD-contain-
ing hemp oil extract. The purpose of this randomized, double-blind, placebo-controlled,
parallel-group study was to determine the effects of a commercially available Hemp Oil
extract (PlusCBD
TM
Extra Strength Hemp Extract) on physical and mental stress resili-
ence, heart rate variability from normal daily physical & mental free-living conditions,
body composition, and blood-based indicators of health. Key findings from this project
revealed an increase in HDL cholesterol from Hemp supplementation and improve-
ments in a limited number of the visual analog scales that were used to assess the psy-
chometric impact of Hemp supplementation on affect and stress responses. Several
other variables exhibited statistical tendencies for group responses to be different after
Hemp supplementation and revealed modest within-group changes as a result of the
intervention. Additionally, another key finding is that supplementation was well toler-
ated as assessed by the general lack of change exhibited by adverse event, side-effect
profiles, hemodynamic and blood-based markers of hepato-renal function, health and
safety (Tables 3 and 4).
Table 9. Continued.
Variables N
Week 0
(Visit 1)
Week 6
(Visit 4)
Within-Group Between-Group
p-value ES 95% CI
p-value
(ES)
PLA 32 22.7 ± 8.8 25.2 ± 10.2 0.08 0.26 0.52
ALT (U/L)
Hemp 33 23.6 ± 14.3 21.2 ± 9.8 0.08 0.20 (9.43, 0.52) 0.08
PLA 32 22.7 ± 12.7 24.8 ± 16.7 0.34 0.14 0.44
Total Cholesterol (mg/dL)
Hemp 33 176.4 ± 33.1 181.9 ± 28.9 0.12 0.18 (1.7, 20.2) 0.09
PLA 32 184.3 ± 29.7 180.5 ± 35.1 0.39 0.12 0.42
Triglycerides (mg/dL)
Hemp 33 89.3 ± 52.8 89.8 ± 58.2 0.94 0.01 (26.2, 24.9) 0.96
PLA 32 92.2 ± 63.5 93.4 ± 64.3 0.92 0.02 0.01
HDL Cholesterol (mg/dL)
Hemp 33 54.0 ± 12.3 57.5 ± 13.3 0.005 0.27 (1.66, 8.24) 0.004
PLA 32 60.3 ± 18.0 58.7 ± 17.5 0.21 0.09 0.75
VLDL Cholesterol (mg/dL)
Hemp 33 17.8 ± 10.5 18.0 ± 11.7 0.91 0.01 (3.62, 5.37) 0.70
PLA 32 18.4 ± 12.7 17.7 ± 8.8 0.70 0.07 0.10
LDL Cholesterol (mg/dL)
Hemp 33 104.5 ± 27.3 106.5 ± 26.8 0.52 0.07 (5.2, 13.0) 0.40
PLA 32 105.6 ± 30.3 103.7 ± 31.3 0.58 0.06 0.21
Uric Acid (mg/dL)
Hemp 33 5.18 ± 1.06 4.84 ± 1.19 0.02 0.30 (0.61, 0.15) 0.23
PLA 32 5.27 ± 1.52 5.16 ± 1.29 0.45 0.08 0.30
Chromogranin A
Hemp 33 0.52 ± 0.46 0.61 ± 0.58 0.44 0.16 (0.14, 0.36) 0.38
PLA 32 0.59 ± 0.47 0.57 ± 0.47 0.66 0.05 0.22
JOURNAL OF DIETARY SUPPLEMENTS 17
While multiple reports have highlighted the ability of cannabis and hashish to impact
cognition and stress, the impact of changes in heart rate variability (HRV) after hemp
oil or CBD supplementation remain largely undetermined. Nervous system control is
mediated by integrated control of the sympathetic and parasympathetic nervous systems
with control over autonomic activity being predominantly controlled by the parasympa-
thetic branch. Heart rate variability (HRV) assessment has evolved into an accepted
measure of autonomic nervous system activity and consist of high and low-frequency
bands while the ratio between these two bands has also been used in HRV assessments.
Usui et al. (Usui and Nishida 2017) used the Stroop color task test to assess changes in
HRV metrics such as the high and very low frequency bands as well as the ratio
between low and high frequency bands. A similar approach was used in the present
study with significant improvements in Stroop performance occurring in both groups
with no between-group differences (Table 7) for Stroop performance or HRV between
groups. Findings from this paper supported the notion that the high frequency and
low-to-high frequency ratio could operate as a quick recovery component while the
very low frequency component could function as an indicator of slow recovery. Low
frequency power did tend to respond differently between groups (Table 6) with a sig-
nificant HRV reduction occurring only in the PLA group (p¼0.05, ES ¼0.19), and a
non-significant mean HRV value increase in Hemp group. Other reductions occurred in
the PLA group during the Stroop (HRV high frequency and RMSSD), but the overall
changes between groups were not statistically significant (Table 6). Although our study
may have been underpowered to detect a significant group x time difference, these
within group observations are compelling, and merit further study in the future.
Phytocannabinoids, including CBD found in the Hemp extract may modulate the auto-
nomic nervous system via endocannabinoid signaling, as there is a well-established role
for the ECS in the regulating sympathetic vs. parasympathetic tone and hypothalamic-
pituitary-adrenal (HPA) axis (OSullivan et al. 2012). Future interrogation of the influ-
ence of Hemp extract supplementation on HRV may require not only a larger sample
size, but also a stronger stressful stimulus to create more robust perturbations in the
autonomic nervous system, and hence HRV values.
Several visual analog scales were assessed to evaluate various aspects of affect, mood,
and mental state before and after supplementation. Overall, no statistically significant
Table 10. Dietary intake.
Variables N
Week 0
(Visit 1)
Week 6
(Visit 4)
Within-Group Between-Group
p-value ES 95% CI
p-value
(ES)
Caloric Intake (kcals/day)
Hemp 33 1451 ± 501 1413 ± 517 0.52 0.07 (264, 110) 0.41
PLA 32 1604 ± 651 1643 ± 494 0.60 0.07 0.20
Carbohydrate Intake (grams/day)
Hemp 33 146 ± 59 144 ± 65 0.71 0.03 (35, 14) 0.39
PLA 32 162 ± 60 170 ± 59 0.44 0.13 0.21
Fat Intake (grams/day)
Hemp 33 60.7 ± 27.5 60.4 ± 29.8 0.92 0.01 (10.3, 10.7) 0.97
PLA 32 67.3 ± 34.1 66.8 ± 26.4 0.90 0.02 0.01
Protein Intake (grams/day)
Hemp 33 73.9 ± 25.0 73.5 ± 35.2 0.91 0.01 (18.0, 5.0) 0.27
PLA 32 85.5 ± 45.3 91.5 ± 49.1 0.13 0.13 0.28
18 H. L. LOPEZ ET AL.
changes were observed between groups for all of the visual analog and Brief Resilience
Scales that were assessed (Table 5). Similar to HRV, VAS and BRS changes occurred in
individual groups that were not reciprocated by the other supplementation group. For
example, the PLA group reported a statistically significant decrement (p¼0.001, ES ¼
0.67, Table 6) in VAS scores for Ability to Cope with Stresswhile the Hemp group
noted a statistically significant improvement in I Get Pleasure from LifeVAS scores
(p¼0.007), Sleep Quality(p¼0.005, ES ¼0.54) and Sleep Quantity(p¼0.01, ES ¼
0.58). While non-significant, the changes for Ability to Cope with Stress(p¼0.07, 95%
CI: 0.73, 19.1, ES ¼0.46) and I Get Pleasure from Life(p¼0.06, 95% CI: 0.26,
14.8, ES ¼0.48) both exhibited a statistical tendency for Hemp to exhibit greater
changes the PLA. The general trend of these statistical tendencies are consistent with
the substantial body of evidence in both animal models of anxiety, psychosocial, and
physiologic stress as well as in healthy human subjects clearly suggest an anxiolytic and
stress resistance effect of CBD (Appiah-Kusi et al. 2020; de Mello Schier et al. 2014;
Schier et al. 2012). Moreover, the products used in prior studies with human volunteers
consisted of isolated CBD as a constituent molecule, at much higher doses than the
composition of Hemp extract utilized in the present study (Bergamaschi et al. 2011;
Zuardi et al. 2017). Additionally, both groups reported strong and consistent improve-
ments in VAS scores for I Feel Relaxedand I Feel Rested Upon Wakingas well as
the average reported score for the Brief Resilience Scale. Because both groups changed
in the same direction to similar magnitudes, no between-group differences were high-
lighted for these outcomes.
