Quantitative methods in exercie a power primer. Nitrates NO3- are exercide components of Eating disorders that are converted in the human body to nitrites NO2- and then Nitric Oxide NO. Article CAS PubMed Google Scholar Krustrup P, Söderlund K, Relu MU, Ferguson RA, Bangsbo J.

Beetroot juice and improved exercise performance -

Black bandages covered the device to eliminate background light [ 26 ]. To measure leg stiffness the validated Runmatic app v2.

The Runmatic app was used to record leg stiffness during the first minute of each steady state on the incremental test i. At the completion of each steady state step of the incremental running test i. Before starting the incremental test, participants were given instruction to point to a number on a sheet held by a researcher corresponding to their RPE at each step during the trial.

Participants were told that 1 means no exertion at all, and 10 means maximal exertion. Data is presented as mean with standard deviations. Probabilities were also determined to establish whether true differences were lower, similar or higher than the smallest worthwhile change 0.

Statistical analyses were performed using a custom spreadsheet [ 28 ] and SPSS 24 for Mac. However, likely to very likely improvements in EG vs.

See Fig 2 , Table 2 and Table 3 for more details. Lower scores i. The results of our study showed low to trivial improvements in different physiological measures such as running economy at different paces after days of nitrate-rich beetroot juice supplementation in elite distance runners.

These results are in line with other experiments conducted with elite distance athletes, in which submaximal exercise economy was not altered after the supplementation period either [ 20 , 29 ].

A novel contribution from our study was the analysis of the potential benefits of beetroot juice ingestion on different biomechanical variables that has been linked to running performance in the literature, such as leg stiffness [ 23 , 30 ].

For example, leg stiffness has showed to be negatively associated with running economy, since, in theory, a stiffer lower limb would store a higher amount of elastic energy that would assist in the force production during running and, therefore, it would reduce the cost of the exercise [ 9 , 23 ].

Results in our study showed trivial effects of beetroot juice supplementation in leg stiffness, similar to submaximal running economy. Contrary to the results discussed above, likely to very likely large improvements were observed both in the rate of perceived exertion and the time to exhaustion in the experimental group in comparison with the placebo group after the supplementation period, where athletes who consumed nitrate-rich beetroot juice endured more time before voluntary stopping the incremental test on the treadmill.

One potential explanation for this observation is the moderate increase in the SmO 2 of the vastus lateralis of the athletes from the experimental group in comparison with the control group after the supplementation period.

Participants who consumed nitrate-rich beetroot supplementation had larger percent of oxygen saturation in their muscles than their counterparts during exercise.

That could have limited the accumulation of fatigue-related metabolites and reduce the depletion of PCr, which, in the end, could increase the time to exhaustion. NIRS is used for providing information about performance and oxygen muscle function and to evaluate response before, during and after exercise.

In our study, the initial-during-end-exercise SmO 2 was greater in experimental group. That finding is related with recovery capacity and improvements in performance status delaying the fatigue [ 31 ]. Then, greater availability of O 2 could have impacted RPE and, finally it could have led to the observed increase in the time to exhaustion in the experimental group.

For example, several studies have observed that increased oxygen availability via hyperoxia enhances exercise performance [ 32 ]. Nevertheless, further investigations are needed to better understand the mechanisms by which beetroot juice supplementation could reduce RPE and increase time to exhaustion in an incremental test on a treadmill in elite distance runners.

Summarizing, results in our study are in line with other investigations that observed no significant increases in running economy after a supplementation period with nitrate-rich beetroot juice in elite athletes [ 20 , 29 ]; however, the large increase in the time to exhaustion, the reduction in RPE and the increase in the oxygen saturation of the vastus lateralis during exercise, observed after 15 consecutive days of nitrate-rich beetroot supplementation provide novel information about the effects of this nutritional aid.

Further investigations are required to confirm the findings of the present study, as well as to better understand the mechanisms behind these potential benefits for elite distance runners.

However, time to exhaustion in an incremental test on a treadmill, as well as rate of perceived exertion at different running paces were meaningfully higher and lower in the experimental group in comparison with the placebo group, respectively.

These results might be explained in part by a higher muscle oxygen saturation observed in the vastus lateralis of the runners from the experimental group in comparison with the control group.

