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  <front>
    <journal-meta>
      <journal-id journal-id-type="publisher-id">ijcs</journal-id>
      <journal-title-group>
        <journal-title>International Journal of Cardiovascular Sciences</journal-title>
        <abbrev-journal-title abbrev-type="publisher">Int. J. Cardiovasc. Sci.</abbrev-journal-title>
      </journal-title-group>
      <issn pub-type="ppub">2359-4802</issn>
      <issn pub-type="epub">2359-5647</issn>
      <publisher>
        <publisher-name>Sociedade Brasileira de Cardiologia</publisher-name>
      </publisher>
    </journal-meta>
    <article-meta>
      <article-id pub-id-type="publisher-id" specific-use="scielo-v3">4VmfZhKfFkzJ7tsJb97MpYK</article-id>
      <article-id pub-id-type="publisher-id" specific-use="scielo-v2">S2359-56472018000300274</article-id>
      <article-id pub-id-type="doi">10.5935/2359-4802.20180023</article-id>
      <article-categories>
        <subj-group subj-group-type="heading">
          <subject>Original Articles</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title>Acute Effect of Manual Lymphatic Drainage on Natriuresis and Lipolysis in Young Women</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <name>
            <surname>Camargo</surname>
            <given-names>&#201;rica A. M</given-names>
          </name>
          <xref ref-type="aff" rid="aff1">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Borghi</surname>
            <given-names>Filipy</given-names>
          </name>
          <xref ref-type="aff" rid="aff1">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Souza</surname>
            <given-names>Agl&#233;cio Luiz de</given-names>
          </name>
          <xref ref-type="aff" rid="aff2">2</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Marcorin</surname>
            <given-names>Denise Maria</given-names>
          </name>
          <xref ref-type="aff" rid="aff3">3</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Rodrigues</surname>
            <given-names>Luana de Lima</given-names>
          </name>
          <xref ref-type="aff" rid="aff3">3</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Crege</surname>
            <given-names>Danilo Roberto Xavier de Oliveira</given-names>
          </name>
          <xref ref-type="aff" rid="aff4">4</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Ishizu</surname>
            <given-names>Larissa Yuri</given-names>
          </name>
          <xref ref-type="aff" rid="aff1">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Silva</surname>
            <given-names>Priscila Cristina da</given-names>
          </name>
          <xref ref-type="aff" rid="aff1">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Grassi-Kassisse</surname>
            <given-names>Dora Maria</given-names>
          </name>
          <xref ref-type="aff" rid="aff1">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Pires-de-Campos</surname>
            <given-names>Maria Silvia Mariani</given-names>
          </name>
          <xref ref-type="aff" rid="aff3">3</xref>
          <xref ref-type="corresp" rid="c1"/>
        </contrib>
      </contrib-group>
      <aff id="aff1">
        <label>1</label>
        <institution content-type="orgname">Universidade Estadual de Campinas</institution>
        <institution content-type="orgdiv1">Instituto de Biologia</institution>
        <addr-line>
          <city>S&#227;o Paulo</city>
          <state>SP</state>
        </addr-line>
        <country country="BR">Brazil</country>
        <institution content-type="original">Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), S&#227;o Paulo, SP - Brazil</institution>
      </aff>
      <aff id="aff2">
        <label>2</label>
        <institution content-type="orgname">Universidade Estadual de Campinas</institution>
        <institution content-type="orgdiv1">Faculdade de Ci&#234;ncias M&#233;dicas</institution>
        <addr-line>
          <city>S&#227;o Paulo</city>
          <state>SP</state>
        </addr-line>
        <country country="BR">Brazil</country>
        <institution content-type="original">Faculdade de Ci&#234;ncias M&#233;dicas, Universidade Estadual de Campinas (UNICAMP), S&#227;o Paulo, SP - Brazil</institution>
      </aff>
      <aff id="aff3">
        <label>3</label>
        <institution content-type="orgname">Universidade Metodista de Piracicaba</institution>
        <institution content-type="orgdiv1">Faculdade de Ci&#234;ncias da Sa&#250;de</institution>
        <addr-line>
          <city>S&#227;o Paulo</city>
          <state>SP</state>
        </addr-line>
        <country country="BR">Brazil</country>
        <institution content-type="original">Faculdade de Ci&#234;ncias da Sa&#250;de (FACIS), Universidade Metodista de Piracicaba (UNIMEP), S&#227;o Paulo, SP - Brazil</institution>
      </aff>
      <aff id="aff4">
        <label>4</label>
        <institution content-type="orgname">Centro Universit&#225;rio Padre Anchieta</institution>
        <addr-line>
          <city>Jundia&#237;</city>
          <state>SP</state>
        </addr-line>
        <country country="BR">Brazil</country>
        <institution content-type="original">Centro Universit&#225;rio Padre Anchieta, Jundia&#237;, SP - Brasil</institution>
      </aff>
      <author-notes>
        <corresp id="c1"><bold>Mailing Address: Maria Silvia Mariani Pires-de-Campos</bold>. Rua do Ros&#225;rio, 1260. Postal Code: 13400-000, Piracicaba, S&#227;o Paulo, SP - Brazil. E-mail: <email>mscampos@unimep.br</email>
				</corresp>
        <fn fn-type="con">
          <p>
            <bold>Author contributions</bold>
          </p>
          <p>Conception and design of the research: Grassi-Kassisse DM, Pires-de-Campos MSM. Acquisition of data: Camargo EAM, Souza AL, Marcorin DM, Rodrigues LL, Crege DRXO, Ishizu LY. Analysis and interpretation of the data: Camargo EAM, Borghi F, Grassi-Kassisse DM, Pires-de-Campos MSM. Statistical analysis: Camargo EAM, Borghi F, Grassi-Kassisse DM, Pires-de-Campos MSM. Obtaining financing: Grassi-Kassisse DM, Pires-de-Campos MSM. Writing of the manuscript: Camargo EAM, Borghi F, Silva PC, Grassi-Kassisse DM, Pires-de-Campos MSM. Critical revision of the manuscript for intellectual content: Camargo EAM, Borghi F, Silva PC, Grassi-Kassisse DM, Pires-de-Campos MSM. Supervision / as the major investigador: Camargo EAM, Grassi-Kassisse DM, Pires-de-Campos MSM.</p>
        </fn>
        <fn fn-type="conflict">
          <p>
            <bold>Potential Conflict of Interest</bold>
          </p>
          <p>No potential conflict of interest relevant to this article was reported.</p>
        </fn>
      </author-notes>
      <pub-date pub-type="epub-ppub">
        <season>May-Jun</season>
        <year>2018</year>
      </pub-date>
      <volume>31</volume>
      <issue>3</issue>
      <fpage>274</fpage>
      <lpage>281</lpage>
      <history>
        <date date-type="received">
          <day>18</day>
          <month>05</month>
          <year>2017</year>
        </date>
        <date date-type="rev-recd">
          <day>16</day>
          <month>10</month>
          <year>2017</year>
        </date>
        <date date-type="accepted">
          <day>13</day>
          <month>11</month>
          <year>2017</year>
        </date>
      </history>
      <permissions>
        <license license-type="open-access" xlink:href="https://creativecommons.org/licenses/by/4.0/" xml:lang="en">
          <license-p>This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p>
        </license>
      </permissions>
      <abstract>
        <title>Abstract</title>
        <sec>
          <title>Background:</title>
          <p>The importance of scientific validation of supporting techniques to various treatments is unquestionable. In this context, the influence of manual lymphatic drainage (MLD) on natriuresis and lipolysis and its interaction with oral contraceptives still need to be investigated.</p>
        </sec>
        <sec>
          <title>Objectives:</title>
          <p>To evaluate the acute effect of MLD on natriuresis and lipolysis in young women using or not oral contraceptives.</p>
        </sec>
        <sec>
          <title>Methods:</title>
          <p>Twenty-nine non-users of oral contraceptives and 29 oral contraceptive users, self-reported healthy, sedentary, normal weight women were enrolled. Analyses were conducted on two different days - control (C), without therapeutic intervention and MLD day. Four urine samples were collected at 60-minute intervals. MLD was performed in lower limbs and abdomen for 45 min following the Leduc method. Urinary flow rate and urinary sodium, glycerol and atrial natriuretic peptide excretion were analyzed. Data normality was tested by the Shapiro-Wilk test. Data without normal distribution were expressed as median and interquartile range (25%-75%), while normally distributed data were expressed as mean &#177; standard error. Mann-Whitney test was used for unpaired data and Wilcoxon test for paired data. Data with normal distribution were evaluated by the unpaired t-Student test. Statistical significance was set at 5%.</p>
        </sec>
        <sec>
          <title>Results:</title>
          <p>One MLD session had an acute effect on both groups, increasing natriuresis in non-users of oral contraceptives and glycerol and atrial natriuretic peptide excretion in oral contraceptive users.</p>
        </sec>
        <sec>
          <title>Conclusion:</title>
          <p>Oral contraceptives influence the effect of MLD on natriuresis.</p>
        </sec>
      </abstract>
      <kwd-group xml:lang="en">
        <title>Keywords:</title>
        <kwd>Musculoskeletal Manipulations</kwd>
        <kwd>Natriuresis</kwd>
        <kwd>Lipolysis</kwd>
        <kwd>Lymphatic System</kwd>
      </kwd-group>
      <funding-group>
        <award-group>
          <funding-source>FAPESP</funding-source>
          <award-id>2013/20510-2</award-id>
        </award-group>
      </funding-group>
    </article-meta>
  </front>
  <body>
    <sec sec-type="intro">
      <title>Introduction</title>
      <p>Manual lymphatic drainage (MLD) was created by the Danish physician Dr. Emil Vodder in 1936 as a supporting therapy, later established as the gold standard for the treatment of lymphedema.<xref ref-type="bibr" rid="B1">1</xref> However, based on beauty standards currently imposed by society, many women undergo some types of treatments that have no scientific basis for their well-being. In this context, MLD has become a popular procedure among healthy individuals to reduce body size, and performed by individuals without knowledge about lymphatic system physiology or pathophysiology.</p>
      <p>Lymphatic system has a crucial role in maintaining fluid balance in the body, macromolecular homeostasis, lipid absorption and immune function. However, the effect of MLD on other systems has been poorly investigated.<xref ref-type="bibr" rid="B1">1</xref>
			</p>
      <p>In women, water and electrolyte balance is influenced by ovarian hormones, whose receptors are located in reproductive tissues and in those involved in body fluid regulation, such as hypothalamus, cardiovascular system and renal tubules. These hormones affect capillary dynamics, by increasing renal sodium reabsorption and influencing plasma volume.<xref ref-type="bibr" rid="B2">2</xref><sup>,</sup><xref ref-type="bibr" rid="B3">3</xref> Stachenfeld &amp; Taylor<xref ref-type="bibr" rid="B4">4</xref> showed that estradiol increased plasma volume by its effect on capillary endothelial permeability, decreasing the outflow of proteins and water, without affecting extracellular fluid volume. Estrogen can affect capillary filtration and permeability, be it by direct action on capillary endothelium or indirectly by compounds like the atrial natriuretic peptide (ANP) and nitric oxide.<xref ref-type="bibr" rid="B4">4</xref>
			</p>
      <p>ANP is an important hormone stored in secretory granules of atrial cells, contributing to water-electrolyte balance. Its secretion depends on atrial distension caused by an increase in venous return, mechanical distension or hypernatremia, which lead to increased natriuresis combined with osmotic diuresis and vasodilation, relaxation of vascular smooth muscle of small arteries, arterioles and metarterioles, and hypotension. Besides, ANP inhibits renin, aldosterone and vasopressin release; it exerts a strong vasodilating action in the kidneys, contributing to increased blood flow and glomerular filtration rate, resulting in natriuresis and increased urine flow.<xref ref-type="bibr" rid="B5">5</xref> In adipocytes, ANP induces lipolysis via cyclic guanosine monophosphate (cGMP), resulting in activation of protein kinase G type I (GK-I), and degradation of triglycerides by perilipin A and hormone-sensitive lipase (HSL).<xref ref-type="bibr" rid="B6">6</xref>
