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<!DOCTYPE article SYSTEM "archivearticle3.dtd">
<article dtd-version="3.0" article-type="Invited Methods Article - JoVE Produced Video">
  <front>
    <journal-meta>
      <journal-id journal-id-type="publisher">jove</journal-id>
      <journal-id journal-id-type="email">editorial-office@jove.com</journal-id>
      <journal-title-group>
        <journal-title>Journal of Visualized Experiments</journal-title>
      </journal-title-group>
      <issn pub-type="ppub">1940-087X</issn>
      <issn pub-type="epub"></issn>
    </journal-meta>
    <article-meta>
      <article-id pub-id-type="publisher-id">JoVE58507</article-id>
      <article-id pub-id-type="manuscript">JoVE58507</article-id>
      <article-categories>
        <subj-group subj-group-type="Article Type">
          <subject>Invited Methods Article - JoVE Produced Video</subject>
        </subj-group>
        <subj-group subj-group-type="Category">
          <subject>JoVE Medicine</subject>
        </subj-group>
        <subj-group subj-group-type="Classification">
          <subject>Infection</subject>
          <subject>Sepsis</subject>
          <subject>Shock, Septic</subject>
          <subject>Pathologic Processes</subject>
          <subject>Inflammation</subject>
          <subject>Systemic Inflammatory Response Syndrome</subject>
          <subject>Sepsis</subject>
          <subject>Diseases</subject>
          <subject>Bacterial Infections and Mycoses</subject>
          <subject>Pathological Conditions, Signs and Symptoms</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title>ENDOTOXIN ACTIVITY ASSAY FOR THE DETECTION OF WHOLE BLOOD ENDOTOXEMIA IN CRITICALLY ILL PATIENTS</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author" corresp="yes">
          <role content-type="1" />
          <name>
            <surname>Pinciroli</surname>
            <given-names>Riccardo</given-names>
            <prefix>Dr</prefix>
          </name>
          <degrees>MD</degrees>
          <email>riccardo.pinciroli@unimib.it</email>
          <xref ref-type="aff" rid="aff1" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="2" />
          <name>
            <surname>Checchi</surname>
            <given-names>Simone</given-names>
          </name>
          <email>simone.checchi@gmail.com</email>
          <xref ref-type="aff" rid="aff2" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="3" />
          <name>
            <surname>Bottiroli</surname>
            <given-names>Maurizio</given-names>
          </name>
          <email>maurizio.bottiroli@ospedaleniguarda.it</email>
          <xref ref-type="aff" rid="aff3" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="4" />
          <name>
            <surname>Monti</surname>
            <given-names>Gianpaola</given-names>
          </name>
          <email>gianpaola.monti@ospedaleniguarda.it</email>
          <xref ref-type="aff" rid="aff4" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="5" />
          <name>
            <surname>Casella</surname>
            <given-names>Giampaolo</given-names>
          </name>
          <email>giampaolo.casella@ospedaleniguarda.it</email>
          <xref ref-type="aff" rid="aff5" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="6" />
          <name>
            <surname>Fumagalli</surname>
            <given-names>Roberto</given-names>
          </name>
          <email>roberto.fumagalli@unimib.it</email>
          <xref ref-type="aff" rid="aff6" />
        </contrib>
        <contrib contrib-type="editor">
          <name>
            <surname>Navani</surname>
            <given-names>Dipesh</given-names>
          </name>
          <degrees>Ph.D.</degrees>
          <email>dipesh.n@jove.com</email>
          <xref ref-type="aff" rid="aff7" />
        </contrib>
        <aff id="aff1">
          <institution>Universita degli Studi di Milano Facolta di Medicina e Chirurgia</institution>
          <institution content-type="position">Assistant Professor of Anesthesia</institution>
          <addr-line content-type="addrline1">Via Cadore 48</addr-line>
          <country>ITALY</country>
          <addr-line content-type="city">Monza</addr-line>
          <addr-line content-type="state">MB</addr-line>
          <addr-line content-type="zipcode">20900</addr-line>
          <phone content-type="primary">+393391232304</phone>
          <institution>Department of Anesthesia and Critical Care, Niguarda Hospital, Milan, Italy; 
University of Milan-Bicocca Medical School, Monza, Italy</institution>
        </aff>
        <aff id="aff2">
          <institution>Universita degli Studi di Milano-Bicocca Scuola di Medicina e Chirurgia</institution>
          <addr-line content-type="addrline1">Via Cadore, 48</addr-line>
          <country>ITALY</country>
          <addr-line content-type="city">Monza</addr-line>
          <addr-line content-type="state">MB</addr-line>
          <addr-line content-type="zipcode">20900</addr-line>
          <institution>University of Milan-Bicocca Medical School</institution>
        </aff>
        <aff id="aff3">
          <institution>Azienda Socio Sanitaria Territoriale Grande Ospedale Metropolitano Niguarda</institution>
          <institution content-type="dept">Anesthesia and Critical Care</institution>
          <addr-line content-type="addrline1">P.zza Ospedale Maggiore, 3.</addr-line>
          <country>ITALY</country>
          <addr-line content-type="city">Milan</addr-line>
          <addr-line content-type="state">MI</addr-line>
          <addr-line content-type="zipcode">20148</addr-line>
          <institution>Department of Anesthesia and Critical Care, Niguarda Hospital, Milan, Italy</institution>
        </aff>
        <aff id="aff4">
          <institution>Azienda Socio Sanitaria Territoriale Grande Ospedale Metropolitano Niguarda</institution>
          <addr-line content-type="addrline1">P.zza Ospedale Maggiore, 3</addr-line>
          <country>ITALY</country>
          <addr-line content-type="city">Milan</addr-line>
