This study aims to examine the effects of acute aerobic exercise at low- and high-intensity on plasma concentrations of cleaved RAGE (cRAGE) and endogenous secretory RAGE (esRAGE) in young, healthy adults. The researchers hypothesize that a high-intensity bout of aerobic exercise will produce a greater increase in both forms of soluble RAGE compared to a low-intensity bout. Ten participants will perform acute bouts of exercise at 50% and 85% of their VO2max, matched for caloric expenditure. Blood samples will be collected before and after each bout to analyze cRAGE and esRAGE levels using ELISA kits. A two-way repeated measures ANOVA will evaluate differences between trials

1. Karia Coleman Jr.
KN 501
February 14, 2014

2. Brief Overview of article related to my Master’s Project
Brief Q&A about the article
Overview and Discussion of Master’s Project

3. Francesca Santilli; Natale Vazzana; Pierpaolo Iodice;
Stefano Lattanzio; Rossella Liani; Rosa Grazia
Bellomo; Gianfranco Lessiani; Francesca Perego;
Raoul Saggini; Giovanni Davì
Thrombosis and Haemostasis 2013
IF = 6.1

4. Is exercise good for you?
It depends on:
◦ Physio / Disease state
 Healthy
 Heart/Vascular Disease / Diabetes
◦ Training state
 Sedentary
 Physically Active
◦ Intensity of Exercise
 Low/Mod/High

5. Vigorous: exercise is usually defined as an absolute exercise
work rate of at least 6 metabolic equivalents (METs), which is
historically assumed to equal an oxygen uptake (VO2) of 21 mL ·
kg-1 · min-1;
 high intensity, unaccustomed exercise activities such as
jogging
- Thompson et al 2007 Circulation
 Regular Habitual: daily exercise physical conditioning
- Thompson et al 2007 Circulation

6.  The greatest amounts/intensities of Ex has greatest
benefits on HDL
 Kraus et al N Eng J Med 2002
 Strenuous Ex positively effects platelet activation
 Kestin et al Circulation 1993
 Regular habitual Ex improves haemostatic profile
 Wang et al Arterioscler Thromb Vasc Biol 2005
 Ex-induced gains in aerobic fitness decrease Ox stress
 Campbell et al Med Sci Sports Exerc 2010
 Chronic AE increase sRAGE
 Choi et al J Clin Endocrinol Metab 2012

7.  2 commonly known forms of sRAGE
◦ Cleaved RAGE (cRAGE)
 extracellular domain of full-length RAGE cleaved by
metalloproteinases
◦ Endogenous Secretory RAGE (esRAGE)
 mRNA variant lacks the COOH-terminal; secreted
extracellularly).
 Both soluble forms of RAGE have the capacity to bind to
AGE ligands attenuating cellular dysfunction and
inflammation

8. Protective Pro-Inflammatory
RAGE

9.  Hypothesis?
High-intensity-high-amount exercise program for two-
months in sedentary, low- or intermediate risk people
may exert beneficial effects on platelet activation
markers by modulating possible intermediate
determinants, including lipoprotein profile, lipid
peroxidation and the RAGE pathway

10.  N=22 15M/7F
 Age 51 (48 – 66) yr
 Sedentary
 BMI 25 (24.1 – 26.4) kg/m2
 T2DM n=0
 Hypertension n=9
 Beta Blockers n=1
 Statins n=0
 Framingham Risk Score 6.0 (3.0 – 14.8)

11. Exercise Training protocol
 Baseline plasma, serum, urine, VO2MAX
 1 mo Run-in
◦ (55% VO2MAX/45min  75% VO2MAX/55min)
 AE 2x/wk/8wk (direct supervision)
◦ Cycle ergometer, 55min/d, 75% VO2max
 Post 8wk plasma, serum, urine, VO2MAX

12. ◦ P-Selectin
 Platelet derived protein; promotes platelet aggregation
 Platelet:Platelet binding
◦ U-11-dehydro-TXB2
 Platelet activation
◦ 8-iso-PGF
 Measures Lipid peroxidation
◦ HDL
◦ esRAGE
 As a marker of oxidative stress – related inflammation

18.  HIHA AE is associated
with long-term changes in
platelet activation in this
population
 Maybe mediated by
effect on oxidative
stress-mediated
inflammation
 Each study only
examined 1 form of
sRAGE
 Varying intensities not
examined
 Is the intensity high? 75%
VO2MAX?

20.  2 commonly known forms of sRAGE
◦ Cleaved RAGE (cRAGE)
 extracellular domain of full-length RAGE cleaved by
metalloproteinases
◦ Endogenous Secretory RAGE (esRAGE)
 mRNA variant lacks the COOH-terminal; secreted
extracellularly).
 Both soluble forms of RAGE have the capacity to bind to
AGE ligands attenuating cellular dysfunction and
inflammation

21. Protective Pro-Inflammatory
RAGE

22. Santilli et al
Throm and Haem 2013
Choi et al JCEM 2012

23. Non-Exercise Control N=38; Exercise N=37
BMI ~ 26, Age ~55 yrs
Aerobic Exercise Training ↑ sRAGE ~ 15%
Choi et al., JCEM 2012
Haus et al ; Unpublished
65 y; BMI 35 kg/m2; sedentary
60min/d; 5d/wk for 12 wks at 85% HRmax

24. sRAGE 27% sRAGE 29%
Danzig et al .Physiol Res 2009
Con n=22, 30 y
CAD n=21, 65 y
BMI: N/A

25. To examine the effects of acute aerobic exercise at low
and high intensity on plasma concentrations of cRAGE
and esRAGE in a young lean healthy population
We hypothesize that the high intensity bout of AE will
produce a greater increase in both forms of sRAGE
(cRAGE & esRAGE) compared to the low intensity bout.
Hypothesis

26.  n=10
 Equal number of males and females
 Age ≤ 35yr
 BMI ≤ 27
 Sedentary
 No Chronic Disease
 No Malignancies in past 5 yr
 No Smoking
 No High Alcohol consumption ≥ 5/wk

27. 3 d dietary log
Blood Draws
50% VO2Max
85% VO2Max
VO2Max
50%
85%
50%
85%VO2Max
T= - 3 T=0 T=4 T=7 T=11 T=14
Study Design
Subject Enrollment
IFC
Randomization
DEXA
Visit 1 Visit 2 Visit 3 Visit 4
1 d diet standard.

28.  Metabolic Controls
 3 day dietary log
 1day diet standadization
 No unaccustomed Ex 72 hr
prior visits
 No alcohol 48 hr prior visits
 No caffeine 24 hr prior visits
 Fasted minimum 8 hrs
 Ex Protocol
 VO2MAX via modified Bruce TM
◦ Cosmed System
 AE Treadmill
◦ 50% VO2MAX
◦ 85% VO2MAX
◦ Cosmed System
 Match Work between trials
◦ Kcal Expenditure

29. Matched Work via Caloric Expenditure
VO2MAX
Total Kcals
85%VO2MAX
Matched Kcals
to VO2max
(X) Minutes
50%VO2MAX
Matched Kcals
to VO2max
(y) Minutes

30. Primary Secondary
 sRAGE
◦ R&D Systems Inc
ELISA Kit
 esRAGE
 B-Bridge International
ELISA Kit
cRAGE = sRAGE (total) – esRAGE
 DEXA
 VO2MAX
 Complete Metabolic Panel
 HBA1c
 Complete Blood Counts
 Dietary Logs
 Phys Act Recall

31.  Two-way ANOVA (trial x time) with repeated
measures
 Bivariate correlation analysis with baseline descriptive
variables
 MEAN±SEM, p<0.05