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<label for="email-citation-format" class="action-panel-label">
Format:
</label>
<select id="email-citation-format" name="citation-format" class="action-panel-selector email-citation-format">
<option selected="selected" value="summary">Summary</option>
<option value="summary-text">Summary (text)</option>
<option value="abstract">Abstract</option>
<option value="abstract-text">Abstract (text)</option>
</select>
</div>
<div class="include-supplemental-container">
<input type="checkbox" aria-label="Include MeSH and other data" name="include-supplemental" id="email-include-supplemental" class="email-include-supplemental">
<label for="email-include-supplemental" class="email-include-supplemental-label">MeSH and other data</label>
</div>
<div class="form-field recaptcha ">
<div class="g-recaptcha" id="id-recaptcha" data-sitekey="6LfsWHMdAAAAAClKbtOpjQ2pMjgsGxvv7NdZW9uI"></div>
</div>
<div id="captcha-error-message" class="usa-input-error-message captcha-validation-message" role="alert"></div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="send">
Send email
</button>
<button class="action-panel-cancel"
aria-label="Close 'Email citations' panel"
ref="linksrc=close_email_panel"
aria-controls="email-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="cancel">
Cancel
</button>
</div>
<input type="hidden" name="email-search-details" value="" />
<input type="hidden" name="email-search-details-hash" value="0e42663a6c3bd85498fcb88798998fed7bfdc45d457db35281e41afe13cc0524" />
</form>
</div>
</div>
<div id="collections-action-panel"
class="collections-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-collections-open-panel-enabled="false"
data-collections-open-panel-url-hash="#open-collections-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to Collections
</h3>
<form id="collections-action-panel-form"
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-existing-collections-url="/list-existing-collections/"
data-add-to-existing-collection-url="/add-to-existing-collection/"
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-myncbi-max-collection-name-length="100"
data-add-to-collection-max-amount="1000"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/">
<input type="hidden" name="csrfmiddlewaretoken" value="loJlFycgzbl21sWdSFxiRQt4ImZlyElTCbtqioFC9M8wVT23vYaklYZIgqHxrfTd">
<div class="choice-group" role="radiogroup">
<ul class="radio-group-items">
<li>
<input type="radio"
id="collections-action-panel-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_new">
<label for="collections-action-panel-new">Create a new collection</label>
</li>
<li>
<input type="radio"
id="collections-action-panel-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_existing">
<label for="collections-action-panel-existing">Add to an existing collection</label>
</li>
</ul>
</div>
<div class="controls-wrapper">
<div class="action-panel-control-wrap new-collections-controls">
<label for="collections-action-panel-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label>
<input
type="text"
name="add-to-new-collection"
id="collections-action-panel-add-to-new"
class="collections-action-add-to-new"
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
maxlength="100"
data-ga-category="save_share"
data-ga-action="create_collection"
data-ga-label="non_favorties_collection">
<div class="collections-new-name-too-long usa-input-error-message selection-validation-message">
Name must be less than 100 characters
</div>
</div>
<div class="action-panel-control-wrap existing-collections-controls">
<label for="collections-action-panel-add-to-existing" class="action-panel-label">
Choose a collection:
</label>
<select id="collections-action-panel-add-to-existing"
class="action-panel-selector collections-action-add-to-existing"
data-ga-category="save_share"
data-ga-action="select_collection"
data-ga-label="($('#collections-action-add-to-existing').val() === 'Favorites') ? 'Favorites' : 'non_favorites_collection'">
</select>
<div class="collections-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your collection due to an error<br>
<a href="#">Please try again</a>
</div>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="add">
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to Collections' panel"
ref="linksrc=close_collections_panel"
aria-controls="collections-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="bibliography-action-panel"
class="bibliography-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-bibliography-open-panel-enabled="false"
data-bibliography-open-panel-url-hash="#open-bibliography-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to My Bibliography
</h3>
<form id="bibliography-action-panel-form"
class="bibliography-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-add-to-bibliography-max-amount="100"
data-add-to-bibliography-batch-size="10"
data-bibliography-delegates-url="/list-bibliography-delegates/"
data-add-to-bibliography-url="/add-to-bibliography/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-mybib-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/mybibliography/">