          <addr-line content-type="state">MI</addr-line>
          <addr-line content-type="zipcode">20148</addr-line>
          <institution>Department of Anesthesia and Critical Care, Niguarda Hospital, Milan, Italy</institution>
        </aff>
        <aff id="aff5">
          <institution>Azienda Socio Sanitaria Territoriale Grande Ospedale Metropolitano Niguarda</institution>
          <addr-line content-type="addrline1">P.zza Ospedale Maggiore, 3</addr-line>
          <country>ITALY</country>
          <addr-line content-type="city">Milan</addr-line>
          <addr-line content-type="state">MI</addr-line>
          <addr-line content-type="zipcode">20148</addr-line>
          <institution>Department of Anesthesia and Critical Care, Niguarda Hospital, Milan, Italy</institution>
        </aff>
        <aff id="aff6">
          <institution>Azienda Socio Sanitaria Territoriale Grande Ospedale Metropolitano Niguarda</institution>
          <addr-line content-type="addrline1">P.zza Ospedale Maggiore, 3</addr-line>
          <country>ITALY</country>
          <addr-line content-type="city">Milan</addr-line>
          <addr-line content-type="zipcode">20148</addr-line>
          <institution>Department of Anesthesia and Critical Care, Niguarda Hospital, Milan, Italy; University of Milan-Bicocca Medical School</institution>
        </aff>
        <aff id="aff7">
          <institution>JoVE</institution>
          <addr-line>1 Alewife Center</addr-line>
          <addr-line>Suite 200</addr-line>
          <addr-line>Cambridge, MA  02140</addr-line>
          <country>UNITED STATES</country>
        </aff>
      </contrib-group>
      <history>
        <date date-type="received">
          <day>23</day>
          <month>05</month>
          <year>2018</year>
        </date>
        <date date-type="rev-recd">
          <day>02</day>
          <month>01</month>
          <year>2019</year>
        </date>
        <date date-type="rev-recd">
          <day>12</day>
          <month>12</month>
          <year>2018</year>
        </date>
        <date date-type="accepted">
          <day>04</day>
          <month>01</month>
          <year>2019</year>
        </date>
      </history>
      <abstract>
        <p>The Lipopolysaccharide, also known as endotoxin, while being a fundamental component of gram-negative bacteria, plays a crucial role in the development of sepsis and septic shock. The early identification of an infectious process, which is rapidly evolving to a critical illness, might prompt a quicker and more intensive treatment, thereby potentially leading to better patient outcomes. The Endotoxin Activity Assay (EAA) and can be used at the bedside as a reliable biomarker of systemic endotoxemia. The detection of elevated endotoxin activity levels has been repeatedly shown to be associated with an increased diseased severity in patients with sepsis and septic shock. The assay is quick and easy to perform. Briefly, after sampling, a small aliquot of whole blood is rapidly incubated with complexes of an anti-endotoxin antibody and LPS. Endotoxin activity is measured as the relative oxidative burst of primed neutrophils as detected by chemioluminescence. The assay's output is expressed on a scale from 0 (absent) to 1 (maximal) and categorized as "low" (&lt; 0.4 units), "intermediate" (0.4-0.59 units), or "high" (≥ 0.6 units). The detailed methodology and rationale for the implementation of the EAA assay are reported in this manuscript.</p>
      </abstract>
      <kwd-group>
        <kwd>Endotoxin Activity</kwd>
        <kwd>Sepsis</kwd>
        <kwd>Shock</kwd>
        <kwd>lipopolysaccharide</kwd>
        <kwd>Chemioluminescence</kwd>
        <kwd>Neutrophils</kwd>
      </kwd-group>
      <funding-group />
      <counts>
        <fig-count count="0" />
      </counts>
      <custom-meta-group>
        <custom-meta>
          <meta-name>Please indicate whether this article will be Standard Access or Open Access. </meta-name>
          <meta-value>Standard Access (US$2,400);</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Please indicate the &lt;b&gt; city, state/province, and country&lt;/b&gt; where this article will be &lt;b&gt;filmed&lt;/b&gt;. Please do not use abbreviations.</meta-name>
          <meta-value>Servizio di Anestesia e Rianimazione 1, Dipartimento EAS, ASST Grande Ospedale Metropolitano Niguarda. P.zza Ospedale Maggiore, 3. 20162 Milan, Italy.</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Black and White Image Count</meta-name>
          <meta-value>0</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Color Image Count</meta-name>
          <meta-value>0</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Acquisitions Editor</meta-name>
          <meta-value>ronald myers</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Assistant who entered lead</meta-name>
          <meta-value>n/a</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>ALA Access Type</meta-name>
          <meta-value>JPV Standard Access ($2400)</meta-value>
        </custom-meta>
      </custom-meta-group>
    </article-meta>
  </front>
  <body>
    <fig>
      <label>Figure 1</label>
      <graphic xlink:href="Fig.1-EA distribution.tiff" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <table-wrap>
      <label>Materials Table</label>
      <graphic xlink:href="JoVE_EAA_SystemMaterials1.xls" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
    <fig>
      <label>Figure 2</label>
      <graphic xlink:href="Fig.2-EA distribution.tiff" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 1</label>
      <graphic xlink:href="Figure 1.tiff" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure</label>
      <graphic xlink:href="Fig1.tif" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <table-wrap>
      <label>Table 1</label>
      <graphic xlink:href="Table_1.xlsx" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
    <table-wrap>
      <label>Table of Materials</label>
      <graphic xlink:href="JoVE_EAA_SystemMaterials1.xlsx" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
  </body>
</article>