UCSD - EPARC Labs Yoga Study
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Transcript of UCSD - EPARC Labs Yoga Study
Project Yeah
Research Questions: Slide 1• Does 3 months of yoga improve fitness, depression, cardiovascular
indicators, weight, etc. in previously sedentary individuals?• Do experienced yoga practitioners have better fitness, depression,
weight, bp than non-exercisers?• Does 3 months of yoga increase parasympathetic tone in novices?• Do experienced yoga users have more parasympathetic tone than
non-exercisers?
Research Questions: Slide 2• Does 3 months of yoga reduce inflammation in systemic, molecular,
and cellular measures in novices?• Do experienced yoga practitioners have less inflammation than non-
exercisers?• Is PNS tone associated with inflammation in systemic, molecular
and cellular measures?
Design• 1. Prospective 3 month intervention for non-exercisers randomized
to yoga or aerobic walking. Comparison of pre-post, and between groups.
• 2. Cross sectional study comparing experienced yoga practitioners with non-exercisers
Inclusion criteria: Non-exercisers
• Female• 25-45 y/o• Not currently engaged in regular physical
activity
Inclusion Criteria: Yoga experts
• Female• 20-45 y/o• Have practiced yoga for 3+ years• Currently practicing at least twice a week
Exclusion criteria
• Alcohol abuse• Drug use/smoking• Prescription drugs considered on case by case• Heart disease• Postmenopausal• Pregnant• Most chronic diseases• Major mental health diseases
Recruitment strategies
• Flyers• Outreach• Listserve• Outreach to yoga studios• Incentives• Total number of ‘touches’ to recruit the
participants: 152 for intervention and 21 for experienced (control) group
RecruitmentPHASE 1: 152 people contacted to recruit 34
– 23% success rate– 21 outside age range (9 old, 12 young)– 50 had schedule conflicts– 11 had exclusionary diseases– 33 “other” (included drug user, male, unwilling to donate blood, pregnant)
Of the 36 recruited, in cohort 1 (n=19)3 no-shows, 1 refused to give blood, 8 randomized to yoga—1 dropped after cited scheduling conflicts7 randomized to walking—4 dropped after (1 moved away, 3 said they thought they would get to do yoga)
In cohort 2 (n=15) 4 no shows All put in yoga group—4 later drops all cited schedule
PHASE 2: Experienced yogi’s 21 people contacted to recruit 10 (details of why not eligible not kept)
IRB Process
• First submission• Consents approved and stamped• Surveys approved• Flyers approved• Revised submission• Process repeated (4 times—including
extension)
Variables
• Demographic and Social History• Behavior• Complementary and Alternative Medicine use • Fitness measures• Biological measures• Cardiac measures• Biological blood work• Inflammatory markers
Basic Biometrics
• Weight• Height• BMI
Demographic & Social History
• Age• Gender• Education• Income• Social status• Medications• Reproductive history• Chronic diseases
Fitness and Cardiovascular Measures
• 3 minute step test • Blood Pressure and Heart Rate at rest, peak, and
during recovery measured by EKG
• Heart Rate Variability gathered via Zephyr Bioharness IV
Behavior & Lifestyle
• Exercise• Smoking history• Drug use• Physical Activity
• Questionnaire• Accelerometer (pre and post)
• Sedentariness
Use of Complementary & Alternative Medicine
• Nutritional modalities• Mind body modalities• Movement modalities• Supplements
Blood based markers of Inflammation
• Complete Blood Count (CBC) Complete Metabolic Panel (CMP) HgA1C: Measure of long-term blood
glucose control. C-Reactive Protein (CRP) Cortisol serum
Cellular and Molecular Markers of Inflammation
Stat3 – cell growth and apoptosis
1:http://www.cellsignal.com/productListing?Ntt=stat3
• required for murine fetal development.
• possesses oncogenic potential
• anti-apoptotic activities.1
Nuclear membrane
Surface receptor
Cell Surface
Participants
• Pre-Post novice yoga completers: 13• 14 completers, but one participant removed from
analysis due to development of diabetes during intervention period.
• Pre-Post novice walkers completers: 3• Pre-novices non-completers: 9
• 5 yoga and 4 walking• Experienced yoga practitioners: 10
YOGA INTERVENTION
• Participants practiced of a blend of Ashtanga and Hatha forms of yoga with a strong focus on breathing throughout the practice.
• Participants were asked to attend 2 classes and practice at least once a week on their own using the provided video. –The video was a 20 minute practice. –Classes lasted for 60 minutes.