No statistically significant differences were observed for any of the body composition,
and hemodynamic variables as well as the majority of the blood-based markers of health
and safety. While not determined to be statistically significant between groups, changes
in lean, fat-free, and total scanned mass did reveal decreases in these values only in the
PLA group. However, with no meaningful changes in diet, exercise, or physical activity
and no changes observed in body mass (Table 4), body mass index (Table 4), dietary
intake (Table 10), and VAS scores for Appetiteand Cravings for Sweets(Table 6), it
is challenging to interpret these outcomes. No between-group changes were observed in
adverse events, resting heart rate, and diastolic blood pressure while changes in systolic
blood pressure tended to be different due to a decrease in the PLA group. The magni-
tude of change, however, was minimal (<3 mm Hg) and not deemed clinically relevant
with both values remaining within accepted normative reference values. The within
group changes with moderate effect size observed in the Hemp group only for decreased
uric acid levels over the duration of the study remain compelling, despite not reaching
a significant between group interaction. Elevated uric acid levels have been independ-
ently predictive and correlated to conditions of chronic systemic inflammation, includ-
ing but not limited to: cardiovascular conditions, hypertension, type 2 diabetes, chronic
renal disease, and inflammatory arthritis (Benn et al. 2018; Essex et al. 2017; Kleber
et al. 2015; Kuwabara et al. 2017; Sluijs et al. 2015). The work of Kono et al. (Kono
et al. 2010) and others (Braga et al. 2017) suggests that uric acid itself may promote
and exacerbate inflammatory responses via NLRP3 Inflammasome/ASC/Caspase-1 acti-
vation. While direct causation has not yet been firmly established between hyperurice-
mia and these inflammatory conditions, and the current study did not delineate
JOURNAL OF DIETARY SUPPLEMENTS 19
potential confounders of changes in plasma urate conditions such as dietary intake of
purines, or urate handling, excretion, and turnover profiles of subjects, future studies
may interrogate this observation more thoroughly (Benn et al. 2018). Changes in AST
(p¼0.04, 95% CI: 8.1, 0.18, ES ¼0.44) were also found to be different between
the groups, while changes in bilirubin (p¼0.07, 95% CI: 0.01, 0.22, ES ¼0.46) and
ALT (p¼0.08, 95% CI: 9.43, 0.52), ES ¼0.44) tended to be different between
groups. Taken together, the safety profile data in this study are noteworthy, albeit pre-
liminary, given the documented concern over potential hepatotoxicity or drug-induced
liver injury from much higher therapeutic doses (>10 to 20 mg/kg body weight CBD)
of a plant-derived pharmaceutical preparation of purified CBD isolate (Devinsky et al.
2018; Thiele et al. 2018). Future studies on commercial hemp-derived CBD products
should continue to evaluate hepatorenal markers with greater frequency of testing to
expand on the limited body of research.
Increased HDL cholesterol in the Hemp supplementation group were the most mean-
ingful change with a statistically significant between-group difference being observed
(p¼0.004, 95% CI: 1.66, 8.24, ES ¼0.75). It is important to highlight that a six-point
difference was observed at baseline between the groups (Hemp: 54.0 ± 12.3 vs. PLA:
60.3 ± 18.0), but these values were not considered to be statistically significant (p¼0.11,
avg. mean difference ¼6.2 ± 3.8, 95% CI: 13.8, 1.4) and that the final value for
Hemp approached the final value for PLA.
In comparison to the extremely limited and relevant human research in this area, our
results for HDL cholesterol do seem to be contrasted with the results of Kaul et al.
(Kaul et al. 2008) who supplemented healthy men and women for 12 weeks with two
grams per day of an omega-6 polyunsaturated fatty acid-rich hemp seed oil. In this
hempseed oil investigation, no significant within group or between group changes were
observed for total cholesterol, LDL, HDL, and triglycerides. Results from the present
study indicated a statistically significantly increase in HDL cholesterol levels after Hemp
supplementation (p¼0.004, 95% CI: 1.66, 8.24, ES ¼0.75, see Table 9). Similar to the
present study, participants in this study were instructed to maintain their normal diet
and to minimally maintain and attempt to increase their completion of moderate phys-
ical activity throughout the study period. A key difference between the studies is the
composition of what was actually supplemented, as participants in the Kaul investiga-
tion supplemented with two grams per day of an omega-6 polyunsaturated fatty acid-
rich hemp seed oil, which is devoid of the terpenophenolic and phytocannabinoid
constituents found in the CBD-containing Hemp extract central to this investigation.
The role of ECS, and its seemingly expansive role in adiposity, appetite, energy balance
and peripheral lipogenesis, lipoprotein and lipid metabolism provides important context
to interpret the increases in HDL observed in the present study with Hemp supplemen-
tation (Di Marzo and Silvestri 2019). CB1 receptor activation via increased endocanna-
binoid tone or high-fat hyperenergetic diet augments de novo lipogenesis and impairs
fatty acid oxidation in hepatocytes and other peripheral tissues such as adipose and
muscle (Osei-Hyiaman et al. 2005). Whereas, CB1 antagonism in humans improves
atherogenic dyslipidemia by increasing HDL cholesterol concentration, particle number
and size, apolipoprotein-A1 and reducing triglycerides (Despres et al. 2009), which
20 H. L. LOPEZ ET AL.
provides a plausible mechanism for Hemp supplementation increasing HDL concentra-
tion via influencing ECS tone.
One of the biggest challenges with interpreting our findings was the lack of relevant
and comparative human findings available in the literature. As more data is likely to
become available, the overall impact of our findings will continue to take shape.
Regardless, it is important to highlight that a key strength of our study was the random-
ized, double-blind, placebo-controlled design that was employed. Overall, our design
was deemed to be adequately powered as the overall magnitude of observed effects were
low to moderate between groups (many reported effect sizes were 0.20 0.50).
However, as an initial scoping interrogation of this CBD/Phytocannabinoid-rich Hemp
extract, our data highlight that certain outcome measures would require greater sample
size, or inclusion criteria stratification to maximize uniformity among study subjects
and/or independent variables to create a greater physiologic stress as a result of the pro-
homeostatic action of the ECS. In this respect, we adequately screened all study
participants to exclude people who may have been suffering from some form of clinical
condition and further affirm our recruited sample was representative of a healthy popu-
lation of men and women between 18 55 years of age.
Several limitations should be highlighted so proper conclusions are drawn surround-
ing the outcomes of this study. First, the supplementation protocol used in this study
was a six-week, single-daily dose supplementation regimen of a commercially available
Hemp oil extract and our findings should only be considered within this conceptual
framework. Certainly, outcomes may differ with any of the common changes to the
supplementation regimen that are common to these types of studies (total single dose,
number of doses per day, timing of doses relative to data collection instruments and
methods, concentration, type and combination of phytocannabinoids). In this respect,
future studies may explore how the administration route of CBD extracts are delivered
(oral, with food, sublingual, transdermal, supplement timing, etc.). While the subject of
pharmacogenetics and pharmacogenomics of cannabinoids is still emerging, future stud-
ies of phytocannabinoid-rich hemp supplementation should account for the impact of
polymorphisms in genes encoding proteins involved in the action, signaling, metabolism
and transport of both phytocannabinoids and endocannabinoids. With respect to the
CBD-containing Hemp extract of the present study, while many candidate genes exist
due to the complexity of cannabinoid physiology, we would anticipate that the receptor
genes CNR1, TRPV1, GPR55 and biotransformation-metabolism genes CYP3A4,
CYP2C19, CYP2C9 and FAAH are most salient to help explain individual variability in
efficacy (Hryhorowicz et al. 2018). Intensive pharmocogenetic-oriented studies may lead
to personalizing hemp extracts with a specific phytocannabinoid/terpene profile to
match a unique pharmacogenetic/omic signature for optimizing therapeutic potential
and minimizing adverse reactions. Future studies may consider increased frequency of
testing for the purposes of additional pharmacokinetic and/or transient changes in bio-
chemical and hematologic markers. Also, our study results are limited to the population
we recruited. Primary interests in this study were to examine the impact of complex
hemp oil extract rich in CBD, other phytocannabinoids, terpenes, polyphenols, toco-
pherols/tocotrienols, multiple saturated and unsaturated fatty acid species in an over-
weight, otherwise healthy population, but it certainly remains probable that our results
JOURNAL OF DIETARY SUPPLEMENTS 21
will be different if samples are recruited from populations who are suffering from any
combination of metabolic, inflammatory, psychological, emotional, social stress or anx-
iety, irrespective of clinical diagnoses.
