2016 ISSN Research Presentations | Exercise & Sport Nutrition

Clearwater, FL, June 11, 2016

Seven days of high and low dose creatine nitrate supplementation I: Hepatorenal, glucose and muscle enzyme function

R Dalton, R Sowinski, T Grubic, PB Collins, A Coletta, A Reyes, B Sanchez, M Koozehchain, YP Jung, C Rasmussen, P Murano, CP Earnest, M Greenwood, RB Kreider

Exercise & Sport Nutrition Lab, Texas A&M University, College Station, TX, USA
Institute for Obesity and Program Evaluation, Texas A&M University, College Station, TX, USA
Nutrabolt, Bryan, TX, USA

Background
Creatine and nitrates are popular dietary supplements. While both have been examined in singularity, little is known regarding their co-ingestion relative to performance, side effects and safety. The purpose of this study was to examine the safety and efficacy of a creatine nitrate dietary supplement.

Methods
In a double-blind, crossover, randomized and placebo-controlled manner; 28 apparently healthy and recreationally active men and women (18 men, 10 women, 21.6±3.7 yr, 20.4±10.6% fat, 24.7±2.9 kg/m2) ingested daily supplements for 7-d consisting of a dextrose flavored placebo (PLA); a low dose of creatine nitrate (Low, 3g) and a high dose of creatine nitrate (6g). Participants repeated the experiment with the alternate supplements randomly with a 7 day washout period between each. Participants had each blood donation after 8+ hours fasting on days 1, 2, 6, and 7 for each supplement. Data were analyzed by repeated measure 4 x 3 MANOVA with Time and Group as factors using Greenhouse-Geisser as appropriate and are presented as mean±SD or mean change from baseline ± 95% CI.

Results
Significant time effects (p<0.001) were observed for blood urea nitrogen (BUN), alkaline phosphatase (ALP), LDL cholesterol (LDL), triglycerides (TRI), and glucose (GLU) (p<.01). Significant group x time interactions were found for CRE (p<.05). No significant differences for time, group, or interactions were found for aspartate amino transferase, and alanine amino transferase, for total cholesterol, HDL cholesterol, creatine kinase, and lactate dehydrogenase (p>0.05). Respective blood values at Day1, Day2, Day6, and Day7 are: BUN (5.07±1.29, 5.00±1.47, 5.29±1.42, 5.26±1.49 mmol/L) with Day2 lower than days 3 & 4, ALP (80.43±22.19, 77.85±20.14, 81.10±21.35, 79.60±20.77 U/L) with Day2 lower than days 1 & 3, LDL (2.61±1.32, 2.53±1.17, 2.29±0.96, 2.35±0.96 mmol/L) with Day3 lower than days 1, 2, & 4, TRI (0.88±0.39, 0.81±0.31, 0.91±0.42, 0.83±0.33 mmol/L) with Day2 lower than days 1 & 3, GLU (5.05±0.48, 4.89±0.40, 4.96±0.54, 5.03±0.44 mmol/L) with Day2 lower than days 1&4, and mean changes for CRE (Day2 PLA: 1.60, 95% CI -2.00, 5.18, Day2 Low: -1.67, 95% CI -5.26, 1.92, Day2 High: 4.25, 95% CI 0.66, 7.84, Day6 PLA: -0.53, 95% CI -5.38, 4.33, Day6 Low: -3.37, 95% CI 8.22, 1.49, Day6 High: 7.58, 95% CI 2.72, 12.43, Day7 PLA: -1.13, 95% CI -5.32, 3.05, Day7 Low: -5.83, 95% CI -10.01, -1.65, Day7 High: 4.45, 95% CI 0.27, 8.64 µmol/L). CRE was found to be higher at Day2 for High compared Low and at Day6 and Day7 for High vs Low & PLA, CRE increased over time for group High compared to PLA and Low, which did not change.

Conclusion
Ingesting a creatine nitrate supplement demonstrated minor time x group interaction differences in CRE. However, these changes did not exceed normal clinical limits. This and the results observed for other clinical markers associated with hepatorenal and muscle enzyme function demonstrate that creatine nitrate appears to be safe when ingested for seven days.

