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The effect of ElevATP™on whole blood ATP levels: a single dose, cross over clinical study.

  • January 2014
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Tania Reyes at FutureCeuticals Inc.
Tania Reyes
  • FutureCeuticals Inc.
C. Shu
C. Shu
C. Shu
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Ruby Argumedo at FutureCeuticals
Ruby Argumedo
  • FutureCeuticals
B. Nemzer
B. Nemzer
B. Nemzer
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Abstract and Figures

Purpose: elevATP™, a supplement containing plant-derived inorganic microelements and apple polyphenols, was previously shown to increase endogenous whole blood ATP levels in healthy human subjects. In this report, we tested the supplement in a larger cohort and assessed the effect of the supplement in muscle. Methods: Twenty healthy, fasted, and resting adult human subjects participated in this acute, placebo-controlled, single-dose crossover clinical study. Oral placebo was administered on the first day of testing followed by a single, 150 mg dose of elevATP™ on the second day. Blood was collected immediately prior to treatment, 60 and 120 minutes after ingestion. Whole blood ATP, plasma ATP, hemoglobin, blood lactate, and blood glucose levels were collected. A muscle biopsy was performed on one resting study subject before, and 60 and 120 minutes after, a single dose of elevATP™. Results: elevATP™ increased whole blood levels of ATP by 40% after 60 minutes (p<0.0001) and by 28% after 120 min (p=0.0009) versus baseline, pre-supplementation levels. ATP plasma levels did not increase after elevATP™ administration under these experimental conditions. Intramuscular ATP levels from biopsy of one patient increased significantly at 60 and 120 minutes after ingestion of elevATP™ and reached higher levels than ATP measured in whole blood. Conclusions: These results indicate that elevATP™ increases intracellular ATP in blood cells, confirming results from a previous study, and suggest that it may increase ATP in muscle tissue. Further clinical testing is needed to confirm tissue- and organ-specific changes in ATP levels following ingestion of elevATP™.
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Journal of Aging Research & Clinical Practice©
THE EFFECT OF ELEVATP™ ON WHOLE BLOOD ATP LEVELS:
A SINGLE DOSE, CROSSOVER CLINICAL STUDY
T. Reyes-Izquierdo1, C. Shu1, R. Argumedo1, B. Nemzer2, Z. Pietrzkowski1
Introduction
Adenosine 5’-triphosphate (ATP) is the active source of
energy within cells and participates in numerous
physiological processes. Extracellular ATP and ADP
influence platelet aggregation, vascular tone, and
nervous, cardiac and muscle tissue function (1). ATP is
critical for cell-to-cell communication and participates in
immune cell coordination (2-5). As humans age,
intracellular ATP levels decrease and the ability to
generate ATP is diminished (6-8). Recent evidence also
suggests that aging humans have lower plasma and
erythrocyte-mediated release of ATP, most notably
during periods of increased skeletal muscle blood flow
(9). This may impair vasodilation and oxygen delivery to
the tissues (9).
The clinical effects of oral ATP administration have
been mixed (10, 11). Researchers have failed to
demonstrate that orally administered ATP supplements
increase blood ATP levels (11, 12). This is likely due to
the poor bioavailability of oral ATP (13), since
intravenous administration of ATP has been reported to
increase ATP levels (14). Long term ATP
supplementation may induce intestinal nucleoside
transporters in humans, thereby increasing absorption
(15). However, enteric-coated and distal-releasing ATP
supplements have also failed to increase blood ATP
levels (12). Because of the unpredictable mechanism of
ATP dosing using direct ATP supplementation,
researchers, including our group, are exploring strategies
to increase endogenous ATP levels.
ElevATP™ is an ancient peat-based bioinorganic
material blended with apple extract polyphenols, as
previously described (16). Our pilot study demonstrated
that a single dose of elevATP™ increased whole blood
ATP levels in human subjects without changing reactive
oxygen species, glucose, or lactate levels in the blood (16).
This study extends that work using a crossover design in
a larger group of subjects. We sought to confirm the
results of our pilot study and to evaluate plasma levels of
ATP as well. In addition, biopsy of muscle tissue was
collected from one individual before and after a single
dose of elevATP™ in order to determine intramuscular
ATP levels.
1. Applied BioClinical Inc., 16259 Laguna Canyon Rd, Irvine, CA, USA 92618; 2.
FutureCeuticals Inc., 2692 N. State Rt. 1-17., Momence, IL, USA 60954
Corresponding Author: Tania Reyes-Izquierdo, 16259 Laguna Canyon Rd, Irvine CA,
92618 USA, Phone +1 949 502 4496, Fax +1 949 502 4987, Email:
tania@abclinicaldiscovery.com
1
Abstract: Purpose: elevATP™, a supplement containing plant-derived inorganic microelements and apple polyphenols, was
previously shown to increase endogenous whole blood ATP levels in healthy human subjects. In this report, we tested the
supplement in a larger cohort and assessed the effect of the supplement in muscle. Methods: Twenty healthy, fasted, and resting
adult human subjects participated in this acute, placebo-controlled, single-dose crossover clinical study. Oral placebo was
administered on the first day of testing followed by a single, 150 mg dose of elevATP™ on the second day. Blood was collected
immediately prior to treatment, 60 and 120 minutes after ingestion. Whole blood ATP, plasma ATP, hemoglobin, blood lactate, and
blood glucose levels were collected. A muscle biopsy was performed on one resting study subject before, and 60 and 120 minutes
after, a single dose of elevATP™. Results: elevATP™ increased whole blood levels of ATP by 40% after 60 minutes (p<0.0001) and
by 28% after 120 min (p=0.0009) versus baseline, pre-supplementation levels. ATP plasma levels did not increase after elevATP™
administration under these experimental conditions. Intramuscular ATP levels from biopsy of one patient increased significantly at
60 and 120 minutes after ingestion of elevATP™ and reached higher levels than ATP measured in whole blood. Conclusions: These
results indicate that elevATP™ increases intracellular ATP in blood cells, confirming results from a previous study, and suggest
that it may increase ATP in muscle tissue. Further clinical testing is needed to confirm tissue- and organ-specific changes in ATP
levels following ingestion of elevATP™.
Key words: Blood ATP, plasma ATP, micronutrients, polyphenols.
Received November 15, 2013
Accepted for publication November 20, 2013
REYES-IZQUIERDO_04 LORD_c 04/12/13 08:41 Page1
Materials and Methods
elevATP™ was provided by FutureCeuticals, Inc.,
Momence, IL USA. Dulbecco's phosphate buffered saline
(PBS), phenyl methane-sulfonyl-fluoride (PMSF),
dimethyl sulfoxide (DMSO), leupeptin, EDTA, NaCl,
nitrobenzyl thioinosine (NBTI), KCl, tricine, forskolin,
isobutylmethylxanthine (IBMX) and water were
purchased from Sigma Chem. Co. (St Louis, MO, USA).
ATP stabilizing solution was prepared as described by
Gorman et al. (17) (118 mmol NaCl, 5 mmol KCl, 40 mmol
tricine buffer, 4.15 mmol EDTA, 5 nmol NBTI, 10 µmol
forskolin and 100 µmol IBMX, at pH 7.4 adjusted with 2
mol/L KOH).
