Wetenschappelijk onderzoek Dr Joe Dispenza
Wetenschappelijk onderzoek
Dr Joe Dispenza heeft in samenwerking met Universiteiten onderzoek gedaan naar meditatie en het autoimmuun systeem.
Wetenschappelijk onderzoek hier te lezen.
Brain, Behavior, & Immunity - Health 32 (2023) 100675
Available online 7 August 2023
2666-3546/Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Meditation-induced bloodborne factors as an adjuvant treatment to
COVID-19 disease
Juan P. Zuniga-Hertz a,b,1
, Ramamurthy Chitteti a,b,1
, Joe Dispenza c
, Raphael Cuomo b
,
Jacqueline A. Bonds a,b
, Elena L. Kopp a,b
, Sierra Simpson b
, Jonathan Okerblom b
,
Svetlana Maurya d
, Brinda K. Rana e
, Atsushi Miyonahara b
, Ingrid R. Niesman f
,
Jacqueline Maree g
, Gianna Belza a,b
, Hillari D. Hamilton c
, Carla Stanton c
, David J. Gonzalez d,h
,
Michelle A. Poirier g
, Tobias Moeller-Bertram g
, Hemal H. Patel a,b,*
a Veterans Affairs San Diego Healthcare System, 3350 La Jolla Village Drive, San Diego, CA, 92161, USA b Department of Anesthesiology, University of California, San Diego, La Jolla, CA, 92093, USA c Encephalon, Inc., Rainier, WA, 98576, USA d Department of Pharmacology, University of California, San Diego, La Jolla, CA, 92093, USA e Department of Psychiatry, University of California, San Diego, La Jolla, CA, 92093, USA f San Diego State University, Electron Microscope Facility, 5500 Campanile Dr, San Diego, CA, 92182, USA g VitaMed Research, 44630 Monterey Ave., Palm Desert, CA, 92260, USA h Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California, San Diego, La Jolla, CA, 92093, USA
ARTICLE INFO
Keywords:
Meditation
Immunity
Adoptive blood transfer
serpin
SARS-CoV-2
Pseudovirus
COVID-19 disease
ABSTRACT
The COVID-19 pandemic has resulted in significant morbidity and mortality worldwide. Management of the
pandemic has relied mainly on SARS-CoV-2 vaccines, while alternative approaches such as meditation, shown to
improve immunity, have been largely unexplored. Here, we probe the relationship between meditation and
COVID-19 disease and directly test the impact of meditation on the induction of a blood environment that
modulates viral infection. We found a significant inverse correlation between length of meditation practice and
SARS-CoV-2 infection as well as accelerated resolution of symptomology of those infected. A meditation “dosing”
effect was also observed. In cultured human lung cells, blood from experienced meditators induced factors that
prevented entry of pseudotyped viruses for SARS-CoV-2 spike protein of both the wild-type Wuhan-1 virus and
the Delta variant. We identified and validated SERPINA5, a serine protease inhibitor, as one possible protein
factor in the blood of meditators that is necessary and sufficient for limiting pseudovirus entry into cells. In
summary, we conclude that meditation can enhance resiliency to viral infection and may serve as a possible
adjuvant therapy in the management of the COVID-19 pandemic.
- Introduction
Emergence and spread of the SARS-CoV-2 pandemic has resulted in
over 500 million cases of COVID-19 worldwide, leading to over 6 million
deaths (University). Spikes, surges, waves, and variants have continued
to upend the normalcy of life over the past two years, impacting work,
school, social interactions, travel, and the economy across the globe.
Though vaccines are mainstays in well-resourced countries, there is a
significant minority of individuals that will not participate in such
interventions due to political, ideological, trust, ethnicity, and socioeconomic inequity issues in resourced countries and fear of vaccine
safety and lack of availability in low and middle income countries
(Glazik et al., 2021; Omer et al., 2021). Several experimental compounds have been proposed for the treatment of COVID-19, including
hydroxychloroquine, remdesivir, lopinavir, interferon-β1, ivermectin,
convalescent plasma; however, many of these treatments result in more
harm than overall benefit (Bishara et al., 2020; Buonfrate et al., 2022;
Consortium et al., 2021; Ortigoza et al., 2021). No safe, effective,
* Corresponding author. Department of Anesthesiology, University of California, VA San Diego Healthcare System, #125, 3350 La Jolla Village Dr., San Diego, CA,
92161, USA.