Conclusions
Six weeks of supplementing with a commercially available, independent conclusion
GRAS (Generally Recognized As Safe) CBD-containing hemp oil extract led to improve-
ments in HDL cholesterol in overweight, otherwise healthy men and women. Stochastic
changes were noted in clinical biomarkers of safety. When combined with no serious
adverse events, nor side effect patterns being reported between groups throughout the
study trial, the CBD-containing Hemp extract supplement was well tolerated with no
concerns for safety within the conditions of this study. Body composition, cognitive
processing, and recovery do not appear to be impacted by supplementation. No
between-group differences were evident for any of the heart rate variability measures.
With respect to VAS outcomes, while some variables exhibited statistically significant
increases or decreases in the measured values in either the Hemp or PLA groups, the
overall pattern of these changes are consistent with modulation of the ECS in areas of
mood, sleep, life pleasure, and stress responses. Furthermore, several VAS variables
exhibited statistically significant improvements in both groups as a result of supplemen-
tation. Overall, these findings suggest that supplementation with this Hemp extract at
the provided dosage in the men and women studied exhibited improvements in HDL
cholesterol, tended to support psychometric measures of perceived sleep quantity and
stress response, perceived life pleasure, and is well tolerated in healthy human subjects.
Acknowledgements
The supplement (PlusCBD Extra Strength Hemp Extract Oil
TM
), placebo, and funding for this
project was received through a restricted external grant from CV Sciences, Inc. The authors
would like to thank the study participants who completed the study protocol. Publication of these
results should not be considered as an endorsement of any product used in this study by the
Center for Applied Health Sciences or any of the organizations where the authors are affiliated.
Declaration of interests
Lopez was previously remunerated with stock options as a consultant for CV Sciences, serving on
the scientific advisory board from 2014 through 2018. Lopez is an officer and member of The
Center for Applied Health Sciences, a privately held contract research organization that has
received external funding from companies that do business in the dietary supplement, natural
products, medical foods and functional foods and beverages industries. He is the co-founder and
member of Supplement Safety Solutions, LLC., serving as an independent consultant for regula-
tory compliance, safety surveillance and Nutravigilance to companies in the dietary supplement
and functional foods industry, but not the sponsor of the current research. Lopez is also co-
inventor on multiple patent applications within the field of dietary supplements, applied nutrition
and bioactive compounds. Raub, Cesareo, Kedia, and Sandrock all report no conflicts of interest.
Kerksick has no conflict in terms of financial or business interests related to this product.
Kerksick has received external grant funding from companies that do business in the nutrition
and sports nutrition sectors. He has received compensation to speak and prepare scientific
22 H. L. LOPEZ ET AL.
manuscripts including white papers and marketing copy on topics related to sports nutrition. He
has and continues to serve in advisory roles to various sport nutrition and nutrition companies.
Ziegenfuss is an officer and member of The Center for Applied Health Sciences, a privately held
contract research organization that has received external funding from companies that do busi-
ness in the dietary supplement, natural products, medical foods and functional foods and bever-
ages industries. Ziegenfuss was previously remunerated with stock options as a consultant for CV
Sciences, serving on the scientific advisory board from 2014 through 2018. Ziegenfuss has
received grants and contracts to conduct research on dietary supplements; has served as a paid
consultant for industry; has received honoraria for speaking at conferences and writing articles
about functional foods and dietary supplements; receives royalties from the sale of several sports
nutrition products (none related to the product examined in the present study); and has served
as an expert witness on behalf of the plaintiff and defense in cases involving dietary supplements.
Ziegenfuss is also co-inventor on multiple patent applications within the field of dietary supple-
ments, applied nutrition and bioactive compounds.
Authorscontributions
HLL and TNZ designed the study, secured funding for the project, and assisted with manuscript
preparation. BR, KC, and JS carried out subject recruitment, data collection, coordination of the
study and compliance. AWK provided medical oversight and screened subjects. CMK performed
the data analyses/interpretation and prepared the manuscript. All authors read and approved the
final manuscript.
Funding
Funding was provided by CV Sciences, Inc. (10070 Barnes Canyon Rd. San Diego, CA 92121)
through a restricted grant to The Center for Applied Health Sciences. Outside of initial discus-
sions, the sponsor played no part in designing the study. Further, the sponsor had no part in col-
lecting the data, analyzing the data, or preparing the manuscript for publication.
Notes on contributors
Hector L. Lopez, MD, MS is CMO and Founding Partner of The Center for Applied Health
Sciences and Supplement Safety Solutions, USA. Sports Medicine, Regenerative Medicine,
Endocrinology & Metabolism. Interested in nutritional biosciences, and the effects of naturally
occurring bioactives on health, human performance and longevity.
Kyle R. Cesareo, MS, CCRC is a Clinical Research Assistant, The Center for Applied Health
Sciences. He is also a Certified Strength & Conditioning Specialist with interests in sports per-
formance, dietary supplements and nutrition. Ohio, USA.
Betsy Raub, RN, BSN is a clinical research nurse and lab coordinator at The Center for Applied
Health Sciences. Additionally, with 20 years of critical care experience, she continues to remain
relevant at the bedside at Aultman Alliance Community Hospital, Ohio, USA.
A. William Kedia, MD is a Medical Advisor to The Center for Applied Health Sciences,
Ohio, USA.
Jennifer E. Sandrock, MS, RD, CCRC is the COO of The Center for Applied Health Sciences.
She is also the Registered Dietitian and Clinical Research Coordinator for all studies conducted
at CAHS.
Chad M. Kerksick, PhD is an Associate Professor of Exercise Science in the Exercise Science
department in the School of Health Sciences at Lindenwood University and Director of the
JOURNAL OF DIETARY SUPPLEMENTS 23
Exercise and Performance Nutrition Laboratory. His primary research interests include sport
nutrition as well as the biochemical, cellular and molecular adaptations relative to various forms
of exercise and nutrition interventions.
Tim N. Ziegenfuss, PhD is the CEO of The Center for Applied Health Sciences and past
President (2009-2011) and Fellow of the International Society of Sports Nutrition. His primary
research interests are applied physiology, dietary supplements, nutrition, and sports medicine.
References
Appiah-Kusi E, Petros N, Wilson R, Colizzi M, Bossong MG, Valmaggia L, Mondelli V, McGuire
P, Bhattacharyya S. 2020. Effects of short-term cannabidiol treatment on response to social
stress in subjects at clinical high risk of developing psychosis. Psychopharmacology (Berl).
237(4):11211130. doi:10.1007/s00213-019-05442-6.
Benn CL, Dua P, Gurrell R, Loudon P, Pike A, Storer RI, Vangjeli C. 2018. Physiology of hyper-
uricemia and urate-lowering treatments. Front Med (Lausanne)). 5:160. doi:10.3389/fmed.2018.
00160.
Bergamaschi MM, Queiroz RH, Chagas MH, de Oliveira DC, De Martinis BS, Kapczinski F,
Quevedo J, Roesler R, Schroder N, Nardi AE, et al. 2011. Cannabidiol reduces the anxiety
induced by simulated public speaking in treatment-naïve social phobia patients .
Neuropsychopharmacology. 36(6):12191226. doi:10.1038/npp.2011.6.
Braga TT, Forni MF, Correa-Costa M, Ramos RN, Barbuto JA, Branco P, Castoldi A, Hiyane MI,
Davanso MR, Latz E, et al. 2017. Soluble Uric Acid Activates the NLRP3 Inflammasome. Sci
Rep. 7:39884., doi:10.1038/srep39884.
Campos AC, Fogaca MV, Sonego AB, Guimaraes FS. 2016. Cannabidiol, neuroprotection and
neuropsychiatric disorders. Pharmacol Res. 112:119127. doi:10.1016/j.phrs.2016.01.033.
Cuka S, Dvornik S, Drazenovic K, Mihic J. 2001. Evaluation of the Dade Behring Dimension RxL
clinical chemistry analyzer. Clin Lab. 47(1-2):3540.
de Mello Schier AR, de Oliveira Ribeiro NP, Coutinho DS, Machado S, Arias-Carrion O, Crippa
JA, Zuardi AW, Nardi AE, Silva AC. 2014. Antidepressant-like and anxiolytic-like effects of
cannabidiol: a chemical compound of Cannabis sativa. CNS Neurol Disord Drug Targets.
13(6):953960. doi:10.2174/1871527313666140612114838.