Low protein binding microtubes were obtained from
Eppendorf (Hauppauge, NY, USA) and RC DC Protein
Assay Kit II was obtained from Bio-Rad (Palo Alto, CA,
USA). ATP-luciferase assays were obtained from
Calbiochem (San Diego, CA, USA). Heparin capillary
blood collection tubes were obtained from Safe-T-Fill®
(Ram Scientific Inc. Yonkers, NY). A portable gas meter
and CG8+ cartridges were obtained from Abbott
Laboratories (Abbott Park, IL, USA). Total hemoglobin
quantification ELISA kits were obtained from
MyBiosource (San Diego, CA, USA). Accutrend® Lactate
Point of Care and BM-Lactate Strips® were obtained from
Roche (Mannheim, Germany). Accu-Chek® Compact
Plus glucometer and Accu-Chek® test strips were
obtained from Roche Diagnostics (Indianapolis, IN, USA).
Clinical Study
Inclusion and Exclusion Criteria
This clinical case study was conducted according to
guidelines laid out in the Declaration of Helsinki. All
procedures involving human subjects were approved by
the Institutional Review Board at Vita Clinical S.A.
Avenida Circunvalacion Norte #135, Guadalajara, JAL,
Mexico 44 270 (study protocol no. ABC-13-09-ATP).
Twenty subjects were selected to participate. They were
generally healthy, and free of rhinitis, influenza, and
other acute infections. 12 female and 8 male subjects were
selected, with ages ranging from 22 to 35 years and BMI
ranging from 24.1 to 30 kg/m². Exclusion criteria
included diagnosis of diabetes mellitus, allergies to
dietary products, use of anti-inflammatory drugs,
analgesics, statins, diabetic drugs, anti-allergy medicines,
multivitamins, and use of supplements within 15 days of
the start of the study. All participants gave written,
informed consent before any experimental procedure was
performed.
Blood Collection
Enrolled participants were instructed not to eat for 12 h
prior to the initial blood draw. Resting subjects were
given an empty capsule as placebo on Day 1 of the study
and 150 mg of encapsulated elevATP™ on Day 2. 250 mL
of water was administered with the capsules each day.
Two hundred microliters of blood was collected by finger
puncture and placed in Safe-T-Fill® Capillary blood
collection tubes (Ram Scientific Inc. Yonkers, NY). Blood
samples were collected immediately prior to test capsule
administration and at 60 and 120 minutes after ingestion.
Plasma Collection for ATP analysis
One hundred µl of blood was transferred to low-
protein binding tubes (Eppendorf, Hauppauge, NY, USA)
immediately after collection. An equal volume of ATP
stabilizing solution (17) was added to each tube. Tubes
were gently mixed by inversion and centrifuged at 13,000
g for 3 min to pellet cells. Supernatant was transferred to
a clean tube and spun again at 13,000 g for 3 min. The
supernatant was then snap frozen and stored at -80°C
prior to ATP analysis.
ATP Detection and Quantification
Blood ATP or plasma ATP concentration were
determined using ATP Assay Kits (Calbiochem, San
Diego, CA, USA) with a modification to the original
method, as previously described[18]. Briefly, 10 μL of
lysed blood or plasma was loaded onto a white plate
(Corning® Fisher Scientific, Waltham, MA, USA). 100 µL
of ATP nucleotide-releasing buffer containing 1 µL
luciferase enzyme mix was added and the plate
immediately placed on a illuminometer (LMaX,
Molecular Devices; Sunnyvale CA, USA). Readings were
performed for 15 min at 3 min intervals, at 470 nm.
Relative Light Units (RLU) were recorded and ATP
concentrations were determined using a standard ATP
curve.
Hemoglobin Measurement
Hemoglobin levels in plasma were determined using a
double sandwich ELISA (MyBiosource, San Diego, CA,
USA), concentration was determined comparing to a
standard curve, according to the manufacturer’s
instructions. Plasma samples collected with the ATP
stabilizing solution were used for this analysis.
Lactate and Glucose Detection
Blood lactate was measured using an Accutrend®
Lactate Point of Care (Roche, Mannheim, Germany) and
BM-Lactate Strips® (Roche, Mannheim, Germany).
Fifteen µL of blood was loaded onto the strip and lactate
levels were read according to the manufacturer’s
instructions. Glucose was measured using an Accu-
EFFECT OF ELEVATP ON BLOOD ATP LEVELS.
2
REYES-IZQUIERDO_04 LORD_c 04/12/13 08:41 Page2
JOURNAL OF AGING RESEARCH AND CLINICAL PRACTICE©
3
Chek® Compact Plus glucometer (Roche Diagnostics,
Indianapolis, IN, USA) and Accu-Chek® test strips
(Roche Diagnostics, Indianapolis, IN, USA). Glucose was
read according to the manufacturer’s instructions. Lactate
and glucose levels were determined at every collection
time point.
Statistical Analysis
For each result obtained from the described assays,
each subject was normalized to their own value
measured at baseline (T0), before ingestion of elevATP™
or placebo. Levels of each assay at 60 (T60) and 120 (T120)
minutes after treatment were compared within
experimental groups to the baseline and between
experimental groups using a paired t-statistic test.
Descriptive analyses were run in GraphPad® to derive
the mean and standard deviation of each group.
Muscle Biopsy and Surgical Procedure
For the muscle biopsy, one twenty two-year old
healthy subject, with a BMI of 24.5 was recruited,
following the same selection criteria as described for the
clinical crossover study. This clinical case study was
conducted according to guidelines laid out in the
Declaration of Helsinki. This procedure was approved by
the Institutional Review Board at Vita Clinical S.A.
Avenida Circunvalacion Norte #135, Guadalajara, JAL,
Mexico 44 270 (study protocol no. ABC-NCI-13-01-ATP-
Mus1). The study subject was selected from the group of
subjects enrolled for whole blood ATP measurement, as
described above. This subject was fasted and resting
during this experiment. Biopsy was performed using
aseptic technique. An antiseptic solution (Isodine) was
applied to the medial region of the arm, over the right
biceps. An 18 g needle was used to infiltrate the skin with
5 cc lidocaine. A skin incision was made using a 3 mm
skin biopsy punch. Subcutaneous tissue was bluntly
divided, allowing resection of 3 mm2 of biceps muscle
using Metzenbaum scissors. Muscle tissue from the
biceps was collected before, and also 60 and 120 minutes
after, ingestion of elevATP™. Muscle tissue was
deposited in a 50 ml conical tube and frozen using liquid
nitrogen prior to further processing needed for
measuring of ATP.
Measurements of ATP in muscle tissue
Frozen muscle tissue was added to a glass tissue
grinder (Fisher Scientific, Chino, CA, USA) containing
200 μL ice cold ATP stabilizing solution, as previously
described by Gorman et al. (17). Tissue was mechanically
ground and transferred to a low-protein binding
microtube (Eppendorf, Hauppauge, NY, USA). The
sample was centrifuged for 5 min at 10,000 g and
supernatant was used for ATP quantification. ATP
concentration was determined using an ATP Assay Kit
(Calbiochem, San Diego, CA, USA) with a modification to
the original method, as previously described (18).
Hemoglobin levels were also determined in muscle
tissue lysates, using a double sandwich ELISA
(MyBiosource, San Diego, CA, USA), according to the
manufacturer’s instructions. Tissue samples
homogenized in ATP stabilizing solution described by
Gorman et al. (17) were used for this analysis.
Results
Twenty healthy subjects were recruited for this
placebo-controlled, crossover study. Subjects fasted
overnight and were then given an empty capsule as
placebo (Day 1). Blood was collected at baseline (before
treatment) and 60 min (T60) and 120 min (T120) after
treatment. Subjects fasted overnight, prior to Day 2, when
a single capsule containing 150 mg of elevATP™ was
administered to each subject. Blood was obtained as
previously described. Blood ATP and glucose, and
plasma ATP and hemoglobin levels were also
determined.