			</p>
      <p>Nelson et al.<xref ref-type="bibr" rid="B7">7</xref> examined plasma concentration of glycerol and its urinary excretion in 12 young, healthy, trained men. The authors found that plasma glycerol peak concentration coincided with its peak in urinary excretion. Krupek et al.<xref ref-type="bibr" rid="B8">8</xref> evaluated urinary glycerol before and after twelve sessions of MLD in 3 young healthy women. Urinary glycerol was not increased in this study.</p>
      <p>Data on the effects of MLD on natriuresis and lipolysis are scarce and conflicting. Our aim was to assess the acute effect of MLD on natriuresis and lipolysis in young women using or not oral contraceptives.</p>
    </sec>
    <sec sec-type="materials|methods">
      <title>Material and methods</title>
      <p>According to the regulations of researches involving human subjects (resolution number 466/2012, Brazilian National Health Council), the study was approved by the Research Ethics Committee of the School of Medical Sciences/University of Campinas (CAAE: 24537613.2.0000.5404), Brazilian Registry of Clinical Trials -45c8br. All participants signed the informed consent form.</p>
      <p>Sample size was calculated based on a pilot study. GraphPad Statmate software, version 2 was used for calculation of 95% confidence interval and power of 80%. Thus, for statistical power of the sample be considered relevant, 30 volunteers per group would be necessary. This calculation was performed by Prof. Maria Imaculada de Lima Monteselo, specialist in the area. Twenty-nine women, non-users of oral contraceptives (nOCPu 21.5 &#177; 0.6 years BMI 21.3 &#177; 0.5 kg/m<sup>2</sup>) and 29 oral contraceptive users (OCPu 21.4 &#177; 0.5 years, BMI 21.8 &#177; 0.4 kg/m<sup>2</sup>), sedentary (international physical activity questionnaire, IPAQ-v8),<xref ref-type="bibr" rid="B9">9</xref> self-reported as healthy, and taking no medications except for OCP were enrolled. Women of the nOCPu group were included in luteal phase of menstrual cycle, whereas women of the OCPu group in the "rest period" from the OCP.</p>
      <p>All procedures were performed with volunteers at rest, in supine position, without fluid ingestion in a temperature-controlled room (22-24&#186;C) and relative humidity between 40 and 60% in the morning. For maintenance of resting metabolism, all volunteers received a standardized diet composed of one Brazilian nut, one nut, two apricots and six almonds without salt or sugar added.</p>
      <sec>
        <title>MLD procedure</title>
        <p>MLD was performed by the same physiotherapist as proposed by Leduc and Leduc10 Volunteers underwent a 45-minute session on abdomen and lower limbs.<xref ref-type="bibr" rid="B11">11</xref> Participants had previously received instructions for the procedure.</p>
        <p>First, in the abdomen, MLD of axillary lymph nodes was done by 10 strokes (clearing motions). Anterior abdominal wall was drained to two directions - the region above the umbilicus was drained toward the axilla, whereas the region below umbilicus was drained toward the inguinal lymph nodes - and thereby the inguinal lymph nodes were drained before the lower abdominal region. Following drainage of axillary and inguinal regions, 10 movements in each region were performed, beginning from the region closer to corresponding lymph nodes toward distal region. And finally, another movement in the opposite direction, i.e., from the distal toward proximal lymph nodes followed by 10 strokes on axillary lymph nodes were performed.</p>
        <p>In the lower limbs, drainage of inguinal lymph nodes was started by 10 strokes to evacuate the lymph. With the hands contacting the skin, a pressure was put to promote the flow of the lymph towards internal iliac lymph nodes, followed by 10 strokes in each region, from proximal to distal thigh to direct the flow of the lymph to the internal saphenous vein. Ten strokes were performed on the knees, aimed at cleaning popliteal lymph nodes, by dividing the area in two regions (upper and lower). Then, drainage of legs and thighs was performed, promoting lymph to drain toward the anterointernal part of the leg. In the ankles, 10 strokes in the retromalleolar region were made, directing the lymph to the leg, which was repeated on the feet. At the end of the procedure, a clearing motion was performed from the foot to the upper part of the leg, followed by 10 motions on popliteal lymph nodes and one motion toward upper thigh. The procedure was finished by 10 strokes on inguinal lymph nodes.</p>
      </sec>
      <sec>
        <title>Urine collection and assessment of urine composition</title>
        <p>Urine was collected at four-time points with 60-minute intervals - -60, 0, 60 and 120 minutes. Urine samples were collected using 1000mL beakers in a water closet available by the experiment room. Volunteers were instructed to completely empty the bladder for correct assessment of urine flow. The sample collected at -60min aimed at excluding potential influence of dietary or climate factors on the results. This protocol was followed on two days - one (control) day without therapeutic intervention and one day of MLD, performed between urine collection at 0 and 60 minutes<xref ref-type="bibr" rid="B11">11</xref> (<xref ref-type="fig" rid="f1">Figure 1</xref>).</p>
        <p>
          <fig id="f1">
            <label>Figure 1</label>
            <caption>
              <title>Urine collection schedule on day without therapeutic intervention (Control) and on day with manual lymphatic drainage (MLD).</title>
            </caption>
            <graphic xlink:href="2359-5647-ijcs-31-03-0274-80023-gf01.jpg"/></fig>
        </p>
        <p>Urinary excretion of the following compounds was calculated from urinary flow rate (mL/min): sodium, by titration with silver nitrate (mM/min), glycerol (mmol/min) by colorimetric assay (LABORCLIN, SP); and ANP (pg/min) by ELISA (USCNLife Science Inc., Houston, EUA).</p>
      </sec>
      <sec>
        <title>Statistical analysis</title>
        <p>Statistical analysis was performed using GraphPadPrism 5.0 software (Inc, La Jolla, CA, USA). Normal distribution was tested by the Shapiro-Wilk test. Data without normal distribution were expressed as median and interquartile range (25%-75%), whereas those with normal distribution as mean &#177; standard error. For data without normal distribution, the Mann-Whitney test and Wilcoxon test were used for unpaired (different "n") and paired (same "n") data, respectively, and normally distributed data were analyzed by the unpaired t-test. Statistical significance was set at 5%.</p>
      </sec>
    </sec>
    <sec sec-type="results">
      <title>Results</title>
      <p>In within-group analysis, on both days (control and MLD), initial values (0 min) in the nOCPu group showed increased urinary flow rate without changes in sodium or glycerol excretion. The same analysis showed that there were no statistical differences in OCPu between days. However, between-group comparison showed that, at the beginning of the study, urinary flow rate on both days and sodium excretion on control day were lower in OCPu than nOCPu (<xref ref-type="table" rid="t1">Table 1</xref>).</p>
      <table-wrap id="t1">
        <label>Table 1</label>
        <caption>
          <title>Urinary composition in young women, users (OCPu) or not (nOCPu) of oral contraceptives on manual lymphatic drainage (MLD) day or control (C) day at baseline</title>
        </caption>
        <table frame="hsides" rules="groups">
          <colgroup>
            <col width="11%"/>
            <col width="11%"/>
            <col width="11%"/>
            <col width="11%"/>
            <col width="11%"/>
            <col width="11%"/>
            <col width="11%"/>
            <col width="11%"/>
            <col width="11%"/>
          </colgroup>
          <thead>
            <tr>
              <th align="left" rowspan="2" style="background-color:#C5C6C8">Urinary flow rate</th>
              <th align="center" colspan="2" style="background-color:#C5C6C8">nOCPu</th>
              <th align="center" colspan="2" style="background-color:#C5C6C8">OCPu</th>
              <th align="center" colspan="2" style="background-color:#C5C6C8">Between-group analysis </th>
              <th align="center" colspan="2" style="background-color:#C5C6C8">Within-group analysis </th>
            </tr>
            <tr>
              <th align="center" style="background-color:#C5C6C8">C</th>
              <th align="center" style="background-color:#C5C6C8">MLD</th>
              <th align="center" style="background-color:#C5C6C8">C</th>
              <th align="center" style="background-color:#C5C6C8">MLD</th>
              <th align="center" style="background-color:#C5C6C8">P*<break/>C</th>
              <th align="center" style="background-color:#C5C6C8">P<sup>#</sup><break/>MLD</th>
              <th align="center" style="background-color:#C5C6C8">P<sup>$</sup><break/>nOCPu</th>
              <th align="center" style="background-color:#C5C6C8">P<sup>$</sup><break/>OCPu</th>
            </tr>
          </thead>
          <tbody>
            <tr>
              <td align="left">Urine<break/>(mL/min)</td>
              <td align="center">0.70(0.47-1.35)</td>
              <td align="center">1.20(0.70-1.93)$</td>
              <td align="center">0.51(0.33-0.75)*</td>
              <td align="center">0.70(0.40-1.00)<sup>#</sup></td>
              <td align="center">0.048</td>
              <td align="center">0.006</td>
              <td align="center">0.034</td>
              <td align="center">0.177</td>
            </tr>
            <tr>
              <td align="left">Sodium (mM/min)</td>
              <td align="center">140(110.50-184.20)</td>
              <td align="center">185.40&#177;100.00</td>
              <td align="center">113.30(78.05-149.50)*</td>
              <td align="center">161.10&#177;87.10</td>
              <td align="center">0.027</td>
              <td align="center">0.329</td>
              <td align="center">0.414</td>
              <td align="center">0.097</td>
            </tr>
            <tr>
              <td align="left">Glycerol (&#181;mol/min)</td>
              <td align="center">0.01(0.007-0.035)</td>
              <td align="center">0.014(0.007-0.02)</td>
              <td align="center">0.04(0.01-0.15)*</td>
              <td align="center">0.09(0.01-0.14)<sup>#</sup></td>
              <td align="center">0.014</td>
              <td align="center">0.004</td>
              <td align="center">0.595</td>
              <td align="center">0.945</td>
            </tr>
            <tr>
              <td align="left">ANP<break/>(pg/min)</td>
              <td align="center">0.79(0.33-2.71)</td>
              <td align="center">1.91(0.57-3.94)</td>
              <td align="center">2.45(1.21-2.86)</td>
              <td align="center">2.38(1.63-4.92)</td>
              <td align="center">0.064</td>
              <td align="center">0.146</td>
              <td align="center">0.214</td>
              <td align="center">0.324</td>
            </tr>
          </tbody>
        </table>
        <table-wrap-foot>
          <fn id="TFN1">
            <p>Normal distribution was tested by the Shapiro-Wilk test. Data without normal distribution were expressed as median and interquartile range (25%-75%), whereas those with normal distribution as mean &#177; standard error. For normally distributed data, the Mann-Whitney test was used for unpaired data (different "n") and the Wilcoxon test for paired data (same "n"). The unpaired t-test was used for data that failed to pass the normality test. Significance level was set at 5%. ANP: atrial natriuretic peptide.</p>
          </fn>
        </table-wrap-foot>
      </table-wrap>
      <p>Urinary flow remained unchanged in nOCPu on both days. However, on control day, OCPs showed an increase in this parameter over time. On MLD day, although urinary flow increased from 0 to 60min, no difference was observed between 120 min and 60 minutes (<xref ref-type="table" rid="t2">Table 2</xref>).</p>
      <table-wrap id="t2">
        <label>Table 2</label>
        <caption>
          <title>Urinary flow rate in young women, users (OCPu) or not (nOCPu) of oral contraceptives (OCPu) on manual lymphatic drainage (MLD) day or control (C) day at 0, 60 and 120 minutes</title>
        </caption>
        <table frame="hsides" rules="groups">
          <colgroup>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
          </colgroup>
          <thead>
            <tr>
              <th align="left" style="background-color:#C5C6C8">
							</th>
              <th align="center" style="background-color:#C5C6C8">
							</th>
              <th align="center" style="background-color:#C5C6C8">0</th>
              <th align="center" style="background-color:#C5C6C8">60</th>