<input type="hidden" name="csrfmiddlewaretoken" value="loJlFycgzbl21sWdSFxiRQt4ImZlyElTCbtqioFC9M8wVT23vYaklYZIgqHxrfTd">
<div class="action-panel-control-wrap bibliographies-controls">
<div class="choice-group">
<ul class="bibliographies-action-add radio-group-items">
<li>
<input name="bibliography" id="my-bibliography" class="my-bibliography" type="radio" checked/>
<label for="my-bibliography">My Bibliography</label>
</li>
</ul>
</div>
</div>
<div class="bibliographies-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your delegates due to an error<br>
<a href="#">Please try again</a>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore>
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to bibliography' panel"
ref="linksrc=close_bibliography_panel"
aria-controls="bibliography-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="mybib"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="saved-search-action-panel" class="saved-search-action-panel action-panel " aria-hidden="true"
data-saved-search-open-panel-enabled="false"
data-saved-search-open-panel-url-hash="#open-saved-search-panel">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your saved search
</h2>
<form id="saved-search-action-panel-form"
class="saved-search-action-panel-form action-panel-content action-form"
data-create-saved-search-url="/create-saved-search/"
data-try-search-terms-url="/try-search-term/"
data-saved-search-root-url="https://www.ncbi.nlm.nih.gov/myncbi/searches/">
<input type="hidden" name="csrfmiddlewaretoken" value="loJlFycgzbl21sWdSFxiRQt4ImZlyElTCbtqioFC9M8wVT23vYaklYZIgqHxrfTd">
<div class="action-panel-control-wrap">
<label for="saved-search-name" class="action-panel-label saved-search-name-label required-field-asterisk">
Name of saved search:
</label>
<input maxlength="200"
type="text"
name="saved-search-name"
id="saved-search-name"
class="saved-search-name"
value=""
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-term" class="action-panel-label required-field-asterisk">
Search terms:
</label>
<textarea name="saved-search-term" id="saved-search-term" class="saved-search-term" required></textarea>
</div>
<div class="test-search-term-wrap">
<a href="#" class="try-search-term">Test search terms</a>
</div>
<div class="choice-group action-panel-extra-margin-top">
<span class="action-panel-label" id="fieldset-label">
Would you like email updates of new search results?
</span>
<fieldset id="saved-search-alert" aria-describedby="fieldset-label">
<legend class="usa-sr-only">Saved Search Alert Radio Buttons</legend>
<ul class="radio-group-items">
<li>
<input type="radio" id="saved-search-alert-yes" class="saved-search-alert-yes" name="saved-search-alert" value="yes" checked>
<label for="saved-search-alert-yes" class="action-panel-label">Yes</label>
</li>
<li>
<input aria-label="No radio input" type="radio" id="saved-search-alert-no" class="saved-search-alert-no" name="saved-search-alert" value="no">
<label for="saved-search-alert-no" class="action-panel-label">No</label>
</li>
</ul>
</fieldset>
</div>
<div class="alert-schedule-wrap">
<div class="action-panel-control-wrap">
<label class="action-panel-label">
Email:
</label>
<span aria-label="Email address" id="saved-search-email" class="action-panel-label"><span class="action-panel-label-bold"></span> (<a class="myncbi-account-settings" href="https://www.ncbi.nlm.nih.gov/account/settings/">change</a>)</span>
</div>
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="saved-search-frequency" class="action-panel-label">
Frequency:
</label>
<select id="saved-search-frequency" class="no-border-panel-selector saved-search-frequency">
<option value="monthly">Monthly</option>
<option value="weekly">Weekly</option>
<option value="daily">Daily</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-monthly-additional">
<label for="saved-search-monthly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-monthly-on-day" class="no-border-panel-selector">
<option value="Sunday">The first Sunday</option>
<option value="Monday">The first Monday</option>
<option value="Tuesday">The first Tuesday</option>
<option value="Wednesday">The first Wednesday</option>
<option value="Thursday">The first Thursday</option>
<option value="Friday">The first Friday</option>
<option value="Saturday">The first Saturday</option>
<option value="day">The first day</option>
<option value="weekday">The first weekday</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-weekly-additional">
<label for="saved-search-weekly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-weekly-on-day" class="no-border-panel-selector saved-search-weekly-on-day">
<option value="Sunday">Sunday</option>
<option value="Monday">Monday</option>
<option value="Tuesday">Tuesday</option>
<option value="Wednesday">Wednesday</option>
<option value="Thursday">Thursday</option>
<option value="Friday">Friday</option>
<option value="Saturday">Saturday</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-report" class="action-panel-label">
Report format:
</label>
<select id="saved-search-report" class="no-border-panel-selector saved-search-report">
<option value="DocSum">Summary</option>
<option value="DocSumText">Summary (text)</option>
<option value="Abstract">Abstract</option>
<option value="AbstractText">Abstract (text)</option>
<option value="MEDLINE">PubMed</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-amount" class="action-panel-label">
Send at most:
</label>
<select id="saved-search-amount" class="no-border-panel-selector saved-search-amount">
<option value="1">1 item</option>
<option value="5" selected>5 items</option>
<option value="10">10 items</option>
<option value="20">20 items</option>
<option value="50">50 items</option>
<option value="100">100 items</option>
<option value="200">200 items</option>
</select>
</div>
<div>
<input type="checkbox" id="saved-search-send-if-no-result" class="saved-search-send-if-no-result" name="saved-search-send-if-no-result">
<label for="saved-search-send-if-no-result" class="action-panel-label smaller-checkbox">
Send even when there aren't any new results
</label>
</div>
<div class="action-panel-control-wrap option-text-in-email-wrap">
<label for="saved-search-email-text" class="action-panel-label">
Optional text in email:
</label>
<textarea name="saved-search-email-text"
id="saved-search-email-text"
class="saved-search-email-text"></textarea>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Saving..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Save
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your saved search' panel"
ref="linksrc=close_saved_search_panel"
aria-controls="saved-search-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="citation-manager-action-panel" class="citation-manager-action-panel action-panel" aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Create a file for external citation management software
</h2>
<form id="citation-manager-action-panel-form"
class="action-panel-content action-form"
action="/results-export-ids/"
data-by-search-action="/results-export-search-data/"
data-by-ids-action="/results-export-ids/"
method="post"
data-by-search-method="post"
data-by-ids-method="post">
<input type="hidden" name="csrfmiddlewaretoken" value="loJlFycgzbl21sWdSFxiRQt4ImZlyElTCbtqioFC9M8wVT23vYaklYZIgqHxrfTd">
<input name="results-format" type="hidden" value="pubmed"/>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="save">
Create file
</button>
<button class="action-panel-cancel"
aria-label="Close 'Send citations to citation manager' panel"
ref="linksrc=close_citation_manager_panel"
aria-controls="citation-manager-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="rss-action-panel" class="rss-action-panel action-panel " aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your RSS Feed
</h2>
<form id="rss-action-panel-form"
class="rss-action-panel-form action-panel-content action-form"
data-create-rss-feed-url="/create-rss-feed-url/"
data-search-form-term-value="">
<input type="hidden" name="csrfmiddlewaretoken" value="loJlFycgzbl21sWdSFxiRQt4ImZlyElTCbtqioFC9M8wVT23vYaklYZIgqHxrfTd">
<div class="action-panel-control-wrap">
<label for="rss-name" class="action-panel-label required-field-asterisk">
Name of RSS Feed:
</label>
<input maxlength="200"
placeholder="Name"
type="text"
name="rss-name"
id="rss-name"
class="rss-name"
value=''
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="rss-limit-wrap">
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="rss-limit" class="action-panel-label">
Number of items displayed:
</label>
<select id="rss-limit" class="no-border-panel-selector rss-limit">
<option value="5">5</option>
<option value="10">10</option>
<option value="15" selected="selected">15</option>
<option value="20">20</option>
<option value="50">50</option>
<option value="100">100</option>
</select>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Creating..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Create RSS
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your RSS' panel"
ref="linksrc=close_rss_panel"
aria-controls="rss-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
<div class="action-panel-control-wrap rss-link-copy-wrap">
<label for="rss-link" class="usa-sr-only">RSS Link</label>
<input placeholder="Your RSS Feed Link" type="text" name="rss-link" id="rss-link" class="rss-link" title="RSS Link">
<button
type="button"
disabled
class="rss-link-copy-button disabled"
data-ga-category="save_share"
data-ga-action="rss"
data-ga-label="copy">
Copy
</button>
</div>
</form>
</div>
</div>
</div>
</div>
<div class="article-page" id="article-page" data-article-pmid="25760458">
<aside class="page-sidebar">
<div class="inner-wrap">
<div class="full-text-links">
<div class="full-view">
<h3 class="title">
Full text links
</h3>
<div class="full-text-links-list">
<a class="link-item
dialog-focus"
href="https://dx.plos.org/10.1371/journal.pone.0118615"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=4656&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=25760458&amp;db=pubmed&amp;nlmid=101285081"