E-mail address: hepatel@ucsd.edu (H.H. Patel). 1 These authors share equal first authorship.
Contents lists available at ScienceDirect
Brain, Behavior, & Immunity - Health
journal homepage: www.editorialmanager.com/bbih/default.aspx
https://doi.org/10.1016/j.bbih.2023.100675
Received 5 December 2022; Received in revised form 12 July 2023; Accepted 6 August 2023
Brain, Behavior, & Immunity - Health 32 (2023) 100675 - easily-applied approach with significant potential to limit initial infection or symptom severity has emerged.
Alternative and complementary approaches such as meditation can
improve the stress response and have a range of health benefits, presumably by creating a new tissue and blood environment that enhances
health and resilience. However, the latter aspect has not been directly
tested. Using such practices generally produces an abundance of benefit
with little to no harm potential. While there are many different schools
and forms of meditation, such as mindfulness meditation, yoga, and
breathing exercises, they all involve mental training with a goal of
transcendence of ordinary consciousness (Ospina et al., 2007). Guided
meditation is particularly beneficial because the very nature of instruction tends to relate to some specific purpose, such as healing or
self-improvement (Ospina et al., 2007). Meditative practices are associated with improvements in mental health conditions such as anxiety
and depression and physical symptoms like pain (Bohlmeijer et al.,
2010; Shapiro, 2009) and can have positive effects on specific markers of
inflammation, cell-mediated immunity, antibody responses, and biological aging (Black et al., 2019). Recent studies carried out in both
healthy and clinical populations have begun to shed light on the molecular changes associated with meditation and other mind-body practices (Gomutbutra et al., 2020; Buric et al., 2017). Collectively, these
studies have revealed that mindfulness-based practices result in downregulation of NF-kB, a key regulator of inflammation (Buric et al., 2017),
and upregulation of BDNF, a neuronal growth factor implicated in
neuronal plasticity (Gomutbutra et al., 2020), and in IRF-1, a gene
important in viral defense mechanisms (Feng et al., 2021).
Recently, hypotheses have been proposed to suggest the potential for
meditation to modulate SARS-CoV-2 and COVID-19, elegantly reviewed
by Mills, Chopra, and colleagues (Bushell et al., 2020). To this end,
Chandran et al. have explored the effect of a vegan diet and an intensive
8-day meditation workshop on gene expression (Chandran et al., 2021).
Data from this genomic study followed by multilevel bioinformatics
analysis demonstrated that this meditative practice leads to an upregulation of 220 genes associated with immune response and that this
meditation-specific network is significantly down-regulated in
COVID-19 patients. These data support the hypothesis that meditation
can serve as an effective complementary adjuvant therapy for COVID-19
disease.
Here, in an effort to better characterize the impact of meditation on
COVID-19, we administered a survey to members of a meditation
community who had previously attended a 7-day advanced meditation
workshop. To systematically identify possible factors that may be
induced by meditative practices employed by this community, we
collected blood plasma from attendees of three comparable meditation
workshops. Using cultured mammalian lung cells, we examined the effect of this plasma on the infectivity of SARS-CoV-2 pseudovirus,
including several variants of concern (VOC). Finally, we carried out
proteomics analysis on the plasma to identify potential factors induced
by meditation that may be specifically involved in protection against
viral infectivity.
2. Materials and methods
2.1. Survey data collection
After Institutional Review Board (IRB) review by the Western IRB
(now WCG-IRB), an internet survey platform was utilized to administer a
COVID-19-related survey online. Data were collected in accordance with
ethical guidelines pertaining to the use of human subjects. This survey
instrument was designed to assess mediation-related behaviors and
COVID-19 outcomes in a selective meditation community, as well as
potential confounders and mediators of these exposures and outcomes.
In addition, participants were asked about their general health status,
history of positive or negative COVID-19 diagnosis, symptoms experienced (if any), and current vaccination status. Meditation-related
behavioral covariates included length of experience with meditation
practice and frequency of meditation practice. Variables generated for
COVID-19 outcomes included self-assessed COVID-19 infection, COVID19 symptoms, duration of COVID-19 infection, COVID-19 vaccination
status, and experience of long-term symptoms. Data were also collected
on respondent demographics (race/ethnicity, sex, age), self-attribution
as a current or former smoker, and medical comorbidities (cancer,
stroke, diabetes, heart disease, vascular disease, lung disease, kidney
disease, liver disease, autoimmune disease, neurological disorder, and
obesity). Additionally, data were available for the respondent location.