Despres JP, Ross R, Boka G, Almeras N, Lemieux I, Investigators AD-L. 2009. Effect of rimona-
bant on the high-triglyceride/low-HDL-cholesterol dyslipidemia, intraabdominal adiposity, and
liver fat: the ADAGIO-Lipids trial. ATVB. 29(3):416423. doi:10.1161/ATVBAHA.108.176362.
Devinsky O, Patel AD, Cross JH, Villanueva V, Wirrell EC, Privitera M, Greenwood SM, Roberts
C, Checketts D, VanLandingham KE, et al. 2018. Effect of Cannabidiol on Drop Seizures in
the Lennox-Gastaut Syndrome. N Engl J Med. 378(20):18881897. doi:10.1056/
NEJMoa1714631.
Di Marzo V, Silvestri C. 2019. Lifestyle and metabolic syndrome: contribution of the endocanna-
binoidome. Nutrients. 11(8):1956. doi:10.3390/nu11081956.
Essex MN, Hopps M, Bienen EJ, Udall M, Mardekian J, Makinson GT. 2017. Evaluation of the
Relationship Between Serum Uric Acid Levels and Cardiovascular Events in Patients With
Gout: A Retrospective Analysis Using Electronic Medical Record Data. J Clin Rheumatol.
23(3):160166. doi:10.1097/RHU.0000000000000496.
Hillard CJ. 2000. Endocannabinoids and vascular function. J Pharmacol Exp Ther. 294(1):2732.
doi:10.1124/jpet.103.054270.
Hryhorowicz S, Walczak M, Zakerska-Banaszak O, Słomski R, Skrzypczak-Zieli
nska M. 2018.
Pharmacogenetics of cannabinoids. Eur J Drug Metab Pharmacokinet. 43(1):112. doi:10.1007/
s13318-017-0416-z.
Kaul N, Kreml R, Austria JA, Richard MN, Edel AL, Dibrov E, Hirono S, Zettler ME, Pierce GN.
2008. A comparison of fish oil, flaxseed oil and hempseed oil supplementation on selected
parameters of cardiovascular health in healthy volunteers. J Am Coll Nutr. 27(1):5158. doi:10.
1080/07315724.2008.10719674.
24 H. L. LOPEZ ET AL.
Kleber ME, Delgado G, Grammer TB, Silbernagel G, Huang J, Kramer BK, Ritz E, Marz W.
2015. Uric acid and cardiovascular events: a mendelian randomization study. J Am Soc
Nephrol. 26(11):28312838. doi:10.1681/ASN.2014070660.
Kono H, Chen CJ, Ontiveros F, Rock KL. 2010. Uric acid promotes an acute inflammatory
response to sterile cell death in mice. J Clin Invest. 120(6):19391949. doi:10.1172/JCI40124.
Kuwabara M, Niwa K, Hisatome I, Nakagawa T, Roncal-Jimenez CA, Andres-Hernando A,
Bjornstad P, Jensen T, Sato Y, Milagres T, et al. 2017. Asymptomatic hyperuricemia without
comorbidities predicts cardiometabolic diseases: five-year japanese cohort study. Hypertension.
69(6):10361044. doi:10.1161/HYPERTENSIONAHA.116.08998.
Lee KA, Hicks G, Nino-Murcia G. 1991. Validity and reliability of a scale to assess fatigue.
Psychiatry Res. 36(3):291298. doi:10.1016/0165-1781(91)90027-M.
Maroon J, Bost J. 2018. Review of the neurological benefits of phytocannabinoids. Surg Neurol
Int. 9:91. doi:10.4103/sni.sni_45_18.
Mechoulam R, Braun P, Gaoni Y. 1967. A stereospecific synthesis of (-)-delta 1- and (-)-delta
1(6)-tetrahydrocannabinols. J Am Chem Soc. 89(17):45524554. doi:10.1021/ja00993a072.
Mechoulam R, Shvo Y, Hashish I. 1963. The structure of cannabidiol. Tetrahedron. 19(12):
20732078. doi:10.1016/0040-4020(63)85022-X.
Morales P, Hurst DP, Reggio PH. 2017. Molecular Targets of the Phytocannabinoids: A Complex
Picture. Prog Chem Org Nat Prod. 103:103131. doi:10.1007/978-3-319-45541-9_4.
Osei-Hyiaman D, DePetrillo M, Pacher P, Liu J, Radaeva S, Batkai S, Harvey-White J, Mackie K,
Offertaler L, Wang L, et al. 2005. Endocannabinoid activation at hepatic CB1 receptors stimu-
lates fatty acid synthesis and contributes to diet-induced obesity. J Clin Invest. 115(5):
12981305. doi:10.1172/JCI200523057.
OSullivan SE, Kendall PJ, Kendall DA. 2012. Endocannabinoids and the cardiovascular response
to stress. J Psychopharmacol (Oxford)). 26(1):7182. doi:10.1177/0269881111408457.
Pertwee RG. 1997. Pharmacology of cannabinoid CB1 and CB2 receptors. Pharmacol Ther. 74(2):
129180. doi:10.1016/S0163-7258(97)82001-3.
Schier AR, Ribeiro NP, Silva AC, Hallak JE, Crippa JA, Nardi AE, Zuardi AW. 2012.
Cannabidiol, a Cannabis sativa constituent, as an anxiolytic drug. Braz J Psychiatry. 34 (Suppl
1):S104S10. doi:10.1016/S1516-4446(12)70057-0.
Schultes R. 1973. Man and marijuana. Natural History. 82:59.
Schultes RE. 1969. Hallucinogens of plant origin. Science. 163(3864):245254. doi:10.1126/science.
163.3864.245.
Sluijs I, Holmes MV, van der Schouw YT, Beulens JW, Asselbergs FW, Huerta JM, Palmer TM,
Arriola L, Balkau B, Barricarte A, et al. 2015. A mendelian randomization study of circulating
uric acid and type 2 diabetes. Diabetes. 64(8):30283036. doi:10.2337/db14-0742.
Smith BW, Dalen J, Wiggins K, Tooley E, Christopher P, Bernard J. 2008. The brief resilience
scale: assessing the ability to bounce back. Int J Behav Med. 15(3):194200. doi:10.1080/
10705500802222972.
Stroop J. 1935. Studies of interference in serial verbal reactions. J Exp Psychol. 18(6):643662.
doi:10.1037/h0054651.
Thiele EA, Marsh ED, French JA, Mazurkiewicz-Beldzinska M, Benbadis SR, Joshi C, Lyons PD,
Taylor A, Roberts C, Sommerville K, et al. 2018. Cannabidiol in patients with seizures associ-
ated with Lennox-Gastaut syndrome (GWPCARE4): a randomised, double-blind, placebo-
controlled phase 3 trial. Lancet. 391(10125):10851096., doi:10.1016/S0140-6736(18)30136-3.
Usui H, Nishida Y. 2017. The very low-frequency band of heart rate variability represents the
slow recovery component after a mental stress task. PLoS One. 12(8):e0182611doi:10.1371/
journal.pone.0182611.
van der Knaap RHP, Kwekkeboom DJ, Ramakers CRB, de Rijke YB. 2015. Evaluation of a new
immunoassay for chromogranin A measurement on the Kryptor system. Pract Lab Med. 1:
511. doi:10.1016/j.plabm.2015.03.002.
Ziegenfuss TN, Habowski SM, Sandrock JE, Kedia AW, Kerksick CM, Lopez HL. 2017. A two-
part approach to examine the effects of theacrine (TeaCrineV
R) supplementation on oxygen
JOURNAL OF DIETARY SUPPLEMENTS 25
consumption, hemodynamic responses, and subjective measures of cognitive and psychometric
parameters . J Diet Suppl. 14(1):924. doi:10.1080/19390211.2016.1178678.
Ziegenfuss TN, Hofheins JE, Mendel RW, Landis J, Anderson RA. 2006. Effects of a water-soluble
cinnamon extract on body composition and features of the metabolic syndrome in pre-diabetic
men and women. J Int Soc Sports Nutr. 3(2):4553. doi:10.1186/1550-2783-3-2-45.
Ziegenfuss TN, Kedia AW, Sandrock JE, Raub BJ, Kerksick CM, Lopez HL. 2018. Effects of an
aqueous extract of withania somnifera on strength training adaptations and recovery: the
STAR trial. Nutrients. 10(11):1807. doi:10.3390/nu10111807.
Ziegenfuss TN, Lopez HL, Sandrock JE, Kedia AW, Habowski S, Kerksick C. 2017. Effect of a
multi-nutrient over-the-counter supplement on changes in metabolic rate and markers of lip-
olysis. J Diet Suppl. 14(3):288302. doi:10.1080/19390211.2016.1215366.