title="See full text options at Public Library of Science"
data-ga-category="full_text"
data-ga-action="Public Library of Science"
data-ga-label="25760458"
><img src="https://cdn.ncbi.nlm.nih.gov/corehtml/query/egifs/https:--journals.plos.org-resource-img-external-pone_120x30.png" alt="Public Library of Science full text link"><span class="text">
Public Library of Science
</span></a><a class="link-item
pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/25760458/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=25760458&amp;db=pubmed&amp;nlmid=101285081"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="25760458"
><span class="text">
Free PMC article
</span></a>
</div>
</div>
<div class="short-view">
<a href="#" class="full-text-links-button full-text-links-dialog-trigger">
Full text links
</a>
</div>
</div>
<div class="actions-buttons sidebar"><h3 class="title">Actions</h3><div class="inner-wrap"><button class="citation-button citation-dialog-trigger"
aria-label="Open dialog with citation text in different styles"
data-ga-category="save_share"
data-ga-action="cite"
data-ga-label="open"
data-all-citations-url="/25760458/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/25760458/export/"><span class="button-label">Cite</span></button><link type="text/css" href="ncbi-overlay-block/src/overlay-block.css"><div class="collections-button-container" data-article-id="25760458" data-article-db="pubmed"><button class="collections-button collections-dialog-trigger"
aria-label="Save article in MyNCBI collections."
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_button"
data-collections-open-dialog-enabled="false"
data-collections-open-dialog-url="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpubmed.ncbi.nlm.nih.gov%2F25760458%2F%23open-collections-dialog"
data-in-collections="false"><span class="button-label">Collections</span></button><div class="overlay" role="dialog"><div id="collections-action-dialog"
class="dialog collections-dialog"
aria-hidden="true"><div class="title">Add to Collections</div><div class="collections-action-panel action-panel"><form id="collections-action-dialog-form"
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-existing-collections-url="/list-existing-collections/"
data-add-to-existing-collection-url="/add-to-existing-collection/"
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
data-myncbi-max-collection-name-length="100"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/"><input type="hidden" name="csrfmiddlewaretoken" value="loJlFycgzbl21sWdSFxiRQt4ImZlyElTCbtqioFC9M8wVT23vYaklYZIgqHxrfTd"><div class="choice-group" role="radiogroup"><ul class="radio-group-items"><li><input type="radio"
id="collections-action-dialog-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_new"><label for="collections-action-dialog-new">Create a new collection</label></li><li><input type="radio"
id="collections-action-dialog-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_existing"><label for="collections-action-dialog-existing">Add to an existing collection</label></li></ul></div><div class="controls-wrapper"><div class="action-panel-control-wrap new-collections-controls"><label for="collections-action-dialog-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label><input
type="text"
name="add-to-new-collection"
id="collections-action-dialog-add-to-new"
class="collections-action-add-to-new"
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
maxlength="100"
data-ga-category="collections_button"
data-ga-action="create_collection"
data-ga-label="non_favorties_collection"><div class="collections-new-name-too-long usa-input-error-message selection-validation-message">
Name must be less than 100 characters
</div></div><div class="action-panel-control-wrap existing-collections-controls"><label for="collections-action-dialog-add-to-existing" class="action-panel-label">
Choose a collection:
</label><select id="collections-action-dialog-add-to-existing"
class="action-panel-selector collections-action-add-to-existing"
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Abstract
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<p>
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Background:
</strong>
The caterpillar of the moth Premolis semirufa, commonly named pararama, is found in the Brazilian Amazon region. Accidental contact with the caterpillar bristles causes an intense itching sensation, followed by symptoms of an acute inflammation, which last for three to seven days after the first incident. After multiple accidents a chronic inflammatory reaction, called "Pararamose", characterized by articular synovial membrane thickening with joint deformities common to chronic synovitis, frequently occurs. Although complement mediated inflammation may aid the host defense, inappropriate or excessive activation of the complement system and generation of anaphylatoxins can lead to inflammatory disorder and pathologies. The aim of the present study was to evaluate, in vitro, whether the Premolis semirufa's bristles extract could interfere with the human complement system.