Consent to participate in meditation-related research was obtained
through the survey instrument itself.
Available subject sampling was used to disseminate surveys to a
cohort of 22,459 individuals that had previously attended a 7-day
meditation-related workshop. Therefore, the sampling frame for
surveying was a population of individuals who had attended this 7-day
meditation workshop. Stratified sampling was not conducted, as the
purpose of the survey was to understand the relationship between
meditation-related behaviors and outcomes related to COVID-19. The
purpose of surveying was not to understand variations in these behaviors and outcomes across subgroups of meditators, though differences in
the relationship between these factors was assessed through stratification during the data analysis phase. Quota groups and exclusion criteria
were not programmed into the survey instrument.
2.2. Survey data analysis
Subject-Level Analysis: Bivariate regressions were conducted to
assess whether COVID-19 outcomes differed significantly for respondents across meditation-related behaviors. In addition, multivariable logistic regression models were computed to predict COVID-19
infection. Backwards selection using the Wald statistic was performed,
and Nagelkerke’s R2 and AIC were computed in a stepwise fashion to
assess model fit. All analyses were done in SPSS version 27 (IBM:
Armonk, New York).
Geospatial Analysis: Geospatial data were used to stratify respondents to determine if attendees of the workshop from different areas
exhibited different COVID-19-related outcomes in response to meditation. The purpose of this analysis was not to obtain representative statistics of behaviors or disease outcomes from different geospatial areas,
but rather to understand whether subject-level relationships between
meditation behaviors and COVID-19 outcomes were retained when
workshop participants were grouped into geospatial strata.
Country, first-level subnational unit (e.g., states/provinces), and city
of residence were manually entered by respondents. These were
manually reviewed by the authors for misspellings, concatenated into
one string per respondent, and processed using the Google Maps API to
generate latitude and longitude coordinates. These were plotted on a
projected coordinate plane and manually reviewed for face validity,
after which they were aggregated to polygons in a shapefile of geospatial
units. In order to ensure robust statistical analysis, geospatial units were
grouped such that they had a minimum of 10 respondents (n ≥ 10),
which is considered a sufficiently high sample size to minimize sampling
bias while also allowing for meaningful comparisons between the
groups. Aggregation was conducted first at the country-level, and then at
the first-level subnational entity for countries with n > 200 survey responses (United States), and finally at the county level for first-level
subnational entities with n > 200 survey responses (California, Florida). Percentages for meditation-related behaviors and COVID-19 outcomes were computed for each geospatial unit. Linear regression was
conducted to determine the bivariate relationship between meditationrelated behaviors and COVID-19-related outcomes across geospatial
strata among participants who attended this meditation workshop.
Statistical power was not available for multivariable modeling at the
ecological level, though these were conducted at the respondent level.
All data used for both subject-level and geospatial analyses were
J.P. Zuniga-Hertz et al.
Brain, Behavior, & Immunity - Health 32 (2023) 100675
3
obtained from the survey; no secondary data relating to COVID-19 rates
or meditation behaviors were utilized in this analysis. All geospatial
processing was done in ArcGIS Desktop version 10.7 (Esri: Redlands,
California).
2.3. Participant selection and meditation workshop
Novice and experienced healthy meditators and non-meditator
controls were recruited, following informed consent, from three meditation retreats held within the United States (n = 117, 6 did not complete
all biological collections and were excluded from the study). A total of
45 novice and 43 experienced meditators as well as 23 controls
completed all study requirements and were included in the molecular
studies. All experimental protocols were housed at VitaMed Research
(Palm Desert, CA) and approved by the Western Institutional Review
Board (WIRB; now WCG-IRB). All clinical records related to these
studies are housed at VitaMed Research as mandated by federal laws.
Research subject recruitment was random after a basic questionnaire set
was utilized to determine eligibility criteria. The only inclusion criteria
were having a current meditation practice incorporating the guided
specialized meditation practice used at the retreats, being over 21 years
of age, and in generally good health. Experienced meditators were
identified as having followed this guided specialized meditation practice
for more than 6 months and having attended a previous 7-day meditation retreat; novice meditators were identified as having followed the
technique for less than 6 months and having not attended a previous
retreat; control subjects were identified as having no meditation experience using this particular meditation practice and were vacationing
guests traveling with workshop participants. The 7-day advanced
meditation retreat consisted of approximately 25 h of instruction and 35
h of meditation focused around Kundilini techniques and containing
elements of focused attention, non-dual awareness, and loving kindness/
compassion meditation with a significant breathwork component.
