<?xml version="1.0" encoding="utf-8"?>
<!DOCTYPE article PUBLIC "-//NLM//DTD Journal Publishing DTD v2.3 20070202//EN" "journalpublishing.dtd">
<article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" article-type="research-article" dtd-version="2.3" xml:lang="EN">
<front>
<journal-meta>
<journal-id journal-id-type="publisher-id">Front. Nutr.</journal-id>
<journal-title>Frontiers in Nutrition</journal-title>
<abbrev-journal-title abbrev-type="pubmed">Front. Nutr.</abbrev-journal-title>
<issn pub-type="epub">2296-861X</issn>
<publisher>
<publisher-name>Frontiers Media S.A.</publisher-name>
</publisher>
</journal-meta>
<article-meta>
<article-id pub-id-type="doi">10.3389/fnut.2024.1345922</article-id>
<article-categories>
<subj-group subj-group-type="heading">
<subject>Nutrition</subject>
<subj-group>
<subject>Original Research</subject>
</subj-group>
</subj-group>
</article-categories>
<title-group>
<article-title>Effect of five hours of mixed exercise on urinary nitrogen excretion in healthy moderate-to-well-trained young adults</article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname>Clauss</surname>
<given-names>Matthieu</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
<uri xlink:href="https://loop.frontiersin.org/people/1113418/overview"/>
<role content-type="https://credit.niso.org/contributor-roles/conceptualization/"/>
<role content-type="https://credit.niso.org/contributor-roles/formal-analysis/"/>
<role content-type="https://credit.niso.org/contributor-roles/investigation/"/>
<role content-type="https://credit.niso.org/contributor-roles/methodology/"/>
<role content-type="https://credit.niso.org/contributor-roles/writing-original-draft/"/>
<role content-type="https://credit.niso.org/contributor-roles/writing-review-editing/"/>
</contrib>
<contrib contrib-type="author" equal-contrib="yes">
<name>
<surname>Burkhardt</surname>
<given-names>Meike</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
<xref ref-type="aff" rid="aff2"><sup>2</sup></xref>
<xref ref-type="author-notes" rid="fn0001"><sup>&#x2020;</sup></xref>
<uri xlink:href="https://loop.frontiersin.org/people/2650904/overview"/>
<role content-type="https://credit.niso.org/contributor-roles/formal-analysis/"/>
<role content-type="https://credit.niso.org/contributor-roles/investigation/"/>
<role content-type="https://credit.niso.org/contributor-roles/writing-review-editing/"/>
</contrib>
<contrib contrib-type="author" equal-contrib="yes">
<name>
<surname>W&#x00F6;ber</surname>
<given-names>Sophie</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
<xref ref-type="aff" rid="aff2"><sup>2</sup></xref>
<xref ref-type="author-notes" rid="fn0001"><sup>&#x2020;</sup></xref>
<uri xlink:href="https://loop.frontiersin.org/people/2590313/overview"/>
<role content-type="https://credit.niso.org/contributor-roles/formal-analysis/"/>
<role content-type="https://credit.niso.org/contributor-roles/investigation/"/>
<role content-type="https://credit.niso.org/contributor-roles/writing-review-editing/"/>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Sk&#x00E5;lhegg</surname>
<given-names>Bj&#x00F8;rn Steen</given-names>
</name>
<xref ref-type="aff" rid="aff3"><sup>3</sup></xref>
<uri xlink:href="https://loop.frontiersin.org/people/414460/overview"/>
<role content-type="https://credit.niso.org/contributor-roles/conceptualization/"/>
<role content-type="https://credit.niso.org/contributor-roles/methodology/"/>
<role content-type="https://credit.niso.org/contributor-roles/supervision/"/>
<role content-type="https://credit.niso.org/contributor-roles/writing-review-editing/"/>
</contrib>
<contrib contrib-type="author" corresp="yes">
<name>
<surname>Jensen</surname>
<given-names>J&#x00F8;rgen</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
<xref ref-type="corresp" rid="c001"><sup>&#x002A;</sup></xref>
<uri xlink:href="https://loop.frontiersin.org/people/25500/overview"/>
<role content-type="https://credit.niso.org/contributor-roles/conceptualization/"/>
<role content-type="https://credit.niso.org/contributor-roles/funding-acquisition/"/>
<role content-type="https://credit.niso.org/contributor-roles/methodology/"/>
<role content-type="https://credit.niso.org/contributor-roles/supervision/"/>
<role content-type="https://credit.niso.org/contributor-roles/writing-review-editing/"/>
</contrib>
</contrib-group>
<aff id="aff1"><sup>1</sup><institution>Department of Physical Performance, Norwegian School of Sport Sciences</institution>, <addr-line>Oslo</addr-line>, <country>Norway</country></aff>
<aff id="aff2"><sup>2</sup><institution>Department of Sport and Sport Science, University of Freiburg</institution>, <addr-line>Freiburg</addr-line>, <country>Germany</country></aff>
<aff id="aff3"><sup>3</sup><institution>Department of Nutrition, Division for Molecular Nutrition, University of Oslo</institution>, <addr-line>Oslo</addr-line>, <country>Norway</country></aff>
<author-notes>
<fn fn-type="edited-by" id="fn0002">
<p>Edited by: Mark Elisabeth Willems, University of Chichester, United Kingdom</p>
</fn>
<fn fn-type="edited-by" id="fn0003">
<p>Reviewed by: Carol Johnston, Arizona State University, United States</p>
<p>Ratko Peric, OrthoSport Banja Luka, Bosnia and Herzegovina</p>
</fn>
<corresp id="c001">&#x002A;Correspondence: J&#x00F8;rgen Jensen, <email>jorgen.jensen@nih.no</email></corresp>
<fn fn-type="equal" id="fn0001">
<p><sup>&#x2020;</sup>These authors have contributed equally to this work</p>
</fn>
</author-notes>
<pub-date pub-type="epub">
<day>21</day>
<month>02</month>
<year>2024</year>
</pub-date>
<pub-date pub-type="collection">
<year>2024</year>
</pub-date>
<volume>11</volume>
<elocation-id>1345922</elocation-id>
<history>
<date date-type="received">
<day>28</day>
<month>11</month>
<year>2023</year>
</date>
<date date-type="accepted">
<day>01</day>
<month>02</month>
<year>2024</year>
</date>
</history>
<permissions>
<copyright-statement>Copyright &#x00A9; 2024 Clauss, Burkhardt, W&#x00F6;ber, Sk&#x00E5;lhegg and Jensen.</copyright-statement>
<copyright-year>2024</copyright-year>
<copyright-holder>Clauss, Burkhardt, W&#x00F6;ber, Sk&#x00E5;lhegg and Jensen</copyright-holder>
<license xlink:href="http://creativecommons.org/licenses/by/4.0/">
<p>This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.</p>
</license>
</permissions>
<abstract>
<sec>
<title>Introduction</title>
<p>Carbohydrates and fats are the primary energy substrates during exercise, but proteins can also contribute. When proteins are degraded in the body, the amino groups are mainly converted to urea and excreted. Therefore, nitrogen excretion has been used as a marker of protein degradation, but a clear conclusion has yet to be reached on the effect of exercise on nitrogen excretion. Thus, we tested whether exercise increases nitrogen excretion.</p>
</sec>
<sec>
<title>Methods</title>
<p>Fifteen young, healthy, moderate-to-well-trained participants (4 females, 11 males, VO<sub>2max</sub> 54.4&#x2009;&#x00B1;&#x2009;1.7&#x2009;mL&#x00B7;kg<sup>&#x2212;1</sup>&#x00B7;min<sup>&#x2212;1</sup>; mean &#x00B1; SEM) participated in a randomized, balanced cross-over design investigation consisting of 1 day with 5 h of exercise (exercise day, EX) and 1 day with no exercise (control day, CON). The participants recorded their dietary intake the day before from 16:00 and throughout the intervention day. They then repeated these dietary intakes on the second trial day. A standardized lunch was provided on both days. In addition, participants were allowed to consume almost protein-free snacks in EX to ensure the same energy balance during both trial days. Urine was collected throughout the whole testing period, and urinary 3-methylhistidine (3-MH) excretion was measured to examine muscular catabolism. The sweat rate was calculated during the exercise period.</p>
</sec>
<sec>
<title>Results and discussion</title>
<p>The urinary nitrogen and 3-MH excretions did not differ significantly between EX and CON (<italic>p</italic>&#x2009;=&#x2009;0.764 and <italic>p</italic>&#x2009;=&#x2009;0.953). The sweat rate was 2.55&#x2009;&#x00B1;&#x2009;0.25 L in EX and 0.14&#x2009;&#x00B1;&#x2009;0.15 L in CON (<italic>p</italic>&#x2009;&#x003C;&#x2009;0.001), and by estimating sweat nitrogen excretion, total nitrogen excretion was shown to differ with exercise. Our results showed that 5 hours of mixed exercise did not significantly impact urinary nitrogen and 3-MH excretions in healthy moderate-to-well-trained young adults.</p>
</sec>
</abstract>
<kwd-group>
<kwd>urinary nitrogen excretion</kwd>
<kwd>sweat nitrogen excretion</kwd>
<kwd>protein metabolism</kwd>
<kwd>endurance exercise</kwd>
<kwd>3-methylhistidine excretion</kwd>
</kwd-group>
<counts>
<fig-count count="6"/>
<table-count count="3"/>
<equation-count count="2"/>
<ref-count count="79"/>
<page-count count="11"/>
<word-count count="8698"/>
</counts>
<custom-meta-wrap>
<custom-meta>
<meta-name>section-at-acceptance</meta-name>
<meta-value>Sport and Exercise Nutrition</meta-value>
</custom-meta>
</custom-meta-wrap>
</article-meta>
</front>
<body>
<sec sec-type="intro" id="sec1">
<label>1</label>
<title>Introduction</title>
<p>The view of proteins as an energy substrate has changed over time. In 1842, Justus von Liebig suggested that protein was the &#x201C;only true nutrient, providing both the machinery of the body and the fuel for its work&#x201D; (<xref ref-type="bibr" rid="ref1">1</xref>). Not long after Liebig&#x2019;s claim, several studies proved this conclusion wrong (<xref ref-type="bibr" rid="ref2">2</xref>). Nevertheless, von Liebig&#x2019;s claim led to many studies combining exercise and the measurement of nitrogen excretion via urine collection during the late 19<sup>th</sup> century. Many of these studies showed rises in nitrogen excretion after physical work or exercise (see <xref rid="SM1" ref-type="supplementary-material">Supplementary Table S1</xref>). However, nitrogen excretion was seldom quantified reliably, nor was the data collection accompanied by a standardized diet or a control trial within the same subjects (<xref ref-type="bibr" rid="ref3">3</xref>&#x2013;<xref ref-type="bibr" rid="ref12">12</xref>). This makes the comparison of studies challenging, as nitrogen excretion is highly correlated with protein (nitrogen) intake from food (<xref ref-type="bibr" rid="ref13">13</xref>). It is important to note that dietary proteins are the only exogenous nitrogen source that can regulate the body&#x2019;s nitrogen metabolism. In addition, nitrogen excretion decreases as energy intake increases at a fixed nitrogen intake (<xref ref-type="bibr" rid="ref13">13</xref>). Furthermore, few participants were included in the studies and no specific energy expenditures were calculated during the work/exercise periods. Consequently, no clear conclusion has been reached on the effect of exercise on urinary nitrogen excretion.</p>
<p>Since then, some newer studies have investigated the effect of exercise on urinary nitrogen excretion in more reliable designs. Urine was collected on an exercise day, including a running or cycling exercise, and compared to urine collected on a resting day (<xref ref-type="bibr" rid="ref14">14</xref>&#x2013;<xref ref-type="bibr" rid="ref17">17</xref>). Urinary nitrogen excretion was either significantly higher or tended to be higher during the exercise day versus the control day. However, another study with a similar cycling exercise did not measure any difference in urinary nitrogen excretion between the exercise and the control day (<xref ref-type="bibr" rid="ref18">18</xref>). Thus, a clear conclusion remains to be reached on the effect of exercise on urinary nitrogen excretion. As nitrogen excretion provides a reasonable estimate of whole-body protein degradation, arriving at a conclusion would allow several practical applications.</p>
<p>During exercise, nitrogen can also be excreted in sweat. However, the amount of sweat nitrogen excreted varies greatly. Some studies have shown as low as &#x2248;50&#x2009;mg&#x00B7;h<sup>&#x2212;1</sup> of nitrogen excreted in sweat (<xref ref-type="bibr" rid="ref19">19</xref>), some &#x2248;250&#x2009;mg&#x00B7;h<sup>&#x2212;1</sup> (<xref ref-type="bibr" rid="ref17">17</xref>, <xref ref-type="bibr" rid="ref20">20</xref>), some &#x2248;500&#x2009;mg&#x00B7;h<sup>&#x2212;1</sup> (<xref ref-type="bibr" rid="ref21">21</xref>), and others reported &#x2248;600&#x2009;mg&#x00B7;h<sup>&#x2212;1</sup> (<xref ref-type="bibr" rid="ref22">22</xref>, <xref ref-type="bibr" rid="ref23">23</xref>), and up to &#x2248;1700&#x2009;mg&#x00B7;h<sup>&#x2212;1</sup> (<xref ref-type="bibr" rid="ref24">24</xref>). Despite the large disparity, most values fall within the same concentration range of approximately 0.4 to 1.2&#x2009;mg/mL. Differences in temperature and dietary nitrogen intake are the main factors explaining the variance within this range (<xref ref-type="bibr" rid="ref19">19</xref>, <xref ref-type="bibr" rid="ref25">25</xref>). A significant linear relationship exists between nitrogen intake and sweat nitrogen loss at a fixed temperature (<xref ref-type="bibr" rid="ref20">20</xref>, <xref ref-type="bibr" rid="ref25">25</xref>). Therefore, it is possible to estimate sweat nitrogen loss if both factors are known, and the sweat rate is measured.</p>
<p>A further <italic>in-vivo</italic> non-invasive method for quantifying skeletal myofibrillar protein catabolism is the measurement of urinary 3-methylhistidine (3-MH) excretion (<xref ref-type="bibr" rid="ref26">26</xref>, <xref ref-type="bibr" rid="ref27">27</xref>). The amino acid 3-MH is formed by adding a methyl group to specific histidine residues in the peptide chains of actin in all muscles and myosin in white muscle fibers (<xref ref-type="bibr" rid="ref28">28</xref>&#x2013;<xref ref-type="bibr" rid="ref30">30</xref>). About 80% of the excreted 3-MH in adults is derived from actin and 20% from myosin (<xref ref-type="bibr" rid="ref31">31</xref>). When muscle protein is degraded, 3-MH is neither reused for protein synthesis (<xref ref-type="bibr" rid="ref32">32</xref>) nor metabolized (<xref ref-type="bibr" rid="ref33">33</xref>) and is excreted in the urine (<xref ref-type="bibr" rid="ref34">34</xref>). Because plasma levels of 3-MH are low and renal clearance is high, urinary excretion reliably reflects myofibrillar degradation rates.</p>
<p>Previous studies have shown that when participants cycle to exhaustion, they need to ingest a large quantity of protein to be in nitrogen balance (<xref ref-type="bibr" rid="ref35">35</xref>&#x2013;<xref ref-type="bibr" rid="ref37">37</xref>). This indicates increased nitrogen excretion, but it remains to be proven. Therefore, in this study, participants completed a randomized, balanced cross-over design of two trial days. One day consisted of 5&#x2009;h of exercise (EX), and the other comprised 5&#x2009;h of rest and no exercise of any form (CON). Urine samples were collected during the entire testing period. Based on this, we hypothesized that the exercise day would increase urinary nitrogen and 3-MH excretions.</p>
</sec>
<sec sec-type="materials|methods" id="sec2">
<label>2</label>
<title>Materials and methods</title>
<sec id="sec3">
<label>2.1</label>
<title>Participants</title>