RESULTS
Characteristics of Participants at Baseline All (n=26) Yoga C (n=13) Yoga NC (n=5) Aerobic C (n=3) Aerobic NC (n=4)
Age 31.75 5.72 33.75 4.47 28.67 3.09 25.33 0.58 25.00 n/aHt (cm) 164.60 7.41 167.96 7.28 162.52 8.18 162.63 5.94 160.15 5.80
Wt (lbs) 149.54 33.33 150.19 40.38 156.12 21.52 127.67 29.30 142.50 68.43
BMI 25.24 6.37 25.35 7.28 27.25 5.94 21.77 3.42 25.11 3.84
HR Rest (bpm) 77.57 11.32 75.31 10.36 73.00 12.30 89.00 10.58 69.00 n/aHR Peak (bpm) 164.87 11.31 161.54 12.55 167.00 5.29 162.00 2.00 180.00 n/a
%MVPA 3.58 2.19 3.85 2.31 3.92 1.90 3.37 2.11 n/a n/a
%Sed 67.32 10.14 66.07 8.92 69.18 19.35 73.85 1.83 n/a n/a
pNN50 11.82 9.95 10.88 8.82 15.58 9.95 5.09 2.83 n/a n/a
* Values are reported as means and standard deviations
Univariate: yoga novices (pre/post)Variable Pre-
(13-14)Post- (13-14)
Mean SD Mean SD p-value
SBP rest 118 5.8 116 7.9 0.912
Weight 150 40.3 150 42.5 0.00
SBP_pk 117.7 5.9 115.5 7.5 0.515
HR rest 75 10.3 71 10.0 0.102
HR peak 161.5 12.5 155.5 16.3 0.002
HR rec1 123 18.4 117 17.1 0.034
HR rec2 103 16.3 99.3 17.2 0.001
pNN50 10.88 8.82 9.72 6.43 0.340
A1C 5.2 0.25 5.4 0.24 0.043
*Calculated using T-testClinical Significance: • Rest HR down 4 bpm• Peak HR dwon 6 bpm (statistically
significant too)
May also be meaningful
• Sedentary time (self report) down 8%
OTHER MARKERS OF INFLAMMATIONPre-yoga (14) Post-yogaMean SD Mean SD p-value
Cortisol (12) 14.25 5.40 16.44 6.50 0.198
CRP (6) 0.640 0.625 0.403 0.471 0.443
WBC (13) 6.3 1.87 5.86 1.234 0.297
55
60
65
70
75
80
85
90
95
100HR REST (PRE_POST)
TIME POINT (LEFT PRE, RIGHT POST)
HR (B
PM)
120
130
140
150
160
170
180
190
HR PEAK
TIME POINT (LEFT =PRE RIGHT=POST)
HR (B
PM)
Characteristics of Yogis
Advanced Yogis (n=10)Mean SD
Age 37.50 6.84Ht (cm) 165.90 10.31Wt (lbs) 140.39 16.75BMI 23.26 2.68
Bivariate: novice vs experiencedVariable Cases (non exercise)
19-20Controls (yogi) 9 p-value
Mean SD Mean SD
SBP (rest) 117.7 5.879 110.2 6.36 0.010
SBP (rec-2 min)
121.8 3.4 118.2 8.45 0.194
HR (rest) 75.31 10.35 66.22 6.3 0.030
HR (peak) 161.5 12.09 152.11 10.73 0.082
HR (rec-2 min) 103.4 16.32 91.11 15.77 0.094
pNN50 11.53 8.77 16.93 12.91 0.132
Clinical Significance: • HR Recovery less by 17 bpm at min 1 & 2• Less time doing sedentary activity (accelerometer 4%, self-
report 14.5%)• Pnn50 50% greater in experienced (11.5% vs 16.9%)
*Calculated using non-paired T-test
System Level ConclusionsModest changes in key variables in 12 weeks, but encouraging trends
Large differences in key variables between experienced and non
CELLULAR & MOLECULAR TESTING
Goal: Develop a molecular workflow to analyze the immune response in human blood to Yoga versus other exercise.
For this project• Feasibility• Generate preliminary data on which later grant applications
can be based• Generate Bio-repository for future gene expression studies
Analysis
Cell Signaling
Cell responsiveness to Nicotine and IL-6
• Nicotine- classic agonist for a7 nACHR
• IL-6- classic agonist for STAT3 pathway
Analysis
Cell Signaling
Biorepository
Cell responsiveness to Nicotine and IL-6
Magnetic bead column separation Quality Control
Monocyte and T Cell count
• Nicotine- classic agonist for a7 nACHR
• IL-6- classic agonist for STAT3 pathway
RNA analysis through RT-PCR
Flow Cytometry
Baseline
Post-Exercise
Draw Antibody Staining
Cell Activation Analysis
2
How:1. Cell Signaling
Flow Cytometry
BaselinePost-Exercise
Blood draw Antibody Staining
Magnetic Bead Separation
Cell Type and Activation Analysis
RNA Analysis
10 2
How:
2. Biorepository
Cell Signaling• Stat3 – cell growth and apoptosis
1:http://www.cellsignal.com/productListing?Ntt=stat3
• required for murine fetal development.