Anyhow, one of the most performance-related variables in distance running i. These results could have potential practical application for elite distance runners seeking nutritional strategies to improve their running performance.

Browse Subject Areas? Click through the PLOS taxonomy to find articles in your field. Article Authors Metrics Comments Media Coverage Reader Comments Figures. Abstract Purpose Nitrate-rich beetroot juice supplementation has been extensively used to increase exercise economy in different populations.

Funding: The authors received no specific funding for this work. Introduction Performance at the highest level of competition in endurance events depends on several physiological, psychological and biomechanical parameters [ 1 — 5 ].

Download: PPT. Procedures Beetroot juice supplementation protocol. Physiological parameters Incremental running test. O2 saturation measurement. Biomechanical parameters To measure leg stiffness the validated Runmatic app v2.

Psychological parameters At the completion of each steady state step of the incremental running test i. Statistical analyses Data is presented as mean with standard deviations. Fig 2. Table 2.

Mean values and standard deviations for the experimental and placebo groups before Pre and after Post the supplementation period. Table 3. Pre-post differences on the studied variables in the placebo vs experimental conditions.

Discussion The results of our study showed low to trivial improvements in different physiological measures such as running economy at different paces after days of nitrate-rich beetroot juice supplementation in elite distance runners.

Supporting information. S1 File. CONSORT checklist of information to include in a randomized controlled trial. s DOC. S2 File. Study protocol as approved by the ethics committee. s PDF. References 1. Alway SE, McCrory JL, Kearcher K, Vickers A, Frear B, Gilleland DL, et al.

Resveratrol Enhances Exercise-Induced Cellular and Functional Adaptations of Skeletal Muscle in Older Men and Women. Journals Gerontol Ser A.

Rabadán M, Díaz V, Calderón FJ, Benito PJ, Peinado AB, Maffulli N. Physiological determinants of speciality of elite middle- and long-distance runners.

J Sports Sci. McCormick A, Meijen C, Marcora S. Psychological Determinants of Whole-Body Endurance Performance. Sport Med. Santos-Concejero J, Billaut F, Grobler L, Oliván J, Noakes TD, Tucker R. Maintained cerebral oxygenation during maximal self-paced exercise in elite Kenyan runners.

J Appl Physiol. Coleman DR, Cannavan D, Horne S, Blazevich AJ. Leg stiffness in human running: Comparison of estimates derived from previously published models to direct kinematic-kinetic measures.

J Biomech. Hoogkamer W, Kipp S, Frank JH, Farina EM, Luo G, Kram R. A Comparison of the Energetic Cost of Running in Marathon Racing Shoes.

Bacon AP, Carter RE, Ogle EA, Joyner MJ. VO2max trainability and high intensity interval training in humans: a meta-analysis. PLoS One. Vuorimaa T, Vasankari T, Rusko H. Comparison of physiological strain and muscular performance of athletes during two intermittent running exercises at the velocity associated with VO2max.

Int J Sports Med. Germany, Federal Republic of; ; 96— Moore IS. Is There an Economical Running Technique? A Review of Modifiable Biomechanical Factors Affecting Running Economy.

Barnes KR, Kilding AE. Running economy: measurement, norms, and determining factors. Sport Med—Open. Balsalobre-Fernández C, Santos-Concejero J, Grivas G V. Effects of Strength Training on Running Economy in Highly Trained Runners.

J Strength Cond Res. Beattie K, Carson BP, Lyons M, Rossiter A, Kenny IC. The Effect of Strength Training on Performance Indicators in Distance Runners.

Volek JS, Noakes T, Phinney SD. Nitrates NO3- are inorganic components of vegetables that are converted in the human body to nitrites NO2- and then Nitric Oxide NO. NO is a gaseous signaling molecule that results in various critical physiological functions, such as improved blood pressure, neurotransmission, enhanced blood flow to muscles, improved muscle efficiency and better sexual health.

Natural sources of nitrates include celery, arugula, radishes, and spinach. But perhaps the best-known source of nitrates for endurance athletes is the humble beet. This red root vegetable is increasingly found in supplements that promise to deliver a punch of nitrates for better athletic performance — but how does the NO effect work?

RELATED: 5 Winter Produce Picks to Include In Your Diet. The consumption of nitrates and subsequent formation of NO is considered to be an essential part of human functioning.