<p>Fifteen young, healthy, moderate-to-well-trained adults (4 women and 11 men) were recruited for the study. Their characteristics are described in <xref ref-type="table" rid="tab1">Table 1</xref>. Participants were informed about the study before providing their written informed consent. In addition, all participants completed a health questionnaire to rule out potential risk factors. The Norwegian School of Sport Sciences&#x2019; Ethics Committee (Application 190&#x2013;170,621) and the Norwegian Centre for Research Data (Reference number 473323) approved the study, which conformed to the standards set by the latest revision of the Declaration of Helsinki.</p>
<table-wrap position="float" id="tab1">
<label>Table 1</label>
<caption>
<p>Age, anthropometric data, and VO<sub>2max</sub> in pretests.</p>
</caption>
<table frame="hsides" rules="groups">
<thead>
<tr>
<th align="left" valign="middle">Parameter</th>
<th align="center" valign="middle">Women and men (<italic>n</italic> =&#x2009;15)</th>
<th align="center" valign="middle">Women (<italic>n</italic> =&#x2009;4)</th>
<th align="center" valign="middle">Men (<italic>n</italic> =&#x2009;11)</th>
</tr>
</thead>
<tbody>
<tr>
<td align="left" valign="middle">Age (years)</td>
<td align="char" valign="middle" char="&#x00B1;">24.4 &#x00B1; 0.7</td>
<td align="char" valign="middle" char="&#x00B1;">23.1 &#x00B1; 1.2</td>
<td align="char" valign="middle" char="&#x00B1;">24.8 &#x00B1; 0.8</td>
</tr>
<tr>
<td align="left" valign="middle">Body weight (kg)</td>
<td align="char" valign="middle" char="&#x00B1;">73.6 &#x00B1; 1.8</td>
<td align="char" valign="middle" char="&#x00B1;">69.9 &#x00B1; 4.7</td>
<td align="char" valign="middle" char="&#x00B1;">74.9 &#x00B1; 1.8</td>
</tr>
<tr>
<td align="left" valign="middle">Height (m)</td>
<td align="char" valign="middle" char="&#x00B1;">1.78 &#x00B1; 0.02</td>
<td align="char" valign="middle" char="&#x00B1;">1.71 &#x00B1; 0.03</td>
<td align="char" valign="middle" char="&#x00B1;">1.80 &#x00B1; 0.02<sup>&#x002A;</sup></td>
</tr>
<tr>
<td align="left" valign="middle">BMI (kg&#x00B7;m<sup>&#x2212;2</sup>)</td>
<td align="char" valign="middle" char="&#x00B1;">23.2 &#x00B1; 0.4</td>
<td align="char" valign="middle" char="&#x00B1;">23.7 &#x00B1; 0.9</td>
<td align="char" valign="middle" char="&#x00B1;">23.0 &#x00B1; 0.4</td>
</tr>
<tr>
<td align="left" valign="middle">Lean mass (kg)</td>
<td align="char" valign="middle" char="&#x00B1;">57.1 &#x00B1; 1.6</td>
<td align="char" valign="middle" char="&#x00B1;">48.7 &#x00B1; 2.0</td>
<td align="char" valign="middle" char="&#x00B1;">60.2 &#x00B1; 1.0<sup>&#x002A;</sup></td>
</tr>
<tr>
<td align="left" valign="middle">Fat mass (kg)</td>
<td align="char" valign="middle" char="&#x00B1;">14.9 &#x00B1; 1.3</td>
<td align="char" valign="middle" char="&#x00B1;">19.8 &#x00B1; 2.4</td>
<td align="char" valign="middle" char="&#x00B1;">13.1 &#x00B1; 1.2<sup>&#x002A;</sup></td>
</tr>
<tr>
<td align="left" valign="middle">Fat mass (%)</td>
<td align="char" valign="middle" char="&#x00B1;">20.2 &#x00B1; 1.7</td>
<td align="char" valign="middle" char="&#x00B1;">28.0 &#x00B1; 2.2</td>
<td align="char" valign="middle" char="&#x00B1;">17.3 &#x00B1; 1.3<sup>&#x002A;</sup></td>
</tr>
<tr>
<td align="left" valign="middle">VO<sub>2max</sub> (L&#x00B7;min<sup>&#x2212;1</sup>)</td>
<td align="char" valign="middle" char="&#x00B1;">4.0 &#x00B1; 0.1</td>
<td align="char" valign="middle" char="&#x00B1;">3.3 &#x00B1; 0.3</td>
<td align="char" valign="middle" char="&#x00B1;">4.3 &#x00B1; 0.1<sup>&#x002A;</sup></td>
</tr>
<tr>
<td align="left" valign="middle">VO<sub>2max</sub> (mL&#x00B7;kg<sup>&#x2212;1</sup>&#x00B7;min<sup>&#x2212;1</sup>)</td>
<td align="char" valign="middle" char="&#x00B1;">54.4 &#x00B1; 1.7</td>
<td align="char" valign="middle" char="&#x00B1;">47.5 &#x00B1; 3.1</td>
<td align="char" valign="middle" char="&#x00B1;">57.0 &#x00B1; 1.4<sup>&#x002A;</sup></td>
</tr>
<tr>
<td align="left" valign="middle">HR<sub>max</sub> (beat&#x00B7;min<sup>&#x2212;1</sup>)</td>
<td align="char" valign="middle" char="&#x00B1;">195.1 &#x00B1; 1.9</td>
<td align="char" valign="middle" char="&#x00B1;">197.8 &#x00B1; 5.2</td>
<td align="char" valign="middle" char="&#x00B1;">194.2 &#x00B1; 1.9</td>
</tr>
</tbody>
</table>
<table-wrap-foot>
<p>Data are given as average&#x2009;&#x00B1;&#x2009;SEM. <sup>&#x002A;</sup><italic>p</italic> &#x003C;&#x2009;0.05 compared to women. BMI body mass index; HR<sub>max</sub> maximum heart rate; VO<sub>2max</sub> maximum oxygen uptake.</p>
</table-wrap-foot>
</table-wrap>
</sec>
<sec id="sec4">
<label>2.2</label>
<title>Study design</title>
<p>This study was performed in a randomized, balanced cross-over design (<xref ref-type="fig" rid="fig1">Figure 1</xref>). The study consisted of two trial days: one exercise (EX) and one control day without physical activity (CON), preceded by a screening day to determine physiological parameters and measure body composition. The same samples were collected in EX and CON. Dietary intakes were identical, except participants were allowed to consume almost protein-free snacks during EX to ensure the same energy balance during the two trial days.</p>
<fig position="float" id="fig1">
<label>Figure 1</label>
<caption>
<p>Design of the interventions. The protocol was completed in a randomized, balanced crossover experimental design. The study consisted of two trial days: one exercise (EX) and one control day (CON). During CON, no physical activity was performed during the trial day. The same diet was consumed the day before and during the two trial days, apart from snacks in EX. The urine collection periods are indicated in the figure.</p>
</caption>
<graphic xlink:href="fnut-11-1345922-g001.tif"/>
</fig>
</sec>
<sec id="sec5">
<label>2.3</label>
<title>Screening day</title>
<sec id="sec6">
<label>2.3.1</label>
<title>Incremental test</title>
<p>Participants performed an incremental test on a cycle ergometer (Lode Excalibur, Lode, Groningen; The Netherlands), during which the relationship between oxygen uptake and heart rate was established. This test was performed in a controlled environment under similar ambient conditions (18&#x2013;19&#x00B0;C). The saddle and handlebar positions were individually adjusted. The test consisted of 3 to 5 steps of 5&#x2009;min, with the load increasing by 25&#x2009;W for each step. The starting load was selected based on the height and body weight of the participant, expected VO<sub>2max</sub>, and previous cycling experience. The pedaling frequency was 75&#x2009;rpm, which had to be maintained throughout the test. Oxygen uptake was measured online (Oxycon Pro; Jager Instruments, Hoechberg, Germany) over 150&#x2009;s (from ~2.5&#x2013;5&#x2009;min at each intensity level), and capillary blood samples were taken for lactate analysis (Biosen C-Line Lactate analyzer, EKF Diagnostics, United Kingdom). Heart rate (Polar RS800CX, Kempele, Finland) was measured continuously throughout the incremental test. The incremental test was terminated after 5 steps or when the blood lactate concentration exceeded 4&#x2009;mM. Linear regression analysis from this incremental test was used to establish a relationship between oxygen uptake and heart rate.</p>
</sec>
<sec id="sec7">
<label>2.3.2</label>
<title>Maximum oxygen uptake test</title>
<p>After 10&#x2009;min of rest following the previous incremental test, maximal oxygen uptake (VO<sub>2max</sub>) was measured using the same equipment. Participants started the VO<sub>2max</sub> test at the second to last step of the incremental test, and intensity was increased by 25&#x2009;W steps every 60&#x2009;s until voluntary exhaustion. The pedaling frequency was 75&#x2009;rpm. Oxygen uptake was measured continuously, and the mean of the two highest measurements was defined as maximal oxygen uptake (VO<sub>2max</sub>). Heart rate (Polar RS800CX, Kempele, Finland) was measured continuously throughout the maximum oxygen uptake test. The maximum heart rate (HR<sub>max</sub>) was the highest heart rate reached during this test.</p>
</sec>
<sec id="sec8">
<label>2.3.3</label>
<title>Body composition</title>
<p>Body composition was measured using a whole body Dual-Energy-X-ray Absorptiometry (Scanex Medical Systems AS, GE Lunar iDXA, GE Healthcare, program ENCORE 18). Fat, lean, bone (bone mineral content), and total body mass were determined.</p>
</sec>
</sec>
<sec id="sec9">
<label>2.4</label>
<title>Trial days</title>
<p>Participants recorded their dietary intake from 16:00 to bedtime the day before the first trial day and were asked to follow the same diet before the second trial day. No exercise was allowed within the last 24&#x2009;h before the trial days. Participants were randomly assigned to either EX or CON first. When conducting EX first, a minimum interval of 7&#x2009;days was set before completing CON to ensure proper recovery. When conducting CON first, a minimum interval of 2&#x2009;days was set before completing EX.</p>
<sec id="sec10">
<label>2.4.1</label>
<title>Exercise day (EX)</title>
<p>During EX, participants performed 5&#x2009;h of endurance-based exercise: 1&#x2009;h of running, 2&#x2009;h of body weight strength circuit training, 1&#x2009;h of cardio endurance workout, and 1&#x2009;h of cycling to exhaustion on a stationary spinning bike. The exercise intensity for these exercises was selected by the participants in a certain intensity range to ensure participants could successfully adhere to the prescribed exercise protocol. Monitoring of the intensity was conducted through both the percentage of maximum heart rate (%HR<sub>max</sub>) and the participants&#x2019; rating of perceived exertion (RPE).</p>
<p>Participants reported to the laboratory at 8:00. Each participant&#x2019;s body weight was measured as well as resting blood glucose concentration (HemoCue Glucose 201 RT Analyzer; HemoCue AB, &#x00C4;ngelholm, Sweden). The resting heart rate was measured after 5&#x2009;min of sitting quietly. Participants then prepared for the exercise.</p>
<p>At 9:00, the participants performed 1&#x2009;h of outdoor running. Throughout the whole EX, heart rate (Polar RS400, Kempele, Finland) was measured continuously, and the Rate of Perceived Exertion (RPE) (<xref ref-type="bibr" rid="ref38">38</xref>) was reported at regular intervals. The running exercise was performed at 80&#x2013;90% HR<sub>max</sub>, with an RPE of 11&#x2013;16. Capillary blood glucose concentration was measured directly after running.</p>
<p>At 10:00, participants began 2&#x2009;h of body weight strength circuit training consisting of 6 circuits of four exercises (4 rounds of 30&#x2009;s intervals, 30&#x2009;s break; 16&#x2009;min per circuit) with a 4&#x2009;min break between each circuit. Circuit 1 consisted of push-ups, crunches, squats, and paddles. Circuit 2 consisted of lunges, sit-ups diagonal, dips, and planks. Circuit 3 involved squat jumps, abs-bug, mountain climber, and shoulder push-ups. Circuit 4 consisted of drop jumps, shoulder press with small weights, Russian twist, and rowing bent forward with small weights. Circuit 5 comprised side lunges, curls with small weights, scissor abs, and swimming abs. Circuit 6 consisted of skipping, step jumps alternating, burpees, and side planks. The body weight strength circuit training was performed at 60&#x2013;75% HR<sub>max</sub>, with an RPE of 11&#x2013;16. Capillary blood glucose concentration was measured directly after completion.</p>
<p>At 12:00, participants were served a standardized low-protein lunch. They could eat as much as they wanted from the food items provided: white bread (Loff; First Price, Norway), apple, jam (Bringeb&#x00E6;rsyltet&#x00F8;y; First Price, Norway), and vegan cheese (Go&#x2019;Vegan Original; Synn&#x00F8;ve, Norway). The calculated macronutrient content of this meal was carbohydrate 1.31&#x2009;&#x00B1;&#x2009;0.10&#x2009;g&#x00B7;kg<sup>&#x2212;1</sup>, protein 0.21&#x2009;&#x00B1;&#x2009;0.01&#x2009;g&#x00B7;kg<sup>&#x2212;1</sup>, fat 0.51&#x2009;&#x00B1;&#x2009;0.06&#x2009;g&#x00B7;kg<sup>&#x2212;1</sup>, calorie intake 807.1&#x2009;&#x00B1;&#x2009;52.1&#x2009;kcal.</p>
<p>At 14:00, capillary blood glucose concentration was measured. Participants started 1&#x2009;h of cardio endurance training. They could choose to distribute their time between cycling, running, and rowing on a rowing machine. The 1&#x2009;h of cardio endurance training was performed at 60&#x2013;80% HR<sub>max</sub>, with an RPE of 9&#x2013;14. Capillary blood glucose concentration was measured directly after the cardio endurance training.</p>
<p>At 15:00, participants started 1&#x2009;hour of cycling to exhaustion on a stationary spinning bike (M3i Indoor Bike, Keiser, California, United States). The cycling protocol consisted of 10&#x2009;min warm-up (50&#x2013;60% HR<sub>max</sub>); 3&#x00D7;3 min at 80% HR<sub>max</sub> interspersed with 3&#x2009;min at 60&#x2013;70% HR<sub>max</sub>; 3&#x00D7;30 s at 90% HR<sub>max</sub> interspersed with 30&#x2009;s at 70% HR<sub>max</sub>; 3&#x00D7;2 min at 90% HR<sub>max</sub> interspersed with 2&#x2009;min at 70% HR<sub>max</sub>; 3&#x00D7;30 s at 90&#x2013;95% HR<sub>max</sub> interspersed with 30&#x2009;s at 70&#x2013;80% HR<sub>max</sub>; 5&#x2009;min cool-down at 50&#x2013;60% HR<sub>max</sub>. The final exercise block of cycling was performed to exhaustion, with a target heart rate of 90&#x2013;95% HR<sub>max</sub> and RPE of 18&#x2013;20. Capillary blood glucose concentration was measured at the end of the cycling to exhaustion. The participants&#x2019; body weight was measured directly at the end of the cycling to exhaustion.</p>
</sec>
<sec id="sec11">
<label>2.4.2</label>
<title>Dietary intervention</title>
<p>Participants were asked to record their breakfast, lunch, snacks, dinner, and evening meal, as well as all drinks of the trial day and breakfast the next day. A dietary registration form was distributed to the participants for this purpose, and participants were encouraged to take photos of their food intake to improve the reproducibility of intake quantities. They were then asked to consume the same food and drinks on the second trial day. They had to check off on the dietary registration form the items ingested. Any differences in nutritional intake during the second visit had to be reported and accurately described on the dietary registration form. No participant reported a difference in dietary intake. The total calorie intake, carbohydrate, protein, and fat intakes were calculated using the Kostholdsplanleggeren program (Norwegian Directorate of Health, Norwegian Food Safety Authority, Norway). <xref ref-type="table" rid="tab2">Table 2</xref> presents the dietary intake during the trial days.</p>
<table-wrap position="float" id="tab2">
<label>Table 2</label>
<caption>
<p>Energy intake and content of macronutrients for the different standardized meals and supplements during the protocol.</p>
</caption>
<table frame="hsides" rules="groups">
<thead>
<tr>
<th/>
<th align="center" valign="middle">Energy intake (kcal)</th>
<th align="center" valign="middle">Carbohydrate intake (g&#x22C5;kg<sup>&#x2212;1</sup>)</th>
<th align="center" valign="middle">Protein intake (g&#x22C5;kg<sup>&#x2212;1</sup>)</th>
<th align="center" valign="middle">Fat intake (g&#x22C5;kg<sup>&#x2212;1</sup>)</th>
</tr>
</thead>
<tbody>
<tr>
<td align="left" valign="middle">Breakfast</td>
<td align="char" valign="middle" char="&#x00B1;">570.3 &#x00B1; 81.3</td>