              <th align="center" style="background-color:#C5C6C8">120</th>
              <th align="center" style="background-color:#C5C6C8">P**<break/>60 vs 0</th>
              <th align="center" style="background-color:#C5C6C8">P<sup>#</sup><break/>120 vs 0</th>
              <th align="center" style="background-color:#C5C6C8">P<sup>&amp;</sup><break/>120 vs 60</th>
            </tr>
          </thead>
          <tbody>
            <tr>
              <td align="left" rowspan="2">nOCPu</td>
              <td align="center">C</td>
              <td align="center">0.70(0.47-1.35)</td>
              <td align="center">1.00(0.70-1.31)</td>
              <td align="center">1.00(0.69-1.47)</td>
              <td align="center">0.06</td>
              <td align="center">0.32</td>
              <td align="center">0.38</td>
            </tr>
            <tr>
              <td align="center">MLD</td>
              <td align="center">0.70(0.49-1.90)</td>
              <td align="center">1.20(0.70-1.93)</td>
              <td align="center">1.20(0.59-1.40)</td>
              <td align="center">0.08</td>
              <td align="center">0.86</td>
              <td align="center">0.15</td>
            </tr>
            <tr>
              <td align="left" rowspan="2">OCPu</td>
              <td align="center">C</td>
              <td align="center">0.51(0.33-0.75)*</td>
              <td align="center">0.83(0.58-1.47)**</td>
              <td align="center">1.18(0.84-1.80)<sup>#&amp;</sup></td>
              <td align="center">0.0001</td>
              <td align="center">0.0001</td>
              <td align="center">0.01</td>
            </tr>
            <tr>
              <td align="center">MLD</td>
              <td align="center">0.70(0.40-1.00)</td>
              <td align="center">1.30(0.69-2.25)**</td>
              <td align="center">1.40(0.96-1.80)<sup>#</sup></td>
              <td align="center">0.0001</td>
              <td align="center">0.0009</td>
              <td align="center">0.56</td>
            </tr>
            <tr>
              <td align="left" rowspan="2">Between-group analysis</td>
              <td align="center">P* C</td>
              <td align="center">0.04</td>
              <td align="center">0.51</td>
              <td align="center">0.19</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
            </tr>
            <tr>
              <td align="center">P MLD</td>
              <td align="center">0.27</td>
              <td align="center">0.75</td>
              <td align="center">0.07</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
            </tr>
            <tr>
              <td align="left" rowspan="2">Within-group analysis</td>
              <td align="center">nOCPu</td>
              <td align="center">0.18</td>
              <td align="center">0.32</td>
              <td align="center">0.91</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
            </tr>
            <tr>
              <td align="center">OCPu</td>
              <td align="center">0.29</td>
              <td align="center">0.13</td>
              <td align="center">0.94</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
            </tr>
          </tbody>
        </table>
        <table-wrap-foot>
          <fn id="TFN2">
            <p>Normal distribution was tested by the Shapiro-Wilk test. Data without normal distribution were expressed as median and interquartile range (25%-75%), whereas those with normal distribution as mean &#177; standard error. For normally distributed data, the Mann-Whitney test was used for unpaired data (different "n") and the Wilcoxon test for paired data (same "n"). The unpaired t-test was used for data that failed to pass the normality test. Significance level was set at 5%.</p>
          </fn>
        </table-wrap-foot>
      </table-wrap>
      <p>Urinary excretion of sodium increased in nOCPu group on control day only at 120 minutes; however, on MLD day, this increase occurred at 60 min, indicating an acute effect of the therapy on sodium in the body. At 0 on control day, sodium urinary excretion was lower in OCPu as compared with the nOCPu group. An increase in this parameter was observed on both study days, suggesting that MLD had no effect on this variable (<xref ref-type="table" rid="t3">Table 3</xref>).</p>
      <table-wrap id="t3">
        <label>Table 3</label>
        <caption>
          <title>Urinary sodium excretion in young women, users (OCPu) or not (nOCPu) of oral contraceptives on manual lymphatic drainage (MLD) day or control (C) day at 0, 60 and 120 minutes</title>
        </caption>
        <table frame="hsides" rules="groups">
          <colgroup>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
          </colgroup>
          <thead>
            <tr>
              <th align="left" style="background-color:#C5C6C8">
							</th>
              <th align="center" style="background-color:#C5C6C8">
							</th>
              <th align="center" style="background-color:#C5C6C8">0</th>
              <th align="center" style="background-color:#C5C6C8">60</th>
              <th align="center" style="background-color:#C5C6C8">120</th>
              <th align="center" style="background-color:#C5C6C8">P**<break/>60 vs 0</th>
              <th align="center" style="background-color:#C5C6C8">P<sup>#</sup><break/>120 vs 0</th>
              <th align="center" style="background-color:#C5C6C8">P<sup>&amp;</sup><break/>120 vs 60</th>
            </tr>
          </thead>
          <tbody>
            <tr>
              <td align="left" rowspan="2">nOCPu</td>
              <td align="center">C</td>
              <td align="center">140(110.50-184.20)</td>
              <td align="center">202.90&#177;113.40</td>
              <td align="center">194.70(148.50-244.20)#</td>
              <td align="center">0.07</td>
              <td align="center">0.04</td>
              <td align="center">0.97</td>
            </tr>
            <tr>
              <td align="center">MLD</td>
              <td align="center">185.40&#177;100.00</td>
              <td align="center">200.00(109.70-329.40)**</td>
              <td align="center">203.00&#177;93.13</td>
              <td align="center">0.02</td>
              <td align="center">0.26</td>
              <td align="center">0.24</td>
            </tr>
            <tr>
              <td align="left" rowspan="2">OCPu</td>
              <td align="center">C</td>
              <td align="center">113.30(78.05-149.50)*</td>
              <td align="center">198.20&#177;87.13**</td>
              <td align="center">210.00(166.90-333.90)<sup>#&amp;</sup></td>
              <td align="center">0.0001</td>
              <td align="center">0.0001</td>
              <td align="center">0.001</td>
            </tr>
            <tr>
              <td align="center">MLD</td>
              <td align="center">161.10&#177;87.10</td>
              <td align="center">199.80(138.70-313.60)**</td>
              <td align="center">237.60(193.70-331.60)<sup>#</sup></td>
              <td align="center">0.0001</td>
              <td align="center">0.0001</td>
              <td align="center">0.08</td>
            </tr>
            <tr>
              <td align="left" rowspan="2">Between-group analysis</td>
              <td align="center">P* C</td>
              <td align="center">0.02</td>
              <td align="center">0.86</td>
              <td align="center">0.10</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
            </tr>
            <tr>
              <td align="center">P MLD</td>
              <td align="center">0.39</td>
              <td align="center">0.82</td>
              <td align="center">0.06</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
            </tr>
            <tr>
              <td align="left" rowspan="2">Within-group analysis</td>
              <td align="center">nOCPu</td>
              <td align="center">0.53</td>
              <td align="center">0.21</td>
              <td align="center">0.90</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
            </tr>
            <tr>
              <td align="center">OCPu</td>
              <td align="center">0.05</td>
              <td align="center">0.19</td>
              <td align="center">0.91</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
            </tr>
          </tbody>
        </table>
        <table-wrap-foot>
          <fn id="TFN3">
            <p>Normal distribution was tested by the Shapiro-Wilk test. Data without normal distribution were expressed as median and interquartile range (25%-75%), whereas those with normal distribution as mean &#177; standard error. For normally distributed data, the Mann-Whitney test was used for unpaired data (different "n") and the Wilcoxon test for paired data (same "n"). The unpaired t-test was used for data that failed to pass the normality test. Significance level was set at 5%.</p>
          </fn>
        </table-wrap-foot>
      </table-wrap>
      <p>Glycerol excretion did not change in nOCPu on both study days, and on control day, values of this variable were greater at 0 for OCPu as compared with nOCPu. Although glycerol excretion increased over time in the OCPu group on control day, it was greater in the OCPu group (<xref ref-type="table" rid="t4">Table 4</xref>).</p>
      <table-wrap id="t4">
        <label>Table 4</label>
        <caption>
          <title>Urinary glycerol excretion in young women, users (OCPu) or not (nOCPu) of oral contraceptives on manual lymphatic drainage (MLD) day or control (C) day at 0, 60 and 120 minutes</title>
        </caption>
        <table frame="hsides" rules="groups">
          <colgroup>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
            <col width="12%"/>
          </colgroup>
          <thead>
            <tr>
              <th align="left" style="background-color:#C5C6C8">
							</th>
              <th align="center" style="background-color:#C5C6C8">
							</th>
              <th align="center" style="background-color:#C5C6C8">0</th>
              <th align="center" style="background-color:#C5C6C8">60</th>
              <th align="center" style="background-color:#C5C6C8">120</th>
              <th align="center" style="background-color:#C5C6C8">P**<break/>60 vs 0</th>
              <th align="center" style="background-color:#C5C6C8">P<sup>#</sup><break/>120 vs 0</th>
              <th align="center" style="background-color:#C5C6C8">P<sup>&amp;</sup><break/>120 vs 0</th>
            </tr>
          </thead>
          <tbody>
            <tr>
              <td align="left" rowspan="2">nOCPu</td>
              <td align="center">C</td>
              <td align="center">0.02(0.01-0.03)</td>
              <td align="center">0.01(0.01-0.06)</td>
              <td align="center">0.02(0.01-0.07)</td>
              <td align="center">0.91</td>
              <td align="center">0.62</td>
              <td align="center">0.52</td>
            </tr>
            <tr>
              <td align="center">MLD</td>
              <td align="center">0.01(0.01-0.02)</td>
              <td align="center">0.03(0.01-0.08)</td>
              <td align="center">0.02(0.01-0.07)</td>
              <td align="center">0.10</td>
              <td align="center">0.19</td>
              <td align="center">0.80</td>
            </tr>
            <tr>
              <td align="left" rowspan="2">OCPu</td>
              <td align="center">C</td>
              <td align="center">0.05(0.02-0.15)*</td>
              <td align="center">0.10(0.04-0.31)*</td>
              <td align="center">0.17(0.06-0.56)*<sup>#</sup></td>
              <td align="center">0.05</td>
              <td align="center">0.01</td>
              <td align="center">0.23</td>
            </tr>
            <tr>
              <td align="center">MLD</td>
              <td align="center">0.09(0.01-0.15)%</td>
              <td align="center">0.12(0.03-0.32)%</td>
              <td align="center">0.17(0.07-0.44)<sup>%#</sup></td>
              <td align="center">0.10</td>
              <td align="center">0.01</td>
              <td align="center">0.38</td>
            </tr>
            <tr>
              <td align="left" rowspan="2">Between-group analysis</td>
              <td align="center">P* C</td>
              <td align="center">0.01</td>
              <td align="center">0.0001</td>
              <td align="center">0.0001</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
            </tr>
            <tr>
              <td align="center">P MLD</td>
              <td align="center">0.0049</td>
              <td align="center">0.0012</td>
              <td align="center">0.0001</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
            </tr>
            <tr>
              <td align="left" rowspan="2">Within-group analysis</td>
              <td align="center">nOCPu</td>
              <td align="center">0.59</td>