</p>
<p>
<strong class="sub-title">
Results:
</strong>
The bristles extract was able to inhibit the haemolytic activity of the alternative pathway, as well as the activation of the lectin pathway, but had no effect on the classical pathway, and this inhibition seemed to be caused by activation and consumption of complement components. The extract induced the production of significant amounts of all three anaphylatoxins, C3a, C4a and C5a, promoted direct cleavage of C3, C4 and C5 and induced a significant generation of terminal complement complexes in normal human serum. By using molecular exclusion chromatography, a serine protease of 82 kDa, which activates complement, was isolated from P. semirufa bristles extract. The protease, named here as Ps82, reduced the haemolytic activity of the alternative and classical pathways and inhibited the lectin pathway. In addition, Ps82 induced the cleavage of C3, C4 and C5 and the generation of C3a and C4a in normal human serum and it was capable to cleave human purified C5 and generate C5a. The use of Phenanthroline, metalloprotease inhibitor, in the reactions did not significantly interfere with the activity of the Ps82, whereas the presence of PMSF, serine protease inhibitor, totally blocked the activity.
</p>
<p>
<strong class="sub-title">
Conclusion:
</strong>
These data show that a serine protease present in the Premolis semirufa's bristles extract has the ability to activate the complement system, which may contribute to the inflammatory process presented in humans after envenomation.
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<p xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:p1="http://pubmed.gov/pub-one"><bold>Competing Interests: </bold>The authors have declared that no competing interests exist.</p>
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Figures
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<p> Fig 1. Activity of the <i> Premolis semirufas… </i> </p>
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<p> Fig 1. Activity of the <i> Premolis semirufas </i> bristles extract on the alternative and lectin pathways. </p>
</strong>
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<strong class="figure-label">
Fig 1. Activity of the <i>Premolis semirufas</i> bristles extract on the alternative and lectin pathways.
</strong>
<div class="figure-caption-contents"><b>[A]</b> Samples (50 μL) of normal human serum (NHS) were pre-incubated with increasing concentrations of the bristles extract or PBS, at 37°C for 30 min, and the residual haemolytic activity of the mixtures was measured. <b>[B]</b> The residual lectin pathway complement activity was assessed by incubation of NHS with the bristles extract (175 μg/mL), in the presence or absence of 1,10-Phenanthroline (10 mM), at 37°C for 30 min, and added to plates coated with mannan (100 μg/mL). Data are representative for three separate experiments, performed in duplicate, and the results are expressed as percentage of AP<sub>50</sub> reduction (%) ± SD (for A) and percentage of C4b deposition (%) ± SD (for B). (*) <i>p</i>&lt;0.05, (**) <i>p</i> &lt; 0.01 and (***) <i>p</i> &lt; 0.001: significant differences between the mean values obtained with PBS and the treatments.</div>
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<p> Fig 2. Human serum anaphylatoxins generation induced… </p>
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<strong class="figure-label">
<p> Fig 2. Human serum anaphylatoxins generation induced by the <i> Premolis semirufas </i> bristles extract. </p>
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<div class="figure-caption-contents"><p> Samples of… </p></div>
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<figcaption id="figure-caption-1" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
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Fig 2. Human serum anaphylatoxins generation induced by the <i>Premolis semirufas</i> bristles extract.