Nitrates are a direct catalyst for a process called vasodilation. Vasodilation is the widening of blood vessels in the body and this function is paramount to human health, as the process itself is utilized by the body to deliver oxygenated blood to areas that are in greatest need. Increased vasodilation also acts as a transporter for glucose, fats, and other nutrients throughout the body.

The effects of NO can be seen all over the body including in brain function neurotransmission , wound healing, hemoglobin delivery of oxygen in the blood, blood flow, blood clotting, muscle contractility, glucose sensitivity, and mitochondrial function.

The daily consumption goal of nitrates is recommended at 3. When we regularly participate in exercise, training, and sport we invariably become more efficient at the same level of intensity and therefore have a perceived reduction in effort. This takes into account mechanical, neurological, and physiological processes.

Consumption of nitrate-containing foods and supplements has been shown to improve the efficiency of working muscles to reduce the work required to achieve the same result. In the context of exercise, it means your athletic capacity can be improved.

Breaking it down even further, it means you can go further and longer at the same intensity without fatigue — something pretty much everyone wants when it comes to sports with an aerobic element.

Nitrate supplementation has been shown to reduce the cellular-level adenosine triphosphate ATP and PhosphoCreatine PCr cost associated with producing muscular contractions, effectively reducing the amount of PCr utilized for the given work completed.

Subsequently, nitrates enhance muscle contractile efficiency , in addition to improving the efficiency of the cellular metabolism for energy production oxidative phosphorylation.

In other words, you can perform more contractions with less energy, thus improving your efficiency. Studies have demonstrated that increased dietary nitrate intake for five to seven consecutive days resulted in more than a fourfold increase in saliva and plasma nitrate, nitrite and subsequent nitric oxide.

Through the study of nitrate-rich supplements meeting the criteria of mg nitrates, improvements have been shown in the reduced cost of oxygen uptake and improved efficiency in walking, running, rowing, cycling, and repeated sprint exercises.

This improved efficiency is not limited to any particular demographic, with benefits seen in all age groups studied , including untrained males, trained males and females, healthy men and women, along with patients suffering from peripheral artery disease.

The benefits of nitrate supplements can assist young and old, male and female and also those individuals who are attempting to adopt a healthier lifestyle. It should be noted that if you are an elite or highly trained athlete then the results may not be as impressive.

Research investigating the effect on professional cyclists has been equivocal and this may be explained by the enhanced physiological system of these athletes and their pre-existing high levels of nitric oxide that is already produced endogenously inside the body.

RELATED: Ask Stacy: Should I Take Supplements?

Beetrolt the impoved effects of foods that are thought to Beetroot juice and improved exercise performance a beneficial effect on aerobic performance, researchers found exercisw beetroot, grapes, improvev cherries, Beetroot juice and improved exercise performance pine bark extract, Beehroot contribute to nitric oxide availability in the Enhance skin texture, boost endurance jucie performance. Assessing Tailored weight management from studies involving participants from 25 different countries, the meta-analysis evaluated the effect of Bertroot nitrate-rich foods typically green leafy vegetablesfoods that contain polyphenols such as berries, cherries and cocoaand L-Citrulline found in watermelon on exercise endurance performance. The study found that the nitrate levels contained in beetroot, which have been shown to boost blood flow and increase the delivery of nutrients and oxygen to muscles during exercise, helped athletes perform better more quickly. Similarly, the polyphenols in grapes, cherries and pine bark extract helped protect nitrate from degradation in the body, boosting stamina. And, despite the ability of L-citrulline to boost nitric oxide production in the body, consuming watermelon high in L-citrulline did not boost exercise performance. Contacts for interview: Professor Jon Buckley E : Jon. Buckley unisa.

Video

Why Beets Are Amazing For Exercise \u0026 Performance! 💪

Beetroot juice and improved exercise performance -

Nitric oxide dilates blood vessels, which delivers oxygen more quickly to the muscles, so energy can be burned to fuel the exercising muscles. The Australian Institute of Sport AIS has assessed beetroot and classified it as a Group A supplement.

This means there is strong scientific evidence for use in specific situations in sport. The AIS advises beetroot supplementation can be beneficial for exercise, training and competitive events that lasts 4—30 minutes and in team sports with intermittent exercise.