Figure 1
Effect of elevATP™ on blood ATP levels. elevATP™
significantly increased blood ATP levels by 40% at T60
(p<0.0001) and 28% at T120 (p=0.0009) over initial
baseline T0 values. Data are presented
as Mean +/- SE. n=20
A single dose of 150 mg elevATP significantly
increased blood ATP by 40% at 60 minutes (p<0.0001)
and 28% at 120 min (p=0.0009) as compared to baseline
ATP level at T0 (Figure 1).
Plasma ATP levels were measured using 10 µL of
plasma in a luciferase-based assay. There was no
significant increase in ATP level at T60 (p=0.83) or T120
(p=0.69) in patients treated with elevATP™ (Figure 2).
No change in plasma ATP level was seen after treatment
with placebo.
Hemoglobin levels were determined in all plasma
samples in order to ensure that mechanical disruption of
erythrocytes did not affect plasma ATP levels. The
placebo group had an increase in plasma hemoglobin of
REYES-IZQUIERDO_04 LORD_c 04/12/13 08:41 Page3
34% at T60 and 37% at T120 on day 1, compared to the T0
baseline (Figure 3). On day 2, after treatment with
elevATP™, there was an increase in plasma hemoglobin
level of 4% at T60 and 28% at T120, compared to the new
T0 baseline. There were no statistically significant
differences in placebo and elevATP™ treatments at T60
(p=0.29) and T120 (p=0.76).
Figure 2
Plasma ATP levels after treatment with elevATP™. Data
is presented as Mean +/- SE, Data are presented as %
change over baseline T0. n=20
Blood glucose levels were monitored after treatment
with placebo (Day 1) and after elevATP™ (Day 2), as
previously described. There were no significant
differences in blood glucose levels between treatments at
T60 (p=0.57) or T120 (p=0.59) in the 20 patients examined.
Blood lactate levels remained unchanged after placebo
(Day 1) or elevATP™ (Day 2) administration. The
difference between treatments was not significant either
at T60 (p=0.61) or T120 (p=0.44).
Figure 3
Hemoglobin levels after treatment with elevATP™. There
was a 4% increase at T60 and 28% increase at T120. There
were no statistical differences when compared to placebo.
Data are presented as Mean +/- SE, n=20
Biceps muscle levels of ATP were determined before
and after administration of elevATP™. At T0 (prior to
treatment), 210 pg ATP per mg protein was detected
(Figure 4). ATP level increased in muscle biopsy tissue to
590 pg/mg protein at 60 minutes and 910 pg/mg protein
at 120 minutes. Hemoglobin was also quantified in
muscle biopsy lysates in order to ensure that mechanical
disruption of the tissue did not affect ATP levels (Figure
5). Hemoglobin levels did not increase after treatment.
Figure 4
ATP levels in muscle tissue after treatment with
elevATP™. ATP levels increased significantly after
treatment. Data is presented as mean +/- SE of 4
determinations
Figure 5
Hemoglobin levels in muscle tissue lysates before and
after treatment with elevATP™. Hemoglobin increased
19% over baseline at T60 and 22% at T120. Data is
presented as Mean +/- SD of 3 determinations
Discussion
These findings provide further support for the
assertion that oral administration of elevATP™ increases
whole blood levels of ATP in healthy humans. The
increases reported here are in agreement with our
previous report (16). elevATP™ appears to selectively
and acutely increase ATP levels within the cellular
component of blood. In contrast, ATP levels in cell-free
plasma remained unchanged following elevATP™
ingestion. We verified the integrity of erythrocyte cell
membranes by quantifying hemoglobin concentrations in
plasma. This suggests that ATP did not originate from
ruptured red blood cells. Likewise, we found no changes
in plasma ATP levels after treatment with elevATP™,
suggesting that it is unlikely that elevATP™ affects
extracellular ATP levels.
Red blood cells constitute the largest cellular
4
EFFECT OF ELEVATP ON BLOOD ATP LEVELS.
REYES-IZQUIERDO_04 LORD_c 04/12/13 08:41 Page4
component of blood, comprising approximately 40 to 50%
of the blood volume. Red blood cells are a major carrier of
ATP (19-21), with intracellular concentrations reaching
the millimolar range (17, 22). ATP production in
erythrocytes takes place via glycolysis, rather than within
mitochondria (19). We previously reported that ingestion
of elevATP™ by fasting and resting study subjects did
not alter blood lactate levels, which could be generated
by increased glycolysis and ATP production in red cells
under these experimental conditions (16). These results
suggest that red blood cells did not contribute to
elevation of whole blood ATP level.
Fresh whole blood was collected and immediately
lysed to measure total ATP. This included ATP
originating from all types of cells present in the blood
collected. The lack of changes in blood glucose and lactate
levels suggests that ATP originated from blood cells with
mitochondria, such as platelets and while blood cells.
Platelets, unlike red blood cells, do contain mitochondria.
Platelets contain roughly 40 nmol of ATP and ADP per
mg of platelet protein (23). Platelets can be stimulated
through thrombin activation to achieve low micromolar
concentrations of ATP and ADP (24).
Further support for increased intracellular organ ATP
formation came from our muscle biopsy case study. As
presented here, muscle tissue, which is rich in
mitochondria, exhibited a substantial increase in ATP
levels after elevATP™ ingestion. Additional work in a
larger number of volunteers is in preparation to confirm
the stimulatory effect of elevATP™ on intramuscular
ATP levels in resting subjects. Our previous work
showed that elevATP™ did not increase reactive oxygen
species, despite increasing ATP levels (16). The
relationship between elevATP™ and mitochondrial
Complex IV and/or V activity is currently under
investigation in our laboratory as well as establishing
ATP/phosphocreatine ration before and after ingestion of
elevATP
In conclusion, we have thus confirmed the ability of
elevATP™ to increase ATP levels in whole blood. We
have further shown that elevATP does not increase ATP
levels in plasma. The precise blood cell type or types that
are sensitive to the action of elevATP™ is not known. A
clinical case experiment demonstrated that ingestion of a
single dose of elevATP™ resulted in a significant increase
in intramuscular ATP under resting conditions. This
result suggests that ingestion of elevATP™ may increase
ATP level in other tissues. Further clinical testing is
justified and is needed to confirm this preliminary result.
Investigating whether elevATP™ may improve muscle
performance and endurance in young and aged
individuals and such experimentation is also justified and
is currently in preparation.
Acknowledgments: All authors declare that they have no conflicts of interest.
The present study was funded by Futureceuticals, Inc. We express our gratitude to
John Hunter and Brad Evers (FutureCeuticals, Inc.) for their comments and
suggestions in the preparation of this article. We would like to thank Michael
Sapko for his help in editing the manuscript.