<td align="char" valign="middle" char="&#x00B1;">1.18 &#x00B1; 0.20</td>
<td align="char" valign="middle" char="&#x00B1;">0.23 &#x00B1; 0.02</td>
<td align="char" valign="middle" char="&#x00B1;">0.22 &#x00B1; 0.04</td>
</tr>
<tr>
<td align="left" valign="middle">Lunch</td>
<td align="char" valign="middle" char="&#x00B1;">807.1 &#x00B1; 52.1</td>
<td align="char" valign="middle" char="&#x00B1;">1.31 &#x00B1; 0.10</td>
<td align="char" valign="middle" char="&#x00B1;">0.21 &#x00B1; 0.01</td>
<td align="char" valign="middle" char="&#x00B1;">0.51 &#x00B1; 0.06</td>
</tr>
<tr>
<td align="left" valign="middle">Snacks (during EX)</td>
<td align="char" valign="middle" char="&#x00B1;">421.9 &#x00B1; 51.8</td>
<td align="char" valign="middle" char="&#x00B1;">1.09 &#x00B1; 0.14</td>
<td align="char" valign="middle" char="&#x00B1;">0.07 &#x00B1; 0.01</td>
<td align="char" valign="middle" char="&#x00B1;">0.10 &#x00B1; 0.01</td>
</tr>
<tr>
<td align="left" valign="middle">Dinner</td>
<td align="char" valign="middle" char="&#x00B1;">776.7 &#x00B1; 117.6</td>
<td align="char" valign="middle" char="&#x00B1;">0.81 &#x00B1; 0.11</td>
<td align="char" valign="middle" char="&#x00B1;">0.62 &#x00B1; 0.11</td>
<td align="char" valign="middle" char="&#x00B1;">0.48 &#x00B1; 0.13</td>
</tr>
<tr>
<td align="left" valign="middle">Evening meal</td>
<td align="char" valign="middle" char="&#x00B1;">626.7 &#x00B1; 138.7</td>
<td align="char" valign="middle" char="&#x00B1;">1.06 &#x00B1; 0.21</td>
<td align="char" valign="middle" char="&#x00B1;">0.32 &#x00B1; 0.10</td>
<td align="char" valign="middle" char="&#x00B1;">0.31 &#x00B1; 0.08</td>
</tr>
<tr>
<td align="left" valign="middle">Breakfast the next day</td>
<td align="char" valign="middle" char="&#x00B1;">531.4 &#x00B1; 73.7</td>
<td align="char" valign="middle" char="&#x00B1;">1.03 &#x00B1; 0.14</td>
<td align="char" valign="middle" char="&#x00B1;">0.22 &#x00B1; 0.02</td>
<td align="char" valign="middle" char="&#x00B1;">0.23 &#x00B1; 0.05</td>
</tr>
<tr>
<td align="left" valign="middle">Total CON</td>
<td align="char" valign="middle" char="&#x00B1;">3312.2 &#x00B1; 284.2</td>
<td align="char" valign="middle" char="&#x00B1;">5.39 &#x00B1; 0.44</td>
<td align="char" valign="middle" char="&#x00B1;">1.60 &#x00B1; 0.13</td>
<td align="char" valign="middle" char="&#x00B1;">1.75 &#x00B1; 0.22</td>
</tr>
<tr>
<td align="left" valign="middle">Total EX</td>
<td align="char" valign="middle" char="&#x00B1;">3734.1 &#x00B1; 301.6</td>
<td align="char" valign="middle" char="&#x00B1;">6.48 &#x00B1; 0.48</td>
<td align="char" valign="middle" char="&#x00B1;">1.67 &#x00B1; 0.14</td>
<td align="char" valign="middle" char="&#x00B1;">1.85 &#x00B1; 0.22</td>
</tr>
</tbody>
</table>
<table-wrap-foot>
<p>The total macronutrient amount in CON and EX is calculated from all food intake during the urine collection period.</p>
</table-wrap-foot>
</table-wrap>
<p>Water intake was <italic>ad libitum</italic> during both trial days, and the volume was recorded.</p>
<p>During EX, participants were allowed to eat the high-carbohydrate snacks provided: banana, cereal bar (Mellombar; Eldorado, Norway), and milk chocolate (Kokesjokolade; Eldorado, Norway). They could also drink as much of a carbohydrate drink as they wanted. The drink was prepared as a 10% solution containing 50% glucose (GPR RECTAPUR&#x00AE;, VWR Chemicals, Radnor, Pennsylvania, USA) and 50% maltodextrin (Dextri-maltose&#x00AE;, MP Biomedicals, Santa Ana, California, USA), and flavored with 100&#x2009;g&#x00B7;L<sup>&#x2212;1</sup> of drink flavoring (Fun light, Stabburet, Norway). These snacks and drinks were given to participants to ensure the same energy balance during the two trial days. The calculated macronutrient content of the snacks was carbohydrate 1.09&#x2009;&#x00B1;&#x2009;0.14&#x2009;g&#x00B7;kg<sup>&#x2212;1</sup>, protein 0.07&#x2009;&#x00B1;&#x2009;0.01&#x2009;g&#x00B7;kg<sup>&#x2212;1</sup>, fat 0.10&#x2009;&#x00B1;&#x2009;0.01&#x2009;g&#x00B7;kg<sup>&#x2212;1</sup>, calorie intake 421.9&#x2009;&#x00B1;&#x2009;51.8&#x2009;kcal.</p>
<p>Nitrogen intake was calculated assuming a nitrogen-protein constant of 6.25 (<xref ref-type="bibr" rid="ref39">39</xref>).</p>
</sec>
<sec id="sec12">
<label>2.4.3</label>
<title>Individual energy expenditure</title>
<p>The individual energy expenditure during exercise in EX was calculated from the heart rate data measured during the exercise. The oxygen uptake was then calculated from the heart rate data using the linear regression between the heart rate and the oxygen uptake established during the incremental test on the screening day. The energy expenditure was then calculated as oxygen uptake (L&#x00B7;min<sup>&#x2212;1</sup>) &#x00D7; time exercise (min) &#x00D7; 20.15 (kJ&#x00B7;L<sup>&#x2212;1</sup>) (<xref ref-type="bibr" rid="ref40">40</xref>).</p>
<p>The resting energy expenditure in CON was calculated as (<xref ref-type="bibr" rid="ref41">41</xref>):</p>
<disp-formula id="E1">
<mml:math id="M1">
<mml:mtable>
<mml:mtr>
<mml:mtd>
<mml:mi mathvariant="normal">Resting</mml:mi>
<mml:mspace width="0.25em"/>
<mml:mi mathvariant="normal">energy</mml:mi>
<mml:mspace width="0.25em"/>
<mml:mi mathvariant="normal">expenditure</mml:mi>
<mml:mo>=</mml:mo>
<mml:mn>9.99</mml:mn>
<mml:mspace width="0.25em"/>
<mml:mo>&#x00D7;</mml:mo>
<mml:mspace width="0.25em"/>
<mml:mi mathvariant="normal">body</mml:mi>
<mml:mspace width="0.25em"/>
<mml:mi mathvariant="normal">weight</mml:mi>
<mml:mo>+</mml:mo>
<mml:mn>6.25</mml:mn>
<mml:mspace width="0.25em"/>
<mml:mo>&#x00D7;</mml:mo>
<mml:mspace width="0.25em"/>
<mml:mi mathvariant="normal">height</mml:mi>
</mml:mtd>
</mml:mtr>
<mml:mtr>
<mml:mtd>
<mml:mspace width="0.25em"/>
<mml:mo>&#x2212;</mml:mo>
<mml:mn>4.92</mml:mn>
<mml:mspace width="0.25em"/>
<mml:mo>&#x00D7;</mml:mo>
<mml:mspace width="0.25em"/>
<mml:mi mathvariant="normal">age</mml:mi>
<mml:mo>+</mml:mo>
<mml:mn>166</mml:mn>
<mml:mspace width="0.25em"/>
<mml:mo>&#x00D7;</mml:mo>
<mml:mspace width="0.25em"/>
<mml:mi mathvariant="normal">sex</mml:mi>
<mml:mspace width="0.25em"/>
</mml:mtd>
</mml:mtr>
<mml:mtr>
<mml:mtd>
<mml:mspace width="0.25em"/>
<mml:mfenced open="(" close=")">
<mml:mrow>
<mml:mi mathvariant="normal">males</mml:mi>
<mml:mo>,</mml:mo>
<mml:mspace width="0.25em"/>
<mml:mn>1</mml:mn>
<mml:mo>;</mml:mo>
<mml:mspace width="0.25em"/>
<mml:mi mathvariant="normal">females</mml:mi>
<mml:mo>,</mml:mo>
<mml:mspace width="0.25em"/>
<mml:mn>0</mml:mn>
</mml:mrow>
</mml:mfenced>
<mml:mo>&#x2212;</mml:mo>
<mml:mn>161</mml:mn>
</mml:mtd>
</mml:mtr>
</mml:mtable>
</mml:math>
</disp-formula>
</sec>
</sec>
<sec id="sec13">
<label>2.5</label>
<title>Urinary nitrogen</title>
<p>All urine was collected in plastic containers in three consecutive batches: morning urine on the trial day (0:00 to 8:00; 8&#x2009;h), the rest of the day on the trial day (8:00 to 24:00; 16&#x2009;h), and the morning urine on the next day (0:00 to 8:00; 8&#x2009;h). The urine volume was measured for each period, and one 15-mL sample and two 2-mL samples were frozen at &#x2212;20&#x00B0;C for analysis. Urinary nitrogen was measured using the Kjeldahl method (<xref ref-type="bibr" rid="ref42">42</xref>) and corrected by urine density.</p>
</sec>
<sec id="sec14">
<label>2.6</label>
<title>Sweat rate</title>
<p>The sweat rate during the exercise period was calculated using a modified equation (<xref ref-type="bibr" rid="ref24">24</xref>, <xref ref-type="bibr" rid="ref43">43</xref>):</p>
<disp-formula id="E2">
<mml:math id="M2">
<mml:mtable>
<mml:mtr>
<mml:mtd>
<mml:mi mathvariant="normal">Sweat</mml:mi>
<mml:mspace width="0.25em"/>
<mml:mi mathvariant="normal">rate</mml:mi>
<mml:mspace width="0.25em"/>
<mml:mfenced open="(" close=")">
<mml:mi mathvariant="normal">L</mml:mi>
</mml:mfenced>
<mml:mo>=</mml:mo>
<mml:mi mathvariant="normal">B</mml:mi>
<mml:mi mathvariant="normal">W</mml:mi>
<mml:mi mathvariant="normal">i</mml:mi>
<mml:mo>&#x2212;</mml:mo>
<mml:mi mathvariant="normal">B</mml:mi>
<mml:mi mathvariant="normal">W</mml:mi>
<mml:mi mathvariant="normal">f</mml:mi>
<mml:mo>+</mml:mo>
<mml:mi mathvariant="normal">WatIn</mml:mi>
<mml:mo>&#x2212;</mml:mo>
<mml:mi mathvariant="normal">A</mml:mi>
<mml:mi mathvariant="normal">R</mml:mi>
<mml:mi mathvariant="normal">W</mml:mi>
<mml:mspace width="0.25em"/>
<mml:mi mathvariant="normal">at</mml:mi>
<mml:mspace width="0.25em"/>
<mml:mi mathvariant="normal">Loss</mml:mi>
</mml:mtd>
</mml:mtr>
<mml:mtr>
<mml:mtd>
<mml:mo>&#x2212;</mml:mo>
<mml:mi mathvariant="normal">UrVol</mml:mi>
<mml:mspace width="0.25em"/>
<mml:mo>+</mml:mo>
<mml:mspace width="0.25em"/>
<mml:mi mathvariant="normal">WFoodIn</mml:mi>
<mml:mspace width="0.25em"/>
<mml:mo>&#x2212;</mml:mo>
<mml:mspace width="0.25em"/>
<mml:mfenced open="(" close=")">
<mml:mrow>
<mml:mi mathvariant="normal">E</mml:mi>
<mml:mi mathvariant="normal">E</mml:mi>
</mml:mrow>
</mml:mfenced>
<mml:mo stretchy="true">/</mml:mo>
<mml:mn>7700</mml:mn>
</mml:mtd>
</mml:mtr>
</mml:mtable>
</mml:math>
</disp-formula>
<p>With BWi the initial body weight (kg), BWf the final body weight (kg), WatIn the water intake (L), ARW at Loss the assumed respiratory water loss (L), UrVol the urine volume (L), WFoodIn the weight of food intake (kg) and EE the energy expenditure during the exercise period (kcal).</p>
<p>Sweat volume and mass were considered equivalent (i.e., 1&#x2009;mL&#x2009;=&#x2009;1&#x2009;g) and expressed as a total and hourly rate.</p>
</sec>
<sec id="sec15">
<label>2.7</label>
<title>3-Methylhistidine in urine</title>
<p>The urine from the day of the trial day (8:00 to 24:00; 16&#x2009;h) and the morning urine from the next day (0:00 to 8:00; 8&#x2009;h) were mixed in proportions relative to the volume of urine during these periods. The concentration of urinary 3-MH was then quantified in these pooled urine samples using an ELISA kit (BioSite ELISA, Nordic BioSite, Norway). The quantity of 3-MH excreted during these 24&#x2009;h was then calculated by multiplying the measured 3-MH concentration by the urine volume.</p>
</sec>
<sec id="sec16">
<label>2.8</label>
<title>Statistics</title>
<p>The results were analyzed using Prism 9 (GraphPad Software, LLC, San Diego, California, United States). Two-way ANOVA (time x condition) with repeated measurements was conducted to analyze the data. After identifying a significant effect, <italic>post hoc</italic> analyses were performed with Bonferroni corrections. When some data were missing, mixed-effect analyses were used. The significance level was set to <italic>p</italic>&#x2009;&#x2264;&#x2009;0.05. Statistical trends are defined as <italic>p</italic>-values between 0.05 and 0.10. Data are presented as mean&#x2009;&#x00B1;&#x2009;SEM in text, figures, and tables.</p>
</sec>
</sec>
<sec sec-type="results" id="sec17">
<label>3</label>
<title>Results</title>
<sec id="sec18">
<label>3.1</label>
<title>Energy expenditure</title>
<p>A description of the energy intake during the study is shown in <xref ref-type="table" rid="tab2">Table 2</xref>. The total energy intake in CON was 2780.8&#x2009;&#x00B1;&#x2009;242.1&#x2009;kcal during the intervention day and 3312.2&#x2009;&#x00B1;&#x2009;284.2&#x2009;kcal during the urine collection period (including breakfast the next day). Therefore, the participants had a positive energy balance in CON during the intervention day of 1092.6&#x2009;&#x00B1;&#x2009;228.2&#x2009;kcal.</p>
<p>In EX, the total energy expenditure during the 5-h exercise period was 3289.9&#x2009;&#x00B1;&#x2009;188.8&#x2009;kcal or 44.6&#x2009;&#x00B1;&#x2009;2.2&#x2009;kcal&#x00B7;kg<sup>&#x2212;1</sup>, meaning that during EX, participants had an additional energy expenditure of 2938.2&#x2009;&#x00B1;&#x2009;182.5&#x2009;kcal compared to the resting energy expenditure measured in CON. The participants were allowed to eat additional almost protein-free snacks during EX to compensate for this additional energy expenditure. They consumed 421.9&#x2009;&#x00B1;&#x2009;51.8&#x2009;kcal extra, thus, participants had a negative energy balance during EX of &#x2212;1423.6&#x2009;&#x00B1;&#x2009;219.5&#x2009;kcal.</p>
</sec>
<sec id="sec19">
<label>3.2</label>
<title>Urinary nitrogen excretion</title>
<p>Urinary nitrogen excretion during the 24-h study period was 12.45&#x2009;&#x00B1;&#x2009;0.88&#x2009;g (168.5&#x2009;&#x00B1;&#x2009;10.1&#x2009;mg&#x00B7;kg<sup>&#x2212;1</sup>) in CON and 12.61&#x2009;&#x00B1;&#x2009;0.98&#x2009;g (170.0&#x2009;&#x00B1;&#x2009;11.0&#x2009;mg&#x00B7;kg<sup>&#x2212;1</sup>) in EX (<xref ref-type="fig" rid="fig2">Figure 2A</xref>) and did not differ significantly between trial days (<italic>p</italic>&#x2009;=&#x2009;0.764). Urinary nitrogen excretion was also not significantly different when normalized by fat-free mass (<xref ref-type="fig" rid="fig2">Figure 2B</xref>).</p>
<fig position="float" id="fig2">
<label>Figure 2</label>
<caption>
<p>Urinary nitrogen excretion. <bold>(A)</bold> Urinary nitrogen excretion normalized by body weight. <bold>(B)</bold> Urinary nitrogen excretion normalized by fat-free mass. Values are means &#x00B1; SEM.</p>
</caption>
<graphic xlink:href="fnut-11-1345922-g002.tif"/>
</fig>
</sec>
<sec id="sec20">
<label>3.3</label>
<title>Sweat rate</title>
<p>The sweat rate was 2.55&#x2009;&#x00B1;&#x2009;0.25 L during EX (<xref ref-type="table" rid="tab3">Table 3</xref>). This represents an hourly sweating rate of 0.32&#x2009;&#x00B1;&#x2009;0.03&#x2009;L&#x00B7;h<sup>&#x2212;1</sup>. During CON, the sweat rate was 0.14&#x2009;&#x00B1;&#x2009;0.15&#x2009;L, representing an hourly sweating rate of 0.02&#x2009;&#x00B1;&#x2009;0.02&#x2009;L&#x00B7;h<sup>&#x2212;1</sup>.</p>
<table-wrap position="float" id="tab3">
<label>Table 3</label>
<caption>
<p>Sweat rate during the two conditions.</p>
</caption>
<table frame="hsides" rules="groups">
<thead>
<tr>
<th align="left" valign="top">Condition</th>
<th align="center" valign="top" colspan="2">EX</th>
<th align="center" valign="top" colspan="2">CON</th>
</tr>
</thead>
<tbody>
<tr>
<td align="left" valign="top" colspan="5">From 8:00 to 16:00</td>
</tr>
<tr>
<td align="left" valign="top">Initial body weight (kg)</td>
<td align="center" valign="middle" colspan="2">73.8&#x2009;&#x00B1;&#x2009;1.9</td>
<td align="center" valign="middle" colspan="2">74.1&#x2009;&#x00B1;&#x2009;2.0</td>
</tr>
<tr>
<td align="left" valign="top">Final body weight (kg)</td>
<td align="center" valign="middle" colspan="2">73.3&#x2009;&#x00B1;&#x2009;2.0</td>
<td align="center" valign="middle" colspan="2">74.2&#x2009;&#x00B1;&#x2009;2.0</td>
</tr>
<tr>
<td align="left" valign="top">Energy expenditure (kcal)</td>
<td align="center" valign="middle" colspan="2">3,501&#x2009;&#x00B1;&#x2009;193</td>
<td align="center" valign="middle" colspan="2">563&#x2009;&#x00B1;&#x2009;13</td>
</tr>
<tr>
<td/>
<td align="center" valign="middle">Intake</td>
<td align="center" valign="middle">Loss</td>
<td align="center" valign="middle">Intake</td>
<td align="center" valign="middle">Loss</td>
</tr>
<tr>
<td align="left" valign="top">Water intake (L)</td>
<td align="center" valign="middle">3.69&#x2009;&#x00B1;&#x2009;0.39</td>
<td/>
<td align="center" valign="middle">1.45&#x2009;&#x00B1;&#x2009;0.17</td>
<td/>
</tr>
<tr>
<td align="left" valign="top">Urine (L)</td>
<td/>
<td align="center" valign="middle">0.79&#x2009;&#x00B1;&#x2009;0.17</td>
<td/>
<td align="center" valign="middle">1.13&#x2009;&#x00B1;&#x2009;0.13</td>