              <td align="center">0.14</td>
              <td align="center">0.72</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
            </tr>
            <tr>
              <td align="center">OCPu</td>
              <td align="center">0.94</td>
              <td align="center">0.86</td>
              <td align="center">0.91</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
              <td align="center">
							</td>
            </tr>
          </tbody>
        </table>
        <table-wrap-foot>
          <fn id="TFN4">
            <p>Normal distribution was tested by the Shapiro-Wilk test. Data without normal distribution were expressed as median and interquartile range (25%-75%), whereas those with normal distribution as mean &#177; standard error. For normally distributed data, the Mann-Whitney test was used for unpaired data (different "n") and the Wilcoxon test for paired data (same "n"). The unpaired t-test was used for data that failed to pass the normality test. Significance level was set at 5%.</p>
          </fn>
        </table-wrap-foot>
      </table-wrap>
      <p>ANP values in nOCPu at 60 min were higher than in OCPu. MLD increased ANP excretion in OCPu only (<xref ref-type="table" rid="t5">Table 5</xref>).</p>
      <table-wrap id="t5">
        <label>Table 5</label>
        <caption>
          <title>Urinary atrial natriuretic peptide excretion in young women, users (OCPu) or not (nOCPu) of oral contraceptives on manual lymphatic drainage (MLD) day or control (C) day at 0, 60 and 120 minutes</title>
        </caption>
        <table frame="hsides" rules="groups">
          <colgroup>
            <col width="20%"/>
            <col width="20%"/>
            <col width="20%"/>
            <col width="20%"/>
            <col width="20%"/>
          </colgroup>
          <thead>
            <tr>
              <th align="left" style="background-color:#C5C6C8">
							</th>
              <th align="center" style="background-color:#C5C6C8">
							</th>
              <th align="center" style="background-color:#C5C6C8">0</th>
              <th align="center" style="background-color:#C5C6C8">60</th>
              <th align="center" style="background-color:#C5C6C8">P**<break/>60 vs 0</th>
            </tr>
          </thead>
          <tbody>
            <tr>
              <td align="left" rowspan="2">nOCPu</td>
              <td align="center">C</td>
              <td align="center">0.79(0.33-2.71)</td>
              <td align="center">1.10(0.54-3.71)</td>
              <td align="center">0.45</td>
            </tr>
            <tr>
              <td align="center">MLD</td>
              <td align="center">1.91(0.57-3.94)</td>
              <td align="center">1.72(0.88-4.79)</td>
              <td align="center">0.87</td>
            </tr>
            <tr>
              <td align="left" rowspan="2">OCPu</td>
              <td align="center">C</td>
              <td align="center">2.45(1.21-2.86)</td>
              <td align="center">4.05(1.90-6.63)*</td>
              <td align="center">0.05</td>
            </tr>
            <tr>
              <td align="center">MLD</td>
              <td align="center">2.38(1.63-4.92)</td>
              <td align="center">7.67&#177;5.48%**</td>
              <td align="center">0.03</td>
            </tr>
            <tr>
              <td align="left" rowspan="2">Between-group analysis</td>
              <td align="center">P* C</td>
              <td align="center">0.06</td>
              <td align="center">0.0047</td>
              <td align="center">
							</td>
            </tr>
            <tr>
              <td align="center">P% MLD</td>
              <td align="center">0.14</td>
              <td align="center">0.0054</td>
              <td align="center">
							</td>
            </tr>
            <tr>
              <td align="left" rowspan="2">Within-group analysis</td>
              <td align="center">nOCPu</td>
              <td align="center">0.21</td>
              <td align="center">0.24</td>
              <td align="center">
							</td>
            </tr>
            <tr>
              <td align="center">OCPu</td>
              <td align="center">0.32</td>
              <td align="center">0.13</td>
              <td align="center">
							</td>
            </tr>
          </tbody>
        </table>
        <table-wrap-foot>
          <fn id="TFN5">
            <p>Normal distribution was tested by the Shapiro-Wilk test. Data without normal distribution were expressed as median and interquartile range (25%-75%), whereas those with normal distribution as mean &#177; standard error. For normally distributed data, the Mann-Whitney test was used for unpaired data (different "n") and the Wilcoxon test for paired data (same "n"). The unpaired t-test was used for data that failed to pass the normality test. Significance level was set at 5%.</p>
          </fn>
        </table-wrap-foot>
      </table-wrap>
    </sec>
    <sec sec-type="discussion">
      <title>Discussion</title>
      <p>One strength of the protocol proposed in this study was a "control" day, to demonstrate that changes occurring after the MLD were actually caused by the procedure.</p>
      <p>When control day of both groups was compared, no difference in urinary excretion was found. Graugaard-Jensen et al.<xref ref-type="bibr" rid="B12">12</xref> investigated 8 healthy, young women with regular menstrual cycle in low and high estrogen phases. The author showed that hormone levels had no effect on urinary excretion. In addition, although urinary excretion did not change between the two time-points, the authors observed a tendency towards sodium retention when estrogen was high.</p>
      <p>Based on our protocol, nOCPu responded to MLD with increased sodium excretion and nOCPu with increased urinary excretion when compared with nOCPu, suggesting an acute effect of this technique on natriureis, and hence on urine composition.</p>
      <p>Camargo et al.<xref ref-type="bibr" rid="B11">11</xref> showed that one MLD session promoted an increase in urinary excretion with unaltered sodium excretion or urinary osmolarity in OCP non-users. On the other hand, OCP users were not sensitive to the acute effect of the therapy. These findings suggest an underlying hormonal regulation of these mechanisms. Increased estrogen and progesterone levels were found in OCP users, even though the exact mechanism of these hormones on water-electrolyte balance is still unclear.<xref ref-type="bibr" rid="B2">2</xref><sup>,</sup><xref ref-type="bibr" rid="B4">4</xref><sup>,</sup><xref ref-type="bibr" rid="B13">13</xref>
			</p>
      <p>Stachenfeld &amp; Taylor<xref ref-type="bibr" rid="B4">4</xref> investigated the effects of gonadotropin-releasing hormone (GnRH) administration, combined or not with 17 beta-estradiol (transdermal patches, 0.1 mg/day) in young, healthy, non-smoking women. The authors found that 17 beta-estradiol caused a reduction in urinary osmolarity by concentration of vasopressin in the plasma at baseline. Stachenfeld &amp; Keefe<xref ref-type="bibr" rid="B3">3</xref> found that estrogen and progesterone in young, healthy women caused little change in water and sodium regulation, suggesting that these hormones affect allostasis. These investigators reported a reduction in osmotic threshold and increase in plasma vasopressin, without changes in urinary free water, indicating that OCP inhibits renal sensitivity to vasopressin, requiring higher hormone production to obtain the same effects. This is corroborated by our results in OCPu on both days, suggesting that OCP induces greater sensitivity to central osmoreceptors. We believe that resting and temperature-controlled conditions in our study prevented water loss from the body surface, resulting in dilution in the extracellular compartment and inhibitory stimulus of vasopressin, which, in turn, promoted an increase in urinary excretion in the OCPu group.</p>
      <p>Increased venous return is one of the stimulus for ANP release by atrial cells. ANP exerts its biological function by binding into receptors located in renal tubules and in the glomerular zone of the adrenal glands, inhibiting sodium reabsorption by decreased renin secretion, resulting in lower production of angiotensin II, aldosterone and vasopressin.</p>
      <p>Graugaard-Jensen et al.<xref ref-type="bibr" rid="B12">12</xref> reported that changes in endogenous estrogen have no effects on plasma ANP levels. In contrast, in our study, women of the OCPu had higher estrogen levels and greater urinary ANP excretion.</p>
      <p>According to Schlueter et al.,<xref ref-type="bibr" rid="B6">6</xref> ANP lipolytic effect occurs via GMPc formation, which induces triglyceride degradation. However, MLD did not affect urinary excretion of ANP or glycerol, suggesting that venous return promoted by this procedure was not sufficiently effective to stimulate ANP and exert a lipolytic effect readily observable in the urine.</p>
      <p>Our data corroborate those found by Krupek et al.<xref ref-type="bibr" rid="B8">8</xref> These authors reported that 12 sessions of MLD had no effect on urinary glycerol excretion in 3 healthy, young women. However, different from our study, the authors investigated the chronic effect rather than the acute effect of MLD.</p>
      <p>One limitation of our study was the fact that urine collection was carried out in the mornings, which limited the participation of many eligible volunteers. </p>
    </sec>
    <sec sec-type="conclusions">
      <title>Conclusion</title>
      <p>One session of MLD promoted an acute effect on natriuresis in women not taking OCP and glycerol and ANP secretion in OCP users by increasing these parameters.</p>
    </sec>
  </body>
  <back>
    <fn-group>
      <fn fn-type="financial-disclosure">
        <p>
          <bold>Sources of Funding</bold>
        </p>
        <p>This study was funded by CAPES e Fapesp.</p>
      </fn>
      <fn fn-type="other">
        <p>
          <bold>Study Association</bold>
        </p>
        <p>This article is part of the thesis of master submitted by &#201;rica A. M. Camargo, from Universidade de Campinas (UNICAMP).</p>
      </fn>
      <fn fn-type="other">
        <p>
          <bold>Ethics approval and consent to participate</bold>
        </p>
        <p>This study was approved by the Ethics Committee of the Faculdade de Ci&#234;ncias M&#233;dicas da Unicamp under the protocol number CAAE: 24537613.2.0000.5404. All the procedures in this study were in accordance with the 1975 Helsinki Declaration, updated in 2013. Informed consent was obtained from all participants included in the study.</p>
      </fn>
    </fn-group>
    <ack>
      <title>Acknowledgements</title>
      <p>We thank all volunteers who participated in this study; the University of Campinas (UNICAMP) and Methodist University of Piracicaba (UNIMEP) for the facilities; FAPESP (2013/20510-2) and CAPES for the financial support.</p>
    </ack>
    <ref-list>
      <title>References</title>
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  <sub-article article-type="translation" id="s1" xml:lang="pt">
    <front-stub>
      <article-categories>
        <subj-group subj-group-type="heading">
          <subject>Artigos Originais</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title>Efeito Agudo da Drenagem Linf&#225;tica Manual sobre a Natriurese e Lip&#243;lise de Mulheres Jovens</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <name>
            <surname>Camargo</surname>
            <given-names>&#201;rica A. M</given-names>
          </name>
          <xref ref-type="aff" rid="aff5">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Borghi</surname>
            <given-names>Filipy</given-names>
          </name>
          <xref ref-type="aff" rid="aff5">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Souza</surname>
            <given-names>Agl&#233;cio Luiz de</given-names>
          </name>
          <xref ref-type="aff" rid="aff6">2</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Marcorin</surname>
            <given-names>Denise Maria</given-names>
          </name>
          <xref ref-type="aff" rid="aff7">3</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Rodrigues</surname>
            <given-names>Luana de Lima</given-names>
          </name>
          <xref ref-type="aff" rid="aff7">3</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Crege</surname>
            <given-names>Danilo Roberto Xavier de Oliveira</given-names>
          </name>
          <xref ref-type="aff" rid="aff8">4</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Ishizu</surname>
            <given-names>Larissa Yuri</given-names>
          </name>
          <xref ref-type="aff" rid="aff5">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Silva</surname>
            <given-names>Priscila Cristina da</given-names>
          </name>