References
1. Gordon JL. Extracellular ATP: effects, sources and fate. Biochem J
1986;233:309-319
2. Trautmann A. Extracellular ATP in the immune system: more than just a
"danger signal". Sci Signal 2009;2:pe6 doi 10.1126/scisignal.256pe6
3. Haag F, Adriouch S, Brass A, Jung C, Moller S, Scheuplein F, Bannas P,
Seman M, Koch-Nolte F. Extracellular NAD and ATP: Partners in immune
cell modulation. Purinergic Signal 2007;3:71-81 doi 10.1007/s11302-006-9038-
7
4. Miyazawa M, Ito Y, Kosaka N, Nukada Y, Sakaguchi H, Suzuki H,
Nishiyama N. Role of TNF-alpha and extracellular ATP in THP-1 cell
activation following allergen exposure. J Toxicol Sci 2008;33:71-83
5. Schwiebert EMZ, A. Extracellular ATP as a signaling molecule for epithelial
cells. Biochim Biophys Acta 2003;1615:7-32 doi 10.1007/s11302-005-0777-7
6. Jordan ANJ, R.; Abraham, E. H.; Salikhova, A.; Mann, J. K.; Morss, G. M.;
Church, T. S.; Lucia, A.; Earnest, C. P. Effects of oral ATP supplementation on
anaerobic power and muscular strength. Med Sci Sports Exerc 2004;36:983-
990
7. Miyoshi NO, H.; Chock, P. B.; Stadtman, E. R. Age-dependent cell death and
the role of ATP in hydrogen peroxide-induced apoptosis and necrosis. Proc
Natl Acad Sci U S A 2006;103:1727-1731 doi 10.1073/pnas. 0510346103
8. Subasinghe W, Spence DM. Simultaneous determination of cell aging and
ATP release from erythrocytes and its implications in type 2 diabetes. Anal
Chim Acta 2008;618:227-233
9. Kirby BS, Crecelius AR, Voyles WF, Dinenno FA. Impaired skeletal muscle
blood flow control with advancing age in humans: attenuated ATP release
and local vasodilation during erythrocyte deoxygenation. Circ Res
2012;111:220-230 doi 10.1161/circresaha.112.269571
10. Rathmacher JA, Fuller JC, Jr., Baier SM, Abumrad NN, Angus HF, Sharp RL.
Adenosine-5'-triphosphate (ATP) supplementation improves low peak
muscle torque and torque fatigue during repeated high intensity exercise
sets. J Int Soc Sports Nutr 2012;9:48 doi 10.1186/1550-2783-9-48
11. Jordan AN, Jurca R, Abraham EH, Salikhova A, Mann JK, Morss GM, Church
TS, Lucia A, Earnest CP. Effects of oral ATP supplementation on anaerobic
power and muscular strength. Med Sci Sports Exerc 2004;36:983-990
12. Arts IC, Coolen EJ, Bours MJ, Huyghebaert N, Stuart MA, Bast A, Dagnelie
PC. Adenosine 5'-triphosphate (ATP) supplements are not orally
bioavailable: a randomized, placebo-controlled cross-over trial in healthy
humans. J Int Soc Sports Nutr 2012;9:16 doi 10.1186/1550-2783-9-16
13. Kichenin K, Decollogne S, Angignard J, Seman M. Cardiovascular and
pulmonary response to oral administration of ATP in rabbits. J Appl Physiol
2000;88:1962-1968
14. Agteresch HJ, Dagnelie PC, Rietveld T, van den Berg JW, Danser AH, Wilson
JH. Pharmacokinetics of intravenous ATP in cancer patients. Eur J Clin
Pharmacol 2000;56:49-55
15. Pastor-Anglada M, Errasti-Murugarren E, Aymerich I, Casado FJ.
Concentrative nucleoside transporters (CNTs) in epithelia: from absorption
to cell signaling. J Physiol Biochem 2007;63:97-110
16. Reyes-Izquierdo T, Nemzer B, Argumedo R, Shu C, Huynh L, Pietrzkowski
Z. Effect of the dietary supplement ElevATP on blood ATP level: An acute
pilot clinical study. J Aging Res Clin Practice 2013;2:178-184
17. Gorman MW, Feigl EO, Buffington CW. Human plasma ATP concentration.
Clin Chem 2007;53:318-325
18. Reyes-Izquierdo TH, L. E.; Sikorski, R.S.; Nemzer, B.; Pietrzkowski, Z. Acute
effect of HH2o on oxygen consumption rate, intracellular ATP and ROS in
freshly isolated human peripheral blood mononuclear cells. Current Topics
in Nutraceutical Research 2011;9:139-146
19. Ellsworth ML, Forrester T, Ellis CG, Dietrich HH. The erythrocyte as a
regulator of vascular tone. Am J Physiol 1995;269:H2155-2161
20. Sprague RS, Ellsworth ML, Stephenson AH, Kleinhenz ME, Lonigro AJ.
Deformation-induced ATP release from red blood cells requires CFTR
activity. Am J Physiol 1998;275:H1726-1732
21. Sprague RS, Hanson MS, Achilleus D, Bowles EA, Stephenson AH, Sridharan
M, Adderley S, Procknow J, Ellsworth ML. Rabbit erythrocytes release ATP
and dilate skeletal muscle arterioles in the presence of reduced oxygen
tension. Pharmacol Rep 2009;61:183-190
22. Montalbetti N, Leal Denis MF, Pignataro OP, Kobatake E, Lazarowski ER,
Schwarzbaum PJ. Homeostasis of extracellular ATP in human erythrocytes. J
Biol Chem 2011;286:38397-38407 doi 10.1074/jbc.M111.221713
23. Da Prada M, Richards J, Kettler R. Amine storage organelles in platelets.
Platelets in biology and pathology 1981;2:107-145
24. Ingerman CM, Smith JB, Silver MJ. Direct measurement of platelet secretion
in whole blood. Thrombosis research 1979;16:335-344.
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REYES-IZQUIERDO_04 LORD_c 04/12/13 08:41 Page5
... Therefore, an indirect approach for increasing ATP levels may be desirable. Previously, Reyes-Izquierdo and colleagues determined that a 150 mg dose of a blend of ancient peat and apple extracts significantly increased blood ATP compared to placebo in 18 [31] and 20 [32] subjects. In the latter research, blood ATP increased by 40 % at 60 min following ingestion, which dropped to 28 % at 120 min following ingestion. ...
... In the latter research, blood ATP increased by 40 % at 60 min following ingestion, which dropped to 28 % at 120 min following ingestion. A muscle biopsy was conducted in one subject, and ATP levels were observed to increase in muscle tissue by 281 % at 60 min and 433 % at 120 min following ingestion [32]. Preliminary reports from this laboratory also support an increase in blood ATP levels, and suggest this occurs without an increase in reactive oxygen species, which may be associated with increased ATP production [12]. ...
... The blend of ancient peat and apple extracts may be capable of promoting skeletal muscle hypertrophy by increasing whole-blood ATP levels [31,32] with a subsequent augmentation of blood flow. ATP and adenosine have been known to induce vasodilation following release from the erythrocytes via production of nitric oxide and prostacyclin [29,38], and it has been recently demonstrated that exogenous ATP supplementation is capable of increasing exercise-induced blood flow [22]. ...
Twelve weeks supplementation with an extended-release caffeine and ATP-enhancing supplement may improve body composition without affecting hematology in resistance-trained men
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Background Increased ATP levels may enhance training-induced muscle accretion and fat loss, and caffeine is a known ergogenic aid. A novel supplement containing ancient peat and apple extracts has reported enhanced mitochondrial ATP production and it has been coupled with an extended-release caffeine. Therefore, the purpose of this investigation was to determine the effects of this supplement on body composition when used in conjunction with 12 weeks of resistance training. Methods Twenty-one resistance-trained subjects (27.2 ± 5.6y; 173.5 ± 5.7 cm; 82.8 ± 12.0 kg) completed this study. Subjects supplemented daily with either 1 serving of the supplement (TRT), which consisted of 150 mg ancient peat and apple extracts, 180 mg blend of caffeine anhydrous and pterostilbene-bound caffeine, and 38 mg B vitamins, or an equal-volume, visually-identical placebo (PLA) 45 min prior to training or at the same time of day on rest days. Supervised resistance training consisted of 8 weeks of daily undulating periodized training followed by a 2-week overreach and a 2-week taper phase. Body composition was assessed using DEXA and ultrasound at weeks 0, 4, 8, 10, and 12. Vital signs and blood markers were assessed at weeks 0, 8, and 12. Results Significant group x time (p < 0.05) interactions were present for cross-sectional area of the rectus femoris, which increased in TRT (+1.07 cm²) versus PLA (−0.08 cm²), as well as muscle thickness (TRT: +0.49 cm; PLA: +0.04 cm). A significant group x time (p < 0.05) interaction existed for creatinine (TRT: +0.00 mg/dL; PLA: +0.15 mg/dL) and estimated glomerular filtration rate (TRT: −0.70 mL/min/1.73; PLA: −14.6 mL/min/1.73), which remained within clinical ranges, but no other significant observations were observed. Conclusions Supplementation with a combination of extended-release caffeine and ancient peat and apple extracts may enhance resistance training-induced skeletal muscle hypertrophy without adversely affecting blood chemistry.