</tr>
<tr>
<td align="left" valign="top">Respiratory water loss (L)</td>
<td/>
<td align="center" valign="middle">0.36&#x2009;&#x00B1;&#x2009;0.02</td>
<td/>
<td align="center" valign="middle">0.05&#x2009;&#x00B1;&#x2009;0.00</td>
</tr>
<tr>
<td align="left" valign="top">Sweat (L)</td>
<td/>
<td align="center" valign="middle">2.55&#x2009;&#x00B1;&#x2009;0.25</td>
<td/>
<td align="center" valign="middle">0.14&#x2009;&#x00B1;&#x2009;0.15</td>
</tr>
</tbody>
</table>
</table-wrap>
</sec>
<sec id="sec21">
<label>3.4</label>
<title>Urinary 3-methylhistidine excretion</title>
<p>Urinary 3-MH excretion during the 24-h study period was 508.7&#x2009;&#x00B1;&#x2009;33.8&#x2009;&#x03BC;mol (6.91&#x2009;&#x00B1;&#x2009;0.40&#x2009;&#x03BC;mol&#x00B7;kg<sup>&#x2212;1</sup>) in CON and 513.0&#x2009;&#x00B1;&#x2009;80.2&#x2009;&#x03BC;mol (6.91&#x2009;&#x00B1;&#x2009;1.05&#x2009;&#x03BC;mol&#x00B7;kg<sup>&#x2212;1</sup>) in EX (<xref ref-type="fig" rid="fig3">Figure 3A</xref>) and was not significantly different (<italic>p</italic>&#x2009;=&#x2009;0.953). When normalized by fat-free mass, urinary 3-MH excretion did still not differ between trial days: 8.87&#x2009;&#x00B1;&#x2009;0.47&#x2009;&#x03BC;mol&#x00B7;kg<sup>&#x2212;1</sup> FFM in CON and 8.89&#x2009;&#x00B1;&#x2009;1.27&#x2009;&#x03BC;mol&#x00B7;kg<sup>&#x2212;1</sup> FFM in EX (<italic>p</italic>&#x2009;=&#x2009;0.991) (<xref ref-type="fig" rid="fig3">Figure 3B</xref>).</p>
<fig position="float" id="fig3">
<label>Figure 3</label>
<caption>
<p>Urinary 3-MH excretion. <bold>(A)</bold> Urinary 3-MH excretion normalized by body weight. <bold>(B)</bold> Urinary 3-MH excretion normalized by fat-free mass. Values are means &#x00B1; SEM.</p>
</caption>
<graphic xlink:href="fnut-11-1345922-g003.tif"/>
</fig>
</sec>
<sec id="sec22">
<label>3.5</label>
<title>Correlations</title>
<p>Urinary nitrogen excretion was significantly correlated with urinary 3-MH excretion but higher in CON (<italic>r</italic><sup>2</sup>&#x2009;=&#x2009;0.697; <italic>p</italic>&#x2009;&#x003C;&#x2009;0.001) than in EX (<italic>r</italic><sup>2</sup>&#x2009;=&#x2009;0.338; <italic>p</italic> =&#x2009;0,023) (<xref ref-type="fig" rid="fig4">Figure 4</xref>). The two slopes were not significantly different (<italic>p</italic>&#x2009;=&#x2009;0.343).</p>
<fig position="float" id="fig4">
<label>Figure 4</label>
<caption>
<p>Correlation between urinary nitrogen and 3-MH excretions. Square dots are female participants; circular dots are male participants.</p>
</caption>
<graphic xlink:href="fnut-11-1345922-g004.tif"/>
</fig>
<p>Urinary nitrogen excretion was also correlated with total protein intake over the study period in CON (<italic>r<sup>2</sup></italic>&#x2009;=&#x2009;0.249; <italic>p</italic>&#x2009;=&#x2009;0.058) and in EX (<italic>r</italic><sup>2</sup>&#x2009;=&#x2009;0.328; <italic>p</italic>&#x2009;=&#x2009;0.026) (<xref ref-type="fig" rid="fig5">Figure 5</xref>). The two slopes were not significantly different (<italic>p</italic>&#x2009;=&#x2009;0.752).</p>
<fig position="float" id="fig5">
<label>Figure 5</label>
<caption>
<p>Correlation between urinary nitrogen excretion and total protein intake during the urine collection period. Square dots are female participants; circular dots are male participants.</p>
</caption>
<graphic xlink:href="fnut-11-1345922-g005.tif"/>
</fig>
</sec>
<sec id="sec23">
<label>3.6</label>
<title>Blood glucose concentration</title>
<p>The blood glucose concentration decreased during EX and CON with no difference up to the lunch break at 12:00 (<xref ref-type="fig" rid="fig6">Figure 6</xref>). However, at 14:00, 90&#x2009;min after lunch completion, the glucose concentration was significantly elevated in CON compared to EX (<italic>p</italic>&#x2009;=&#x2009;0.001). After that, the blood glucose concentration decreased with no difference between the conditions.</p>
<fig position="float" id="fig6">
<label>Figure 6</label>
<caption>
<p>Blood glucose concentration during the test days. Values are means &#x00B1; SEM.</p>
</caption>
<graphic xlink:href="fnut-11-1345922-g006.tif"/>
</fig>
</sec>
</sec>
<sec sec-type="discussion" id="sec24">
<label>4</label>
<title>Discussion</title>
<p>Urinary nitrogen excretion during the 24-h study period did not significantly differ between EX and CON.</p>
<sec id="sec25">
<label>4.1</label>
<title>Reasons for the lack of difference observed in urinary nitrogen excretion in the present study</title>
<p>Disparities in urinary nitrogen excretion have been reported during exercise compared to a control condition. Most of the cited studies in <xref rid="SM1" ref-type="supplementary-material">Supplementary Table S1</xref> reported increased urinary nitrogen excretion during or after exercise (<xref ref-type="bibr" rid="ref3">3</xref>, <xref ref-type="bibr" rid="ref5">5</xref>&#x2013;<xref ref-type="bibr" rid="ref12">12</xref>, <xref ref-type="bibr" rid="ref14">14</xref>&#x2013;<xref ref-type="bibr" rid="ref17">17</xref>, <xref ref-type="bibr" rid="ref44">44</xref>&#x2013;<xref ref-type="bibr" rid="ref51">51</xref>), not always being significant (<xref ref-type="bibr" rid="ref18">18</xref>, <xref ref-type="bibr" rid="ref52">52</xref>&#x2013;<xref ref-type="bibr" rid="ref56">56</xref>). However, because of the limitations, which included the nitrogen excretion quantification method, no standardized diet, no control trial in the same subjects, small sample sizes, and no specific energy expenditure calculation during the work/exercise periods, no clear conclusion has been reached on the effect of exercise on urinary nitrogen excretion. Newer studies have also tended to indicate that exercise increases urinary nitrogen excretion, but the absence of standardized conditions still limits the conclusions reached.</p>
<sec id="sec26">
<label>4.1.1</label>
<title>The effect of energy balance</title>
<p>In similar conditions to the present study, when participants were in negative energy balance with exercise, Todd et al. (<xref ref-type="bibr" rid="ref57">57</xref>) and Butterfield and Calloway (<xref ref-type="bibr" rid="ref58">58</xref>) found that urinary nitrogen excretion increased with exercise but non-significantly: there was an average 4% increase, ranging from &#x2212;3 to 9%. This is in the order of magnitude of the 1% increase in the present study. However, at a fixed energy balance (when the extra energy expenditure of training was compensated by additional energy intake), exercise decreased urinary nitrogen excretion by 7% on average, ranging from &#x2212;11 to 2% (<xref ref-type="bibr" rid="ref57">57</xref>, <xref ref-type="bibr" rid="ref58">58</xref>). It is well established that increasing energy intake decreases nitrogen excretion at fixed nitrogen intake (<xref ref-type="bibr" rid="ref13">13</xref>, <xref ref-type="bibr" rid="ref59">59</xref>). This shows that the effect of exercise on urinary nitrogen excretion can be caused by a decreased energy balance with exercise. This agrees with studies reporting increased excretion with exercise when energy intake was insufficient (<xref ref-type="bibr" rid="ref3">3</xref>, <xref ref-type="bibr" rid="ref47">47</xref>, <xref ref-type="bibr" rid="ref55">55</xref>, <xref ref-type="bibr" rid="ref56">56</xref>). In the present study, participants were allowed to ingest almost protein-free snacks in EX in an attempt to have a fixed energy balance between the conditions, although the energy balance was still negative in EX. However, the ingestion of snacks in EX could have reduced the effect of exercise on urinary nitrogen excretion, leading to the non-significant difference between conditions.</p>
<p>To our knowledge, the mechanism for decreased urinary nitrogen excretion with exercise while the participants are in energy balance has not yet been established. However, some studies showed that low glycogen levels and low carbohydrate availability can increase nitrogen excretion (<xref ref-type="bibr" rid="ref24">24</xref>, <xref ref-type="bibr" rid="ref35">35</xref>).</p>
</sec>
</sec>
<sec id="sec27">
<label>4.2</label>
<title>Sweat nitrogen excretion</title>
<p>Urine is not the only way for the body to excrete nitrogen. Nitrogen can also be excreted in sweat, feces, or other miscellaneous routes [nails, hair, tooth brushing (<xref ref-type="bibr" rid="ref21">21</xref>)]. Nitrogen excretion through these different compartments is connected. Some studies have previously reported a link between urinary and sweat nitrogen excretion: urinary nitrogen excretion decreased in proportion when sweat nitrogen losses increased (<xref ref-type="bibr" rid="ref19">19</xref>, <xref ref-type="bibr" rid="ref20">20</xref>), as in the studies commented on previously (<xref ref-type="bibr" rid="ref57">57</xref>, <xref ref-type="bibr" rid="ref58">58</xref>). This could also explain why the urinary nitrogen excretion was not significantly different between EX and CON. In similar conditions to the present study, sweat nitrogen excretion increased by 22% on average, ranging from &#x2212;20 to 58% when the participants exercised while in energy balance (<xref ref-type="bibr" rid="ref57">57</xref>, <xref ref-type="bibr" rid="ref58">58</xref>).</p>
<p>In the present study, we calculated the sweat rate and reported an hourly sweating rate of 0.32&#x2009;&#x00B1;&#x2009;0.03&#x2009;L&#x00B7;h<sup>&#x2212;1</sup> during EX. This agrees with sweat rates during exercise reported in similar ambient conditions (<xref ref-type="bibr" rid="ref43">43</xref>). However, we did not measure the sweat nitrogen concentration. Using previously published data, we can estimate this concentration. For this purpose, we used the relationship between sweat nitrogen concentration and nitrogen intake established during exercise performed mid-day under similar thermal conditions (<xref ref-type="bibr" rid="ref20">20</xref>, <xref ref-type="bibr" rid="ref25">25</xref>). We used this estimation because sweat nitrogen concentration has been shown to differ with time of day, temperature, and dietary nitrogen intake, with this last factor having the most significant impact on sweat nitrogen concentration. Based on the regression between sweat nitrogen concentration and protein intake established during exercise conditions (<xref ref-type="bibr" rid="ref20">20</xref>), estimated nitrogen excretion through sweating during exercise was &#x2248;1.3&#x2009;g in EX. The estimated average sweat nitrogen concentration was 0.5&#x2009;mg/mL, consistent with previous studies (<xref ref-type="bibr" rid="ref19">19</xref>, <xref ref-type="bibr" rid="ref21">21</xref>, <xref ref-type="bibr" rid="ref23">23</xref>, <xref ref-type="bibr" rid="ref24">24</xref>).</p>
<p>In addition, resting sweat nitrogen excretion can be estimated from the relationship between resting sweat nitrogen excretion and nitrogen intake (<xref ref-type="bibr" rid="ref21">21</xref>): resting sweat nitrogen excretion (mg&#x00B7;d<sup>&#x2212;1</sup>)&#x2009;=&#x2009;4.8022 x nitrogen intake (g&#x00B7;d<sup>&#x2212;1</sup>)&#x2009;+&#x2009;104. This led to a resting sweat nitrogen excretion of &#x2248;0.2&#x2009;g in CON and EX, with no significant difference between conditions. Finally, fecal nitrogen loss can be estimated from previous studies at 12.41&#x2009;mg&#x00B7;kg<sup>&#x2212;1</sup>&#x00B7;d<sup>&#x2212;1</sup> and miscellaneous nitrogen losses at 1.77&#x2009;mg&#x00B7;kg<sup>&#x2212;1</sup>&#x00B7;d<sup>&#x2212;1</sup> (<xref ref-type="bibr" rid="ref25">25</xref>). The total nitrogen excretion can then be calculated as the sum of the urinary, sweat, fecal, and miscellaneous nitrogen excretions.</p>
</sec>
<sec id="sec28">
<label>4.3</label>
<title>Sweat nitrogen excretion should be included in future studies</title>
<p>Including or excluding sweat nitrogen excretion can dramatically change a study&#x2019;s conclusions. For example, the nitrogen balance technique calculates the difference between nitrogen intake and loss. A positive nitrogen balance shows that the human body stores more nitrogen than it loses: it is in an anabolic state. Similarly, a negative nitrogen balance indicates a catabolic state. Thus, the sign of the nitrogen balance is crucial for any interpretation.</p>
<p>Gontzea et al. (<xref ref-type="bibr" rid="ref19">19</xref>) conducted a study in which six males cycled for 2&#x2009;h daily in laboratory conditions, and all nitrogen excretion pathways were measured. In these well-controlled conditions, with all food intake provided, the authors studied the effect of including nitrogen loss through sweat on nitrogen balance. If sweat nitrogen losses were neglected, 1 in 6 participants had a positive nitrogen balance (average&#x2009;&#x2212;&#x2009;0.827&#x2009;g&#x00B7;d<sup>&#x2212;1</sup>) when exercising with protein intakes between 65 and 75&#x2009;g. When sweat nitrogen losses were considered, all 6 participants had a negative nitrogen balance (average&#x2009;&#x2212;&#x2009;1.721&#x2009;g&#x00B7;d<sup>&#x2212;1</sup>). Similarly, when exercising with a protein intake of between 105 and 115&#x2009;g, if sweat nitrogen losses were neglected, 4 in 6 participants had a positive nitrogen balance (average 0.676&#x2009;g&#x00B7;d<sup>&#x2212;1</sup>). When sweat nitrogen losses were considered, only 2 in 6 participants had a positive nitrogen balance (average&#x2009;&#x2212;&#x2009;0.371&#x2009;g&#x00B7;d<sup>&#x2212;1</sup>). Based on these results, it could be concluded that the protein intake was enough to induce a positive nitrogen balance in the last case if sweat nitrogen losses were neglected. Nevertheless, this conclusion would have been erroneous. Similarly, in another study, ignoring sweat nitrogen losses resulted in a positive nitrogen balance (average 0.62&#x2009;g&#x00B7;d<sup>&#x2212;1</sup>) (<xref ref-type="bibr" rid="ref60">60</xref>). When sweat nitrogen losses were included, the nitrogen balance was negative (average&#x2009;&#x2212;&#x2009;2.01&#x2009;g&#x00B7;d<sup>&#x2212;1</sup>), changing the conclusion of this intervention.</p>
<p>In the present study, the nitrogen balance was positive in the two conditions both when sweat nitrogen losses were considered (CON, 70.8&#x2009;&#x00B1;&#x2009;18.3&#x2009;mg&#x00B7;kg<sup>&#x2212;1</sup>; EX, 63.0&#x2009;&#x00B1;&#x2009;16.8&#x2009;mg&#x00B7;kg<sup>&#x2212;1</sup>) and when they were ignored and only urinary nitrogen losses were considered (CON, 73.4&#x2009;&#x00B1;&#x2009;18.4&#x2009;mg&#x00B7;kg<sup>&#x2212;1</sup>; EX, 83.2&#x2009;&#x00B1;&#x2009;17.8&#x2009;mg&#x00B7;kg<sup>&#x2212;1</sup>). Thus, including sweat nitrogen losses would not have changed the interpretation. However, including them would have considerably increased the total nitrogen excretion in EX. Total nitrogen excretion during the 24-h study period would have been 13.69&#x2009;&#x00B1;&#x2009;0.90&#x2009;g (185.3&#x2009;&#x00B1;&#x2009;10.2&#x2009;mg&#x00B7;kg<sup>&#x2212;1</sup>) in CON and 15.15&#x2009;&#x00B1;&#x2009;1.14&#x2009;g (204.3&#x2009;&#x00B1;&#x2009;12.6&#x2009;mg&#x00B7;kg<sup>&#x2212;1</sup>) in EX, leading to a significant effect of exercise (<italic>p</italic>&#x2009;=&#x2009;0.033). It means that proteins contributed to 1.3% of energy expenditure during exercise. It is, therefore, crucial to include sweat nitrogen losses when quantifying nitrogen excretion. Studies without direct measurements should estimate these losses.</p>