Acknowledgments
This study was supported by Nutrabolt (Bryan, TX) through an unrestricted grant to Texas A&M University.

Seven days of high and low dose creatine nitrate supplementation II: Performance

T Grubic, R Dalton, R Sowinski, PB Collins, A Coletta, A Reyes, B Sanchez, M Koozehchain, YP Jung, C Rasmussen, P Murano, CP Earnest, M Greenwood, RB Kreider

Exercise & Sport Nutrition Lab, Texas A&M University, College Station, TX, USA;
Institute for Obesity and Program Evaluation, Texas A&M University, College Station, TX, USA;
Nutrabolt, Bryan, TX, USA

Background
Creatine and nitrates are popular dietary supplements. While both have been examined in singularity, little is known regarding their co-ingestion relative to performance, side effects and safety. The purpose of this study was to examine the safety and efficacy of a creatine nitrate dietary supplement.

Methods
In a double-blind, crossover, randomized and placebo-controlled manner; 28 apparently healthy and recreationally active men and women (18 men, 10 women, 21.6±3.7 yr, 20.4±10.6% fat, 24.7±2.9 kg/m2) ingested daily supplements for 7-d consisting of a dextrose flavored placebo (PLA); a low dose of creatine nitrate (Low, 3g) and a high dose of creatine nitrate (6g). Participants repeated the experiment with the alternate supplements randomly with a 7 day washout period between each. Performance outcomes were: Bench and Leg Press 1RM, reps to fatigue during the 3rd set of BP and LP 1RM at 70% 1RM repeated 30 mins post supplement on days 1 and 6 and 4 km cycling time trial performance on days 2 and 7. Data were analyzed by repeated measure 4 x 3 x 2 MANOVA with Time, Group, and Gender as factors using Greenhouse-Geisser as appropriate and are presented as mean±SD.

Results
No significant group x time interactions were observed among supplementation groups. Therefore, results for the pooled cohort, separated by gender, are presented. Overall, we observed significant time, gender, and time x gender interaction for bench press and leg press 1RM and bench press lifting reps to fatigue (all, p<0.001), as well as for time to complete the 4-km trial (p<0.01) and average time trial power (p<0.001). A time x gender interaction for only bench press 1RM was observed. Female vs. Male bench press 1RM are: Day1 (89.2±16.9 vs. 203.6±43.8), Day1 Post Supplement (84.2±18.4 vs. 185.9±46.7), Day6 (92.2±19.5 vs. 206.5±45.2), Day 6 Post Supplement (87.3±16.7 vs. 193.0±46.4 lbs). Female vs. Male leg press 1RM: Day1 (630.2±113.0 vs.1047.7±210.9), Day1 Post Supplement (606.3±111.1 vs. 1002.9±215.4), Day6 (652.5±121.1 vs. 1062.9±215.4), Day 6 Post Supplement (611.3±106.4 vs. 1020.4±213.7 lbs). All comparisons are significantly higher for males vs. females. Women decreased less post supplement compared to men for bench press 1RM. Female vs. Male reps to fatigue for the BP are: Day1 (14.9±6.0, vs. 13.7±4.8), Day1 Post Supplement (15.6±4.4 vs. 13.2±4.7), Day6 (15.9±5.2 vs. 14.1±4.9), and Day6 Post Supplement (17.2±4.9 vs. 14.5±5.2). All comparisons are significantly higher for females vs. males. No differences were found for leg press reps to fatigue. Time trial performance was significantly slower for women vs. men, respectively, at all testing time points: Day2 (390.8±79.3, vs. 209.9±35.2), Day7 (390.0±97.9 vs. 203.9±3.6 sec), with men also demonstrating significantly higher average power: Day2 (158.6±26.8, vs. 292.9±54.5), Day7 (161.6±32.3vs. 304.1±59.3 sec).

Conclusion
Ingesting 3 and 6 g/d of creatine nitrate for 7-days had no significant effects on performance measures in comparison to ingesting a placebo. Males were found to be stronger than females, but with significantly lower bench press endurance. Males also performed better on the bicycle tests regarding time to complete the time trial and the average power output associated with the cycling task.

Acknowledgments
This study was supported by Nutrabolt (Bryan, TX) through an unrestricted grant to Texas A&M University.