Zuardi AW, Rodrigues NP, Silva AL, Bernardo SA, Hallak JEC, Guimaraes FS, Crippa JAS. 2017.
Inverted U-shaped dose-response curve of the anxiolytic effect of cannabidiol during public
speaking in real life. Front Pharmacol. 8:259. doi:10.3389/fphar.2017.00259.
26 H. L. LOPEZ ET AL.
... Lopez et al. 32 reported that 6 weeks of oral CBD treatment (15 mg/day) increased blood concentrations of highdensity lipoproteins (HDL; sometimes known as "good cholesterol") in overweight but otherwise healthy individuals. Self-reported improvements in sleep quality and quantity were also measured relative to baseline within the CBD-treated group; however, no statistically significant differences between the CBD and placebo groups were detected. ...
... The very low dose of CBD (15 mg) in healthy overweight participants produced marginally significant improvements in sleep quality and quantity over 6 weeks of dosing compared to baseline, but not compared to placebo. 32 Sleep quality improvements were also evident in patients with insomnia in a study, 29 although only with 160 mg CBD and not with doses of 40 or 80 mg. On the other hand, sleep quality and sleep architecture were not improved in healthy volunteers with 300 mg CBD in the more recent study, which used polysomnography. ...
... The current review also found intriguing but not necessarily compelling evidence for low-dose CBD efficacy in a range of other conditions. This includes the protective effects against GVHD (300 mg CBD) 56 ; positive effects (300 mg CBD) on Parkinsonian tremor 45 ; improved quality of life and reduced psychotic symptoms in patients with Parkinson's disease (300 mg CBD) 36,59 ; and beneficial effects (15 mg CBD) on HDL (cholesterol), 32 although this was not repeated with higher doses of CBD (200 mg CBD). 44 ...
Article
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Global interest in the non-intoxicating cannabis constituent, cannabidiol (CBD), is increasing with claims of therapeutic effects across a diversity of health conditions. At present, there is sufficient clinical trial evidence to support the use of high oral doses of CBD (e.g., 10-50 mg/kg) in treating intractable childhood epilepsies. However, a question remains as to whether "low-dose" CBD products confer any therapeutic benefits. This is an important question to answer, as low-dose CBD products are widely available in many countries, often as nutraceutical formulations. The present review therefore evaluated the efficacy and safety of low oral doses of CBD. The review includes interventional studies that measured the clinical efficacy in any health condition and/or safety and tolerability of oral CBD dosed at less than or equal to 400 mg per day in adult populations (i.e., ≥18 years of age). Studies were excluded if the product administered had a Δ9 -tetrahydrocannabinol content greater than 2.0%. Therapeutic benefits of CBD became more clearly evident at doses greater than or equal to 300 mg. Increased dosing from 60 to 400 mg/day did not appear to be associated with an increased frequency of adverse effects. At doses of 300-400 mg, there is evidence of efficacy with respect to reduced anxiety, as well as anti-addiction effects in drug-dependent individuals. More marginal and less consistent therapeutic effects on insomnia, neurological disorders, and chronic pain were also apparent. Larger more robust clinical trials are needed to confirm the therapeutic potential of lower (i.e., <300 mg/day) oral doses of CBD.
... One of such insects is honey bees. positive effects of hemp extracts have been described many times in relation to diseases such as depression, epilepsy, Alzheimer's, appetite disorders, as an aid in the treatment of cancer and in multiple sclerosis [27][28][29][30][31]. There are also interesting studies in which the hemp extract helped to regenerate damaged brain tissues in rats, damaged as a result of the action of chemical agents [32]. ...
Article
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We examined how CBD extract influences the activity of the immune system in the hemolymph of honey bees in the hive test. The bees were divided into 3 groups: (CSy) bees fed with CBD in sugar syrup with glycerin; (CSt) cotton strip with CBD placed in hive bees fed pure sugar syrup, (C) control bees fed sugar syrup with glycerin. CBD extract increased the total protein concentrations, proteases and their inhibitor activities in each age (the except for acidic protease activities in the 21st and 28th day and alkaline protease inhibitor activities in the 28th day in CSt group) in comparison with group C. In the groups with the extract there was also an increase in the enzymatic marker activities: ALP, AST (decrease on day 28 for CSt), ALT; and non-enzymatic marker concentrations: glucose; triglycerides; cholesterol and creatinine. The urea acid and albumin concentrations were lower in CSy and CSt groups compared to the C group (higher concentration of albumin was displayed by control bees). Higher activities/concentrations of most of biochemical parameters were obtained in the CSy compared to the CSt and C. CBD supplementation can positively influence workers’ immune system.
... Prior studies in humans did not reveal the change in plasma cholesterol in heavy marijuana smokers (Muniyappa et al., 2013;Bruins et al., 2016) or people consuming CBD (Jadoon et al., 2016) with an increase in blood cholesterol in users of CBD-containing hemp oil extract (Lopez et al., 2020). Marijuana smokers had, however, decreased blood levels of HDL-cholesterol (Muniyappa et al., 2013). ...
Article
Spent hemp biomass (SHB), a byproduct of cannabinoids extraction from the production of industrial hemp has not been approved by FDA-CVM since its effects on animal health, performance, and product quality are unknown. Our objective was to investigate the effects of feeding two levels of SHB and a four-week withdrawal period on performance, carcass characteristic, meat quality and hematological parameters in finishing lambs. A total of 35 weaned, Polypay male lambs kept in single pens were randomly assigned to five feeding treatments (n=7) and fed diets containing either no SHB (CON) or SHB at 10% (LH1) or 20% (HH1) for 4 weeks with 4 weeks of clearing period from SHB, or SHB at 10 (LH2) or 20% (HH2) for 8 weeks. Chemical analysis revealed SHB to have nutritive quality similar to alfalfa with no mycotoxin, terpenes, or organic residuals as a result of the extraction process. Feed intake of lambs was negatively affected by 20% SHB in Period 1 but not in Period 2 where feed intake was the greatest in HH1 and LH2. In contrast, none of the performance data, including liveweight gains, were different across the groups and periods. In Period 1, blood glucose, cholesterol, calcium, paraoxonase, and tocopherol were decreased by the level of SHB fed, while bilirubin and alkaline phosphatase (ALP) were increased. In Period 2, concentration in blood of urea, magnesium, bilirubin, ALP, and ferric reducing ability of the plasma (FRAP) were higher in LH2 and HH2 as compared to CON, while β-hydroxybutyrate was lower in HH2. Blood parameters related to liver health, kidney function, immune status, and inflammation were unaffected by feeding SHB. Most carcass and meat quality parameters did not differ across feeding groups either. Except carcass purge loss and meat cook loss were larger in lambs that were fed 20% SHB. Although lower feed intake of lambs that were fed 20% SHB initially in period 1 suggested SHB was not palatable to the lambs, increased feed intake at lower level of inclusion at 10% in Period 2 may point to a positive long-term effect of feeding SHB.
... ng/mL) and similar to the (peak) plasma CBD concentrations observed on the 15 mg CBD treatment (4.7 (0.0-25.7) ng/mL) (when no CBD was present at Baseline). This is important because no RCTs appear to have detected meaningful phenotypic effects of CBD at doses <200 mg (Chagas et al., 2014;Freeman et al., 2020;Jadoon et al., 2016;Linares et al., 2019;Lopez et al., 2020;Naftali et al., 2017;Zuardi et al., 2017). It is therefore unlikely that these low, residual levels of CBD influenced performance. ...
Article
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Background Cannabidiol (CBD), a major cannabinoid of Cannabis sativa, is widely consumed in prescription and non-prescription products. While CBD is generally considered ‘non-intoxicating’, its effects on safety-sensitive tasks are still under scrutiny. Aim We investigated the effects of CBD on driving performance. Methods Healthy adults ( n = 17) completed four treatment sessions involving the oral administration of a placebo, or 15, 300 or 1500 mg CBD in a randomised, double-blind, crossover design. Simulated driving performance was assessed between ~45–75 and ~210–240 min post-treatment (Drives 1 and 2) using a two-part scenario with ‘standard’ and ‘car following’ (CF) components. The primary outcome was standard deviation of lateral position (SDLP), a well-established measure of vehicular control. Cognitive function, subjective experiences and plasma CBD concentrations were also measured. Non-inferiority analyses tested the hypothesis that CBD would not increase SDLP by more than a margin equivalent to a 0.05% blood alcohol concentration (Cohen’s d z = 0.50). Results Non-inferiority was established during the standard component of Drive 1 and CF component of Drive 2 on all CBD treatments and during the standard component of Drive 2 on the 15 and 1500 mg treatments (95% CIs < 0.5). The remaining comparisons to placebo were inconclusive (the 95% CIs included 0 and 0.50). No dose of CBD impaired cognition or induced feelings of intoxication ( ps > 0.05). CBD was unexpectedly found to persist in plasma for prolonged periods of time (e.g. >4 weeks at 1500 mg). Conclusion Acute, oral CBD treatment does not appear to induce feelings of intoxication and is unlikely to impair cognitive function or driving performance (Registration: ACTRN12619001552178).