<p>The magnitude of the effect of exercise on total nitrogen excretion can be compared to results from previous studies (<xref rid="SM1" ref-type="supplementary-material">Supplementary Table S2</xref>) (<xref ref-type="bibr" rid="ref19">19</xref>, <xref ref-type="bibr" rid="ref57">57</xref>, <xref ref-type="bibr" rid="ref58">58</xref>, <xref ref-type="bibr" rid="ref61">61</xref>&#x2013;<xref ref-type="bibr" rid="ref63">63</xref>). Exercise was reported to increase total nitrogen excretion in four of these six studies, with an average increase of 0.23&#x2009;g&#x00B7;d<sup>&#x2212;1</sup>. Of these six studies, the two reporting decreased total nitrogen excretion with exercise also showed an inappropriate nitrogen intake before the intervention days, leading to negative nitrogen balance at baseline (<xref ref-type="bibr" rid="ref62">62</xref>, <xref ref-type="bibr" rid="ref63">63</xref>). It is well-established that nitrogen excretion increases with nitrogen intake (<xref ref-type="bibr" rid="ref13">13</xref>), which is consistent with our data. Therefore, it is expected that some differences in the amount of nitrogen excretion should arise. However, our estimation of 1.5&#x2009;g&#x00B7;d<sup>&#x2212;1</sup> was in the range of variation of previously published studies, which showed levels between &#x2212;1.8 and 2.6&#x2009;g&#x00B7;d<sup>&#x2212;1</sup> (<xref ref-type="bibr" rid="ref19">19</xref>, <xref ref-type="bibr" rid="ref57">57</xref>, <xref ref-type="bibr" rid="ref58">58</xref>, <xref ref-type="bibr" rid="ref61">61</xref>&#x2013;<xref ref-type="bibr" rid="ref63">63</xref>). Such an increase in total nitrogen excretion during EX would mean that exercise increased protein catabolism. However, this does not indicate the location of this degradation.</p>
</sec>
<sec id="sec29">
<label>4.4</label>
<title>Urinary 3-methylhistidine excretion</title>
<p>Urinary 3-MH excretion during the 24-h study period was not significantly different between EX and CON. This would mean no significant increase in skeletal myofibrillar protein catabolism with exercise. Previous studies measuring urinary 3-MH excretion during exercise have reported conflicting results. Most of these studies did not note any significant difference in urinary 3-MH excretion with exercise (<xref ref-type="bibr" rid="ref17">17</xref>, <xref ref-type="bibr" rid="ref64">64</xref>&#x2013;<xref ref-type="bibr" rid="ref71">71</xref>), while some reported an increase (<xref ref-type="bibr" rid="ref72">72</xref>) and other a decrease (<xref ref-type="bibr" rid="ref73">73</xref>, <xref ref-type="bibr" rid="ref74">74</xref>). These studies consisted of acute endurance exercises, mostly of shorter duration than this study. Studies similar in duration to the present study did not find a difference in urinary 3-MH excretion (<xref ref-type="bibr" rid="ref64">64</xref>, <xref ref-type="bibr" rid="ref65">65</xref>, <xref ref-type="bibr" rid="ref67">67</xref>), apart from one showing a decrease (<xref ref-type="bibr" rid="ref74">74</xref>). This latter study included urinary 3-MH excretion measured at different periods on the day of exercise. In this study, urinary 3-MH excretion was not different from the control day, before and after exercise. However, the drop in excretion during exercise meant that the total excretion on the exercise day was significantly lower than on the control day. All this indicates that exercise does not increase urinary 3-MH excretion and therefore, does not affect the catabolism of skeletal myofibrillar proteins.</p>
</sec>
<sec id="sec30">
<label>4.5</label>
<title>Practical considerations</title>
<sec id="sec31">
<label>4.5.1</label>
<title>Daily variations of the urinary nitrogen excretion</title>
<p>The day-to-day variation in urinary nitrogen excretion in resting conditions varies from &#x2248;4% (<xref ref-type="bibr" rid="ref75">75</xref>) to &#x2248;14% (<xref ref-type="bibr" rid="ref76">76</xref>), with most studies reporting around 10% (<xref ref-type="bibr" rid="ref77">77</xref>, <xref ref-type="bibr" rid="ref78">78</xref>). In exercise conditions, the day-to-day variation in urinary nitrogen excretion seems similar, with Gontzea et al. reporting &#x2248;9% (<xref ref-type="bibr" rid="ref19">19</xref>).</p>
<p>One strength of the present study is that it used a randomized, balanced, cross-over design. This design, combined with the same diet before and during both trial days, except for snacks in EX, should have reduced variation. Including 15 participants further reduced the influence of daily variations on the conclusion about the effect of exercise on urinary nitrogen excretion.</p>
</sec>
<sec id="sec32">
<label>4.5.2</label>
<title>Participants&#x2019; habitual protein intake</title>
<p>A potential weakness of the present study was that not all food was supplied during the intervention days. Participants received a standardized lunch. However, they were free to ingest the food of their choice during the rest of the intervention and only had to replicate this intake on the second trial day. This intentionally allowed participants to consume their habitual protein intake during the trial days. It has previously been established that, when changing from high to low protein intake, or vice versa, a few days are necessary before a nitrogen balance is regained: the amino acid oxidative capacity of the organism is determined by the habitual protein intake (<xref ref-type="bibr" rid="ref79">79</xref>).</p>
</sec>
<sec id="sec33">
<label>4.5.3</label>
<title>Nitrogen excretion the day after an exercise period</title>
<p>Some studies suggest that increased nitrogen output continues after the exercise stops (<xref ref-type="bibr" rid="ref53">53</xref>&#x2013;<xref ref-type="bibr" rid="ref55">55</xref>). Consequently, the increase in output may be more significant on the succeeding day (<xref ref-type="bibr" rid="ref49">49</xref>). Similarly, urinary urea excretion decreased immediately after a 100-km run but increased the day after (<xref ref-type="bibr" rid="ref67">67</xref>). In the present study, urinary nitrogen excretion was measured up to 8:00 the day after EX and CON, yet no significant difference was observable within this period. In addition, total production (excretion + retention) increased immediately after a 100-km run and decreased the day after (<xref ref-type="bibr" rid="ref67">67</xref>). It is therefore unlikely that some differences appeared the day after that could have compensated for the effects of exercise.</p>
</sec>
</sec>
</sec>
<sec sec-type="conclusions" id="sec34">
<label>5</label>
<title>Conclusion</title>
<p>Our findings suggest that exercise did not significantly impact urinary nitrogen excretion. It is possible that the effect of exercise on urinary nitrogen excretion was reduced due to the reduction of the energy deficit, which was achieved by consuming almost protein-free snacks during exercise. As the sweat rate was measured in the present study, the sweat nitrogen excretion could be estimated based on previously published sweat nitrogen concentrations. This enabled us to estimate that exercise led to a significant increase in total nitrogen excretion. However, the urinary 3-MH excretion, which is an indicator of myofibrillar protein breakdown, did not change significantly with exercise. We recommend that future studies include measurements of sweat nitrogen excretion or at least an estimate to determine the effect of exercise on nitrogen excretion accurately.</p>
</sec>
<sec sec-type="data-availability" id="sec35">
<title>Data availability statement</title>
<p>The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation<monospace>.</monospace></p>
</sec>
<sec sec-type="ethics-statement" id="sec36">
<title>Ethics statement</title>
<p>The studies involving humans were approved by the Norwegian School of Sport Sciences&#x2019; Ethics Committee (Application 190-170621) and the Norwegian Centre for Research Data (Reference number 473323). The studies were conducted in accordance with the local legislation and institutional requirements. The participants provided their written informed consent to participate in this study.</p>
</sec>
<sec sec-type="author-contributions" id="sec37">
<title>Author contributions</title>
<p>MC: Conceptualization, Formal analysis, Investigation, Methodology, Writing &#x2013; original draft, Writing &#x2013; review &#x0026; editing. MB: Formal analysis, Investigation, Writing &#x2013; review &#x0026; editing. SW: Formal analysis, Investigation, Writing &#x2013; review &#x0026; editing. BS: Conceptualization, Methodology, Supervision, Writing &#x2013; review &#x0026; editing. JJ: Conceptualization, Funding acquisition, Methodology, Supervision, Writing &#x2013; review &#x0026; editing.</p>
</sec>
</body>
<back>
<sec sec-type="funding-information" id="sec38">
<title>Funding</title>
<p>The author(s) declare financial support was received for the research, authorship, and/or publication of this article. Internal funding from the Norwegian School of Sport Sciences supported this investigation.</p>
</sec>
<ack>
<p>The authors would like to thank the volunteers for participating and cooperating during the study. The authors would also like to thank Siri Taxer&#x00E5;s Dalen for her help in this study.</p>
</ack>
<sec sec-type="COI-statement" id="sec39">
<title>Conflict of interest</title>
<p>The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.</p>
<p>The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision.</p>
</sec>
<sec id="sec100" sec-type="disclaimer">
<title>Publisher&#x2019;s note</title>
<p>All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.</p>
</sec>
<sec sec-type="supplementary-material" id="sec40">
<title>Supplementary material</title>
<p>The Supplementary material for this article can be found online at: <ext-link xlink:href="https://www.frontiersin.org/articles/10.3389/fnut.2024.1345922/full#supplementary-material" ext-link-type="uri">https://www.frontiersin.org/articles/10.3389/fnut.2024.1345922/full#supplementary-material</ext-link></p>
<supplementary-material xlink:href="Table_1.DOCX" id="SM1" mimetype="application/vnd.openxmlformats-officedocument.wordprocessingml.document" xmlns:xlink="http://www.w3.org/1999/xlink"/>
<supplementary-material xlink:href="Table_2.DOCX" id="SM2" mimetype="application/vnd.openxmlformats-officedocument.wordprocessingml.document" xmlns:xlink="http://www.w3.org/1999/xlink"/>
</sec>
<ref-list>
<title>References</title>
<ref id="ref1">
<label>1.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>von Liebig</surname>
<given-names>JF</given-names>
</name>
</person-group>. <source>Animal chemistry, or organic chemistry in its applications to physiology and pathology</source>. <publisher-loc>Cambridge</publisher-loc>: <publisher-name>Taylor and Walton</publisher-name>. (<year>1842</year>). <fpage>412 p.</fpage></citation>
</ref>
<ref id="ref2">
<label>2.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Fick</surname>
<given-names>A</given-names>
</name> <name>
<surname>Wislicenus</surname>
<given-names>J</given-names>
</name></person-group>. <article-title>On the origin of muscular power</article-title>. <source>Lond Edinb Dubl Phil Mag</source>. (<year>1866</year>) <volume>31</volume>:<fpage>485</fpage>&#x2013;<lpage>503</lpage>. doi: <pub-id pub-id-type="doi">10.1080/14786446608644105</pub-id></citation>
</ref>
<ref id="ref3">
<label>3.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Argutinsky</surname>
<given-names>P</given-names>
</name>
</person-group>. <article-title>Muskelarbeit und Stickstoffumsatz [muscle work and nitrogen turnover]</article-title>. <source>Pfl&#x00FC;ger Arch</source>. (<year>1890</year>) <volume>46</volume>:<fpage>552</fpage>&#x2013;<lpage>80</lpage>. doi: <pub-id pub-id-type="doi">10.1007/BF01789543</pub-id></citation>
</ref>
<ref id="ref4">
<label>4.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Cathcart</surname>
<given-names>EP</given-names>
</name>
</person-group>. <article-title>The influence of muscle work on protein metabolism</article-title>. <source>Physiol Rev</source>. (<year>1925</year>) <volume>5</volume>:<fpage>225</fpage>&#x2013;<lpage>43</lpage>. doi: <pub-id pub-id-type="doi">10.1152/physrev.1925.5.2.225</pub-id></citation>
</ref>
<ref id="ref5">
<label>5.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Pavy</surname>
<given-names>FW</given-names>
</name>
</person-group>. <article-title>The effect of prolonged muscular exercise on the system</article-title>. <source>Lancet</source>. (<year>1876</year>) <volume>107</volume>:<fpage>353</fpage>&#x2013;<lpage>6</lpage>. doi: <pub-id pub-id-type="doi">10.1016/S0140-6736(02)46443-4</pub-id></citation>
</ref>
<ref id="ref6">
<label>6.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Pavy</surname>
<given-names>FW</given-names>
</name>
</person-group>. <article-title>The effect of prolonged muscular exercise on the system</article-title>. <source>Lancet</source>. (<year>1876</year>) <volume>107</volume>:<fpage>392</fpage>&#x2013;<lpage>4</lpage>. doi: <pub-id pub-id-type="doi">10.1016/S0140-6736(02)32021-X</pub-id></citation>
</ref>
<ref id="ref7">
<label>7.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Pavy</surname>
<given-names>FW</given-names>
</name>
</person-group>. <article-title>The effect of prolonged muscular exercise on the system</article-title>. <source>Lancet</source>. (<year>1876</year>) <volume>107</volume>:<fpage>429</fpage>&#x2013;<lpage>31</lpage>. doi: <pub-id pub-id-type="doi">10.1016/S0140-6736(02)32051-8</pub-id></citation>
</ref>
<ref id="ref8">
<label>8.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Pavy</surname>
<given-names>FW</given-names>
</name>
</person-group>. <article-title>The effect of prolonged muscular exercise on the system</article-title>. <source>Lancet</source>. (<year>1876</year>) <volume>107</volume>:<fpage>466</fpage>&#x2013;<lpage>8</lpage>. doi: <pub-id pub-id-type="doi">10.1016/S0140-6736(02)21242-8</pub-id></citation>
</ref>
<ref id="ref9">
<label>9.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Pavy</surname>
<given-names>FW</given-names>
</name>
</person-group>. <article-title>The effect of prolonged muscular exercise upon the urine in relation to the source of muscular power</article-title>. <source>Lancet</source>. (<year>1876</year>) <volume>108</volume>:<fpage>741</fpage>&#x2013;<lpage>3</lpage>. doi: <pub-id pub-id-type="doi">10.1016/S0140-6736(02)31881-6</pub-id></citation>
</ref>
<ref id="ref10">
<label>10.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Pavy</surname>
<given-names>FW</given-names>
</name>
</person-group>. <article-title>The effect of prolonged muscular exercise upon the urine in relation to the source of muscular power</article-title>. <source>Lancet</source>. (<year>1876</year>) <volume>108</volume>:<fpage>815</fpage>&#x2013;<lpage>8</lpage>. doi: <pub-id pub-id-type="doi">10.1016/S0140-6736(02)49561-X</pub-id></citation>
</ref>
<ref id="ref11">
<label>11.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Pavy</surname>
<given-names>FW</given-names>
</name>
</person-group>. <article-title>The effect of prolonged muscular exercise upon the urine in relation to the source of muscular power</article-title>. <source>Lancet</source>. (<year>1876</year>) <volume>108</volume>:<fpage>848</fpage>&#x2013;<lpage>50</lpage>. doi: <pub-id pub-id-type="doi">10.1016/S0140-6736(02)49585-2</pub-id></citation>