</strong>
<div class="figure-caption-contents">Samples of NHS (50 μL) were incubated with the bristles extract (175 μg/mL) or PBS, in the presence or absence of 10 mM 1,10-Phenanthroline (Phen), at 37°C for 30 min. The generation of the anaphylatoxins (C3a, C4a and C5a) was measured using the <i>Human Anaphylatoxin Cytometric Bead Array</i> (CBA). Data are representative for two separate experiments, performed in duplicate, and the results are expressed as concentration of each anaphylatoxin per mL of human serum (ng/mL) ± SD. (*) <i>p</i> &lt; 0.05 and (**) <i>p</i> &lt; 0.01: significant differences between the mean values obtained with the buffer and between the treatments.</div>
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<p> Fig 3. Terminal Complement Complex (TCC) generation… </p>
</strong>
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<strong class="figure-label">
<p> Fig 3. Terminal Complement Complex (TCC) generation induced by the <i> Premolis semirufa </i> <i> s </i> bristles… </p>
</strong>
<div class="figure-caption-contents"><p> Samples… </p></div>
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<strong class="figure-label">
Fig 3. Terminal Complement Complex (TCC) generation induced by the <i>Premolis semirufa</i><i>s</i> bristles extract.
</strong>
<div class="figure-caption-contents">Samples of NHS (50 μL) were incubated with the bristles extract (175 μg/mL) or PBS, in the presence or absence of 10 mM 1,10-Phenanthroline (Phen), at 37°C for 30 min, and SC5b-9 complex present in human serum samples was measured using the MicroVue SC5b-9 Plus EIA Kit. Data are representative for two separate experiments, performed in duplicate, and the results are expressed as concentration of SC5b-9 complex per mL of human serum (ng/mL) ± SD. (***) <i>p</i> &lt; 0.001: significant differences between the mean values obtained with the buffer and between the treatments.</div>
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<img class="figure-thumb" itemprop="thumbnail"
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<p> Fig 4. Proteolytic action of the <i> Premolis… </i> </p>
</strong>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig 4. Proteolytic action of the <i> Premolis semirufas </i> bristles extract on purified human C-components C3,… </p>
</strong>
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<figcaption id="figure-caption-3" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig 4. Proteolytic action of the <i>Premolis semirufas</i> bristles extract on purified human C-components C3, C4 and C5.
</strong>
<div class="figure-caption-contents">Samples of the bristles extract (2.0 μg for C3 and C4 or 3.0 μg for C5) were incubated, in the absence or presence of 10 mM PMSF or 1,10-Phenanthroline (Phen), with human C3 (3 μg) <b>[A]</b>, human C4 (3 μg) <b>[B]</b> and human C5 (3 μg) <b>[C]</b> at 37°C for 1 h. Proteolytic activity was examined on 10% polyacrylamide gel under reducing conditions and stained by silver. In the 1<sup>st</sup> lanes of gels: electrophoretic separation of purified components incubated with PBS as a positive control; 2<sup>nd</sup> lanes: incubation of purified components with the extract; 3<sup>rd</sup> lanes: incubation of the mixture with PMSF; 4<sup>th</sup> lanes: incubation of the mixture with Phenanthroline and 5<sup>th</sup> lanes: electrophoretic separation of the extract. α (115 kDa) and β (75 kDa) for C3; α (93 kDa), β (75 kDa) and γ (33 kDa) for C4 and α (115 kDa) and β (75 kDa) for C5.</div>
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<p> Fig 5. Chromatography of the <i> Premolis semirufas… </i> </p>
</strong>
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<strong class="figure-label">
<p> Fig 5. Chromatography of the <i> Premolis semirufas </i> bristles extract and isolation of the protease responsible… </p>
</strong>
</div>
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<strong class="figure-label">
Fig 5. Chromatography of the <i>Premolis semirufas</i> bristles extract and isolation of the protease responsible for complement activation.