... In 1 trial, different types of inflorescences were administered to evaluate the most efficacious ratio of THC to CBD concentrations against pain. (Pretzsch, et al., 2019b;Solowij, et al., 2019;Van de Donk, et al., 2019;Abrams, et al., 2020;Farokhnia, et al., 2020;Liu, et al., 2020;Lopez, et al., 2020;Thompson, et al., 2020;Naftali, et al., 2021a;Anderson, et al., 2021;Aran, et al., 2021;Naftali, et al., 2021b) ...
Article
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Cannabis has long been regarded as a recreational substance in the Western world. The recent marketing authorization of some medicinal products of industrial origin and the introduction onto the market of inflorescences for medical use mean that medical doctors can now prescribe Cannabis-based medicines in those countries which allow it. Nevertheless, there is still considerable controversy on this topic in the scientific community. In particular, this controversy concerns: the plant species to be used; the pathologies that can be treated and consequently the efficacy and safety of use; the routes of administration; the methods of preparation; the type and dosage of cannabinoids to be used; and, the active molecules of interest. As such, although medical Cannabis has been historically used, the results of currently completed and internationally published studies are inconclusive and often discordant. In light of these considerations, the aim of this work is to analyse the current legislation in countries that allow the use of medical Cannabis, in relation to the impact that this legislation has had on clinical trials. First of all, a literature search has been performed (PubMed and SciFinder) on clinical trials which involved the administration of Cannabis for medical use over the last 3 years. Of the numerous studies extrapolated from the literature, only about 43 reported data on clinical trials on medical Cannabis, with these mainly being performed in Australia, Brazil, Canada, Denmark, Germany, Israel, Netherlands, Switzerland, the United Kingdom and the United States of America. Once the reference countries were identified, an evaluation of the legislation in relation to Cannabis for medical use in each was carried out via the consultation of the pertinent scientific literature, but also of official government documentation and that of local regulatory authorities. This analysis provided us with an overview of the different legislation in these countries and, consequently, allowed us to analyse, with greater awareness, the results of the clinical trials published in the last 3 years in order to obtain general interest indications in the prosecution of scientific research in this area.
... With the increase in consumer consumption of hemp extracts there has been an increased interest in and demand for determining the safety of these extracts. There are recently published studies in both humans and laboratory animals evaluating the safety of orally consumed hemp extracts, however with the variation in the composition of these extracts, comparison of the information must be conducted in tandem with a detailed evaluation of the extract composition [5][6][7]. The bioactivity or potential for hemp extracts to cause toxicity may be influenced by method of manufacture or slight differences in the chemical profile. ...
Article
Full-text available
VOHO Hemp Oil (Verdant Nature LLC (in collaboration with HempFusion)) is an extract of the aerial parts of hemp ( Cannabis sativa L) manufactured using a supercritical CO 2 extraction process. The results of four safety studies are reported here including a bacterial reverse mutation assay, an in vivo mammalian micronucleus study, a maximum tolerated dose study in rats and a 90-day repeat dose subchronic toxicity study in rats. VOHO Hemp oil can contain up to 30% phytocannabinoids and less than 0.2% is tetrahydrocannabinol (THC). VOHO Hemp Oil was found to be non-mutagenic in the bacterial reverse mutation assay and was negative for inducing micronuclei in the rat bone marrow micronucleus assay. The maximum tolerated dose in male and female Wistar rats was 2250 mg/kg bw/day. A 90-day repeat dose study was conducted in male and female Wistar rats according to OECD Guideline 408 and included a 21-day recovery period. The doses used in the study were 0, 25, 90 and 324 mg/kg bw per day in the main study, and in the recovery phase a control and 324 mg/kg bw/day group were included. One mortality was reported during the study, a high dose female, and test substance-related adverse clinical signs were reported in the high dose group. Other test substance-related changes noted in the high dose group included changes in body weights, activated partial thromboplastin time (APTT) values, and in absolute and relative organ weights. Based on the results of the study, the no observed adverse effect level (NOAEL) for VOHO Hemp Oil was determined to be 90 mg/kg bw/day in both male and female Wistar rats.
... However, during Period 1, the decrease in cholesterol and glucose in lambs fed 10% SHB cannot be explained by feed intake since their feed intake was similar to those were fed the control diet. Prior studies in humans did not reveal change of plasma cholesterol in heavy marijuana smokers (Muniyappa et al., 2013;Bruins et al., 2016) or people consuming CBD (Jadoon et al., 2016) with an increase in blood cholesterol in users of CBD-containing hemp oil extract (Lopez et al., 2020). Marijuana smokers had however a decreased blood levels of HDLcholesterol (Muniyappa et al., 2013). ...
Article
The 2018 Farm Bill removed hemp (Cannabis sativa) from the Controlled Substances Act, classifying it as an agricultural product. The process of cannabidiol extraction from hemp yields large quantities of spent hemp biomass (SHB) that may potentially be included in animal diets. However, the use of SHB in animal diets has not been approved by FDA yet since its effect on animal health, production and product quality is still unknown. Thus, a feeding study was carried out to investigate the effects of varying levels of SHB and a four-week withdrawal period on feed intake and liveweight gains of weaned lambs. A total of 35 weaned, male Polypay lambs kept in single pens were randomly assigned to five feeding treatments (n=7) and fed diets containing either no SHB (CON) or SHB at 10% (LH1) or 20% (HH1) for 4 weeks with 4 weeks withdrawal from SHB, or SHB at 10 (LH2) or 20% (HH2) for 8 weeks. The nutritive analysis of the SHB indicated a high-quality feed, with 20% (DM) crude protein and 27% NDF. Dry matter (DM) intake of lambs was negatively affected by 20% SHB during the first period. In the second period, DM intake was larger in lambs fed 10% SHB vs. CON, with the largest feed intake observed in HH1 lambs. In contrast, none of the performance data, including liveweight gains, were different across the groups and periods. Feeding 20% SHB decreased plasma cholesterol, NEFA, BHBA, Ca, and Cl and increased urea and Mg, while 10% SHB increased glucose, cholesterol, and NEFA. Our findings indicated that 10% SHB can be included in ruminant diets without causing any detrimental effect on performance with a possible positive effect on feed intake. The long-term feeding of 20% SHB strongly affects the metabolism.
... Scheme 1. Use of cannabis extract. Hemp in various forms has been tested in the treatment of many neurological, psychological, cancer, viral, and metabolic diseases [11,[14][15][16][17]. ...
Article
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In the study, we assessed the effect of hemp extract on activities of resistance parameters and the metabolic compound concentration in adult workers’ hemolymph. Bees were divided into the following groups: (1) control group fed with mixture of sugar and water-glycerine solution, (2) experimental group with pure sugar syrup and inside with cotton strips soaked with hemp extract, (3) experimental group with a mixture of sugar syrup with hemp extract. Hemp extracts caused an increase in the protein concentrations and reduced the protease activities regardless of the administration method. The protease inhibitor activities were decreased only in the group that received hemp extract on the strips. The biomarker activities (ALP, ALT, AST) increased from the control group and workers feeding extract in syrup and decreased in workers supplemented with the extract on strips. In young, 2-day-old workers, the glucose concentration was higher in the groups feeding with the extract than in the control. Hemp extract influenced an increase in urea concentrations in workers’ hemolymph in comparison with the control. The hemp supplementation positively influences the immune system of workers, and the appropriate method of administration may be adapted to the health problems of bees.