          <xref ref-type="aff" rid="aff5">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Grassi-Kassisse</surname>
            <given-names>Dora Maria</given-names>
          </name>
          <xref ref-type="aff" rid="aff5">1</xref>
        </contrib>
        <contrib contrib-type="author">
          <name>
            <surname>Pires-de-Campos</surname>
            <given-names>Maria Silvia Mariani</given-names>
          </name>
          <xref ref-type="aff" rid="aff7">3</xref>
          <xref ref-type="corresp" rid="c2"/>
        </contrib>
      </contrib-group>
      <aff id="aff5">
        <label>1</label>
        <institution content-type="original">Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), S&#227;o Paulo, SP - Brasil</institution>
      </aff>
      <aff id="aff6">
        <label>2</label>
        <institution content-type="original">Faculdade de Ci&#234;ncias M&#233;dicas, Universidade Estadual de Campinas (UNICAMP), S&#227;o Paulo, SP - Brasil</institution>
      </aff>
      <aff id="aff7">
        <label>3</label>
        <institution content-type="original">Faculdade de Ci&#234;ncias da Sa&#250;de (FACIS), Universidade Metodista de Piracicaba (UNIMEP), S&#227;o Paulo, SP - Brasil</institution>
      </aff>
      <aff id="aff8">
        <label>4</label>
        <institution content-type="original">Centro Universit&#225;rio Padre Anchieta, Jundia&#237;, SP- Brasil</institution>
      </aff>
      <author-notes>
        <corresp id="c2"><bold>Correspond&#234;ncia: Maria Silvia Mariani Pires-de-Campos.</bold> Rua do Ros&#225;rio, 1260. CEP: 13400-000, Piracicaba, S&#227;o Paulo, SP - Brasil. E-mail: <email>mscampos@unimep.br</email>
				</corresp>
        <fn fn-type="con">
          <p>
            <bold>Contribui&#231;&#227;o dos autores</bold>
          </p>
          <p>Concep&#231;&#227;o e desenho da pesquisa: Grassi-Kassisse DM, Pires-de-Campos MSM. Obten&#231;&#227;o de dados: Camargo EAM, Souza AL, Marcorin DM, Rodrigues LL, Crege DRXO, Ishizu LY. An&#225;lise e interpreta&#231;&#227;o dos dados: Camargo EAM, Borghi F, Grassi-Kassisse DM, Pires-de-Campos MSM. An&#225;lise estat&#237;stica: Camargo EAM, Borghi F, Grassi-Kassisse DM, Pires-de-Campos MSM. Obten&#231;&#227;o de financiamento: Grassi-Kassisse DM, Pires-de-Campos MSM. Reda&#231;&#227;o do manuscrito: Camargo EAM, Borghi F, Silva PC, Grassi-Kassisse DM, Pires-de-Campos MSM. Revis&#227;o cr&#237;tica do manuscrito quanto ao conte&#250;do intelectual importante: Camargo EAM, Borghi F, Silva PC, Grassi-Kassisse DM, Pires-de-Campos MSM. Supervis&#227;o / como investigador principal: Camargo EAM, Grassi-Kassisse DM, Pires-de-Campos MSM.</p>
        </fn>
        <fn fn-type="conflict">
          <p>
            <bold>Potencial Conflito de Interesse</bold>
          </p>
          <p>Declaro n&#227;o haver conflito de interesses pertinentes.</p>
        </fn>
      </author-notes>
      <abstract>
        <title>Resumo</title>
        <sec>
          <title>Fundamentos:</title>
          <p>A import&#226;ncia da valida&#231;&#227;o cient&#237;fica de t&#233;cnicas coadjuvantes a diversos tratamentos de sa&#250;de &#233; inquestion&#225;vel. Desta forma, a influ&#234;ncia da drenagem linf&#225;tica manual (DLM) na natriurese e na lip&#243;lise sob intera&#231;&#227;o de anticoncepcional oral precisa ser investigada.</p>
        </sec>
        <sec>
          <title>Objetivos:</title>
          <p>Avaliar o efeito agudo da DLM sobre a natriurese e lip&#243;lise de mulheres jovens usu&#225;rias ou n&#227;o de anticoncepcional oral.</p>
        </sec>
        <sec>
          <title>M&#233;todo:</title>
          <p>Participaram 29 mulheres n&#227;o usu&#225;rias de anticoncepcional oral e 29 usu&#225;rias, autodeclaradas saud&#225;veis, sedent&#225;rias e eutr&#243;ficas. As an&#225;lises foram realizadas em dois dias distintos, Controle (C), sem interven&#231;&#227;o terap&#234;utica e DLM. Nos dias C e DLM coletamos 4 amostras de urina com intervalos de 60 min. Seguimos o m&#233;todo de Leduc, abdomem e membros inferiores, durante 45 min. Analisamos o fluxo urin&#225;rio e excre&#231;&#227;o urin&#225;ria de s&#243;dio, glicerol e peptideo natriur&#233;tico atrial. A normalidade dos dados foi analisada pelo teste Shapiro-Wilk. Os dados que n&#227;o obedeceram &#224; normalidade foram apresentados em mediana e intervalo interquartil (25%-75%), enquanto os que obedeceram foram apresentados em m&#233;dia &#177; erro padr&#227;o. O teste de Mann-Whitney foi usado para dados n&#227;o pareados e Wilcoxon para dados pareados. Dados com normalidade foram avaliados pelo teste t-Student n&#227;o pareado. O n&#237;vel de signific&#226;ncia estat&#237;stica adotado foi de 5%.</p>
        </sec>
        <sec>
          <title>Resultados:</title>
          <p>O efeito agudo de uma sess&#227;o de DLM promove aumento da natriurese em mulheres n&#227;o usu&#225;rias de anticoncepcional oral, por outro lado, em usu&#225;rias induz aumento na libera&#231;&#227;o de glicerol e de ANP.</p>
        </sec>
        <sec>
          <title>Conclus&#227;o:</title>
          <p>O uso de anticoncepcional oral interfere no efeito da DLM sobre a natriurese.</p>
        </sec>
      </abstract>
      <kwd-group xml:lang="pt">
        <title>Palavras-chave:</title>
        <kwd>Manipula&#231;&#245;es Musculoesquel&#233;ticas</kwd>
        <kwd>Natriurese</kwd>
        <kwd>Lip&#243;lise</kwd>
        <kwd>Sistema Linf&#225;tico</kwd>
      </kwd-group>
    </front-stub>
    <body>
      <sec sec-type="intro">
        <title>Introdu&#231;&#227;o</title>
        <p>A drenagem linf&#225;tica manual (DLM) foi criada pelo m&#233;dico dinamarqu&#234;s Emil Vodder em 1936 como uma t&#233;cnica coadjuvante, estabelecida como padr&#227;o ouro no tratamento de linfedema.<xref ref-type="bibr" rid="B1">1</xref> Entretanto, atualmente a sociedade dita um padr&#227;o de beleza no qual muitas mulheres submetem-se a tratamentos sem fundamenta&#231;&#227;o cient&#237;fica para se sentirem bem. A partir desta realidade, a DLM tornou-se uma t&#233;cnica popular, realizada por pessoas sem conhecimento sobre a fisiologia e a fisiopatologia do sistema linf&#225;tico e sua rela&#231;&#227;o com demais sistemas no nosso organismo, e amplamente aplicada em indiv&#237;duos saud&#225;veis para redu&#231;&#227;o de medidas.</p>
        <p>Sabe-se que o sistema linf&#225;tico tem papel crucial no equil&#237;brio do flu&#237;do corporal e macromolecular, absor&#231;&#227;o lip&#237;dica e na fun&#231;&#227;o imune. Por&#233;m, o mecanismo fisiol&#243;gico da DLM sobre demais sistemas permanece pouco investigado.<xref ref-type="bibr" rid="B1">1</xref>
				</p>
        <p>O balan&#231;o hidroeletrol&#237;tico nas mulheres &#233; influenciado pelos horm&#244;nios ovarianos. Seus receptores est&#227;o presentes nos tecidos reprodutivos e tamb&#233;m naqueles envolvidos na regula&#231;&#227;o do fluido corporal, como o hipot&#225;lamo, o sistema cardiovascular e os t&#250;bulos renais. Tais horm&#244;nios interferem na din&#226;mica capilar, aumentando a reabsor&#231;&#227;o renal de s&#243;dio e alterando o volume plasm&#225;tico.<xref ref-type="bibr" rid="B2">2</xref><sup>,</sup><xref ref-type="bibr" rid="B3">3</xref> Stachenfeld e Taylor, <xref ref-type="bibr" rid="B4">4</xref> demonstraram que o estradiol aumenta o volume plasm&#225;tico pela a&#231;&#227;o na permeabilidade do endot&#233;lio capilar, diminuindo o escape de prote&#237;nas e &#225;gua, sem modificar o volume do fluido extracelular. Os estrog&#234;nios podem influenciar na filtra&#231;&#227;o e permeabilidade capilar, seja por a&#231;&#227;o direta no endot&#233;lio capilar ou por estimula&#231;&#227;o de subst&#226;ncias como o pept&#237;deo natriur&#233;tico atrial (ANP) e &#243;xido n&#237;trico.<xref ref-type="bibr" rid="B4">4</xref>
				</p>
        <p>O ANP &#233; um importante horm&#244;nio estocado nos gr&#226;nulos secret&#243;rios das c&#233;lulas atriais card&#237;acas, contribuindo para o equil&#237;brio hidroeletrol&#237;tico. Sua libera&#231;&#227;o ocorre pela distens&#227;o atrial decorrente do aumento do retorno venoso, distens&#227;o mec&#226;nica ou hipernatremia, provocando natriurese intensa acompanhada de diurese osm&#243;tica e vasodilata&#231;&#227;o, relaxando a musculatura lisa vascular das pequenas art&#233;rias, arter&#237;olas e meta-arter&#237;olas, causando hipotens&#227;o. Al&#233;m disso, o ANP inibe a libera&#231;&#227;o de renina, aldosterona e vasopressina. Sua a&#231;&#227;o vasodilatadora &#233; potente sobre os rins, contribuindo para o aumento do fluxo sangu&#237;neo e na taxa de filtra&#231;&#227;o glomerular, resultando em natriurese e aumento do fluxo urin&#225;rio.<xref ref-type="bibr" rid="B5">5</xref> No adip&#243;cito, o ANP induz a lip&#243;lise via guanosina monofosfato c&#237;clico (GMPc), que leva &#224; ativa&#231;&#227;o de prote&#237;na quinase dependente de GMPc tipo I (GK-I), e degrada&#231;&#227;o de triglicer&#237;deos por perilipina A e lipase horm&#244;nio-sens&#237;vel (LHS).<xref ref-type="bibr" rid="B6">6</xref>
				</p>
        <p>Nelson et al., <xref ref-type="bibr" rid="B7">7</xref> investigaram em 12 homens jovens, saud&#225;veis e treinados, a concentra&#231;&#227;o plasm&#225;tica de glicerol e sua excre&#231;&#227;o urin&#225;ria. Observaram que o pico da concentra&#231;&#227;o plasm&#225;tica de glicerol coincidia com o seu pico na excre&#231;&#227;o urin&#225;ria. Krupek et al.,<xref ref-type="bibr" rid="B8">8</xref> avaliaram o glicerol urin&#225;rio antes e ap&#243;s doze sess&#245;es de DLM em 3 mulheres jovens e saud&#225;veis. Esse estudo n&#227;o apontou aumento de glicerol na urina.</p>
        <p>Al&#233;m de escassos, os resultados na literatura s&#227;o conflitantes quanto aos efeitos desta t&#233;cnica sobre a natriurese e a lip&#243;lise. Diante do exposto, nosso objetivo foi avaliar o efeito agudo de uma sess&#227;o de DLM sobre a natriurese e lip&#243;lise de mulheres jovens usu&#225;rias ou n&#227;o de anticoncepcional oral.</p>
      </sec>
      <sec sec-type="materials|methods">
        <title>Material e m&#233;todos</title>
        <p>Respeitando as normas de conduta em pesquisa experimental com seres humanos (Resolu&#231;&#227;o 466/2012 do Conselho Nacional da Sa&#250;de), este estudo foi aprovado pelo Comit&#234; de &#201;tica em Pesquisa FCM/UNICAMP (CAAE: 24537613.2.0000.5404), Registro Brasileiro de Ensaios Cl&#237;nicos RBR-45c8br. Todos os participantes assinaram o Termo de Consentimento Livre e Esclarecido.</p>
        <p>Foi realizado o c&#225;lculo amostral para esta popula&#231;&#227;o tomando por base estudo piloto. Utilizou-se o programa GraphPad Statmate vers&#227;o 2 para n&#237;vel de confian&#231;a 95% e poder amostral 80%. Com isso, 30 volunt&#225;rios por grupo s&#227;o necess&#225;rios para que o poder amostral estat&#237;stico pudesse ser considerado relevante. Este estudo estat&#237;stico foi realizado pela Prof&#170; Maria Imaculada de Lima Monteselo, especialista na &#225;rea. Participaram 29 mulheres n&#227;o usu&#225;rias de anticoncepcional oral (MnACO 21,5 &#177; 0,6 anos, IMC 21,3 &#177; 0,5 kg/m<sup>2</sup>) e 29 usu&#225;rias (MACO 21,4 &#177; 0,5 anos, IMC 21,8 &#177; 0,4 kg/m<sup>2</sup>), sedent&#225;rias (Question&#225;rio Internacional de Atividade F&#237;sica, IPAQ-v8),<xref ref-type="bibr" rid="B9">9</xref> autodeclaradas saud&#225;veis e sem uso de medica&#231;&#245;es exceto anticoncepcional oral (ACO). O grupo MnACO participou na fase l&#250;tea do ciclo menstrual, enquanto que MACO na fase de repouso do ACO.</p>
        <p>Todos os procedimentos foram realizados em sala climatizada entre 22 e 24&#176;C e umidade relativa do ar entre 40 e 60%, sempre no per&#237;odo da manh&#227;. Durante o experimento, os volunt&#225;rios permaneceram em repouso em dec&#250;bito dorsal sem ingest&#227;o de l&#237;quido. Para manuten&#231;&#227;o do metabolismo de repouso, foi oferecido entre os intervalos das coletas, uma por&#231;&#227;o padr&#227;o contendo uma castanha do Brasil, uma noz, dois damascos e seis am&#234;ndoas sem adi&#231;&#227;o de sal ou a&#231;&#250;car.</p>
        <sec>
          <title>Procedimento da DLM</title>
          <p>O procedimento seguiu o proposto por Leduc e Leduc,<xref ref-type="bibr" rid="B10">10</xref> e realizado sempre pela mesma fisioterapeuta. As volunt&#225;rias receberam uma sess&#227;o de 45 min, aplicada no abdome e nos membros inferiores.<xref ref-type="bibr" rid="B11">11</xref> As participantes receberam orienta&#231;&#245;es pr&#233;vias para o procedimento.</p>