For performance benefits, the AIS advises the beetroot product be it juice, powder or food should have between —mg of inorganic nitrate in it. Check the label. There are several concentrated juices available on the market. Beetroot contains about mg per g of nitrate, so you need to consume at least g of baked beetroot to get the same effect.

To give the nitrates time to be converted to nitric oxide and absorbed into your bloodstream, you need to consume the product 2—3 hours before training or competition.

You may get added benefits drinking beetroot juice for several days leading up to training or competition. These will kill the bacteria in your mouth needed to convert the nitrate to nitrite. Your urine will turn red, and this will make it difficult to determine if you are dehydrated.

Your poo may also turn red. Some people may experience an upset stomach when consuming beetroot juice. So try drinking it while training to determine if you have any problems. NO is a gaseous signaling molecule that results in various critical physiological functions, such as improved blood pressure, neurotransmission, enhanced blood flow to muscles, improved muscle efficiency and better sexual health.

Natural sources of nitrates include celery, arugula, radishes, and spinach. But perhaps the best-known source of nitrates for endurance athletes is the humble beet. This red root vegetable is increasingly found in supplements that promise to deliver a punch of nitrates for better athletic performance — but how does the NO effect work?

RELATED: 5 Winter Produce Picks to Include In Your Diet. The consumption of nitrates and subsequent formation of NO is considered to be an essential part of human functioning.

Nitrates are a direct catalyst for a process called vasodilation. Vasodilation is the widening of blood vessels in the body and this function is paramount to human health, as the process itself is utilized by the body to deliver oxygenated blood to areas that are in greatest need.

Increased vasodilation also acts as a transporter for glucose, fats, and other nutrients throughout the body. The effects of NO can be seen all over the body including in brain function neurotransmission , wound healing, hemoglobin delivery of oxygen in the blood, blood flow, blood clotting, muscle contractility, glucose sensitivity, and mitochondrial function.

The daily consumption goal of nitrates is recommended at 3. When we regularly participate in exercise, training, and sport we invariably become more efficient at the same level of intensity and therefore have a perceived reduction in effort.

This takes into account mechanical, neurological, and physiological processes. Consumption of nitrate-containing foods and supplements has been shown to improve the efficiency of working muscles to reduce the work required to achieve the same result.

In the context of exercise, it means your athletic capacity can be improved. Breaking it down even further, it means you can go further and longer at the same intensity without fatigue — something pretty much everyone wants when it comes to sports with an aerobic element.

Nitrate supplementation has been shown to reduce the cellular-level adenosine triphosphate ATP and PhosphoCreatine PCr cost associated with producing muscular contractions, effectively reducing the amount of PCr utilized for the given work completed.

Subsequently, nitrates enhance muscle contractile efficiency , in addition to improving the efficiency of the cellular metabolism for energy production oxidative phosphorylation.

The final position was sitting back onto the heels and trying to achieve as deep a squat as possible. Then, participants immediately reversed the motion and ascended back to the up-right position and simultaneously press, or toss, the ball to a target spot about eight feet up the wall.

FBS exercise was performed with free weight. This was executed starting from the upright position with the knees and hips fully extended. Both feet were positioned flat on the floor in parallel or slightly rotated outwards at a distance of approximately shoulder-width apart. Then, the participants flexed the knees and hips eccentric action to descend the barbell in a controlled manner until the top of the thighs reached below the horizontal plane.

From this position, the participants immediately reversed motion and ascended back to the up-right position until the knees and hips were fully extended. Serum cortisol concentrations were analyzed using chemiluminescent microparticle immunoassay by Abbott Architect immunoassay analyzer Abbott Laboratories, Abbott Park, IL, USA [ 48 ].

Blood lactate concentrations were measured by an experienced investigator using the analyzer Lactate ProTM 2 Arkray Factory Inc.

Clean blood samples were obtained from the index finger of the left hand as in previous studies [ 19 , 22 ]. Nonin oximeters have commonly been used for monitoring SpO 2 [ 50 ]. SpO 2 was tested on the index finger of the right hand.

Raw data were stored on the internal memory of the WristOx 2. Lower limb fatigue was determined by a countermovement jump CMJ test. The CMJ test was performed using a force platform Musclelab, Ergotest Technology AS, Langesund, Norway before and after the WOD test following a method described previously [ 44 , 45 ].