</strong>
<div class="figure-caption-contents"><b>[A]</b> Chromatography of 1 mg of the extract on a FPLC-GP-250 Plus system using a molecular exclusion column (Superdex 200 10/300 GL), in 0.05 M ammonium bicarbonate buffer, pH 7.4. <b>[B]</b> SDS-PAGE of fractions screened by their ability to cleave the component C3. <b>[C]</b> SDS-PAGE (12%) of 10 μg of the extract and 0.25 μg of the fraction 10, under reducing conditions, followed by silver staining.</div>
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<strong class="figure-label">
<p> Fig 6. Proteolytic activity of Ps82 analyzed… </p>
</strong>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig 6. Proteolytic activity of Ps82 analyzed by zymography. </p>
</strong>
<div class="figure-caption-contents"><p> Samples of 0.05 μg of Ps82… </p></div>
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<strong class="figure-label">
Fig 6. Proteolytic activity of Ps82 analyzed by zymography.
</strong>
<div class="figure-caption-contents">Samples of 0.05 μg of Ps82 or 0.5 μg of the extract were incubated in the absence (Lane 1) or presence of 10 mM 1,10-Phenanthroline (Lane 2) or PMSF (Lane 3), inhibitors of metallo- and serineproteases, respectively, submitted to electrophoresis and subsequently incubated, at 37°C for 12 h, in 50 mM Tris-HCl, pH 8.3. Following incubation, the gel was stained with 0.2% Coomassie Brilliant Blue R-250 and the gelatinolytic activity was detected as unstained bands on a dark background. The figure represents the negative of the original image.</div>
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<strong class="figure-label">
<p> Fig 7. Action of Ps82 on the… </p>
</strong>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig 7. Action of Ps82 on the complement pathways. </p>
</strong>
<div class="figure-caption-contents"><p> Samples (50 μL) of normal human… </p></div>
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<figcaption id="figure-caption-6" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig 7. Action of Ps82 on the complement pathways.
</strong>
<div class="figure-caption-contents">Samples (50 μL) of normal human serum (NHS) were pre-incubated with 0.25, 0.33 or 0.5 μg/mL of Ps82 or with 175 μg/mL of the <i>Premolis semirufa</i>s bristles extract. The mixtures were pre-incubated for 30 min for the alternative pathway <b>[A]</b> or 1 h for the classical pathway <b>[C]</b> at 37°C, and then assayed for the residual complement activity in haemolytic tests. For lectin pathway <b>[B]</b>, samples of 50 μL of NHS were pre-incubated, in the presence or absence of 10 mM 1,10-Phenanthroline, with 0.5 μg/mL of Ps82 or 175 μg/mL of the <i>Premolis semirufa</i>s bristles extract for 30 min at 37°C and then evaluated for the residual complement activity by ELISA. Alternatively, samples of NHS incubated with Ps82 or the <i>Premolis semirufa</i>s bristles extract were evaluated in conditions for activation of the classical pathway, by the deposition of complement component C4b by ELISA <b>[D]</b>. Data are representative for three separate experiments, performed in duplicate, and the results are expressed as percentage of haemolysis (%) ± SD (for A and C) and percentage of C4b deposition (%) ± SD (for B and D) in relation to the control samples (NHS + buffer) and between the treatments. (*) <i>p</i> &lt; 0.05 (**) <i>p</i> &lt; 0.01 and (***) <i>p</i> &lt; 0.001. The symbol (#) indicates significant differences between the extract and Ps82.</div>
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<p> Fig 8. Human serum anaphylatoxins generation induced… </p>
</strong>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
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<p> Fig 8. Human serum anaphylatoxins generation induced by Ps82. </p>
</strong>
<div class="figure-caption-contents"><p> Samples of NHS (50 μL) were… </p></div>
</div>
<figcaption id="figure-caption-7" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig 8. Human serum anaphylatoxins generation induced by Ps82.