          <p>Inicialmente, no abdome, foi realizada a drenagem dos linfonodos axilares com 10 manobras de evacua&#231;&#227;o. A parede anterior do abdome foi drenada em duas dire&#231;&#245;es: a regi&#227;o supra umbilical foi drenada em dire&#231;&#227;o &#224; fossa axilar e a regi&#227;o infra umbilical em dire&#231;&#227;o aos linfonodos inguinais. Desta forma, antes da realiza&#231;&#227;o da DLM da regi&#227;o inferior do abdome &#233; feita drenagem dos linfonodos inguinais. Ap&#243;s a drenagem das regi&#245;es axilares e inguinais, foram realizadas 10 manobras de evacua&#231;&#227;o por regi&#227;o, sempre iniciando pela regi&#227;o mais pr&#243;xima aos linfonodos correspondentes, indo em dire&#231;&#227;o &#224; regi&#227;o distal e finalizando, novamente, com uma manobra de evacua&#231;&#227;o da regi&#227;o distal &#224; proximal aos linfonodos correspondentes de cada regi&#227;o e 10 manobras de evacua&#231;&#227;o sobre os linfonodos axilares.</p>
          <p>Nos membros inferiores, foi iniciada a drenagem dos linfonodos inguinais pela manobra de evacua&#231;&#227;o. Com as m&#227;os em contato com a pele, pressiona-se a linfa em dire&#231;&#227;o &#224;s cadeias il&#237;acas profundas por 10 vezes. Em seguida, s&#227;o realizadas 10 manobras de evacua&#231;&#227;o por regi&#227;o, iniciando na regi&#227;o proximal da coxa at&#233; a distal, orientando a linfa no trajeto da veia safena interna. No joelho, foram realizadas 10 manobras de evacua&#231;&#227;o dos linfonodos popl&#237;teos, dividindo a regi&#227;o em superior e inferior. Ap&#243;s esta etapa, &#233; realizada a evacua&#231;&#227;o na perna e na coxa, orientando a linfa em dire&#231;&#227;o &#224; face &#226;ntero-interna da perna. No tornozelo foram realizadas 10 manobras de evacua&#231;&#227;o das vias retro-maleolares, direcionando a linfa para a perna, realizando a mesma manobra nos p&#233;s. Na finaliza&#231;&#227;o do procedimento, foi feita uma manobra de evacua&#231;&#227;o do p&#233; at&#233; a raiz da perna, 10 manobras sobre os linfonodos popl&#237;teos seguidos por uma manobra de evacua&#231;&#227;o at&#233; a raiz da coxa. O procedimento foi finalizado com 10 manobras de evacua&#231;&#227;o dos linfonodos inguinais.</p>
        </sec>
        <sec>
          <title>Coleta e avalia&#231;&#227;o da composi&#231;&#227;o urin&#225;ria</title>
          <p>Foram realizadas 4 coletas de urina com intervalos de 60 min, denominadas -60, 0, 60 e 120 min. As coletas foram realizadas em b&#233;queres de 1000 mL em sanit&#225;rio reservado ao lado da sala de experimenta&#231;&#227;o. Foi orientado &#224;s volunt&#225;rias que esvaziassem totalmente a bexiga com o objetivo de que o fluxo unir&#225;rio fosse corretamente avaliado. A coleta -60 min teve como objetivo eliminar poss&#237;veis efeitos alimentares e condi&#231;&#245;es clim&#225;ticas que possam interferir no resultado. Este protocolo foi seguido em dois dias de experimento, sendo um dia controle sem interven&#231;&#227;o terap&#234;utica e outro com drenagem linf&#225;tica manual, relizada entre as coletas de urina 0 e 60 min<xref ref-type="bibr" rid="B11">11</xref> (<xref ref-type="fig" rid="f2">Figura 1</xref>).</p>
          <p>
            <fig id="f2">
              <label>Figura 1</label>
              <caption>
                <title>Esquema de coleta de urina no dia sem interven&#231;&#227;o terap&#234;utica (Controle) e no dia com drenagem linf&#225;tica manual (DLM).</title>
              </caption>
              <graphic xlink:href="2359-5647-ijcs-31-03-0274-80023-gf01-pt.jpg"/></fig>
          </p>
          <p>A partir do fluxo urin&#225;rio (mL/min), foram calculados os fluxos de: s&#243;dio por meio da titula&#231;&#227;o de argentimetria (mM/min), glicerol (mmol/min) por ensaio colorim&#233;trico (LABORCLIN, SP), e ANP (pg/min) por ELISA (USCNLife Science Inc., Houston, EUA).</p>
        </sec>
        <sec>
          <title>An&#225;lise estat&#237;stica</title>
          <p>Foi utilizado o programa GraphPadPrism 5.0 (Inc, La Jolla, CA, USA). A distribui&#231;&#227;o normal foi analisada pelo teste de Shapiro-Wilk. Os dados que n&#227;o obedeceram &#224; normalidade foram apresentados em mediana e intervalo interquartil (25%-75%), enquanto os que obedeceram foram apresentados em m&#233;dia &#177; erro padr&#227;o. Utilizamos o teste Mann-Whitney para dados n&#227;o pareados ("n" diferente) e o teste de Wilcoxon para dados pareados (mesmo n) quando obedeceram &#224; normalidade e t-Student n&#227;o pareado para os dados que obedeceram &#224; normalidade. O n&#237;vel de signific&#226;ncia estat&#237;stica adotado foi de 5%.</p>
        </sec>
      </sec>
      <sec sec-type="results">
        <title>Resultados</title>
        <p>Na an&#225;lise intragrupo dos valores iniciais (0 min) do grupo MnACO, em ambos os dias (C e DLM), houve aumento no fluxo de urina, sem altera&#231;&#227;o nos fluxos de s&#243;dio e de glicerol. A mesma an&#225;lise demonstrou que o grupo MACO n&#227;o apresentou diferen&#231;as estat&#237;sticas nestes indicadores entre os dias C e DLM. Entretanto, na compara&#231;&#227;o entre os grupos, observamos que MACO iniciou os experimentos com menor fluxo de urina em ambos os dias e de s&#243;dio apenas no dia controle quando comparado ao grupo MnACO (<xref ref-type="table" rid="t6">Tabela 1</xref>).</p>
        <table-wrap id="t6">
          <label>Tabela 1</label>
          <caption>
            <title>Valores da composi&#231;&#227;o urin&#225;ria de mulheres jovens usu&#225;rias (MACO) ou n&#227;o de anticoncepcionais orais (MnACO) no dia de drenagem linf&#225;tica manual (DLM) ou no dia controle (C) no tempo zero</title>
          </caption>
          <table frame="hsides" rules="groups">
            <colgroup>
              <col width="11%"/>
              <col width="11%"/>
              <col width="11%"/>
              <col width="11%"/>
              <col width="11%"/>
              <col width="11%"/>
              <col width="11%"/>
              <col width="11%"/>
              <col width="11%"/>
            </colgroup>
            <thead>
              <tr>
                <th align="left" rowspan="2" style="background-color:#C5C6C8">Fluxo</th>
                <th align="center" colspan="2" style="background-color:#C5C6C8">MnACO</th>
                <th align="center" colspan="2" style="background-color:#C5C6C8">MACO</th>
                <th align="center" colspan="2" style="background-color:#C5C6C8">An&#225;lise intergrupo</th>
                <th align="center" colspan="2" style="background-color:#C5C6C8">An&#225;lise<break/>intragrupo</th>
              </tr>
              <tr>
                <th align="center" style="background-color:#C5C6C8">C</th>
                <th align="center" style="background-color:#C5C6C8">DLM</th>
                <th align="center" style="background-color:#C5C6C8">C</th>
                <th align="center" style="background-color:#C5C6C8">DLM</th>
                <th align="center" style="background-color:#C5C6C8">P*<break/>C</th>
                <th align="center" style="background-color:#C5C6C8">P<sup>#</sup><break/>DLM</th>
                <th align="center" style="background-color:#C5C6C8">P<sup>$</sup><break/>MnACO</th>
                <th align="center" style="background-color:#C5C6C8">P<sup>$</sup><break/>MACO</th>
              </tr>
            </thead>
            <tbody>
              <tr>
                <td align="left">Urina<break/>(mL/min)</td>
                <td align="center">0,70(0,47-1,35)</td>
                <td align="center">1,20(0,70-1,93)<sup>$</sup></td>
                <td align="center">0,51(0,33-0,75)*</td>
                <td align="center">0,70(0,40-1,00)<sup>#</sup></td>
                <td align="center">0,048</td>
                <td align="center">0,006</td>
                <td align="center">0,034</td>
                <td align="center">0,177</td>
              </tr>
              <tr>
                <td align="left">S&#243;dio<break/>(mM/min)</td>
                <td align="center">140(110,50-184,20)</td>
                <td align="center">185,40&#177;100,00</td>
                <td align="center">113,30(78,05-149,50)*</td>
                <td align="center">161,10&#177;87,10</td>
                <td align="center">0,027</td>
                <td align="center">0,329</td>
                <td align="center">0,414</td>
                <td align="center">0,097</td>
              </tr>
              <tr>
                <td align="left">Glicerol (&#181;mol/min)</td>
                <td align="center">0,01(0,007-0,035)</td>
                <td align="center">0,014(0,007-0,02)</td>
                <td align="center">0,04(0,01-0,15)*</td>
                <td align="center">0,09(0,01-0,14)<sup>#</sup></td>
                <td align="center">0,014</td>
                <td align="center">0,004</td>
                <td align="center">0,595</td>
                <td align="center">0,945</td>
              </tr>
              <tr>
                <td align="left">ANP<break/>(pg/min)</td>
                <td align="center">0,79(0,33-2,71)</td>
                <td align="center">1,91(0,57-3,94)</td>
                <td align="center">2,45(1,21-2,86)</td>
                <td align="center">2,38(1,63-4,92)</td>
                <td align="center">0,064</td>
                <td align="center">0,146</td>
                <td align="center">0,214</td>
                <td align="center">0,324</td>
              </tr>
            </tbody>
          </table>
          <table-wrap-foot>
            <fn id="TFN6">
              <p>A distribui&#231;&#227;o normal foi analisada pelo teste de Shapiro-Wilk. Os dados que n&#227;o obedeceram &#224; normalidade foram apresentados em mediana e intervalo interquartil (25%-75%), enquanto os que obedeceram foram apresentados em m&#233;dia &#177; erro padr&#227;o. Utilizamos os testes Mann-Whitney para dados n&#227;o pareados ("n" diferente) e Wilcoxon para dados pareados (mesmo "n") quando obedeceram &#224; normalidade e <italic>t-Student</italic> n&#227;o pareado para os dados que obedeceram &#224; normalidade. O n&#237;vel de signific&#226;ncia estat&#237;stica adotado foi de 5%. ANP: pept&#237;deo natriur&#233;tico atrial.</p>
            </fn>
          </table-wrap-foot>
        </table-wrap>
        <p>O fluxo urin&#225;rio n&#227;o foi alterado em MnACO em nenhum dos dias de estudos. Por&#233;m, no dia controle, MACO apresentou aumento ao longo do tempo. No dia DLM, houve tamb&#233;m aumento de fluxo urin&#225;rio ao longo do tempo, entretanto o aumento m&#225;ximo foi atingido na coleta 60 min, isto &#233;, o fluxo observado em 120 min n&#227;o foi diferente de 60 min (<xref ref-type="table" rid="t7">Tabela 2</xref>).</p>
        <table-wrap id="t7">
          <label>Tabela 2</label>
          <caption>
            <title>Valores de fluxo urin&#225;rio dos grupos de mulheres jovens usu&#225;rias (MACO) ou n&#227;o de anticoncepcionais orais (MnACO) no dia de drenagem linf&#225;tica manual (DLM) ou no dia controle (C) nos tempos 0, 60 e 120 minutos</title>
          </caption>
          <table frame="hsides" rules="groups">
            <colgroup>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
            </colgroup>
            <thead>
              <tr>
                <th align="left" style="background-color:#C5C6C8">
								</th>
                <th align="center" style="background-color:#C5C6C8">
								</th>
                <th align="center" style="background-color:#C5C6C8">0</th>
                <th align="center" style="background-color:#C5C6C8">60</th>
                <th align="center" style="background-color:#C5C6C8">120</th>
                <th align="center" style="background-color:#C5C6C8">P**<break/>60 vs 0</th>
                <th align="center" style="background-color:#C5C6C8">P<sup>#</sup><break/>120 vs 0</th>
                <th align="center" style="background-color:#C5C6C8">P<sup>&amp;</sup><break/>120 vs 60</th>
              </tr>
            </thead>
            <tbody>
              <tr>
                <td align="left" rowspan="2">MnACO</td>
                <td align="center">C</td>
                <td align="center">0,70(0,47-1,35)</td>
                <td align="center">1,00(0,70-1,31)</td>
                <td align="center">1,00(0,69-1,47)</td>
                <td align="center">0,06</td>
                <td align="center">0,32</td>
                <td align="center">0,38</td>
              </tr>
              <tr>
                <td align="center">DLM</td>
                <td align="center">0,70(0,49-1,90)</td>
                <td align="center">1,20(0,70-1,93)</td>
                <td align="center">1,20(0,59-1,40)</td>
                <td align="center">0,08</td>
                <td align="center">0,86</td>
                <td align="center">0,15</td>
              </tr>
              <tr>
                <td align="left" rowspan="2">MACO</td>
                <td align="center">C</td>
                <td align="center">0,51(0,33-0,75)*</td>
                <td align="center">0,83(0,58-1,47)**</td>
                <td align="center">1,18(0,84-1,80)<sup>#&amp;</sup></td>
                <td align="center">0,0001</td>