</ref>
<ref id="ref12">
<label>12.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Pavy</surname>
<given-names>FW</given-names>
</name>
</person-group>. <article-title>The effect of prolonged muscular exercise upon the urine in relation to the source of muscular power</article-title>. <source>Lancet</source>. (<year>1876</year>) <volume>108</volume>:<fpage>887</fpage>&#x2013;<lpage>9</lpage>. doi: <pub-id pub-id-type="doi">10.1016/S0140-6736(02)31787-2</pub-id></citation>
</ref>
<ref id="ref13">
<label>13.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Pellett</surname>
<given-names>PL</given-names>
</name> <name>
<surname>Young</surname>
<given-names>VR</given-names>
</name></person-group>. <article-title>The effects of different levels of energy intake on protein metabolism and of different levels of protein intake on energy metabolism: a statistical evaluation from the published literature</article-title>. <source>Protein-Energy Interactions</source>. (<year>1991</year>).</citation>
</ref>
<ref id="ref14">
<label>14.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Tarnopolsky</surname>
<given-names>LJ</given-names>
</name> <name>
<surname>MacDougall</surname>
<given-names>JD</given-names>
</name> <name>
<surname>Atkinson</surname>
<given-names>SA</given-names>
</name> <name>
<surname>Tarnopolsky</surname>
<given-names>MA</given-names>
</name> <name>
<surname>Sutton</surname>
<given-names>JR</given-names>
</name></person-group>. <article-title>Gender differences in substrate for endurance exercise</article-title>. <source>J Appl Physiol</source>. (<year>1990</year>) <volume>68</volume>:<fpage>302</fpage>&#x2013;<lpage>8</lpage>. doi: <pub-id pub-id-type="doi">10.1152/jappl.1990.68.1.302</pub-id></citation>
</ref>
<ref id="ref15">
<label>15.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Tarnopolsky</surname>
<given-names>MA</given-names>
</name> <name>
<surname>Atkinson</surname>
<given-names>SA</given-names>
</name> <name>
<surname>Phillips</surname>
<given-names>SM</given-names>
</name> <name>
<surname>MacDougall</surname>
<given-names>JD</given-names>
</name></person-group>. <article-title>Carbohydrate loading and metabolism during exercise in men and women</article-title>. <source>J Appl Physiol</source>. (<year>1995</year>) <volume>78</volume>:<fpage>1360</fpage>&#x2013;<lpage>8</lpage>. doi: <pub-id pub-id-type="doi">10.1152/jappl.1995.78.4.1360</pub-id>, PMID: <pub-id pub-id-type="pmid">7615443</pub-id></citation>
</ref>
<ref id="ref16">
<label>16.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Tarnopolsky</surname>
<given-names>MA</given-names>
</name> <name>
<surname>Bosman</surname>
<given-names>M</given-names>
</name> <name>
<surname>Macdonald</surname>
<given-names>JR</given-names>
</name> <name>
<surname>Vandeputte</surname>
<given-names>D</given-names>
</name> <name>
<surname>Martin</surname>
<given-names>J</given-names>
</name> <name>
<surname>Roy</surname>
<given-names>BD</given-names>
</name></person-group>. <article-title>Postexercise protein-carbohydrate and carbohydrate supplements increase muscle glycogen in men and women</article-title>. <source>J Appl Physiol</source>. (<year>1997</year>) <volume>83</volume>:<fpage>1877</fpage>&#x2013;<lpage>83</lpage>. doi: <pub-id pub-id-type="doi">10.1152/jappl.1997.83.6.1877</pub-id></citation>
</ref>
<ref id="ref17">
<label>17.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Calles-Escandon</surname>
<given-names>J</given-names>
</name> <name>
<surname>Cunningham</surname>
<given-names>JJ</given-names>
</name> <name>
<surname>Snyder</surname>
<given-names>P</given-names>
</name> <name>
<surname>Jacob</surname>
<given-names>R</given-names>
</name> <name>
<surname>Huszar</surname>
<given-names>G</given-names>
</name> <name>
<surname>Loke</surname>
<given-names>J</given-names>
</name> <etal/></person-group>. <article-title>Influence of exercise on urea, creatinine, and 3-methylhistidine excretion in normal human subjects</article-title>. <source>Am J Phys</source>. (<year>1984</year>) <volume>246</volume>:<fpage>E334</fpage>&#x2013;<lpage>8</lpage>. doi: <pub-id pub-id-type="doi">10.1152/ajpendo.1984.246.4.E334</pub-id>, PMID: <pub-id pub-id-type="pmid">6720887</pub-id></citation>
</ref>
<ref id="ref18">
<label>18.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Broad</surname>
<given-names>EM</given-names>
</name> <name>
<surname>Maughan</surname>
<given-names>RJ</given-names>
</name> <name>
<surname>Galloway</surname>
<given-names>SDR</given-names>
</name></person-group>. <article-title>Carbohydrate, protein, and fat metabolism during exercise after oral carnitine supplementation in humans</article-title>. <source>Int J Sport Nutr Exerc Metab</source>. (<year>2008</year>) <volume>18</volume>:<fpage>567</fpage>&#x2013;<lpage>84</lpage>. doi: <pub-id pub-id-type="doi">10.1123/ijsnem.18.6.567</pub-id>, PMID: <pub-id pub-id-type="pmid">19164828</pub-id></citation>
</ref>
<ref id="ref19">
<label>19.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Gontzea</surname>
<given-names>I</given-names>
</name> <name>
<surname>Sutzesco</surname>
<given-names>P</given-names>
</name> <name>
<surname>Dumitrache</surname>
<given-names>S</given-names>
</name></person-group>. <article-title>Recherches concernant l&#x2019;influence de l&#x2019;activit&#x00E9; musculaire Sur le m&#x00E9;tabolisme azot&#x00E9; et Sur le besoin en prot&#x00E9;ines de l&#x2019;homme [research concerning the influence of muscular activity on nitrogen metabolism and on the human protein requirement]</article-title>. <source>Ann Nutr Alim</source>. (<year>1968</year>) <volume>22</volume>:<fpage>183</fpage>&#x2013;<lpage>238</lpage>.</citation>
</ref>
<ref id="ref20">
<label>20.</label>
<citation citation-type="book"><person-group person-group-type="author"><name>
<surname>Bourges</surname>
<given-names>H</given-names>
</name> <name>
<surname>Young</surname>
<given-names>VR</given-names>
</name> <name>
<surname>Scrimshaw</surname>
<given-names>NS</given-names>
</name></person-group>. <article-title>Nitrogen losses in sweat induced by exercise</article-title> In: <source>Protein-energy-requirement studies in developing countries: Results of international research</source>. <publisher-loc>Tokyo</publisher-loc>: <publisher-name>The United Nations University</publisher-name>. (<year>1984</year>).</citation>
</ref>
<ref id="ref21">
<label>21.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Calloway</surname>
<given-names>DH</given-names>
</name> <name>
<surname>Odell</surname>
<given-names>AC</given-names>
</name> <name>
<surname>Margen</surname>
<given-names>S</given-names>
</name></person-group>. <article-title>Sweat and miscellaneous nitrogen losses in human balance studies</article-title>. <source>J Nutr</source>. (<year>1971</year>) <volume>101</volume>:<fpage>775</fpage>&#x2013;<lpage>86</lpage>. doi: <pub-id pub-id-type="doi">10.1093/jn/101.6.775</pub-id>, PMID: <pub-id pub-id-type="pmid">5578925</pub-id></citation>
</ref>
<ref id="ref22">
<label>22.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Oosthuyse</surname>
<given-names>T</given-names>
</name> <name>
<surname>Avidon</surname>
<given-names>I</given-names>
</name></person-group>. <article-title>Changes in substrate utilisation and protein catabolism during multiday cycling in well-trained cyclists</article-title>. <source>J Sports Sci</source>. (<year>2015</year>) <volume>33</volume>:<fpage>507</fpage>&#x2013;<lpage>17</lpage>. doi: <pub-id pub-id-type="doi">10.1080/02640414.2014.949827</pub-id>, PMID: <pub-id pub-id-type="pmid">25299240</pub-id></citation>
</ref>
<ref id="ref23">
<label>23.</label>
<citation citation-type="book"><person-group person-group-type="author">
<name>
<surname>Cerny</surname>
<given-names>FJ</given-names>
</name>
</person-group>. <article-title>Protein metabolism during two hour ergometer exercise</article-title> In: <person-group person-group-type="editor"><name><surname>Howald</surname> <given-names>H</given-names></name> <name><surname>Poortmans</surname> <given-names>JR</given-names></name></person-group>, editors. <source>Metabolic adaptation to prolonged physical exercise: Proceedings of the second international symposium on biochemistry of exercise</source>. <publisher-loc>Basel</publisher-loc>: <publisher-name>Birkh&#x00E4;user</publisher-name> (<year>1975</year>). <fpage>232</fpage>&#x2013;<lpage>7</lpage>.</citation>
</ref>
<ref id="ref24">
<label>24.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Lemon</surname>
<given-names>PW</given-names>
</name> <name>
<surname>Mullin</surname>
<given-names>JP</given-names>
</name></person-group>. <article-title>Effect of initial muscle glycogen levels on protein catabolism during exercise</article-title>. <source>J Appl Physiol Respir Environ Exerc Physiol</source>. (<year>1980</year>) <volume>48</volume>:<fpage>624</fpage>&#x2013;<lpage>9</lpage>. doi: <pub-id pub-id-type="doi">10.1152/jappl.1980.48.4.624</pub-id></citation>
</ref>
<ref id="ref25">
<label>25.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Rand</surname>
<given-names>WM</given-names>
</name> <name>
<surname>Pellett</surname>
<given-names>PL</given-names>
</name> <name>
<surname>Young</surname>
<given-names>VR</given-names>
</name></person-group>. <article-title>Meta-analysis of nitrogen balance studies for estimating protein requirements in healthy adults</article-title>. <source>Am J Clin Nutr</source>. (<year>2003</year>) <volume>77</volume>:<fpage>109</fpage>&#x2013;<lpage>27</lpage>. doi: <pub-id pub-id-type="doi">10.1093/ajcn/77.1.109</pub-id>, PMID: <pub-id pub-id-type="pmid">12499330</pub-id></citation>
</ref>
<ref id="ref26">
<label>26.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Williamson</surname>
<given-names>DH</given-names>
</name> <name>
<surname>Farrell</surname>
<given-names>R</given-names>
</name> <name>
<surname>Kerr</surname>
<given-names>A</given-names>
</name> <name>
<surname>Smith</surname>
<given-names>R</given-names>
</name></person-group>. <article-title>Muscle-protein catabolism after injury in man, as measured by urinary excretion of 3-methylhistidine</article-title>. <source>Clin Sci</source>. (<year>1977</year>) <volume>52</volume>:<fpage>527</fpage>&#x2013;<lpage>33</lpage>. doi: <pub-id pub-id-type="doi">10.1042/cs0520527</pub-id></citation>
</ref>
<ref id="ref27">
<label>27.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Dohm</surname>
<given-names>GL</given-names>
</name> <name>
<surname>Tapscott</surname>
<given-names>EB</given-names>
</name> <name>
<surname>Kasperek</surname>
<given-names>GJ</given-names>
</name></person-group>. <article-title>Protein degradation during endurance exercise and recovery</article-title>. <source>Med Sci Sports Exerc</source>. (<year>1987</year>) <volume>19</volume>:<fpage>S172???S178</fpage>&#x2013;<lpage>1</lpage>. doi: <pub-id pub-id-type="doi">10.1249/00005768-198710001-00011</pub-id></citation>
</ref>
<ref id="ref28">
<label>28.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Bilmazes</surname>
<given-names>C</given-names>
</name> <name>
<surname>Uauy</surname>
<given-names>R</given-names>
</name> <name>
<surname>Haverberg</surname>
<given-names>LN</given-names>
</name> <name>
<surname>Munro</surname>
<given-names>HN</given-names>
</name> <name>
<surname>Young</surname>
<given-names>VR</given-names>
</name></person-group>. <article-title>Muscle protein breakdown rates in humans based on Ntau-methylhistidine (3-methylhistidine) content of mixed proteins in skeletal muscle and urinary output of Ntau-methylhistidine</article-title>. <source>Metab Clin Exp</source>. (<year>1978</year>) <volume>27</volume>:<fpage>525</fpage>&#x2013;<lpage>30</lpage>. doi: <pub-id pub-id-type="doi">10.1016/0026-0495(78)90018-5</pub-id>, PMID: <pub-id pub-id-type="pmid">642824</pub-id></citation>
</ref>
<ref id="ref29">
<label>29.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Sj&#x00F6;lin</surname>
<given-names>J</given-names>
</name> <name>
<surname>Stjernstr&#x00F6;m</surname>
<given-names>H</given-names>
</name> <name>
<surname>Henneberg</surname>
<given-names>S</given-names>
</name> <name>
<surname>Hambraeus</surname>
<given-names>L</given-names>
</name> <name>
<surname>Friman</surname>
<given-names>G</given-names>
</name></person-group>. <article-title>Evaluation of urinary 3-methylhistidine excretion in infection by measurements of 1-methylhistidine and the creatinine ratios</article-title>. <source>Am J Clin Nutr</source>. (<year>1989</year>) <volume>49</volume>:<fpage>62</fpage>&#x2013;<lpage>70</lpage>. doi: <pub-id pub-id-type="doi">10.1093/ajcn/49.1.62</pub-id>, PMID: <pub-id pub-id-type="pmid">2912013</pub-id></citation>
</ref>
<ref id="ref30">
<label>30.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Wassner</surname>
<given-names>SJ</given-names>
</name> <name>
<surname>Orloff</surname>
<given-names>S</given-names>
</name> <name>
<surname>Holliday</surname>
<given-names>MA</given-names>
</name></person-group>. <article-title>Protein degradation in muscle: response to feeding and fasting in growing rats</article-title>. <source>Am J Phys</source>. (<year>1977</year>) <volume>233</volume>:<fpage>E119</fpage>&#x2013;<lpage>23</lpage>. doi: <pub-id pub-id-type="doi">10.1152/ajpendo.1977.233.2.E119</pub-id>, PMID: <pub-id pub-id-type="pmid">888948</pub-id></citation>
</ref>
<ref id="ref31">
<label>31.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Ballard</surname>
<given-names>FJ</given-names>
</name> <name>
<surname>Tomas</surname>
<given-names>FM</given-names>
</name></person-group>. <article-title>3-Methylhistidine as a measure of skeletal muscle protein breakdown in human subjects: the case for its continued use</article-title>. <source>Clin Sci (Lond)</source>. (<year>1983</year>) <volume>65</volume>:<fpage>209</fpage>&#x2013;<lpage>15</lpage>. doi: <pub-id pub-id-type="doi">10.1042/cs0650209</pub-id>, PMID: <pub-id pub-id-type="pmid">6872455</pub-id></citation>
</ref>
<ref id="ref32">
<label>32.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Young</surname>
<given-names>VR</given-names>
</name> <name>
<surname>Alexis</surname>
<given-names>SD</given-names>
</name> <name>
<surname>Baliga</surname>
<given-names>BS</given-names>
</name> <name>
<surname>Munro</surname>
<given-names>HN</given-names>
</name> <name>
<surname>Muecke</surname>
<given-names>W</given-names>
</name></person-group>. <article-title>Metabolism of administered 3-methylhistidine. Lack of muscle transfer ribonucleic acid charging and quantitative excretion as 3 methylhistidine and its N-acetyl derivative</article-title>. <source>J Biol Chem</source>. (<year>1972</year>) <volume>247</volume>:<fpage>3592</fpage>&#x2013;<lpage>600</lpage>. doi: <pub-id pub-id-type="doi">10.1016/S0021-9258(19)45182-X</pub-id></citation>
</ref>
<ref id="ref33">
<label>33.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Long</surname>
<given-names>CL</given-names>
</name> <name>
<surname>Haverberg</surname>
<given-names>LN</given-names>
</name> <name>
<surname>Young</surname>
<given-names>VR</given-names>
</name> <name>
<surname>Kinney</surname>
<given-names>JM</given-names>
</name> <name>
<surname>Munro</surname>
<given-names>HN</given-names>
</name> <name>
<surname>Geiger</surname>
<given-names>JW</given-names>