... Therefore, supplementation strategies for increasing endogenous ATP levels may be desirable. Previously, oral supplementation with a proprietary blend of ancient peat and apple extracts have been demonstrated to increase intracellular ATP levels in whole blood and intramuscular levels of ATP in resting subjects, suggesting increased activity of bodily processes that lead to endogenous ATP production [8,9]. ...
... Previous research has found oral supplementation with a proprietary blend of ancient peat and apple extracts to increase ATP concentrations in whole blood of resting subjects as well as intramuscular concentrations in one volunteer [9]. Preliminary reports from this laboratory suggest this occurs without an increase in reactive oxygen species, which may be associated with increased ATP production [10]. ...
... Despite these observations, supplementation for indirect ATP enhancement is yet to be evaluated for potential to augment performance in response to resistance training. However, the existing data on ancient peat and apple extracts for increasing both whole blood and muscle ATP levels [8,9] and muscle mass [16] support the plausibility for chronic supplementation yielding positive augmentation of performance following resistance training. Therefore, the purpose of this study is to determine the effects of a proprietary blend of ancient peat and apple extracts on athletic performance. ...
Ancient peat and apple extracts supplementation may improve strength and power adaptations in resistance trained men
Article
Full-text available
Background Increased cellular ATP levels have the potential to enhance athletic performance. A proprietary blend of ancient peat and apple extracts has been supposed to increase ATP production. Therefore, the purpose of this investigation was to determine the effects of this supplement on athletic performance when used during 12 weeks of supervised, periodized resistance training. Methods Twenty-five healthy, resistance-trained, male subjects completed this study. Subjects supplemented once daily with either 1 serving (150 mg) of a proprietary blend of ancient peat and apple extract (TRT) or an equal-volume, visually-identical placebo (PLA) daily. Supervised resistance training consisted of 8 weeks of daily undulating periodized training followed by a 2 week overreach and a 2 week taper phase. Strength was determined using 1-repetition-maximum (1RM) testing in the barbell back squat, bench press (BP), and deadlift exercises. Peak power and peak velocity were determined during BP at 30 % 1RM and vertical jump tests as well as a 30s Wingate test, which also provided relative power (watt:mass) ResultsA group x time interaction was present for squat 1RM, deadlift 1RM, and vertical jump peak power and peak velocity. Squat and deadlift 1RM increased in TRT versus PLA from pre to post. Vertical jump peak velocity increased in TRT versus PLA from pre to week 10 as did vertical jump peak power, which also increased from pre to post. Wingate peak power and watt:mass tended to favor TRT. Conclusions Supplementing with ancient peat and apple extract while participating in periodized resistance training may enhance performance adaptations. Trial RegistrationClinicalTrials.gov registration ID: NCT02819219, retrospectively registered on 6/29/2016
... Previously, oral supplementation with a proprietary blend of ancient peat and apple extracts has been demonstrated to increase intracellular ATP levels in whole blood and muscle, suggesting increased activity of bodily processes that produce or release endogenous ATP (Reyes-Izquierdo et al. 2013. Reyes-Izquierdo et al. (2014) determined that a 150-mg dose of this blend of ancient peat and apple extracts significantly increased ATP in cellular fraction of blood compared with placebo. Specifically, blood ATP was increased by 40% at 60 min following ingestion and dropped to 28% at 120 min following ingestion. ...
... Specifically, blood ATP was increased by 40% at 60 min following ingestion and dropped to 28% at 120 min following ingestion. A muscle biopsy was conducted in 1 resting subject, and ATP levels in muscle tissue were observed to increase 281% at 60 min and 433% at 120 min following ingestion (Reyes-Izquierdo et al. 2014). Preliminary reports from this laboratory suggest this occurs without increases in reactive oxygen species, which may be associated with increased ATP production (Chang et al. 2010). ...
... An improved resistance to fatiguing exercise is also possible via this mechanism, such as with previous research on creatine monohydrate, which increases ATP availability (Racette 2003), and previous research regarding creatine's capabilities for increasing muscle mass has been well established (Buford et al. 2007). However, more thorough research is required before considering this as a viable mechanism because of the fact that previously enhanced intramuscular ATP levels have only been examined in 1 resting subject (Reyes-Izquierdo et al. 2014). Yet, the blend of ancient peat and apple extracts may be advantageous to direct ATP supplementation for these reasons, as direct ATP supplementation may only exert extracellular effects because of rapid degradation to its metabolites (Hochachka et al. 1991;Gorman et al. 2007;Mortensen et al. 2011;Jäger et al. 2014). ...
Supplementation with a Proprietary Blend of Ancient Peat and Apple Extract May Improve Body Composition without Affecting Hematology in Resistance-Trained Men
Article
Full-text available
Adenosine-5'-triphosphate (ATP) is primarily known as a cellular source of energy. Increased ATP levels may have the potential to enhance body composition. A novel, proprietary blend of ancient peat and apple extracts has been reported to increase ATP levels, potentially by enhancing mitochondrial ATP production. Therefore, the purpose of this investigation was to determine the supplement's effects on body composition when consumed during 12 weeks of resistance training. Twenty-five healthy, resistance-trained, male subjects (age, 27.7 ± 4.8 years; height, 176.0 ± 6.5 cm; body mass, 83.2 ± 12.1 kg) completed this study. Subjects supplemented once daily with either 1 serving (150 mg) of a proprietary blend of ancient peat and apple extracts (TRT) or placebo (PLA). Supervised resistance training consisted of 8 weeks of daily undulating periodized training followed by a 2-week overreach and a 2-week taper phase. Body composition was assessed using dual-energy X-ray absorptiometry and ultrasound at weeks 0, 4, 8, 10, and 12. Vital signs and blood markers were assessed at weeks 0, 8, and 12. Significant group × time (p < 0.05) interactions were present for ultrasound-determined cross-sectional area, which increased in TRT (+0.91 cm(2)) versus PLA (-0.08 cm(2)), as well as muscle thickness (TRT: +0.46; PLA: +0.04 cm). A significant group × time (p < 0.05) interaction existed for creatinine (TRT: +0.06; PLA: +0.15 mg/dL), triglycerides (TRT: +24.1; PLA: -20.2 mg/dL), and very-low-density lipoprotein (TRT: +4.9; PLA: -3.9 mg/dL), which remained within clinical ranges. Supplementation with a proprietary blend of ancient peat and apple extracts may enhance resistance training-induced skeletal muscle hypertrophy without affecting fat mass or blood chemistry in healthy males.
... Oral intake of a single dose of ElevATP ™ has been shown to increase whole blood levels of ATP in human subjects without affecting reactive oxygen species or blood glucose and lactate levels [4,15]. Ingestion of single doses of ElevATP TM has also been associated with increased whole blood and muscle levels of ATP [4,15]. ...