Article
Study objectives As cannabis is increasingly used to treat sleep disorders, we performed a systematic review to examine the effects of cannabis on sleep and to guide cannabis prescribers in their recommendations to patients, specifically focusing on dosing. Methods We searched EMBASE, Medline, and Web of Science and identified 4,550 studies for screening. 568 studies were selected for full-text review and 31 were included for analysis. Study results were considered positive based on improvements in sleep architecture or subjective sleep quality. Bias in randomised controlled trials was assessed using Cochrane Risk of Bias tool 2.0. Results Sleep improvements were seen in 7 out of 19 randomised studies and in 7 out of 12 uncontrolled trials. There were no significant differences between the effects of tetrahydrocannabinol and cannabidiol. Cannabis showed most promise at improving sleep in patients with pain-related disorders, as compared to those with neurologic, psychiatric, or sleep disorders, and showed no significant effects on healthy participants’ sleep. While subjective improvements in sleep quality were often observed, diagnostic testing showed no improvements in sleep architecture. Adverse events included headaches, sedation, and dizziness, and occurred more frequently at higher doses, though no serious adverse events were observed. Conclusion High-quality evidence to support cannabis use for sleep remains limited. Heterogeneity in cannabis types, doses, timing of administration, and sleep outcome measures limit the ability to make specific dosing recommendations.
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Rationale Stress is a risk factor for psychosis and treatments which mitigate its harmful effects are needed. Cannabidiol (CBD) has antipsychotic and anxiolytic effects. Objectives We investigated whether CBD would normalise the neuroendocrine and anxiety responses to stress in clinical high risk for psychosis (CHR) patients. Methods Thirty-two CHR patients and 26 healthy controls (HC) took part in the Trier Social Stress Test (TSST) and their serum cortisol, anxiety and stress associated with public speaking were estimated. Half of the CHR participants were on 600 mg/day of CBD (CHR-CBD) and half were on placebo (CHR-P) for 1 week. Results One-way analysis of variance (ANOVA) revealed a significant effect of group (HC, CHR-P, CHR-CBD (p = .005) on cortisol reactivity as well as a significant (p = .003) linear decrease. The change in cortisol associated with experimental stress exposure was greatest in HC controls and least in CHR-P patients, with CHR-CBD patients exhibiting an intermediate response. Planned contrasts revealed that the cortisol reactivity was significantly different in HC compared with CHR-P (p = .003), and in HC compared with CHR-CBD (p = .014), but was not different between CHR-P and CHR-CBD (p = .70). Across the participant groups (CHR-P, CHR-CBD and HC), changes in anxiety and experience of public speaking stress (all p’s < .02) were greatest in the CHR-P and least in the HC, with CHR-CBD participants demonstrating an intermediate level of change. Conclusions Our findings show that it is worthwhile to design further well powered studies which investigate whether CBD may be used to affect cortisol response in clinical high risk for psychosis patients and any effect this may have on symptoms.
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Lifestyle is a well-known environmental factor that plays a major role in facilitating the development of metabolic syndrome or eventually exacerbating its consequences. Various lifestyle factors, especially changes in dietary habits, extreme temperatures, unusual light–dark cycles, substance abuse, and other stressful factors, are also established modifiers of the endocannabinoid system and its extended version, the endocannabinoidome. The endocannabinoidome is a complex lipid signaling system composed of a plethora (>100) of fatty acid-derived mediators and their receptors and anabolic and catabolic enzymes (>50 proteins) which are deeply involved in the control of energy metabolism and its pathological deviations. A strong link between the endocannabinoidome and another major player in metabolism and dysmetabolism, the gut microbiome, is also emerging. Here, we review several examples of how lifestyle modifications (westernized diets, lack or presence of certain nutritional factors, physical exercise, and the use of cannabis) can modulate the propensity to develop metabolic syndrome by modifying the crosstalk between the endocannabinoidome and the gut microbiome and, hence, how lifestyle interventions can provide new therapies against cardiometabolic risk by ensuring correct functioning of both these systems.
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Withania somnifera (Ashwagandha) is an Ayurvedic herb categorized as having “rasayana” (rejuvenator), longevity, and revitalizing properties. Sensoril® is a standardized aqueous extract of the roots and leaves of Withania somnifera. Purpose: To examine the impact of Sensoril® supplementation on strength training adaptations. Methods: Recreationally active men (26.5 ± 6.4 years, 181 ± 6.8 cm, 86.9 ± 12.5 kg, 24.5 ± 6.6% fat) were randomized in a double-blind fashion to placebo (PLA, n = 19) or 500 mg/d Sensoril® (S500, n = 19). Body composition (DEXA), muscular strength, power, and endurance, 7.5 km cycling time trial, and clinical blood chemistries were measured at baseline and after 12 weeks of supplementation and training. Subjects were required to maintain their normal dietary habits and to follow a specific, progressive overload resistance-training program (4-day/week, upper body/lower body split). 2 × 2 mixed factorial ANOVA was used for analysis and statistical significance was set a priori at p ≤ 0.05. Results: Gains in 1-RM squat (S500: +19.1 ± 13.0 kg vs. PLA +10.0 ± 6.2 kg, p = 0.009) and bench press (S500: +12.8 ± 8.2 kg vs. PLA: +8.0 ± 6.0 kg, p = 0.048) were significantly greater in S500. Changes in DEXA-derived android/gynoid ratio (S500: +0.0 ± 0.14 vs. PLA: +0.09 ± 0.1, p = 0.03) also favored S500. No other between-group differences were found for body composition, visual analog scales for recovery and affect, or systemic hemodynamics, however, only the S500 group experienced statistically significant improvements in average squat power, peak bench press power, 7.5 km time trial performance, and perceived recovery scores. Clinical chemistry analysis indicated a slight polycythemia effect in PLA, with no other statistical or clinically relevant changes being noted. Conclusions: A 500 mg dose of an aqueous extract of Ashwagandha improves upper and lower-body strength, supports a favorable distribution of body mass, and was well tolerated clinically in recreationally active men over a 12-week resistance training and supplementation period.
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Gout is the most common form of inflammatory arthritis and is a multifactorial disease typically characterized by hyperuricemia and monosodium urate crystal deposition predominantly in, but not limited to, the joints and the urinary tract. The prevalence of gout and hyperuricemia has increased in developed countries over the past two decades and research into the area has become progressively more active. We review the current field of knowledge with emphasis on active areas of hyperuricemia research including the underlying physiology, genetics and epidemiology, with a focus on studies which suggest association of hyperuricemia with common comorbidities including cardiovascular disease, renal insufficiency, metabolic syndrome and diabetes. Finally, we discuss current therapies and emerging drug discovery efforts aimed at delivering an optimized clinical treatment strategy.
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Background Numerous physical, psychological, and emotional benefits have been attributed to marijuana since its first reported use in 2,600 BC in a Chinese pharmacopoeia. The phytocannabinoids, cannabidiol (CBD), and delta-9-tetrahydrocannabinol (Δ9-THC) are the most studied extracts from cannabis sativa subspecies hemp and marijuana. CBD and Δ9-THC interact uniquely with the endocannabinoid system (ECS). Through direct and indirect actions, intrinsic endocannabinoids and plant-based phytocannabinoids modulate and influence a variety of physiological systems influenced by the ECS. Methods In 1980, Cunha et al. reported anticonvulsant benefits in 7/8 subjects with medically uncontrolled epilepsy using marijuana extracts in a phase I clinical trial. Since then neurological applications have been the major focus of renewed research using medical marijuana and phytocannabinoid extracts. Results Recent neurological uses include adjunctive treatment for malignant brain tumors, Parkinson's disease, Alzheimer's disease, multiple sclerosis, neuropathic pain, and the childhood seizure disorders Lennox-Gastaut and Dravet syndromes. In addition, psychiatric and mood disorders, such as schizophrenia, anxiety, depression, addiction, postconcussion syndrome, and posttraumatic stress disorders are being studied using phytocannabinoids. Conclusions In this review we will provide animal and human research data on the current clinical neurological uses for CBD individually and in combination with Δ9-THC. We will emphasize the neuroprotective, antiinflammatory, and immunomodulatory benefits of phytocannabinoids and their applications in various clinical syndromes.
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The very low-frequency (VLF) band of heart rate variability (HRV) has different characteristics compared with other HRV components. Here we investigated differences in HRV changes after a mental stress task. After the task, the high-frequency (HF) band and ratio of high- to low-frequency bands (LF/HF) immediately returned to baseline. We evaluated the characteristics of VLF band changes after a mental stress task. We hypothesized that the VLF band decreases during the Stroop color word task and there would be a delayed recovery for 2 h after the task (i.e., the VLF change would exhibit a “slow recovery”). Nineteen healthy, young subjects were instructed to rest for 10 min, followed by a Stroop color word task for 20 min. After the task, the subjects were instructed to rest for 120 min. For all subjects, R-R interval data were collected; analysis was performed for VLF, HF, and LF/HF ratio. HRV during the rest time and each 15-min interval of the recovery time were compared. An analysis of the covariance was performed to adjust for the HF band and LF/HF ratio as confounding variables of the VLF component. HF and VLF bands significantly decreased and the LF/HF ratio significantly increased during the task compared with those during rest time. During recovery, the VLF band was significantly decreased compared with the rest time. After the task, the HF band and LF/HF ratio immediately returned to baseline and were not significantly different from the resting values. After adjusting for HF and LF/HF ratio, the VLF band had significantly decreased compared with that during rest. The VLF band is the “slow recovery” component and the HF band and LF/HF ratio are the “quick recovery” components of HRV. This VLF characteristic may clarify the unexplained association of the VLF band in cardiovascular disease prevention.