• possesses oncogenic potential
• anti-apoptotic activities.1
Nuclear membrane
Surface receptor
Cell Surface
Cell Signaling• Stat3 – cell growth and apoptosis
SignalingStat3
Low Signal
High Signal
Low Cell Count
High Cell Count
Median Fluorescent Intensity (MFI)• “middle” of curve• Very standard statistic in literature (>10,000 events)• Less affected by outliers
Fluorescent Intensity
Quantification
*n=6 P=0.0122, Paired t-test
*
Nicotine P-STAT3 response ratio reduced post-YOGA (n=6)
Change in response to nicotine after exercise• Each point on scatterplot expressed as ratio of MFI post Nicotine to
MFI sham• Statistically significant Decrease in STAT3 activation after exercise
Raw data…
Patient ID Baseline Pre Nicotine Pre Ratio Baseline Post Nicotine Post Ratio201-7 3250 4998 1.537846154 993 1130 1.13796576
201-9 1472 2020 1.372282609 551 612 1.110707804
201-11 585 1379 2.357264957 276 549 1.989130435
201-19 1013 1878 1.853899309 729 375 0.514403292
201-21 953 2209 2.317943337 632 747 1.181962025
201-23 1454 3008 2.068775791 2740 2062 0.752554745
Baseline Raw data…
*n=6 P=0.315, Paired t-test
*
*n=6 P=0.0084, Paired t-test
*
IL-6 P-STAT3 response ratio increased post-YOGA (n=6)
Raw data…
Patient ID Baseline Pre IL-6 Pre Ratio Baseline Post IL-6 Post Ratio201-27 2054 5600 2.726387537 1508 6833 4.531167109
201-29 4096 7440 1.81640625 1649 4266 2.587022438
201-33 4063 9689 2.384691115 1704 6429 3.772887324
201-35 1972 4703 2.384888438 1352 6173 4.565828402
201-39 7625 5738 0.75252459 5976 15085 2.524263722
201-41 6508 9969 1.531807007 720 4683 6.504166667
Change in response to IL-6 after exercise• Each point on scatterplot expressed as raw MFI post IL-6• Statistically significant decrease in response ratio and Baseline
Baseline Raw data…
*n=6 P=0.0313, Paired t-test
*
Conclusion 1 of 2
• We have a way to assess the responsiveness of a cell to an agonist that is relevant to the human inflammation signaling before and after exercise
• There is preliminary data to suggest that Yoga might lower STAT3 activation
Analysis
Cell Signaling
Cell responsiveness to Nicotine and IL-6
• Nicotine- classic agonist for a7 nACHR
• IL-6- classic agonist for STAT3 pathway
Biorepository
Magnetic bead column separation Quality Control
Monocyte and T Cell count
RNA analysis through RT-PCR
Flow Cytometry
Baseline
Post-
Exercise
Blood Draw Antibody Staining
Magnetic Bead Separation
Cell Type and Activation Analysis
RNA Analysis
10 2
How:
Gene Expression• Stained for CD14+ monocytes• Average 3.3% of blood• Purification of CD14+ monocytes through magnetic bead separation
for RNA analysis
Biorepository
Conclusion 2 of 2• We can detect gene expression in fractions of peripheral blood
• 10mL is likely a rate limiting amount for magnetic bead separation
Next Steps?• Continue with BioRepository and RT-PCR
• Correlations with Blood draw data and demographics
• Finish analysis of Experienced practitioners
• What else needed for publication?
Other outcomes
Acknowledgements• David Wing – blood collection• Ann-Marie Hageny – cell staining and flow cytometry• Emelie Amburn – RNA prep• Glory BUI – cDNA, PCR and gel• Matthew Eliceiri – Design, Analysis and Summary• Dr. Linda Hill-Study PI• CalIT2-Funding Support
TEAMLinda HillJeanne NicholsAndrew BairdBrian EliceiriAnne Marie HagenyDenise GallagherDavid WingElizabeth FriedmanEmelie AmburnGlory BUIMatthew Eliceiri Walking leaders/Yoga teachersTravis NguyenGraeme BairdJill WaalenLaura Schmalzl
Additional input• Deborah Forster• Ramesh Rao• Paul Mills• Giorgio• Etc, need to list
Analysis challenges
• HRV–zephyr–bliss buzzer
• Multiple variables• Two labs: Hillcrest/Quest• Yoga studies: synergy, variation in type**
Other challenges and limitationsBudgetRecruitmentStaffingBuilding limitations
Next steps• Publications
• Changes in fitness, depression, activity• Changes in inflammation• Challenges with HRV
• Grant proposals