                <td align="center">0,0001</td>
                <td align="center">0,01</td>
              </tr>
              <tr>
                <td align="center">DLM</td>
                <td align="center">0,70(0,40-1,00)</td>
                <td align="center">1,30(0,69-2,25)**</td>
                <td align="center">1,40(0,96-1,80)<sup>#</sup></td>
                <td align="center">0,0001</td>
                <td align="center">0,0009</td>
                <td align="center">0,56</td>
              </tr>
              <tr>
                <td align="left" rowspan="2">An&#225;lise intergrupo</td>
                <td align="center">P* C</td>
                <td align="center">0,04</td>
                <td align="center">0,51</td>
                <td align="center">0,19</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
              </tr>
              <tr>
                <td align="center">P DLM</td>
                <td align="center">0,27</td>
                <td align="center">0,75</td>
                <td align="center">0,07</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
              </tr>
              <tr>
                <td align="left" rowspan="2">An&#225;lise intragrupo</td>
                <td align="center">MnACO</td>
                <td align="center">0,18</td>
                <td align="center">0,32</td>
                <td align="center">0,91</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
              </tr>
              <tr>
                <td align="center">MACO</td>
                <td align="center">0,29</td>
                <td align="center">0,13</td>
                <td align="center">0,94</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
              </tr>
            </tbody>
          </table>
          <table-wrap-foot>
            <fn id="TFN7">
              <p>A distribui&#231;&#227;o normal foi analisada pelo teste de Shapiro-Wilk. Os dados que n&#227;o obedeceram &#224; normalidade foram apresentados em mediana e intervalo interquartil (25%-75%), enquanto os que obedeceram foram apresentados em m&#233;dia &#177; erro padr&#227;o. Utilizamos os testes Mann-Whitney para dados n&#227;o pareados ("n" diferente) e Wilcoxon para dados pareados (mesmo "n") quando obedeceram &#224; normalidade e <italic>t-Student</italic> n&#227;o pareado para os dados que obedeceram &#224; normalidade. O n&#237;vel de signific&#226;ncia estat&#237;stica adotado foi de 5%.</p>
            </fn>
          </table-wrap-foot>
        </table-wrap>
        <p>O fluxo urin&#225;rio de s&#243;dio no grupo MnACO no dia controle aumentou somente no tempo 120, por&#233;m no dia DLM, este aumento foi antecipado para a coleta 60 min, demonstrando o efeito agudo desta terapia sobre o s&#243;dio no organismo. Os valores de fluxo de s&#243;dio foram menores na coleta 0 do dia C para a popula&#231;&#227;o MACO quando comparado com o mesmo dia da popula&#231;&#227;o MnACO. Esta popula&#231;&#227;o apresentou aumento ao longo do tempo nos dois dias de experimento, sugerindo que para esta popula&#231;&#227;o a DLM n&#227;o interferiu nesta vari&#225;vel (<xref ref-type="table" rid="t8">Tabela 3</xref>).</p>
        <table-wrap id="t8">
          <label>Tabela 3</label>
          <caption>
            <title>Valores de excre&#231;&#227;o de s&#243;dio dos grupos de mulheres jovens usu&#225;rias (MACO) ou n&#227;o de anticoncepcionais orais (MnACO) no dia de drenagem linf&#225;tica manual (DLM) ou no dia controle (C) nos tempos 0, 60 e 120 minutos</title>
          </caption>
          <table frame="hsides" rules="groups">
            <colgroup>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
            </colgroup>
            <thead>
              <tr>
                <th align="left" style="background-color:#C5C6C8">
								</th>
                <th align="center" style="background-color:#C5C6C8">
								</th>
                <th align="center" style="background-color:#C5C6C8">0</th>
                <th align="center" style="background-color:#C5C6C8">60</th>
                <th align="center" style="background-color:#C5C6C8">120</th>
                <th align="center" style="background-color:#C5C6C8">P**<break/>60 vs 0 </th>
                <th align="center" style="background-color:#C5C6C8">P<sup>#</sup><break/>120 vs 0</th>
                <th align="center" style="background-color:#C5C6C8">P<sup>&amp;</sup><break/>120 vs 60</th>
              </tr>
            </thead>
            <tbody>
              <tr>
                <td align="left" rowspan="2">MnACO</td>
                <td align="center">C</td>
                <td align="center">140(110,50-184,20)</td>
                <td align="center">202,90&#177;113,40</td>
                <td align="center">194,70(148,50-244,20)#</td>
                <td align="center">0,07</td>
                <td align="center">0,04</td>
                <td align="center">0,97</td>
              </tr>
              <tr>
                <td align="center">DLM</td>
                <td align="center">185,40&#177;100,00</td>
                <td align="center">200,00(109,70-329,40)**</td>
                <td align="center">203,00&#177;93,13</td>
                <td align="center">0,02</td>
                <td align="center">0,26</td>
                <td align="center">0,24</td>
              </tr>
              <tr>
                <td align="left" rowspan="2">MACO</td>
                <td align="center">C</td>
                <td align="center">113,30(78,05-149,50)*</td>
                <td align="center">198,20&#177;87,13**</td>
                <td align="center">210,00(166,90-333,90)<sup>#&amp;</sup></td>
                <td align="center">0,0001</td>
                <td align="center">0,0001</td>
                <td align="center">0,001</td>
              </tr>
              <tr>
                <td align="center">DLM</td>
                <td align="center">161,10&#177;87,10</td>
                <td align="center">199,80(138,70-313,60)**</td>
                <td align="center">237,60(193,70-331,60)<sup>#</sup></td>
                <td align="center">0,0001</td>
                <td align="center">0,0001</td>
                <td align="center">0,08</td>
              </tr>
              <tr>
                <td align="left" rowspan="2">An&#225;lise intergrupo</td>
                <td align="center">P* C</td>
                <td align="center">0,02</td>
                <td align="center">0,86</td>
                <td align="center">0,10</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
              </tr>
              <tr>
                <td align="center">P DLM</td>
                <td align="center">0,39</td>
                <td align="center">0,82</td>
                <td align="center">0,06</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
              </tr>
              <tr>
                <td align="left" rowspan="2">An&#225;lise intragrupo</td>
                <td align="center">MnACO</td>
                <td align="center">0,53</td>
                <td align="center">0,21</td>
                <td align="center">0,90</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
              </tr>
              <tr>
                <td align="center">MACO</td>
                <td align="center">0,05</td>
                <td align="center">0,19</td>
                <td align="center">0,91</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
              </tr>
            </tbody>
          </table>
          <table-wrap-foot>
            <fn id="TFN8">
              <p>A distribui&#231;&#227;o normal foi analisada pelo teste de Shapiro-Wilk. Os dados que n&#227;o obedeceram &#224; normalidade foram apresentados em mediana e intervalo interquartil (25%-75%), enquanto os que obedeceram foram apresentados em m&#233;dia &#177; erro padr&#227;o. Utilizamos os testes Mann-Whitney para dados n&#227;o pareados ("n" diferente) e Wilcoxon para dados pareados (mesmo "n") quando obedeceram &#224; normalidade e <italic>t-Student</italic> n&#227;o pareado para os dados que obedeceram &#224; normalidade. O n&#237;vel de signific&#226;ncia estat&#237;stica adotado foi de 5%.</p>
            </fn>
          </table-wrap-foot>
        </table-wrap>
        <p>O fluxo de glicerol n&#227;o se alterou no grupo MnACO nos dias de experimento. Estes valores foram maiores na coleta 0 do dia C para a popula&#231;&#227;o MACO quando comparado com o mesmo dia da popula&#231;&#227;o MnACO. No grupo MACO, houve aumento no fluxo de glicerol tanto no dia C quanto no dia DLM ao longo do tempo. Contudo, foi observado maior fluxo de glicerol no grupo MACO (<xref ref-type="table" rid="t9">Tabela 4</xref>).</p>
        <table-wrap id="t9">
          <label>Tabela 4</label>
          <caption>
            <title>Valores de fluxo glicerol dos grupos de mulheres jovens usu&#225;rias (MACO) ou n&#227;o de anticoncepcionais orais (MnACO) no dia de drenagem linf&#225;tica manual (DLM) ou no dia controle (C) nos tempos 0, 60 e 120 minutos</title>
          </caption>
          <table frame="hsides" rules="groups">
            <colgroup>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
              <col width="12%"/>
            </colgroup>
            <thead>
              <tr>
                <th align="left" style="background-color:#C5C6C8">
								</th>
                <th align="center" style="background-color:#C5C6C8">
								</th>
                <th align="center" style="background-color:#C5C6C8">0</th>
                <th align="center" style="background-color:#C5C6C8">60</th>
                <th align="center" style="background-color:#C5C6C8">120</th>
                <th align="center" style="background-color:#C5C6C8">P**<break/>60 vs 0</th>
                <th align="center" style="background-color:#C5C6C8">P<sup>#</sup><break/>120 vs 0</th>
                <th align="center" style="background-color:#C5C6C8">P<sup>&amp;</sup><break/>120 vs 0</th>
              </tr>
            </thead>
            <tbody>
              <tr>
                <td align="left" rowspan="2">MnACO</td>
                <td align="center">C</td>
                <td align="center">0,02(0,01-0,03)</td>
                <td align="center">0,01(0,01-0,06)</td>
                <td align="center">0,02(0,01-0,07)</td>
                <td align="center">0,91</td>
                <td align="center">0,62</td>
                <td align="center">0,52</td>
              </tr>
              <tr>
                <td align="center">DLM</td>
                <td align="center">0,01(0,01-0,02)</td>
                <td align="center">0,03(0,01-0,08)</td>
                <td align="center">0,02(0,01-0,07)</td>
                <td align="center">0,10</td>
                <td align="center">0,19</td>
                <td align="center">0,80</td>
              </tr>
              <tr>
                <td align="left" rowspan="2">MACO</td>
                <td align="center">C</td>
                <td align="center">0,05(0,02-0,15)*</td>
                <td align="center">0,10(0,04-0,31)*</td>
                <td align="center">0,17(0,06-0,56)*<sup>#</sup></td>
                <td align="center">0,05</td>
                <td align="center">0,01</td>
                <td align="center">0,23</td>
              </tr>
              <tr>
                <td align="center">DLM</td>
                <td align="center">0,09(0,01-0,15)%</td>
                <td align="center">0,12(0,03-0,32)%</td>
                <td align="center">0,17(0,07-0,44)<sup>%#</sup></td>
                <td align="center">0,10</td>
                <td align="center">0,01</td>
                <td align="center">0,38</td>
              </tr>
              <tr>
                <td align="left" rowspan="2">An&#225;lise intergrupo</td>
                <td align="center">P* C</td>
                <td align="center">0,01</td>
                <td align="center">0,0001</td>
                <td align="center">0,0001</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
              </tr>
              <tr>
                <td align="center">P<sup>%</sup> DLM</td>
                <td align="center">0,0049</td>
                <td align="center">0,0012</td>
                <td align="center">0,0001</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
              </tr>
              <tr>
                <td align="left" rowspan="2">An&#225;lise intragrupo</td>
                <td align="center">MnACO</td>
                <td align="center">0,59</td>
                <td align="center">0,14</td>
                <td align="center">0,72</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
              </tr>
              <tr>
                <td align="center">MACO</td>
                <td align="center">0,94</td>
                <td align="center">0,86</td>
                <td align="center">0,91</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
                <td align="center">