</name></person-group>. <article-title>Metabolism of 3-methylhistidine in man</article-title>. <source>Metab Clin Exp</source>. (<year>1975</year>) <volume>24</volume>:<fpage>929</fpage>&#x2013;<lpage>35</lpage>. doi: <pub-id pub-id-type="doi">10.1016/0026-0495(75)90084-0</pub-id></citation>
</ref>
<ref id="ref34">
<label>34.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Hasselgren</surname>
<given-names>P-O</given-names>
</name> <name>
<surname>Pedersen</surname>
<given-names>P</given-names>
</name> <name>
<surname>Sax</surname>
<given-names>HC</given-names>
</name> <name>
<surname>Warner</surname>
<given-names>BW</given-names>
</name> <name>
<surname>Fischer</surname>
<given-names>JE</given-names>
</name></person-group>. <article-title>Methods for studying protein synthesis and degradation in liver and skeletal muscle</article-title>. <source>J Surg Res</source>. (<year>1988</year>) <volume>45</volume>:<fpage>389</fpage>&#x2013;<lpage>415</lpage>. doi: <pub-id pub-id-type="doi">10.1016/0022-4804(88)90136-9</pub-id></citation>
</ref>
<ref id="ref35">
<label>35.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Rustad</surname>
<given-names>PI</given-names>
</name> <name>
<surname>Sailer</surname>
<given-names>M</given-names>
</name> <name>
<surname>Cumming</surname>
<given-names>KT</given-names>
</name> <name>
<surname>Jeppesen</surname>
<given-names>PB</given-names>
</name> <name>
<surname>Kolnes</surname>
<given-names>KJ</given-names>
</name> <name>
<surname>Sollie</surname>
<given-names>O</given-names>
</name> <etal/></person-group>. <article-title>Intake of protein plus carbohydrate during the first two hours after exhaustive cycling improves performance the following day</article-title>. <source>PLoS One</source>. (<year>2016</year>) <volume>11</volume>:<fpage>e0153229</fpage>. doi: <pub-id pub-id-type="doi">10.1371/journal.pone.0153229</pub-id>, PMID: <pub-id pub-id-type="pmid">27078151</pub-id></citation>
</ref>
<ref id="ref36">
<label>36.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Sollie</surname>
<given-names>O</given-names>
</name> <name>
<surname>Jeppesen</surname>
<given-names>PB</given-names>
</name> <name>
<surname>Tangen</surname>
<given-names>DS</given-names>
</name> <name>
<surname>Jerner&#x00E9;n</surname>
<given-names>F</given-names>
</name> <name>
<surname>Nellemann</surname>
<given-names>B</given-names>
</name> <name>
<surname>Valsdottir</surname>
<given-names>D</given-names>
</name> <etal/></person-group>. <article-title>Protein intake in the early recovery period after exhaustive exercise improves performance the following day</article-title>. <source>J Appl Physiol</source>. (<year>2018</year>) <volume>125</volume>:<fpage>1731</fpage>&#x2013;<lpage>42</lpage>. doi: <pub-id pub-id-type="doi">10.1152/japplphysiol.01132.2017</pub-id>, PMID: <pub-id pub-id-type="pmid">30212306</pub-id></citation>
</ref>
<ref id="ref37">
<label>37.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Clauss</surname>
<given-names>M</given-names>
</name> <name>
<surname>Skattebo</surname>
<given-names>&#x00D8;</given-names>
</name> <name>
<surname>D&#x00E6;hli</surname>
<given-names>MR</given-names>
</name> <name>
<surname>Valsdottir</surname>
<given-names>D</given-names>
</name> <name>
<surname>Bastani</surname>
<given-names>NE</given-names>
</name> <name>
<surname>Johansen</surname>
<given-names>EI</given-names>
</name> <etal/></person-group>. <article-title>Carbohydrate ingestion during prolonged cycling improves next-day time trial performance and alters amino acid concentrations</article-title>. <source>Med Sci Sports Exerc</source>. (<year>2023</year>) <volume>55</volume>:<fpage>2228</fpage>&#x2013;<lpage>40</lpage>. doi: <pub-id pub-id-type="doi">10.1249/MSS.0000000000003264</pub-id>, PMID: <pub-id pub-id-type="pmid">37535337</pub-id></citation>
</ref>
<ref id="ref38">
<label>38.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Borg</surname>
<given-names>GA</given-names>
</name>
</person-group>. <article-title>Perceived exertion: a note on &#x201C;history&#x201D; and methods</article-title>. <source>Med Sci Sports</source>. (<year>1973</year>) <volume>5</volume>:<fpage>90</fpage>&#x2013;<lpage>3</lpage>. doi: <pub-id pub-id-type="doi">10.1249/00005768-197300520-00017</pub-id>, PMID: <pub-id pub-id-type="pmid">4721012</pub-id></citation>
</ref>
<ref id="ref39">
<label>39.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Mariotti</surname>
<given-names>F</given-names>
</name> <name>
<surname>Tom&#x00E9;</surname>
<given-names>D</given-names>
</name> <name>
<surname>Mirand</surname>
<given-names>PP</given-names>
</name></person-group>. <article-title>Converting nitrogen into protein&#x2014; beyond 6.25 and Jones&#x2019; factors</article-title>. <source>Crit Rev Food Sci Nutr</source>. (<year>2008</year>) <volume>48</volume>:<fpage>177</fpage>&#x2013;<lpage>84</lpage>. doi: <pub-id pub-id-type="doi">10.1080/10408390701279749</pub-id>, PMID: <pub-id pub-id-type="pmid">18274971</pub-id></citation>
</ref>
<ref id="ref40">
<label>40.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>P&#x00E9;ronnet</surname>
<given-names>F</given-names>
</name> <name>
<surname>Massicotte</surname>
<given-names>D</given-names>
</name></person-group>. <article-title>Table of nonprotein respiratory quotient: an update</article-title>. <source>Can J Sport Sci</source>. (<year>1991</year>) <volume>16</volume>:<fpage>23</fpage>&#x2013;<lpage>9</lpage>. PMID: <pub-id pub-id-type="pmid">1645211</pub-id></citation>
</ref>
<ref id="ref41">
<label>41.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Mifflin</surname>
<given-names>MD</given-names>
</name> <name>
<surname>St Jeor</surname>
<given-names>ST</given-names>
</name> <name>
<surname>Hill</surname>
<given-names>LA</given-names>
</name> <name>
<surname>Scott</surname>
<given-names>BJ</given-names>
</name> <name>
<surname>Daugherty</surname>
<given-names>SA</given-names>
</name> <name>
<surname>Koh</surname>
<given-names>YO</given-names>
</name></person-group>. <article-title>A new predictive equation for resting energy expenditure in healthy individuals</article-title>. <source>Am J Clin Nutr</source>. (<year>1990</year>) <volume>51</volume>:<fpage>241</fpage>&#x2013;<lpage>7</lpage>. doi: <pub-id pub-id-type="doi">10.1093/ajcn/51.2.241</pub-id></citation>
</ref>
<ref id="ref42">
<label>42.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Kjeldahl</surname>
<given-names>J</given-names>
</name>
</person-group>. <article-title>Neue Methode zur Bestimmung des Stickstoffs in organischen K&#x00F6;rpern [New method for determining nitrogen in organic bodies]</article-title>. <source>Fresenius Z Anal Chem</source>. (<year>1883</year>) <volume>22</volume>:<fpage>366</fpage>&#x2013;<lpage>82</lpage>. doi: <pub-id pub-id-type="doi">10.1007/BF01338151</pub-id></citation>
</ref>
<ref id="ref43">
<label>43.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Cheuvront</surname>
<given-names>SN</given-names>
</name> <name>
<surname>Montain</surname>
<given-names>SJ</given-names>
</name> <name>
<surname>Goodman</surname>
<given-names>DA</given-names>
</name> <name>
<surname>Blanchard</surname>
<given-names>L</given-names>
</name> <name>
<surname>Sawka</surname>
<given-names>MN</given-names>
</name></person-group>. <article-title>Evaluation of the limits to accurate sweat loss prediction during prolonged exercise</article-title>. <source>Eur J Appl Physiol</source>. (<year>2007</year>) <volume>101</volume>:<fpage>215</fpage>&#x2013;<lpage>24</lpage>. doi: <pub-id pub-id-type="doi">10.1007/s00421-007-0492-x</pub-id>, PMID: <pub-id pub-id-type="pmid">17534643</pub-id></citation>
</ref>
<ref id="ref44">
<label>44.</label>
<citation citation-type="journal"><article-title>On the physiological effects of severe and protracted muscular exercise; with special reference to its influence upon the excretion of nitrogen</article-title>. <source>New York Med Jour</source>. (<year>1870</year>) <volume>13</volume>:<fpage>653</fpage>.</citation>
</ref>
<ref id="ref45">
<label>45.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Schenk</surname>
<given-names>F</given-names>
</name>
</person-group>. <article-title>II. &#x00DC;ber den Einfluss der Muskelarbeit auf die Eiweisszersetzung im menschlichen Organismus [About the influence of muscle work on protein breakdown in the human organism]</article-title>. <source>Arch Exp Path Pharm</source>. (<year>1874</year>) <volume>2</volume>:<fpage>21</fpage>&#x2013;<lpage>32</lpage>. doi: <pub-id pub-id-type="doi">10.1007/BF01829507</pub-id></citation>
</ref>
<ref id="ref46">
<label>46.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Hirschfeld</surname>
<given-names>F</given-names>
</name>
</person-group>. <article-title>Beitr&#x00E4;ge zur Ern&#x00E4;hrungslehre des Menschen [Contributions to human nutrition]</article-title>. <source>Archiv f pathol Anat</source>. (<year>1888</year>) <volume>114</volume>:<fpage>301</fpage>&#x2013;<lpage>40</lpage>. doi: <pub-id pub-id-type="doi">10.1007/BF01882632</pub-id></citation>
</ref>
<ref id="ref47">
<label>47.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Krummacher</surname>
<given-names>O</given-names>
</name>
</person-group>. <article-title>&#x00DC;ber den Einfluss der Muskelarbeit auf die Eiweisszersetzung bei gleicher Nahrung [about the influence of muscle work on protein breakdown with the same food]</article-title>. <source>Pfl&#x00FC;ger Arch</source>. (<year>1890</year>) <volume>47</volume>:<fpage>454</fpage>&#x2013;<lpage>68</lpage>. doi: <pub-id pub-id-type="doi">10.1007/BF01789825</pub-id></citation>
</ref>
<ref id="ref48">
<label>48.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Bornstein</surname>
<given-names>K</given-names>
</name>
</person-group>. <article-title>Eiweissmast und Muskelarbeit [Protein pole and muscle work]</article-title>. <source>Pfl&#x00FC;ger Arch</source>. (<year>1901</year>) <volume>83</volume>:<fpage>540</fpage>&#x2013;<lpage>56</lpage>. doi: <pub-id pub-id-type="doi">10.1007/BF01746508</pub-id></citation>
</ref>
<ref id="ref49">
<label>49.</label>
<citation citation-type="book"><person-group person-group-type="author">
<name>
<surname>Atwater</surname>
<given-names>S</given-names>
</name>
</person-group>. <source>The effect of severe and prolonged muscular work on food consumption, digestion, and metabolism</source>. <publisher-loc>Bull - US</publisher-loc>: <publisher-name>Dep Agric</publisher-name> (<year>1901</year>). <fpage>98</fpage> p.</citation>
</ref>
<ref id="ref50">
<label>50.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Campbell</surname>
<given-names>JA</given-names>
</name> <name>
<surname>Webster</surname>
<given-names>TA</given-names>
</name></person-group>. <article-title>Effect of severe muscular work on composition of the urine</article-title>. <source>Biochem J</source>. (<year>1922</year>) <volume>16</volume>:<fpage>106</fpage>&#x2013;<lpage>10</lpage>. doi: <pub-id pub-id-type="doi">10.1042/bj0160106</pub-id>, PMID: <pub-id pub-id-type="pmid">16743053</pub-id></citation>
</ref>
<ref id="ref51">
<label>51.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Campbell</surname>
<given-names>JA</given-names>
</name> <name>
<surname>Webster</surname>
<given-names>TA</given-names>
</name></person-group>. <article-title>Day and night urine during complete rest, laboratory routine, light muscular work and oxygen administration</article-title>. <source>Biochem J</source>. (<year>1921</year>) <volume>15</volume>:<fpage>660</fpage>&#x2013;<lpage>4</lpage>. doi: <pub-id pub-id-type="doi">10.1042/bj0150660</pub-id>, PMID: <pub-id pub-id-type="pmid">16743037</pub-id></citation>
</ref>
<ref id="ref52">
<label>52.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Brietzcke</surname>
<given-names>H</given-names>
</name>
</person-group>. <article-title>Urea and its relation to muscular force</article-title>. <source>Br Foreign Med Chir Rev</source>. (<year>1877</year>) <volume>60</volume>:<fpage>190</fpage>&#x2013;<lpage>200</lpage>. PMID: <pub-id pub-id-type="pmid">30164732</pub-id></citation>
</ref>
<ref id="ref53">
<label>53.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Dunlop</surname>
<given-names>JC</given-names>
</name> <name>
<surname>Paton</surname>
<given-names>DN</given-names>
</name> <name>
<surname>Stockman</surname>
<given-names>R</given-names>
</name> <name>
<surname>Maccadam</surname>
<given-names>I</given-names>
</name></person-group>. <article-title>On the influence of muscular exercise, sweating, and massage, on the metabolism</article-title>. <source>J Physiol</source>. (<year>1897</year>) <volume>22</volume>:<fpage>68</fpage>&#x2013;<lpage>91</lpage>. doi: <pub-id pub-id-type="doi">10.1113/jphysiol.1897.sp000677</pub-id>, PMID: <pub-id pub-id-type="pmid">16992431</pub-id></citation>
</ref>
<ref id="ref54">
<label>54.</label>
<citation citation-type="journal"><article-title>Kaup</article-title>. <source>Z Biol</source> (<year>1902</year>) <volume>43</volume>:<fpage>221</fpage>.</citation>
</ref>
<ref id="ref55">
<label>55.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Kocher</surname>
<given-names>RA</given-names>
</name>
</person-group>. <article-title>&#x00DC;ber die Gro&#x00DF;e des Eiwei&#x00DF;zerfalls bei Fieber und bei Arbeitsleistung [About the extent of protein breakdown during fever and during work]</article-title>. <source>Dtsch Arch Klin Med.</source> (<year>1914</year>) <volume>65</volume>:<fpage>82</fpage>.</citation>
</ref>
<ref id="ref56">
<label>56.</label>
<citation citation-type="book"><person-group person-group-type="author">
<name>
<surname>Boyd</surname>
<given-names>J.</given-names>
</name>
</person-group> <source>A contribution to the metabolism of creatine</source>. (M.D. thesis). <publisher-name>University of Glasgow</publisher-name>. (<year>1914</year>).</citation>
</ref>
<ref id="ref57">
<label>57.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Todd</surname>
<given-names>KS</given-names>
</name> <name>
<surname>Butterfield</surname>
<given-names>GE</given-names>
</name> <name>
<surname>Calloway</surname>
<given-names>DH</given-names>
</name></person-group>. <article-title>Nitrogen balance in men with adequate and deficient energy intake at three levels of work</article-title>. <source>J Nutr</source>. (<year>1984</year>) <volume>114</volume>:<fpage>2107</fpage>&#x2013;<lpage>18</lpage>. doi: <pub-id pub-id-type="doi">10.1093/jn/114.11.2107</pub-id></citation>
</ref>
<ref id="ref58">
<label>58.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Butterfield</surname>
<given-names>GE</given-names>
</name> <name>
<surname>Calloway</surname>
<given-names>DH</given-names>
</name></person-group>. <article-title>Physical activity improves protein utilization in young men</article-title>. <source>BJN</source>. (<year>1984</year>) <volume>51</volume>:<fpage>171</fpage>. doi: <pub-id pub-id-type="doi">10.1079/BJN19840021</pub-id></citation>
</ref>
<ref id="ref59">
<label>59.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Calloway</surname>
<given-names>DH</given-names>
</name> <name>
<surname>Spector</surname>
<given-names>H</given-names>
</name></person-group>. <article-title>Nitrogen balance as related to caloric and protein intake in active young men</article-title>. <source>Am J Clin Nutr</source>. (<year>1954</year>) <volume>2</volume>:<fpage>405</fpage>&#x2013;<lpage>12</lpage>. doi: <pub-id pub-id-type="doi">10.1093/ajcn/2.6.405</pub-id>, PMID: <pub-id pub-id-type="pmid">13207036</pub-id></citation>