... Oral intake of a single dose of ElevATP ™ has been shown to increase whole blood levels of ATP in human subjects without affecting reactive oxygen species or blood glucose and lactate levels [4,15]. Ingestion of single doses of ElevATP TM has also been associated with increased whole blood and muscle levels of ATP [4,15]. In this study, subjects who did not exercise regularly were evaluated and they took more steps, traveled further, and burned more calories after ingesting a single dose of either lot of ElevATP ™ , than after taking a placebo control. ...
The Effect of ElevATP ™ on Exercise Output: A Single Dose, Blinded, Three-Way Cross-Over Study
Article
Full-text available
  • Aug 2016
Background: Adenosine 5’-triphosphate (ATP) serves as a cellular source of energy for metabolic processes. Maintaining high levels of intracellular ATP has the potential to enhance performance. ElevATP™, a proprietary blend of ancient peat and apple extract, has twice been reported to increase intracellular blood levels of ATP in resting subjects after a single dose per os. More recently, resistance-trained males supplemented with ElevATP™ for 12 weeks, have shown increased resistance training-induced skeletal muscle hypertrophy. For this pilot study, we determined the acute effects of a single dose of ElevATP™ on the exercise output of subjects with sedentary lifestyles. Methods and findings: Nine healthy subjects were evaluated before, during, and after 20 minute duration stepping exercise in a blinded three-way crossover study of placebo and a single dose of ElevATP™. Two lots of ElevATP™ (Lot A and Lot B) were each compared to placebo in all subjects. Four (4) female and five (5) male subjects that did not regularly exercise were evaluated. Mean subject age was 27.3 ± 5.0 years and BMI was 28.97 ± 6.6 kg/m². ElevATP™ supplemented subjects (Lot A and Lot B) took a significantly greater number of steps and burned more calories than when treated with placebo. Post-exercise analysis has shown no significant changes in blood lactate or glucose levels between ElevATP™ (Lota A or Lot B) and placebo treatments. Conclusion: All nine subjects experienced an increase in exercise performance after ingesting a single dose of either of the two tested lots of ElevATP™, compared to a placebo control.
... Although these studies' findings are compelling, none have evaluated ancient peat and apple extracts' effects on high-intensity endurance exercise in recreational or trained endurance athletes. In addition, many of the authors associated with the studies described above [26][27][28][29] acknowledged an affiliation with the manufacturer of elevATP™; therefore, more randomized, PL-controlled trials conducted in independent laboratories are necessary to confirm its efficacy for enhancing endurance performance. ...
The Effect of a Multi-Ingredient Pre-Workout Supplement on Time to Fatigue in NCAA Division I Cross-Country Athletes
Article
Full-text available
  • May 2021
This investigation aimed to determine the effect of a multi-ingredient pre-workout supplement (MIPS) on heart rate (HR), perceived exertion (RPE), lactate concentration, and time to fatigue (TTF) during a running task to volitional exhaustion. Eleven NCAA Division I cross-country runners (20 ± 2 year; height: 171 ± 14 cm; weight: 63.5 ± 9.1 kg) participated in this randomized, double-blind, placebo-controlled cross-over study. Bayesian statistical methods were utilized, and parameter estimates were interpreted as statistically significant if the 95% highest-density intervals (HDIs) did not include zero. TTF was increased in the MIPS condition with a posterior Meandiff = 154 ± 4.2 s (95% HDI: −167, 465) and a 0.84 posterior probability that the supplement would increase TTF relative to PL. Blood lactate concentration immediately post-exercise was also higher in the MIPS condition compared to PL with an estimated posterior Meandiff = 3.99 ± 2.1 mmol (95% HDI: −0.16, 7.68). There were no differences in HR or RPE between trials. These findings suggest that a MIPS ingested prior to sustained running at lactate threshold has an 84% chance of increasing TTF in highly trained runners and may allow athletes to handle a higher level of circulating lactate before reaching exhaustion.
... Recently, a MIES, Reckless (Maximum Human Performance [MHP], LLC), has been designed in an effort to take advantage of the potential synergistic effects of the aforemen- tioned ingredients, along with additional ingredients that may acutely increase blood flow (e.g., L-citrulline, L-norvaline, creatine) ( Martin et al., 2017), enhance circulating ATP lev- els (e.g., ancient peat and apple fruit extract; Reyes-Izquierdo, Shu, Argumedo, Nemzer, & Pietrzkowski, 2014), and serve as antioxidants (e.g., Spectra;Nemzer, Fink, & Fink, 2014). This study set out to assess whether this MIES could enhance cognitive performance during an attention-switching task. ...
Effects of a Multi-Ingredient Energy Supplement on Cognitive Performance and Cerebral-Cortical Activation
Article
  • Mar 2018
This study assessed whether a multi-ingredient energy supplement (MIES) could enhance cerebral-cortical activation and cognitive performance during an attention-switching task. Cerebral-cortical activation was recorded in 24 young adults (12 males, 12 females; 22.8 ± 3.8 yrs) via electroencephalography (EEG) both at rest and during the attention-switching task before (pretest) and 30 min after (posttest) consumption of a single serving of a MIES (MIES-1), two servings of a MIES (MIES-2), or a placebo (PL) in a double-blinded, randomized crossover experimental design. EEG upper-alpha power was assessed at rest and during the task, wherein d′ (Z[hit rate]–Z[false alarm rate]) and median reaction time (RT) for correct responses to targets on attention-hold and attention-switch trials were analyzed. For both d′ and RT, the Session (MIES-1, MIES-2, PL) × Time (pretest, posttest) interaction approached statistical significance (p = .07, η²p = 0.106). Exploring these interactions with linear contrasts, a significant linear effect of supplement dose on the linear effect of time was observed (ps ≤.034), suggesting the pretest-to-posttest improvement in sensitivity to task target stimuli (d′) and RT increased as a function of supplement dose. With respect to upper-alpha power, the Session × Time interaction was significant (p < .001, η²p = 0.422). Exploring this interaction with linear contrasts, a significant linear effect of supplement dose on the linear effect of time was observed (p < .001), suggesting pretest-to-posttest increases in cerebral-cortical activation were a function of supplement dose. In conclusion, our findings suggest that MIES can increase cerebral-cortical activation and RT during task performance while increasing sensitivity to target stimuli in a dose-dependent manner.
... Spectra™, a "full-spectrum" antioxidant product comprised of fruit, vegetable and herb extracts, may also act in a synergistic fashion with other ingredients by reducing NO scavenging by reactive oxygen species and improving NO bioavailability [16]. Finally, ancient peat and apple fruit extract has been shown to enhance circulating ATP levels [17,18] which may augment blood flow in a NOS independent manner [19,20]. Indeed, although many PWS marketed to increase the blood flow response to resistance training focus on NO modulation, the contribution of NO to resistance exercise induced hyperemia is not absolute and likely partially includes ATP-sensitive potassium channels [21] which may interact with ingredient(s) found in a PWS (e.g., elevATP®). ...