Seven days of high and low dose creatine nitrate supplementation III: Hemodynamics

R Sowinski, R Dalton, T Grubic, PB Collins, A Coletta, A Reyes, B Sanchez, M Koozehchain, YP Jung, C Rasmussen, P Murano, CP Earnest, M Greenwood, RB Kreider

Methods
In a double-blind, crossover, randomized and placebo-controlled manner; 28 apparently healthy and recreationally active men and women (18 men, 10 women, 21.6±3.7 yr, 20.4±10.6% fat, 24.7±2.9 kg/m2) ingested daily supplements for 7-d consisting of a dextrose flavored placebo (PLA); a low dose of creatine nitrate (Low, 3g) and a high dose of creatine nitrate (6g). Participants repeated the experiment with the alternate supplements randomly with a 7 day washout period between each. Participants had systolic and diastolic blood pressure and heart rate measured after lying supine for 15 minutes and again after 2 minutes lying vertical on a tilt table. This was conducted both pre and 30 minutes post supplement, post resistance exercise on days 1 and 6. Data were analyzed by repeated measure 8 x 3 MANOVA with Time and Group as factors using Greenhouse-Geisser as appropriate and are presented as mean change from Day1 Supine ± 95%CI.

Results
Statistical analysis revealed significant changes over time for DBP and heart rate (HR) (p<.001), but not SBP (p>0.05). No significant group differences or group x time interactions were observed among groups. Therefore, the following reflects pooled cohort time point comparisons. Mean change± 95%CI from Day1 Supine for DBP (mmHg) are: Day 1 vertical (2.8, 95%CI 1.3, 4.3), Day1 Post Supplement, Post Exercise Supine (0.7, 95%CI -0.8, 2.3), Day1 Post Supplement, Post Exercise Vertical (4.0, 95%CI 2.2, 5.8), Day 6 supine (0.8, 95%CI -1.3, 2.9), Day6 vertical (3.8, 95%CI 1.5, 6.2), Day6 Post Supplement, Post Exercise Supine (1.0, 95%CI -1.1, 3.0), and Day6 Post Supplement, Post Exercise Vertical (3.5, 95%CI 1.3, 5.6). Significant differences were found between all supine and vertical DBP measures except between Day1 Vertical and Day6 Supine & Day6 Post Supplement, Post Exercise Supine. Mean change ± 95%CI for HR (bpm) are: Day 1 vertical (11.8, 95%CI 10.0, 13.5), Day1 Post Supplement, Post Exercise Supine (14.5, 95%CI 12.3, 16.6), Day1 Post Supplement, Post Exercise Vertical (24.4, 95%CI 21.8, 27.0), Day 6 supine (-1.4, 95%CI -3.4, 0.5), Day6 vertical (12.4, 95%CI 10.0, 14.8), Day6 Post Supplement, Post Exercise Supine (15.2, 95%CI 12.5, 17.9), and Day6 Post Supplement, Post Exercise Vertical (24.8, 95%CI 21.5, 28.0). Significant differences were found between all time points, Day1 Supine & Day6 Supine, Day1 Vertical & Day6 Vertical, Day1 Post Supplement, Post Exercise Supine & Day6 Vertical, Day1 Post Supplement, Post Exercise Supine & Day6 Post Supplement, Post Exercise Supine, and Day1 Post Supplement, Post Exercise Vertical & Day6 Post Supplement, Post Exercise Vertical.

Conclusion
Changes in body position and exercise increased DBP and HR. However, supplementation of 3 or 6 g/d of creatine nitrate did not significantly affect hemodynamic responses in comparison to a placebo.

Acknowledgments
This study was supported by Nutrabolt (Bryan, TX) through an unrestricted grant to Texas A&M University.

Effect of a multi-ingredient preworkout supplement on cognitive function and perceptions of readiness to perform

MS Koozehchian, PB Collins, R Sowinski, T Grubic, R Dalton, A O’Connor, SY Shin, Y Peter Jung, BK Sanchez, A Coletta, M Cho, A Reyes, C Rasmussen, CP Earnest, PS Murano, M Greenwood, RB Kreider.