Participants carried out 3 jumps separated by a s rest, and the mean values of vertical height recorded in the 3 CMJs were used in the statistical analysis. PL , a two-way analysis of variance ANOVA with repeated measures was applied experimental condition x time.

The magnitude of the response to both experimental conditions was estimated by partial eta-squared η p 2. The scale for classification of η p 2 was 0. Statistical power SP was calculated. All statistical procedures were applied using the software package SPSS version All participants reported consuming BJ or PL supplement at the correct times.

BJ intake was well tolerated by all participants of the study, however, some athletes showed beeturia red urine and red stools.

The participants maintained exercise and dietary habits before each testing visit according to nutritional and exercise guidelines. Number of repetitions performed after beetroot juice BJ and placebo PL intake. The statistical analyses corresponding to lactate, SpO 2 and mechanical fatigue are shown in Table 1.

Abbreviations: BJ: beetroot juice; PL: placebo; arterial oxygen saturation: SpO 2. The main finding of this study was that BJ intake improved WOD performance only after the rest time between exercises.

Interestingly, cortisol levels and muscular fatigue increased, and SpO 2 fell to a greater extent following BJ intake than under PL conditions. There are limited studies available regarding the effect of BJ intake on WOD performance. Kramer et al. The WOD was different from that proposed in this study, however, they did not find improvements in WOD performance.

Both WOD were characterized by a strong involvement of anaerobic metabolism and the need to maintain the effort required through aerobic metabolism. Surprisingly, we only discovered a positive ergogenic effect of BJ when a 3-min rest was established between the WB shots and the FBS exercise.

When WB and FBS exercises were executed without rest, anaerobic conditions of the routine were increased, and no improvements were noted in the number of repetitions.

During rest periods, the capacity to repeat exercise sets at high work rates depends on the rate of phosphocreatine PCr resynthesis by oxidative phosphorylation.

PCr resynthesis is determined through local muscle blood flow and O 2 delivery to skeletal muscle [ 53 , 54 ]. The blood lactate concentrations obtained in this study demonstrated that WOD was performed under high acidification conditions. As an ergogenic aid, BJ intake before training or competition of high intensity intermittent efforts high lactate levels could be an appropriate strategy to delay muscle fatigue and improve exercise performance during rest time between sets.

Further research is warranted to elucidate the potential efficacy of BJ ingestion during recovery time in primarily anaerobic efforts. As expected, cortisol and testosterone levels were markedly increased in both experimental conditions.

CF workouts seem to influence testosterone and cortisol hormonal response [ 39 , 58 ]. However, there is little scientific evidence to suggest that consistent alterations in cortisol and testosterone concentrations occur after BJ intake.

Cortisol levels augmented in both experimental groups in response to high-intensity exercise. It has been stated that cortisol secretion is induced during high-intensity exercise in humans [ 28 ] and is also stimulated by NO concentrations [ 27 ].

We conjecture that a series of exercise- and BJ-dependent multisystemic physiological events were triggered. Cortisol elevation enlarged the availability of all fuel substrates by mobilizing glucose [ 30 , 31 ]. Consequently, BJ improved performance in the first routine and, notwithstanding increasing anaerobic exercise conditions, performance was maintained during the second routine.

Testosterone response may have been conditioned by the cortisol levels observed after the CF workouts. High cortisol levels in response to anaerobic exercise possibly caused a potential for inhibition of testosterone production via steroid inhibition, and thus, cortisol induced a direct suppressive effect on testosterone steroidogenesis [ 61 ].

An absolute ratio less than or equal to 0. The data reported in this study did not indicate a negative catabolic state or changes between the two experimental groups.

Another physiological mechanism associated with the low availability of muscle oxygen during lactic anaerobic efforts could be the decrease in SpO 2. To our knowledge, there are no studies examining the effects of BJ intake on SpO 2 during CF workouts, thus, the arguments are limited to discussing findings related to aerobic and anaerobic efforts.

Several studies reported that BJ did not alter SpO 2 during predominantly aerobic efforts in trained male cyclists [ 64 ] and well-trained runners [ 65 ] under normoxia and hypoxia conditions. Engan et al. Other authors determined that BJ supplementation did not prompt a significant variation in SpO 2 during submaximal static and dynamic apneas [ 67 ].

Arnold et al. indicated that runners who improved time-trial performance responded to hypoxia with greater arterial desaturation [ 65 ]. In our study, CF athletes who improved WOD performance after BJ intake also showed a greater reduction in SpO 2 3.