Effects of a pre-workout supplement on hyperemia following leg extension resistance exercise to failure with different resistance loads
Article
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Background: We sought to determine if a pre-workout supplement (PWS), containing multiple ingredients thought to enhance blood flow, increases hyperemia associated with resistance training compared to placebo (PBO). Given the potential interaction with training loads/time-under-tension, we evaluated the hyperemic response at two different loads to failure. Methods: Thirty males participated in this double-blinded study. At visit 1, participants were randomly assigned to consume PWS (Reckless™) or PBO (maltodextrin and glycine) and performed four sets of leg extensions to failure at 30% or 80% of their 1-RM 45-min thereafter. 1-wk. later (visit 2), participants consumed the same supplement as before, but exercised at the alternate load. Heart rate (HR), blood pressure (BP), femoral artery blood flow, and plasma nitrate/nitrite (NOx) were assessed at baseline (BL), 45-min post-PWS/PBO consumption (PRE), and 5-min following the last set of leg extensions (POST).Vastus lateralisnear infrared spectroscopy (NIRS) was employed during leg extension exercise. Repeated measures ANOVAs were performed with time, supplement, and load as independent variables and Bonferroni correction applied for multiplepost-hoccomparisons. Data are reported as mean ± SD. Results: With the 30% training load compared to 80%, significantly more repetitions were performed (p < 0.05), but there was no difference in total volume load (p > 0.05). NIRS derived minimum oxygenated hemoglobin (O2Hb) was lower in the 80% load condition compared to 30% for all rest intervals between sets of exercise (p < 0.0167). HR and BP did not vary as a function of supplement or load. Femoral artery blood flow at POST was higher independent of exercise load and treatment. However, a time*supplement*load interaction was observed revealing greater femoral artery blood flow with PWS compared to PBO at POST in the 80% (+56.8%;p = 0.006) but not 30% load condition (+12.7%;p = 0.476). Plasma NOx was ~3-fold higher with PWS compared to PBO at PRE and POST (p < 0.001). Conclusions: Compared to PBO, the PWS consumed herein augmented hyperemia following multiple sets to failure at 80% of 1-RM, but not 30%. This specificity may be a product of interaction with local perturbations (e.g., reduced tissue oxygenation levels [minimum O2Hb] in the 80% load condition) and/or muscle fiber recruitment.
Effect of Multi-Ingredient Preworkout Supplementation on Repeated Sprint Performance in Recreationally Active Men and Women
Article
  • Jan 2020
Gonzalez, AM, Pinzone, AG, Bram, J, Salisbury, JL, Lee, S, and Mangine, GT. Effect of multi-ingredient preworkout supplementation on repeated sprint performance in recreationally active men and women. J Strength Cond Res XX(X): 000-000, 2019-The purpose of this investigation was to examine the effects of acute supplementation of a multi-ingredient preworkout supplement (MIPS), containing a proprietary blend of ancient peat and apple extracts, creatine monohydrate, taurine, ribose, and magnesium, on sprint cycling performance. Seventeen recreationally active men and women (23.2 ± 5.9 years; 172.9 ± 14.3 cm; 82.4 ± 14.5 kg) underwent 2 testing sessions administered in a randomized, counterbalanced, double-blind fashion. Subjects were provided either MIPS or placebo (PL) one hour before performing a sprint cycling protocol, which consisted of ten 5-second "all-out" sprints interspersed by 55 seconds of unloaded pedaling. Average power (PAVG), peak power (PPK), average velocity (VAVG), and distance covered were recorded for each sprint. Separate linear mixed models revealed decrements (p < 0.05) compared to the first sprint in PAVG (75-229 W) and PPK (79-209 W) throughout all consecutive sprints after the initial sprint during PL. Likewise, diminished (p ≤ 0.029) VAVG (3.37-6.36 m·s) and distance covered (7.77-9.00 m) were noted after the third and fifth sprints, respectively, during PL. By contrast, during MIPS, only VAVG decreased (2.34-5.87 m·s, p ≤ 0.002) on consecutive sprints after the first sprint, whereas PAVG and PPK were maintained. In addition, a significant decrease (p = 0.045) in distance covered was only observed on the ninth sprint during MIPS. These data suggest that recreational athletes who consumed the MIPS formulation, one hour before a repeated sprinting session on a cycle ergometer, better maintained performance compared with PL.
Effect of the dietary supplement ElevATP on blood ATP level: An acute pilot clinical study
Article
Full-text available
  • Jan 2013
Acute effect of HH2O on oxygen Consumption rate, intracellular ATP and ROS in freshly isolated Human peripheral blood mononuclear cells
Article
Full-text available
Human peripheral blood mononuclear cells (PBMC) were used as a biological model to measure intracellular ATP (iATP) reactive oxygen species (ROS), Oxygen Consumption Rate (OCR) and extracellular acidification rate (ECAR), in response to acute ex vivo treatments with HH2o. "HH2o", a fermented, food-based material commercially marketed as "Mitochroma™ has been identified as a potential modulator of iATP, OCR and ROS in these cells. PBMC were exposed to HH2O (0.1, 0.01 and 0.001% w/v) and Carbonyl-cyanide p-trifluoro-methoxyphenyl-hydrazone (FCCP) or Oligomycin, two known modulators of mitochondrial activity were used as controls. Results showed that treatment of the cells with HH2O resulted in increase of iA TP up to 220% and reduction of iADP by up to 40% while iROS levels remained unchanged. At the same time, OCR level was increased up to 24% compared to initial baseline. These results suggest that HH2O acts similarly to uncoupling protein 2 (UCP2) when induced and that it may have potential use in developing health-promoting products. These results support the use of freshly isolated human PBMCs as an experimental ex vivo model to detect the effects of nutraceutical products on dynamic cell metabolites, such as oxygen consumption, ATP, ADP and ROS.
Adenosine-5'-triphosphate (ATP) supplementation improves low peak muscle torque and torque fatigue during repeated high intensity exercise sets
Article
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Background Intracellular concentrations of adenosine-5’-triphosphate (ATP) are many times greater than extracellular concentrations (1–10 mM versus 10–100 nM, respectively) and cellular release of ATP is tightly controlled. Transient rises in extracellular ATP and its metabolite adenosine have important signaling roles; and acting through purinergic receptors, can increase blood flow and oxygenation of tissues; and act as neurotransmitters. Increased blood flow not only increases substrate availability but may also aid in recovery through removal of metabolic waste products allowing muscles to accomplish more work with less fatigue. The objective of the present study was to determine if supplemental ATP would improve muscle torque, power, work, or fatigue during repeated bouts of high intensity resistance exercise. Methods Sixteen participants (8 male and 8 female; ages: 21–34 years) were enrolled in a double-blinded, placebo-controlled study using a crossover design. The participants received either supplemental ATP (400 mg/d divided into 2 daily doses) or placebo for 15 d. After an overnight fast, participants underwent strength and fatigue testing, consisting of 3 sets of 50 maximal knee extensions performed on a Biodex® leg dynamometer. Results No differences were detected in high peak torque, power, or total work with ATP supplementation; however, low peak torque in set 2 was significantly improved (p < 0.01). Additionally, in set 3, a trend was detected for less torque fatigue with ATP supplementation (p < 0.10). Conclusions Supplementation with 400 mg ATP/d for 15 days tended to reduce muscle fatigue and improved a participant’s ability to maintain a higher force output at the end of an exhaustive exercise bout.
Impaired Skeletal Muscle Blood Flow Control With Advancing Age in Humans: Attenuated ATP Release and Local Vasodilation During Erythrocyte Deoxygenation
Article
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Skeletal muscle blood flow is coupled with the oxygenation state of hemoglobin in young adults, whereby the erythrocyte functions as an oxygen sensor and releases ATP during deoxygenation to evoke vasodilation. Whether this function is impaired in humans of advanced age is unknown. To test the hypothesis that older adults demonstrate impaired muscle blood flow and lower intravascular ATP during conditions of erythrocyte deoxygenation. We showed impaired forearm blood flow responses during 2 conditions of erythrocyte deoxygenation (systemic hypoxia and graded handgrip exercise) with age, which was caused by reduced local vasodilation. In young adults, both hypoxia and exercise significantly increased venous [ATP] and ATP effluent (forearm blood flow×[ATP]) draining the skeletal muscle. In contrast, hypoxia and exercise did not increase venous [ATP] in older adults, and both venous [ATP] and ATP effluent were substantially reduced compared with young people despite similar levels of deoxygenation. Next, we demonstrated that this could not be explained by augmented extracellular ATP hydrolysis in whole blood with age. Finally, we found that deoxygenation-mediated ATP release from isolated erythrocytes was essentially nonexistent in older adults. Skeletal muscle blood flow during conditions of erythrocyte deoxygenation was markedly reduced in aging humans, and reductions in plasma ATP and erythrocyte-mediated ATP release may be a novel mechanism underlying impaired vasodilation and oxygen delivery during hypoxemia with advancing age. Because aging is associated with elevated risk for ischemic cardiovascular disease and exercise intolerance, interventions that target erythrocyte-mediated ATP release may offer therapeutic potential.