								</td>
              </tr>
            </tbody>
          </table>
          <table-wrap-foot>
            <fn id="TFN9">
              <p>A distribui&#231;&#227;o normal foi analisada pelo teste de Shapiro-Wilk. Os dados que n&#227;o obedeceram &#224; normalidade foram apresentados em mediana e intervalo interquartil (25%-75%), enquanto os que obedeceram foram apresentados em m&#233;dia &#177; erro padr&#227;o. Utilizamos os testes Mann-Whitney para dados n&#227;o pareados ("n" diferente) e Wilcoxon para dados pareados (mesmo "n") quando obedeceram &#224; normalidade e <italic>t-Student</italic> n&#227;o pareado para os dados que obedeceram &#224; normalidade. O n&#237;vel de signific&#226;ncia estat&#237;stica adotado foi de 5%.</p>
            </fn>
          </table-wrap-foot>
        </table-wrap>
        <p>Os valores de ANP no grupo MnACO na coleta 60 min foram maiores que nas mesmas coletas da popula&#231;&#227;o MnACO. A DLM aumentou a libera&#231;&#227;o de ANP somente na popula&#231;&#227;o MACO (<xref ref-type="table" rid="t10">Tabela 5</xref>).</p>
        <table-wrap id="t10">
          <label>Tabela 5</label>
          <caption>
            <title>Valores de pept&#237;deo natriur&#233;tico atrial dos grupos de mulheres jovens usu&#225;rias (MACO) ou n&#227;o de anticoncepcionais orais (MnACO) no dia de drenagem linf&#225;tica manual (DLM) ou no dia controle (C) nos tempos 0, 60 e 120 minutos</title>
          </caption>
          <table frame="hsides" rules="groups">
            <colgroup>
              <col width="20%"/>
              <col width="20%"/>
              <col width="20%"/>
              <col width="20%"/>
              <col width="20%"/>
            </colgroup>
            <thead>
              <tr>
                <th align="left" style="background-color:#C5C6C8">
								</th>
                <th align="center" style="background-color:#C5C6C8">
								</th>
                <th align="center" style="background-color:#C5C6C8">0</th>
                <th align="center" style="background-color:#C5C6C8">60</th>
                <th align="center" style="background-color:#C5C6C8">P**<break/>60 vs 0</th>
              </tr>
            </thead>
            <tbody>
              <tr>
                <td align="left" rowspan="2">MnACO</td>
                <td align="center">C</td>
                <td align="center">0,79(0,33-2,71)</td>
                <td align="center">1,10(0,54-3,71)</td>
                <td align="center">0,45</td>
              </tr>
              <tr>
                <td align="center">DLM</td>
                <td align="center">1,91(0,57-3,94)</td>
                <td align="center">1,72(0,88-4,79)</td>
                <td align="center">0,87</td>
              </tr>
              <tr>
                <td align="left" rowspan="2">MACO</td>
                <td align="center">C</td>
                <td align="center">2,45(1,21-2,86)</td>
                <td align="center">4,05(1,90-6,63)*</td>
                <td align="center">0,05</td>
              </tr>
              <tr>
                <td align="center">DLM</td>
                <td align="center">2,38(1,63-4,92)</td>
                <td align="center">7,67&#177;5,48%**</td>
                <td align="center">0,03</td>
              </tr>
              <tr>
                <td align="left" rowspan="2">An&#225;lise intergrupo</td>
                <td align="center">P* C</td>
                <td align="center">0,06</td>
                <td align="center">0,0047</td>
                <td align="center">
								</td>
              </tr>
              <tr>
                <td align="center">P<sup>%</sup> DLM</td>
                <td align="center">0,14</td>
                <td align="center">0,0054</td>
                <td align="center">
								</td>
              </tr>
              <tr>
                <td align="left" rowspan="2">An&#225;lise intragrupo</td>
                <td align="center">MnACO</td>
                <td align="center">0,21</td>
                <td align="center">0,24</td>
                <td align="center">
								</td>
              </tr>
              <tr>
                <td align="center">MACO</td>
                <td align="center">0,32</td>
                <td align="center">0,13</td>
                <td align="center">
								</td>
              </tr>
            </tbody>
          </table>
          <table-wrap-foot>
            <fn id="TFN10">
              <p>A distribui&#231;&#227;o normal foi analisada pelo teste de Shapiro-Wilk. Os dados que n&#227;o obedeceram &#224; normalidade foram apresentados em mediana e intervalo interquartil (25%-75%), enquanto os que obedeceram foram apresentados em m&#233;dia &#177; erro padr&#227;o. Utilizamos os testes Mann-Whitney para dados n&#227;o pareados ("n" diferente) e Wilcoxon para dados pareados (mesmo "n") quando obedeceram &#224; normalidade e <italic>t-Student</italic> n&#227;o pareado para os dados que obedeceram &#224; normalidade. O n&#237;vel de signific&#226;ncia estat&#237;stica adotado foi de 5%.</p>
            </fn>
          </table-wrap-foot>
        </table-wrap>
      </sec>
      <sec sec-type="discussion">
        <title>Discuss&#227;o</title>
        <p>O protocolo proposto nesta investiga&#231;&#227;o teve como diferencial um dia de coleta controle justamente para comprovar que as altera&#231;&#245;es observadas com a DLM s&#227;o de fato promovidas pela t&#233;cnica.</p>
        <p>Ao analisarmos o dia controle de ambos os grupos, n&#227;o houve diferen&#231;as no fluxo urin&#225;rio. Graugaard-Jensen et al., <xref ref-type="bibr" rid="B12">12</xref> estudaram 8 volunt&#225;rias jovens e saud&#225;veis, em momentos de menor e maior n&#237;vel de estr&#243;geno num ciclo menstrual regular, e notou que o n&#237;vel hormonal n&#227;o interferiu no fluxo urin&#225;rio. Ainda no mesmo estudo, esses autores notaram que mesmo n&#227;o havendo altera&#231;&#227;o na excre&#231;&#227;o de s&#243;dio, houve tend&#234;ncia &#224; reten&#231;&#227;o em rela&#231;&#227;o ao maior n&#237;vel de estr&#243;geno.</p>
        <p>Diante do nosso protocolo, o grupo MnACO respondeu &#224; DLM com aumento na elimina&#231;&#227;o de s&#243;dio e as MACO com maior fluxo urin&#225;rio quando comparadas &#224;s MnACO. Isso sugere que o efeito agudo da t&#233;cnica interfere na composi&#231;&#227;o da urina, promovendo natriurese.</p>
        <p>Camargo et al., <xref ref-type="bibr" rid="B11">11</xref> demonstraram que uma sess&#227;o de DLM promoveu aumento do fluxo urin&#225;rio com manuten&#231;&#227;o da concentra&#231;&#227;o de s&#243;dio e da osmolaridade urin&#225;ria em mulheres n&#227;o usu&#225;rias de ACO. Por outro lado, mulheres usu&#225;rias de ACO n&#227;o foram sens&#237;veis ao efeito agudo da terapia. Estes resultados indicam que os mecanismos est&#227;o sob regula&#231;&#227;o hormonal. As concentra&#231;&#245;es de estr&#243;geno e progesterona encontraram-se elevadas em usu&#225;rias de ACO, no entanto o mecanismo exato desses horm&#244;nios sobre o equil&#237;brio hidroeletrol&#237;tico ainda n&#227;o est&#225; bem esclarecido.<xref ref-type="bibr" rid="B2">2</xref><sup>,</sup><xref ref-type="bibr" rid="B4">4</xref><sup>,</sup><xref ref-type="bibr" rid="B13">13</xref>
				</p>
        <p>Stachenfeld e Taylor,<xref ref-type="bibr" rid="B4">4</xref> investigaram a administra&#231;&#227;o de horm&#244;nio liberador de gonadotrofina (GnRH) para supress&#227;o do eixo hipot&#225;lamo-hip&#243;fise-ovariano, associado ou n&#227;o com 17&#946;-estradiol por meio de adesivo transd&#233;rmico na dose de 0,1 mg/dia em mulheres jovens, saud&#225;veis e n&#227;o fumantes. Eles constataram que o 17&#946;-estradiol induziu menor osmolaridade urin&#225;ria com concentra&#231;&#227;o plasm&#225;tica do horm&#244;nio antidiur&#233;tico basal. Stachenfeld e Keefe,<xref ref-type="bibr" rid="B3">3</xref> observaram que o estr&#243;geno e a progesterona em mulheres jovens e saud&#225;veis promoveu m&#237;nimas altera&#231;&#245;es na regula&#231;&#227;o de &#225;gua e s&#243;dio, sugerindo que estes horm&#244;nios alteram o ponto de alostase. Esses pesquisadores relataram a redu&#231;&#227;o no limiar osm&#243;tico e aumento da concentra&#231;&#227;o de horm&#244;nio antidiur&#233;tico plasm&#225;tico, sem diminui&#231;&#227;o da quantidade de &#225;gua livre urin&#225;ria, pressupondo que o ACO induz menor sensibilidade renal ao horm&#244;nio antidiur&#233;tico e o produzem em maior quantidade para observarem os mesmos efeitos. Desta forma, nossos resultados para o grupo MACO em ambos os dias de coleta corroboram a hip&#243;tese desses pesquisadores, que ACO induz maior sensibilidade dos osmorreceptores centrais. Sugerimos que a condi&#231;&#227;o de repouso e de temperatura controlada no nosso estudo promoveu um ambiente com diminui&#231;&#227;o na perda de &#225;gua pela superf&#237;cie corporal, resultando em dilui&#231;&#227;o do compartimento extracelular e consequente est&#237;mulo inibit&#243;rio do horm&#244;nio antidiur&#233;tico, o que promoveu aumento de fluxo urin&#225;rio nas MACO.</p>
        <p>O aumento do retorno venoso &#233; um dos est&#237;mulos para a secre&#231;&#227;o do ANP pelas c&#233;lulas atriais. O ANP exerce sua atividade biol&#243;gica por meio de receptores localizados nos t&#250;bulos renais e na zona glomerulosa da gl&#226;ndula adrenal, inibindo a reabsor&#231;&#227;o de s&#243;dio nas c&#233;lulas do ducto coletor pela inibi&#231;&#227;o da libera&#231;&#227;o de renina, ocasionando menor produ&#231;&#227;o de angiotensina II, aldosterona e horm&#244;nio antidiur&#233;tico.</p>
        <p>Graugaard-Jensen et al.,<xref ref-type="bibr" rid="B12">12</xref> mencionam que a varia&#231;&#227;o de estr&#243;geno end&#243;geno n&#227;o interfere na concentra&#231;&#227;o do ANP plasm&#225;tico. Em nosso estudo, observamos que no grupo MACO, que apresenta maior concentra&#231;&#227;o de estr&#243;geno, houve maior excre&#231;&#227;o de ANP urin&#225;rio, contrapondo esses pesquisadores.</p>
        <p>De acordo com Schlueter et al.,<xref ref-type="bibr" rid="B6">6</xref> o efeito lipol&#237;tico do ANP ocorre a partir da via de forma&#231;&#227;o de GMPc, que leva a degrada&#231;&#227;o de triglicer&#237;deos. Entretanto, a DLM n&#227;o alterou o fluxo urin&#225;rio de ANP e de glicerol, sugerindo que o retorno venoso promovido pelo procedimento n&#227;o foi intenso o suficiente para estimular o ANP e exercer um efeito lipol&#237;tico detect&#225;vel na urina.</p>
        <p>Nossos dados refor&#231;am os de Krupek et al.,<xref ref-type="bibr" rid="B8">8</xref> que n&#227;o observaram altera&#231;&#227;o do glicerol urin&#225;rio em 3 mulheres jovens e saud&#225;veis, ap&#243;s 12 sess&#245;es de DLM. Entretanto, nesse estudo, foi avaliado o efeito cr&#244;nico da t&#233;cnica, que o difere do objetivo do presente estudo.</p>
        <p>A limita&#231;&#227;o do nosso estudou foi o per&#237;odo de coletas, que ocorreram durante o per&#237;odo de 2 manh&#227;s, o que impossibilitou a participa&#231;&#227;o de muitos. Isto foi adicionado no manuscrito para informa&#231;&#227;o aos leitores.</p>
      </sec>
      <sec sec-type="conclusions">
        <title>Conclus&#227;o</title>
        <p>Conclu&#237;mos que o efeito agudo de uma sess&#227;o de DLM promove aumento da natriurese em mulheres n&#227;o usu&#225;rias de ACO, e, por outro lado, em mulheres usu&#225;rias induz aumento na libera&#231;&#227;o de glicerol e de ANP.</p>
      </sec>
    </body>
    <back>
      <fn-group>
        <fn fn-type="financial-disclosure">
          <p>
            <bold>Fontes de Financiamento</bold>
          </p>
          <p>O presente estudo foi financiado por CAPES e Fapesp.</p>
        </fn>
        <fn fn-type="other">
          <p>
            <bold>Vincula&#231;&#227;o Acad&#234;mica</bold>
          </p>
          <p>Este artigo &#233; parte de tese de Doutorado de &#201;rica A. M. Camargo pela Universidade de Campinas (UNICAMP).</p>
        </fn>
        <fn fn-type="other">
          <p>Aprova&#231;&#227;o &#233;tica e consentimento informado</p>
          <p>Este estudo foi aprovado pelo Comit&#234; de &#201;tica da Faculdade de Ci&#234;ncias M&#233;dicas da Unicamp sob o n&#250;mero de protocolo CAAE: 24537613.2.0000.5404. Todos os procedimentos envolvidos nesse estudo est&#227;o de acordo com a Declara&#231;&#227;o de Helsinki de 1975, atualizada em 2013. O consentimento informado foi obtido de todos os participantes inclu&#237;dos no estudo.</p>
        </fn>
      </fn-group>
      <ack>
        <title>Agradecimentos</title>
        <p>A todos os volunt&#225;rios que participaram deste estudo. &#192; Universidade Estadual de Campinas (UNICAMP) e &#224; Universidade Metodista de Piracicaba (UNIMEP), pela infraestrutura. &#192; FAPESP (2013/20510-2) e CAPES pelo apoio financeiro.</p>
      </ack>
    </back>
  </sub-article>
</article>