A decline in SpO 2 is indicative of an intensification in oxygen uptake by the muscle mass involved. The greater number of repetitions carried out in the total WOD after BJ intake likely caused an increase in oxygen consumption in the active muscles and promoted a fall in SpO 2 , demanding the participation of lactic anaerobic metabolism.

A predominantly anaerobic metabolism was evidenced in both experimental conditions by the lactate concentrations detected. Rogatzki et al. demonstrated using a parallel squat exercise in which lactate appears to accumulate in response to a rising number of repetitions with reducing recovery time between sets [ 68 ].

In their study, a workout of muscular endurance was compared with a hypertrophy workout. Higher blood lactate levels were observed during muscular endurance training Δ In our study, a higher number of repetitions was carried out during the first exercise routine after BJ conditions.

In the second routine, recovery time was suppressed, and a similar number of repetitions was completed Δ 0. The total count of repetitions including the first and second routine was significantly higher after BJ intake than in PL condition data not shown.

However, blood lactate levels were 3. On intermittent exercise, other groups have found that lactate pre-exercise vs. The debate over the role that BJ plays in lactate response to exercise is controversial. In our study, lactate concentrations almost doubled or tripled those of the studies mentioned.

The differences observed among studies might be attributed to the type of exercise and the evaluation protocol used in each study. BJ was expected to increase lactate concentrations to a higher magnitude than under PL conditions, however this physiological event did not occur. The fall in SpO 2 and the blood lactate concentrations determined a significant metabolic fatigue in both experimental conditions.

Rises in lactate values provides information about a possible state of muscle fatigue [ 68 ]. Previous findings in CF practitioners have determined that metabolic fatigue is associated with post-exercise muscular fatigue [ 70 ].

Effectively, a higher muscular fatigue was verified after BJ intake Mechanical fatigue evaluated by a CMJ test has been considered as a good indicator of muscle fatigue in resistance exercises and in CF workouts in aerobic and anaerobic conditions [ 70 , 71 ].

We suggest that the physiological benefits vasodilation, increased blood flow, etc. of BJ during the established 3-min rest after WB produced an increase in the number of repetitions in the FBS exercise 1st routine. This improvement in performance probably increased the anaerobic conditions of the muscle mass involved before starting the second routine and, therefore, a greater fall in S P O 2 to supply the oxygen needs of the muscle mass was produced at the end of exercise.

The relationship of these physiological mechanisms resulted in increased muscle fatigue after completing the WOD. This study presents some limitations that should be considered. The sample size should be augmented, because minimal changes are usually observed in well-trained athletes [ 19 ].

An increased cortisol response was observed in nine athletes after BJ intake. BJ intake improved WOD performance only after a rest time between WB and FBS exercises. Performance was maintained when anaerobic conditions of exercise were augmented.

This performance increase in the first routine generated greater hypoxia in the muscle mass involved, possibly conditioning post-exercise performance. This was observed in the fall in SpO 2 and in the increase in muscle fatigue measured at the end of the WOD.

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Photo: Beetfoot Images "], Beetroot juice and improved exercise performance { "nextExceptions": "img, blockquote, exefcise, "nextContainsExceptions": "img, blockquote, a. Beetrooot, a. They are good for you! What mum did not realize, however, was Beetroot juice and improved exercise performance reason why eating your greens may be beneficial: Leafy greens contain a compound called nitrates. Turns out these little gems do have a whole lot of benefits, and mum probably never even knew about them! Nitrates NO3- are inorganic components of vegetables that are converted in the human body to nitrites NO2- and then Nitric Oxide NO. Athletes often seek exfrcise Beetroot juice and improved exercise performance performande Beetroot juice and improved exercise performance to increase performance during exercise. Among various supplements, much attention has been paid to beetroot in recent years. Beetroot Beetgoot a source Importance of breakfast variety carbohydrates, fiber, protein, minerals, and vitamins; also, it is a natural source of nitrate and associated with improved sports performance. Nitrates can the modification of skeletal muscle contractile proteins or calcium handling after translation. The time to reach the peak plasma nitrate is between 1 and 3 h after consumption of a single dose of nitrate. Nitrate is metabolized by conversion to nitrite and subsequently nitric oxide.