Meditations were carried out seated, standing, lying down, and/or
walking. All subjects maintained a similar routine and time schedule and
ate similar breakfasts and lunches during the week. To ensure scientific
rigor and reproducibility, male and female subjects were used
throughout these studies.
2.4. Human plasma collection
Meditation blood plasma samples were collected at Indian Wells 02/
2020, Orlando 04/2021, and Denver 7/2021 retreats. All study subjects
fasted for at least 30 min prior to providing a blood sample. In total, 23
controls, 45 novice, and 43 experienced meditators completed all biological collections and were included in the analyses. Blood was
collected beginning at 12 p.m. and ending at 4 p.m. local time on retreat
Days 0 and 1 (pre-retreat samples) and on Days 7 and 8 (post-retreat
samples). All blood samples were stored at 4 ◦C (on wet ice) for no more
than 1 h after collection. Plasma was isolated by centrifugation at 3000
RPM for 15 min using an E8 Touch tabletop centrifuge, aliquoted into
1.5 ml Eppendorf tubes, and immediately frozen on dry ice. All samples
were stored and shipped to UC San Diego on dry ice.
2.5. Cell culture
Human lung epithelial cells line (A-549, ATCC© CCL-185) were
maintained in culture in F–12K supplemented media (10% FBS, 1% Pen/
Strep) at 37 ◦C, 5% CO2. Cells were maintained at passages 3–8 for all
experiments. For virus infection assays, 50,000 cells/well were cultured
in 96 well plates. For microscopy, 50,000 cells were plated in poly-Dlysin-coated glass-bottom 35 mm dishes. Mouse skeletal muscle cells
(C2C12, ATCC© CRL-1772) were used as a control cell line for viral
infection assays. Cells were maintained in DMEM supplemented media
(10% FBS, 1% Pen/Strep) at 37 ◦C, 5% CO2.
2.6. Viral constructs
Viral constructs were prepared at the UCSD Vector Development
Core Lab as described in Fig. 2a. Briefly, HIV-1 viral constructs were
engineered to express red fluorescence protein under the control of the
pCMV promoter. HIV-1 packaging plasmids were used in combination
with the viral envelope proteins and including the SARS-CoV-2.
SpikeΔ19 protein. The same procedure was used to prepare the variants
of SARS-CoV-2, including the Delta (B.1.617.2; HIV1.RFPCoV-2-SDelta), Beta (B.1351; HIV1.RFPCoV-2-S-Beta), U.K. (B.1.1.7; HIV1.
RFPCoV-2-S-UK), Brazil (P.1; HIV1.RFPCoV-2-S-Brazil), and D614G
(Wuhan-Hu-1 D614G; HIV1.RFPCoV-2-S-D614G) variants.
2.7. Immunofluorescence assays
A-549 (or C2C12 control) cells were seeded in 35 mm glass bottom
plates. After 24 h of culture, cells were treated with human plasma (1%
final concentration in 2 mL culture media). After 1 h of treatment, 1 ×
105 IU/mL of virus was added. Following 24 h of co-incubation, cells
were treated with NucBlue™ LiveReadyProbes™ Reagents (Hoechst
33342) for nuclear staining and observed at 100X in a live cell imaging
microscope (Keyence BZ-X700) coupled with an incubation chamber at
37 ◦C, 5% CO2 and controlled humidity. All sample groups remained
blinded until the completion of data analysis.
2.8. Scanning and transmission electron microscopy
Human lung epithelial cells were grown to confluence in six-well
plates with or without glass coverslips. Cells were fixed with 2.5%
glutaraldehyde, 4% paraformaldehyde in 0.1 M cacodylate buffer. For
SEM analyses, coverslips were washed with 0.1 M cacodylate buffer,
post-fixed with 1% OsO4, dehydrated, and dried overnight with
hexamethyl-D-silazane (HMDS). Coverslips were mounted and coated
with 6 nm platinum before viewing on an FEI Quanta 450 FEG SEM. For
TEM analyses, plates were washed with 0.1 M cacodylate buffer, postfixed with 1% OsO4, en-bloc stained with 1% uranyl acetate, follow