Adenosine 5′-triphosphate (ATP) supplements are not orally bioavailable: a randomized, placebo-controlled cross-over trial in healthy humans
Article
Full-text available
Nutritional supplements designed to increase adenosine 5'-triphosphate (ATP) concentrations are commonly used by athletes as ergogenic aids. ATP is the primary source of energy for the cells, and supplementation may enhance the ability to maintain high ATP turnover during high-intensity exercise. Oral ATP supplements have beneficial effects in some but not all studies examining physical performance. One of the remaining questions is whether orally administered ATP is bioavailable. We investigated whether acute supplementation with oral ATP administered as enteric-coated pellets led to increased concentrations of ATP or its metabolites in the circulation. Eight healthy volunteers participated in a cross-over study. Participants were given in random order single doses of 5000 mg ATP or placebo. To prevent degradation of ATP in the acidic environment of the stomach, the supplement was administered via two types of pH-sensitive, enteric-coated pellets (targeted at release in the proximal or distal small intestine), or via a naso-duodenal tube. Blood ATP and metabolite concentrations were monitored by HPLC for 4.5 h (naso-duodenal tube) or 7 h (pellets) post-administration. Areas under the concentration vs. time curve were calculated and compared by paired-samples t-tests. ATP concentrations in blood did not increase after ATP supplementation via enteric-coated pellets or naso-duodenal tube. In contrast, concentrations of the final catabolic product of ATP, uric acid, were significantly increased compared to placebo by ~50% after administration via proximal-release pellets (P = 0.003) and naso-duodenal tube (P = 0.001), but not after administration via distal-release pellets. A single dose of orally administered ATP is not bioavailable, and this may explain why several studies did not find ergogenic effects of oral ATP supplementation. On the other hand, increases in uric acid after release of ATP in the proximal part of the small intestine suggest that ATP or one of its metabolites is absorbed and metabolized. Uric acid itself may have ergogenic effects, but this needs further study. Also, more studies are needed to determine whether chronic administration of ATP will enhance its oral bioavailability.
Homeostasis of Extracellular ATP in Human Erythrocytes
Article
Full-text available
We explored the intra- and extracellular processes governing the kinetics of extracellular ATP (ATPe) in human erythrocytes stimulated with agents that increase cAMP. Using the luciferin-luciferase reaction in off-line luminometry we found both direct adenylyl cyclase activation by forskolin and indirect activation through β-adrenergic stimulation with isoproterenol-enhanced [ATP]e in a concentration-dependent manner. A mixture (3V) containing a combination of these agents and the phosphodiesterase inhibitor papaverine activated ATP release, leading to a 3-fold increase in [ATP]e, and caused increases in cAMP concentration (3-fold for forskolin + papaverine, and 10-fold for 3V). The pannexin 1 inhibitor carbenoxolone and a pannexin 1 blocking peptide ((10)Panx1) decreased [ATP]e by 75-84%. The residual efflux of ATP resulted from unavoidable mechanical perturbations stimulating a novel, carbenoxolone-insensitive pathway. In real-time luminometry experiments using soluble luciferase, addition of 3V led to an acute increase in [ATP]e to a constant value of ∼1 pmol × (10(6) cells)(-1). A similar treatment using a surface attached luciferase (proA-luc) triggered a rapid accumulation of surface ATP levels to a peak concentration of 2.4 pmol × (10(6) cells)(-1), followed by a slower exponential decay (t(&frac12;) = 3.7 min) to a constant value of 1.3 pmol × (10(6) cells)(-1). Both for soluble luciferase and proA-luc, ATP efflux was fully blocked by carbenoxolone, pointing to a 3V-induced mechanism of ATP release mediated by pannexin 1. Ecto-ATPase activity was extremely low (∼28 fmol × (10(6) cells min)(-1)), but nevertheless physiologically relevant considering the high density of erythrocytes in human blood.
Extracellular ATP in the Immune System: More Than Just a "Danger Signal"
Article
Full-text available
  • Feb 2009
Extracellular adenosine 5'-triphosphate (eATP) is ubiquitously used for cell-to-cell communication. The low concentration of eATP ([eATP]) that exists in a "halo" surrounding resting cells signals the presence of neighboring living cells. Transient increases in [eATP] are used for basic physiological signaling, namely, in the nervous and vascular systems. Larger increases in [eATP] that are associated with cell death serve as a key "danger" signal in inflammatory processes. Two studies now point to roles for ATP in the immune system: providing a costimulatory signal to T cells and driving the differentiation of intestinal T helper 17 (T(H)17) cells.
Amine storage organelles
Article
  • Jan 1981
Rabbit Erythrocytes Release ATP and Dilate Skeletal Muscle Arterioles in the Presence of Reduced Oxygen Tension
Article
In skeletal muscle, oxygen (O(2)) delivery to appropriately meet metabolic need requires mechanisms for detection of the magnitude of O(2) demand and the regulation of O(2) delivery. Erythrocytes, when exposed to a decrease in O(2) tension, release both O(2) and the vasodilator adenosine triphosphate (ATP). The aims of this study were to establish that erythrocytes release ATP in response to reduced O(2) tension and determine if erythrocytes are necessary for the dilation of isolated skeletal muscle arterioles exposed to reduced extraluminal O(2) tension. Rabbit erythrocytes exposed to reduced O(2) tension in a tonometer (n = 5, pO(2) = 27 +/- 3, p < 0.01) released ATP in response to reduced O(2) tension. ATP release increased in proportion to the decrease in O(2) tension. The contribution of erythrocytes to the response of skeletal muscle arterioles to reduced extraluminal O(2) tension was determined using isolated hamster cheek pouch retractor muscle arterioles perfused with buffer (n = 11, mean diameter 52 +/- 3 mum) in the absence and presence of rabbit erythrocytes. Without erythrocytes, arterioles did not dilate when exposed to reduced extraluminal O(2) tension (pO(2) = 32 +/- 4 mmHg). In contrast, when rabbit erythrocytes were present in the perfusate (hematocrit 15%), the same decrease in O(2) tension resulted in a 20 +/- 4% dilation (p < 0.01). These results provide support for the hypothesis that erythrocytes, via their ability to release O(2) along with ATP in response to exposure to reduced O(2) tension, can participate in the matching of O(2) delivery with metabolic need in skeletal muscle.
Direct measurement of platelet secretion in whole blood
Article
Platelet secretion is usually measured using platelet-rich plasma (PRP) or washed platelets resuspended in buffered saline. We report here measurements of the release of adenosine triphosphate (ATP) from platelets in citrated whole blood in response to thrombin and collagen. The amount of ATP released correlates with the platelet content of the whole blood and is partially dependent on prostaglandin synthesis when release is induced by collagen. Further, the release of ATP does not occur in blood from a patient with storage-pool deficient platelets.