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The purpose of this study was to investigate whether the anxiolytic effect of cannabidiol (CBD) in humans follows the same pattern of an inverted U-shaped dose-effect curve observed in many animal studies. Sixty healthy subjects of both sexes aged between 18 and 35 years were randomly assigned to five groups that received placebo, clonazepam (1 mg), and CBD (100, 300, and 900 mg). The subjects were underwent a test of public speaking in a real situation (TPSRS) where each subject had to speak in front of a group formed by the remaining participants. Each subject completed the anxiety and sedation factors of the Visual Analog Mood Scale and had their blood pressure and heart rate recorded. These measures were obtained in five experimental sessions with 12 volunteers each. Each session had four steps at the following times (minutes) after administration of the drug/placebo, as time 0: -5 (baseline), 80 (pre-test), 153 (speech), and 216 (post-speech). Repeated-measures analyses of variance showed that the TPSRS increased the subjective measures of anxiety, heart rate, and blood pressure. Student-Newman-Keuls test comparisons among the groups in each phase showed significant attenuation in anxiety scores relative to the placebo group in the group treated with clonazepam during the speech phase, and in the clonazepam and CBD 300 mg groups in the post-speech phase. Clonazepam was more sedative than CBD 300 and 900 mg and induced a smaller increase in systolic and diastolic blood pressure than CBD 300 mg. The results confirmed that the acute administration of CBD induced anxiolytic effects with a dose-dependent inverted U-shaped curve in healthy subjects, since the subjective anxiety measures were reduced with CBD 300 mg, but not with CBD 100 and 900 mg, in the post-speech phase.
Article
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Although the application of medical marijuana and cannabinoid drugs is controversial, it is a part of modern-day medicine. The list of diseases in which cannabinoids are promoted as a treatment is constantly expanding. Cases of significant improvement in patients with a very poor prognosis of glioma or epilepsy have already been described. However, the occurrence of side effects is still difficult to estimate, and the current knowledge of the therapeutic effects of cannabinoids is still insufficient. In our opinion, the answers to many questions and concerns regarding the medical use of cannabis can be provided by pharmacogenetics. Knowledge based on proteins and molecules involved in the transport, action, and metabolism of cannabinoids in the human organism leads us to predict candidate genes which variations are responsible for the presence of the therapeutic and side effects of medical marijuana and cannabinoid-based drugs. We can divide them into: receptor genes-CNR1, CNR2, TRPV1, and GPR55, transporters-ABCB1, ABCG2, SLC6A, biotransformation, biosynthesis, and bioactivation proteins encoded by CYP3A4, CYP2C19, CYP2C9, CYP2A6, CYP1A1, COMT, FAAH, COX2, ABHD6, ABHD12 genes, and also MAPK14. This review organizes the current knowledge in the context of cannabinoids pharmacogenetics according to individualized medicine and cannabinoid drugs therapy.
Article
Background Cannabidiol has been used for treatment-resistant seizures in patients with severe early-onset epilepsy. We investigated the efficacy and safety of cannabidiol added to a regimen of conventional antiepileptic medication to treat drop seizures in patients with the Lennox–Gastaut syndrome, a severe developmental epileptic encephalopathy. Methods In this double-blind, placebo-controlled trial conducted at 30 clinical centers, we randomly assigned patients with the Lennox–Gastaut syndrome (age range, 2 to 55 years) who had had two or more drop seizures per week during a 28-day baseline period to receive cannabidiol oral solution at a dose of either 20 mg per kilogram of body weight (20-mg cannabidiol group) or 10 mg per kilogram (10-mg cannabidiol group) or matching placebo, administered in two equally divided doses daily for 14 weeks. The primary outcome was the percentage change from baseline in the frequency of drop seizures (average per 28 days) during the treatment period. Results A total of 225 patients were enrolled; 76 patients were assigned to the 20-mg cannabidiol group, 73 to the 10-mg cannabidiol group, and 76 to the placebo group. During the 28-day baseline period, the median number of drop seizures was 85 in all trial groups combined. The median percent reduction from baseline in drop-seizure frequency during the treatment period was 41.9% in the 20-mg cannabidiol group, 37.2% in the 10-mg cannabidiol group, and 17.2% in the placebo group (P=0.005 for the 20-mg cannabidiol group vs. placebo group, and P=0.002 for the 10-mg cannabidiol group vs. placebo group). The most common adverse events among the patients in the cannabidiol groups were somnolence, decreased appetite, and diarrhea; these events occurred more frequently in the higher-dose group. Six patients in the 20-mg cannabidiol group and 1 patient in the 10-mg cannabidiol group discontinued the trial medication because of adverse events and were withdrawn from the trial. Fourteen patients who received cannabidiol (9%) had elevated liver aminotransferase concentrations. Conclusions Among children and adults with the Lennox–Gastaut syndrome, the addition of cannabidiol at a dose of 10 mg or 20 mg per kilogram per day to a conventional antiepileptic regimen resulted in greater reductions in the frequency of drop seizures than placebo. Adverse events with cannabidiol included elevated liver aminotransferase concentrations. (Funded by GW Pharmaceuticals; GWPCARE3 ClinicalTrials.gov number, NCT02224560.)
Article
Background: Patients with Lennox-Gastaut syndrome, a rare, severe form of epileptic encephalopathy, are frequently treatment resistant to available medications. No controlled studies have investigated the use of cannabidiol for patients with seizures associated with Lennox-Gastaut syndrome. We therefore assessed the efficacy and safety of cannabidiol as an add-on anticonvulsant therapy in this population of patients. Methods: In this randomised, double-blind, placebo-controlled trial done at 24 clinical sites in the USA, the Netherlands, and Poland, we investigated the efficacy of cannabidiol as add-on therapy for drop seizures in patients with treatment-resistant Lennox-Gastaut syndrome. Eligible patients (aged 2-55 years) had Lennox-Gastaut syndrome, including a history of slow (<3 Hz) spike-and-wave patterns on electroencephalogram, evidence of more than one type of generalised seizure for at least 6 months, at least two drop seizures per week during the 4-week baseline period, and had not responded to treatment with at least two antiepileptic drugs. Patients were randomly assigned (1:1) using an interactive voice response system, stratified by age group, to receive 20 mg/kg oral cannabidiol daily or matched placebo for 14 weeks. All patients, caregivers, investigators, and individuals assessing data were masked to group assignment. The primary endpoint was percentage change from baseline in monthly frequency of drop seizures during the treatment period, analysed in all patients who received at least one dose of study drug and had post-baseline efficacy data. All randomly assigned patients were included in the safety analyses. This study is registered with ClinicalTrials.gov, number NCT02224690. Findings: Between April 28, 2015, and Oct 15, 2015, we randomly assigned 171 patients to receive cannabidiol (n=86) or placebo (n=85). 14 patients in the cannabidiol group and one in the placebo group discontinued study treatment; all randomly assigned patients received at least one dose of study treatment and had post-baseline efficacy data. The median percentage reduction in monthly drop seizure frequency from baseline was 43·9% (IQR -69·6 to -1·9) in the cannibidiol group and 21·8% (IQR -45·7 to 1·7) in the placebo group. The estimated median difference between the treatment groups was -17·21 (95% CI -30·32 to -4·09; p=0·0135) during the 14-week treatment period. Adverse events occurred in 74 (86%) of 86 patients in the cannabidiol group and 59 (69%) of 85 patients in the placebo group; most were mild or moderate. The most common adverse events were diarrhoea, somnolence, pyrexia, decreased appetite, and vomiting. 12 (14%) patients in the cannabidiol group and one (1%) patient in the placebo group withdrew from the study because of adverse events. One patient (1%) died in the cannabidiol group, but this was considered unrelated to treatment. Interpretation: Add-on cannabidiol is efficacious for the treatment of patients with drop seizures associated with Lennox-Gastaut syndrome and is generally well tolerated. The long-term efficacy and safety of cannabidiol is currently being assessed in the open-label extension of this trial. Funding: GW Pharmaceuticals.