</ref>
<ref id="ref60">
<label>60.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Consolazio</surname>
<given-names>CF</given-names>
</name> <name>
<surname>Nelson</surname>
<given-names>RA</given-names>
</name> <name>
<surname>Matoush</surname>
<given-names>LO</given-names>
</name> <name>
<surname>Harding</surname>
<given-names>RS</given-names>
</name> <name>
<surname>Canham</surname>
<given-names>JE</given-names>
</name></person-group>. <article-title>Nitrogen excretion in sweat and its relation to nitrogen balance requirements</article-title>. <source>J Nutr</source>. (<year>1963</year>) <volume>79</volume>:<fpage>399</fpage>&#x2013;<lpage>406</lpage>. doi: <pub-id pub-id-type="doi">10.1093/jn/79.4.399</pub-id>, PMID: <pub-id pub-id-type="pmid">14022657</pub-id></citation>
</ref>
<ref id="ref61">
<label>61.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Phillips</surname>
<given-names>SM</given-names>
</name> <name>
<surname>Atkinson</surname>
<given-names>SA</given-names>
</name> <name>
<surname>Tarnopolsky</surname>
<given-names>MA</given-names>
</name> <name>
<surname>MacDougall</surname>
<given-names>JD</given-names>
</name></person-group>. <article-title>Gender differences in leucine kinetics and nitrogen balance in endurance athletes</article-title>. <source>J Appl Physiol</source>. (<year>1993</year>) <volume>75</volume>:<fpage>2134</fpage>&#x2013;<lpage>41</lpage>. doi: <pub-id pub-id-type="doi">10.1152/jappl.1993.75.5.2134</pub-id>, PMID: <pub-id pub-id-type="pmid">8307870</pub-id></citation>
</ref>
<ref id="ref62">
<label>62.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Pikosky</surname>
<given-names>MA</given-names>
</name> <name>
<surname>Smith</surname>
<given-names>TJ</given-names>
</name> <name>
<surname>Grediagin</surname>
<given-names>A</given-names>
</name> <name>
<surname>Castaneda-Sceppa</surname>
<given-names>C</given-names>
</name> <name>
<surname>Byerley</surname>
<given-names>L</given-names>
</name> <name>
<surname>Glickman</surname>
<given-names>EL</given-names>
</name> <etal/></person-group>. <article-title>Increased protein maintains nitrogen balance during exercise-induced energy deficit</article-title>. <source>Med Sci Sports Exerc</source>. (<year>2008</year>) <volume>40</volume>:<fpage>505</fpage>&#x2013;<lpage>12</lpage>. doi: <pub-id pub-id-type="doi">10.1249/MSS.0b013e31815f6643</pub-id>, PMID: <pub-id pub-id-type="pmid">18379214</pub-id></citation>
</ref>
<ref id="ref63">
<label>63.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Smith</surname>
<given-names>TJ</given-names>
</name> <name>
<surname>Pikosky</surname>
<given-names>MA</given-names>
</name> <name>
<surname>Grediagin</surname>
<given-names>A</given-names>
</name> <name>
<surname>Castaneda-Sceppa</surname>
<given-names>C</given-names>
</name> <name>
<surname>Byerley</surname>
<given-names>LO</given-names>
</name> <name>
<surname>Glickman</surname>
<given-names>EL</given-names>
</name> <etal/></person-group>. <article-title>Aerobic fitness does not modulate protein metabolism in response to increased exercise: a controlled trial</article-title>. <source>Nutr Metab (Lond)</source>. (<year>2009</year>) <volume>6</volume>:<fpage>28</fpage>. doi: <pub-id pub-id-type="doi">10.1186/1743-7075-6-28</pub-id>, PMID: <pub-id pub-id-type="pmid">19531244</pub-id></citation>
</ref>
<ref id="ref64">
<label>64.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Carraro</surname>
<given-names>F</given-names>
</name> <name>
<surname>Hartl</surname>
<given-names>WH</given-names>
</name> <name>
<surname>Stuart</surname>
<given-names>CA</given-names>
</name> <name>
<surname>Layman</surname>
<given-names>DK</given-names>
</name> <name>
<surname>Jahoor</surname>
<given-names>F</given-names>
</name> <name>
<surname>Wolfe</surname>
<given-names>RR</given-names>
</name></person-group>. <article-title>Whole body and plasma protein synthesis in exercise and recovery in human subjects</article-title>. <source>Am J Phys</source>. (<year>1990</year>) <volume>258</volume>:<fpage>E821</fpage>&#x2013;<lpage>31</lpage>. doi: <pub-id pub-id-type="doi">10.1152/ajpendo.1990.258.5.E821</pub-id></citation>
</ref>
<ref id="ref65">
<label>65.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Carraro</surname>
<given-names>F</given-names>
</name> <name>
<surname>Stuart</surname>
<given-names>CA</given-names>
</name> <name>
<surname>Hartl</surname>
<given-names>WH</given-names>
</name> <name>
<surname>Rosenblatt</surname>
<given-names>J</given-names>
</name> <name>
<surname>Wolfe</surname>
<given-names>RR</given-names>
</name></person-group>. <article-title>Effect of exercise and recovery on muscle protein synthesis in human subjects</article-title>. <source>Am J Phys</source>. (<year>1990</year>) <volume>259</volume>:<fpage>E470</fpage>&#x2013;<lpage>6</lpage>. doi: <pub-id pub-id-type="doi">10.1152/ajpendo.1990.259.4.E470</pub-id></citation>
</ref>
<ref id="ref66">
<label>66.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Colombani</surname>
<given-names>PC</given-names>
</name> <name>
<surname>Kovacs</surname>
<given-names>E</given-names>
</name> <name>
<surname>Frey-Rindova</surname>
<given-names>P</given-names>
</name> <name>
<surname>Frey</surname>
<given-names>W</given-names>
</name> <name>
<surname>Langhans</surname>
<given-names>W</given-names>
</name> <name>
<surname>Arnold</surname>
<given-names>M</given-names>
</name> <etal/></person-group>. <article-title>Metabolic effects of a protein-supplemented carbohydrate drink in marathon runners</article-title>. <source>Int J Sport Nutr</source>. (<year>1999</year>) <volume>9</volume>:<fpage>181</fpage>&#x2013;<lpage>201</lpage>. doi: <pub-id pub-id-type="doi">10.1123/ijsn.9.2.181</pub-id>, PMID: <pub-id pub-id-type="pmid">10362454</pub-id></citation>
</ref>
<ref id="ref67">
<label>67.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>D&#x00E9;combaz</surname>
<given-names>J</given-names>
</name> <name>
<surname>Reinhardt</surname>
<given-names>P</given-names>
</name> <name>
<surname>Anantharaman</surname>
<given-names>K</given-names>
</name> <name>
<surname>von Glutz</surname>
<given-names>G</given-names>
</name> <name>
<surname>Poortmans</surname>
<given-names>JR</given-names>
</name></person-group>. <article-title>Biochemical changes in a 100 km run: free amino acids, urea, and creatinine</article-title>. <source>Eur J Appl Physiol Occup Physiol</source>. (<year>1979</year>) <volume>41</volume>:<fpage>61</fpage>&#x2013;<lpage>72</lpage>. doi: <pub-id pub-id-type="doi">10.1007/bf00424469</pub-id>, PMID: <pub-id pub-id-type="pmid">446472</pub-id></citation>
</ref>
<ref id="ref68">
<label>68.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Dohm</surname>
<given-names>GL</given-names>
</name> <name>
<surname>Israel</surname>
<given-names>RG</given-names>
</name> <name>
<surname>Breedlove</surname>
<given-names>RL</given-names>
</name> <name>
<surname>Williams</surname>
<given-names>RT</given-names>
</name> <name>
<surname>Askew</surname>
<given-names>EW</given-names>
</name></person-group>. <article-title>Biphasic changes in 3-methylhistidine excretion in humans after exercise</article-title>. <source>Am J Physiol Endocrinol Metab</source>. (<year>1985</year>) <volume>248</volume>:<fpage>E588</fpage>&#x2013;<lpage>92</lpage>. doi: <pub-id pub-id-type="doi">10.1152/ajpendo.1985.248.5.E588</pub-id>, PMID: <pub-id pub-id-type="pmid">3993777</pub-id></citation>
</ref>
<ref id="ref69">
<label>69.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Evans</surname>
<given-names>WJ</given-names>
</name> <name>
<surname>Meredith</surname>
<given-names>CN</given-names>
</name> <name>
<surname>Cannon</surname>
<given-names>JG</given-names>
</name> <name>
<surname>Dinarello</surname>
<given-names>CA</given-names>
</name> <name>
<surname>Frontera</surname>
<given-names>WR</given-names>
</name> <name>
<surname>Hughes</surname>
<given-names>VA</given-names>
</name> <etal/></person-group>. <article-title>Metabolic changes following eccentric exercise in trained and untrained men</article-title>. <source>J Appl Physiol (1985)</source>. (<year>1986</year>) <volume>61</volume>:<fpage>1864</fpage>&#x2013;<lpage>8</lpage>. doi: <pub-id pub-id-type="doi">10.1152/jappl.1986.61.5.1864</pub-id></citation>
</ref>
<ref id="ref70">
<label>70.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Fielding</surname>
<given-names>RA</given-names>
</name> <name>
<surname>Meredith</surname>
<given-names>CN</given-names>
</name> <name>
<surname>O&#x2019;Reilly</surname>
<given-names>KP</given-names>
</name> <name>
<surname>Frontera</surname>
<given-names>WR</given-names>
</name> <name>
<surname>Cannon</surname>
<given-names>JG</given-names>
</name> <name>
<surname>Evans</surname>
<given-names>WJ</given-names>
</name></person-group>. <article-title>Enhanced protein breakdown after eccentric exercise in young and older men</article-title>. <source>J Appl Physiol</source>. (<year>1991</year>) <volume>71</volume>:<fpage>674</fpage>&#x2013;<lpage>9</lpage>. doi: <pub-id pub-id-type="doi">10.1152/jappl.1991.71.2.674</pub-id>, PMID: <pub-id pub-id-type="pmid">1938741</pub-id></citation>
</ref>
<ref id="ref71">
<label>71.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Plante</surname>
<given-names>RI</given-names>
</name> <name>
<surname>Houston</surname>
<given-names>ME</given-names>
</name></person-group>. <article-title>Exercise and protein catabolism in women</article-title>. <source>Ann Nutr Metab</source>. (<year>1984</year>) <volume>28</volume>:<fpage>123</fpage>&#x2013;<lpage>9</lpage>. doi: <pub-id pub-id-type="doi">10.1159/000176792</pub-id></citation>
</ref>
<ref id="ref72">
<label>72.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Dohm</surname>
<given-names>GL</given-names>
</name> <name>
<surname>Williams</surname>
<given-names>RT</given-names>
</name> <name>
<surname>Kasperek</surname>
<given-names>GJ</given-names>
</name> <name>
<surname>van Rij</surname>
<given-names>AM</given-names>
</name></person-group>. <article-title>Increased excretion of urea and N tau -methylhistidine by rats and humans after a bout of exercise</article-title>. <source>J Appl Physiol Respir Environ Exerc Physiol</source>. (<year>1982</year>) <volume>52</volume>:<fpage>27</fpage>&#x2013;<lpage>33</lpage>. doi: <pub-id pub-id-type="doi">10.1152/jappl.1982.52.1.27</pub-id></citation>
</ref>
<ref id="ref73">
<label>73.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Radha</surname>
<given-names>E</given-names>
</name> <name>
<surname>Bessman</surname>
<given-names>SP</given-names>
</name></person-group>. <article-title>Effect of exercise on protein degradation: 3-methylhistidine and creatinine excretion</article-title>. <source>Biochem Med</source>. (<year>1983</year>) <volume>29</volume>:<fpage>96</fpage>&#x2013;<lpage>100</lpage>. doi: <pub-id pub-id-type="doi">10.1016/0006-2944(83)90058-3</pub-id>, PMID: <pub-id pub-id-type="pmid">6838503</pub-id></citation>
</ref>
<ref id="ref74">
<label>74.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Rennie</surname>
<given-names>MJ</given-names>
</name> <name>
<surname>Edwards</surname>
<given-names>RH</given-names>
</name> <name>
<surname>Krywawych</surname>
<given-names>S</given-names>
</name> <name>
<surname>Davies</surname>
<given-names>CT</given-names>
</name> <name>
<surname>Halliday</surname>
<given-names>D</given-names>
</name> <name>
<surname>Waterlow</surname>
<given-names>JC</given-names>
</name> <etal/></person-group>. <article-title>Effect of exercise on protein turnover in man</article-title>. <source>Clin Sci</source>. (<year>1981</year>) <volume>61</volume>:<fpage>627</fpage>&#x2013;<lpage>39</lpage>. doi: <pub-id pub-id-type="doi">10.1042/cs0610627</pub-id></citation>
</ref>
<ref id="ref75">
<label>75.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Bodwell</surname>
<given-names>CE</given-names>
</name> <name>
<surname>Schuster</surname>
<given-names>EM</given-names>
</name> <name>
<surname>Kyle</surname>
<given-names>E</given-names>
</name> <name>
<surname>Brooks</surname>
<given-names>B</given-names>
</name> <name>
<surname>Womack</surname>
<given-names>M</given-names>
</name> <name>
<surname>Steele</surname>
<given-names>P</given-names>
</name> <etal/></person-group>. <article-title>Obligatory urinary and fecal nitrogen losses in young women, older men, and young men and the factorial estimation of adult human protein requirements</article-title>. <source>Am J Clin Nutr</source>. (<year>1979</year>) <volume>32</volume>:<fpage>2450</fpage>&#x2013;<lpage>9</lpage>. doi: <pub-id pub-id-type="doi">10.1093/ajcn/32.12.2450</pub-id>, PMID: <pub-id pub-id-type="pmid">506969</pub-id></citation>
</ref>
<ref id="ref76">
<label>76.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Scrimshaw</surname>
<given-names>NS</given-names>
</name> <name>
<surname>Hussein</surname>
<given-names>MA</given-names>
</name> <name>
<surname>Murray</surname>
<given-names>E</given-names>
</name> <name>
<surname>Rand</surname>
<given-names>WM</given-names>
</name> <name>
<surname>Young</surname>
<given-names>VR</given-names>
</name></person-group>. <article-title>Protein requirements of man: variations in obligatory urinary and fecal nitrogen losses in young men</article-title>. <source>J Nutr</source>. (<year>1972</year>) <volume>102</volume>:<fpage>1595</fpage>&#x2013;<lpage>604</lpage>. doi: <pub-id pub-id-type="doi">10.1093/jn/102.12.1595</pub-id>, PMID: <pub-id pub-id-type="pmid">4634993</pub-id></citation>
</ref>
<ref id="ref77">
<label>77.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Rand</surname>
<given-names>WM</given-names>
</name> <name>
<surname>Scrimshaw</surname>
<given-names>NS</given-names>
</name> <name>
<surname>Young</surname>
<given-names>VR</given-names>
</name></person-group>. <article-title>An analysis of temporal patterns in urinary nitrogen excretion of young adults receiving constant diets at two nitrogen intakes for 8 to 11 weeks</article-title>. <source>Am J Clin Nutr</source>. (<year>1979</year>) <volume>32</volume>:<fpage>1408</fpage>&#x2013;<lpage>14</lpage>. doi: <pub-id pub-id-type="doi">10.1093/ajcn/32.7.1408</pub-id>, PMID: <pub-id pub-id-type="pmid">453056</pub-id></citation>
</ref>
<ref id="ref78">
<label>78.</label>
<citation citation-type="journal"><person-group person-group-type="author"><name>
<surname>Huang</surname>
<given-names>PC</given-names>
</name> <name>
<surname>Chong</surname>
<given-names>HE</given-names>
</name> <name>
<surname>Rand</surname>
<given-names>WM</given-names>
</name></person-group>. <article-title>Obligatory urinary and fecal nitrogen losses in young Chinese men</article-title>. <source>J Nutr</source>. (<year>1972</year>) <volume>102</volume>:<fpage>1605</fpage>&#x2013;<lpage>13</lpage>. doi: <pub-id pub-id-type="doi">10.1093/jn/102.12.1605</pub-id>, PMID: <pub-id pub-id-type="pmid">4634994</pub-id></citation>
</ref>
<ref id="ref79">
<label>79.</label>
<citation citation-type="journal"><person-group person-group-type="author">
<name>
<surname>Millward</surname>
<given-names>DJ</given-names>
</name>
</person-group>. <article-title>The metabolic basis of amino acid requirements</article-title>. <source>Protein-Energy Interactions</source>. (<year>1991</year>)</citation>
</ref>
</ref-list>
</back>
</article>