Background
The purpose of this study was to examine the short-term effects of ingesting multi-ingredient preworkout supplements (PWS) and PWS at 1.5 times recommended dose (PWS150) on cognitive function and perceptions of readiness to perform.

Methods
We recruited 16 apparently healthy and recreationally active men (21.56 ± 2.11 yr, 20.51 ± 7.64% fat, 27.28 ± 4.25 kg/m2) to participate in a double-blind, crossover, randomized and placebo-controlled manner. Supplements were (1) a dextrose placebo (PLA, 12 g/d); (2) a PWS supplement containing 1.6 g β-alanine, 1.0 g arginine AKG, 1.0 g creatine nitrate, 250 mg ascorbic acid, 150 mg N-acetyl tyrosine, 150 mg caffeine, 5 mg tetramethyluric acid or (3) PWS at ~150% dosage (PWS150) of the base formula for seven days. Participants were required oral administration of one serving of the supplement mixed in ~8 fl oz. of plain water. On testing days, ingesting followed immediately following 30 minutes resting energy expenditure measurement. During non-testing days of the supplementation week, participants ingested their assigned supplement in two forms: during workout days, they ingested the supplement immediately prior to their workout; during non-workout days, they ingested their assigned supplement around noon. In the supplementation week, participants performed a Stroop-Color cognitive function test (CFT) and rated perceptions of readiness to perform on a visual analogue scale (RTP-VAS) on days one, three, five, and seven. On testing days (days one and seven) participants completed CFT and RTP- VAS approximately 30 minutes following supplementation. On days three and five, they referred to the lab to only perform CFT and RTP-VAS. Participants repeated the experiment after a one week washout period with the alternate supplements in a randomized and counterbalanced manner. Data were analyzed by repeated measure MANOVA or ANOVA and are presented as means ± SD and 95% confidence interval from baseline.

Results
The CFT results indicate a significant interaction between groups for the Word test (p = 0.04). In addition, there was a Time effect between groups for the Word test (p = 0.001), Color test (p = 0.002), and Word-Color test (p = 0.001). A significant change from baseline was seen in cognitive function as determined by the Stroop Word-Color Test (p < 0.05). In the regard, we observed a significant improvement in Word count in supplement groups at day three: PWS (6.56 counts, 95% CI, 1.99, 11.13), PWS150 (4.75 counts, 95% CI, 0.17, 9.32), but not the PLA (3.06 counts, 95% CI, -1.50, 7.63); at day five for supplement groups: PWS (6.56 counts, 95% CI, 1.99, 11.13), PWS150 (4.75 counts, 95% CI, 0.17, 9.32), but not the PLA (3.06 counts, 95% CI, -1.50, 7.63); and at day seven for supplement groups: PWS (6.12 counts, 95% CI, 0.23, 12.01), PWS150 (13.06 counts, 95% CI, 7.17, 18.94), but not the PLA (1.81 counts, 95% CI, -4.07, 7.69); For Color recognition, significant improvements were seen in PWS150 and PLA groups at day seven: PWS150 (8.12 counts, 95% CI, 3.89, 12.35), PLA (4.25 counts, 95% CI, 0.02, 8.47), but not the PWS group (1.93 counts, 95% CI, -2.29, 6.16). For the Word-Color assessment, the improvement was seen only in PWS150 at day five: PWS150 (4.87 counts, 95% CI, 0.22, 9.52), but not the PWS (1.81 counts, 95% CI, -2.83, 6.46), and PLA groups (1.68 counts, 95% CI, -2.96, 6.33); and at day seven for PWS150 and PLA groups: PWS150 (4.87 counts, 95% CI, 0.22, 9.52), PLA (1.81 counts, 95% CI, -4.07, 7.69); but not the PWS (3.06 counts, 95% CI, -2.46, 8.58). No significant interactions or changes from baseline were observed between groups regarding perceived readiness to perform as determined by the VAS (p > 0.05).

Conclusion
These data indicate that ingesting a multi-ingredient PWS results in a statistically significant improvement in measured indices of cognitive function test compared to placebo. Furthermore, a significant dose-dependent difference was also observed, as PWS150 showed a higher impact on cognitive performance compared to PWS.

Acknowledgments
This study was supported by Nutrabolt (Bryan, TX) through an unrestricted grant to Texas A&M University.