</strong>
<div class="figure-caption-contents">Samples of NHS (50 μL) were incubated with Ps82 (0.33 μg/mL) or the <i>Premolis semirufas</i> bristles extract (175 μg/mL), in the presence or absence of 10 mM 1,10-Phenanthroline (Phen), for 30 min at 37°C. The generation of the anaphylatoxins (C3a, C4a and C5a) was measured using the kits "Human C3a ELISA Kit, Human C4a ELISA Kit and Human C5a ELISA Kit". Data are representative for two separate experiments, performed in duplicate, and the results are expressed as concentration of each anaphylatoxin per mL of human serum (ng/mL) ± SD. (*) <i>p</i> &lt; 0.05 (**) <i>p</i> &lt; 0.01 and (***) <i>p</i> &lt; 0.001: significant differences between the mean values obtained with the Buffer and the treatments.</div>
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itemprop="contentUrl"
aria-describedby="figure-caption-8"
role="button"
data-image-width="693"
data-image-height="824"
data-image-alt="Fig 9"
data-pmc-id="PMC4356561"
data-figure-id="pone.0118615.g009">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-8"
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alt="Fig 9" />
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<strong class="figure-label">
<p> Fig 9. Proteolytic action of Ps82 on… </p>
</strong>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig 9. Proteolytic action of Ps82 on purified human C components C3, C4 and C5. </p>
</strong>
</div>
<figcaption id="figure-caption-8" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig 9. Proteolytic action of Ps82 on purified human C components C3, C4 and C5.
</strong>
<div class="figure-caption-contents">Samples of 0.1 μg of Ps82 were incubated, in the absence or presence of 10 mM PMSF or 1,10-Phenanthroline (Phen), with human C3 (3 μg) <b>[A]</b>, human C4 (3 μg) <b>[B]</b> and human C5 (3 μg) <b>[C]</b> at 37°C for 1 h. Proteolytic activity was examined on 10% polyacrylamide gel under reducing conditions and stained by silver. In the 1<sup>st</sup> lanes of gels: electrophoretic separation of purified components incubated with PBS as a positive control; 2<sup>nd</sup> lanes: incubation of purified components with Ps82; 3<sup>rd</sup> lanes: incubation of the mixture with PMSF and 4<sup>th</sup> lanes: incubation of the mixture with Phenanthroline. α (115 kDa) and β (75 kDa) for C3; α (93 kDa), β (75 kDa) and γ (33 kDa) for C4 and α (115 kDa) and β (75 kDa) for C5.</div>
</figcaption>
</figure>
<figure class="figure-item tail"
itemscope itemtype="http://schema.org/ImageObject"
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data-slide-index="9"
data-label-slug="fig-10">
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href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9351/4356561/3316fd1cc9b6/pone.0118615.g010.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-9"
role="button"
data-image-width="700"
data-image-height="359"
data-image-alt="Fig 10"
data-pmc-id="PMC4356561"
data-figure-id="pone.0118615.g010">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-9"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9351/4356561/da1739575e74/pone.0118615.g010.gif"
alt="Fig 10" />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="700">
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<strong class="figure-label">
<p> Fig 10. Anaphylatoxins generation from purified human… </p>
</strong>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> Fig 10. Anaphylatoxins generation from purified human C5 induced by the <i> Premolis semirufas </i> bristles extract… </p>
</strong>
</div>
<figcaption id="figure-caption-9" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
Fig 10. Anaphylatoxins generation from purified human C5 induced by the <i>Premolis semirufas</i> bristles extract and Ps82.
</strong>
<div class="figure-caption-contents">Samples of 3 μg of the extract or 0.1 μg of Ps82 were incubated, in the absence or presence of 10 mM PMSF, with human C5 (3 μg) for 30 min at 37°C. The generation of the anaphylatoxin C5a was measured using the "Human C5a ELISA Kit". Data are representative for two separate experiments, performed in duplicate, and the results are expressed as concentration of anaphylatoxin per mL (ng/mL) ± SD. (**) <i>p</i> &lt; 0.01 and (***) <i>p</i> &lt; 0.001: significant differences between the mean values obtained with the Buffer and the treatments.</div>
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Villas-Boas IM, Gonçalves-de-Andrade RM, Pidde-Queiroz G, Assaf SL, Portaro FC, Sant'Anna OA, et al. (2012) Premolis semirufa (Walker, 1856) envenomation, disease affecting rubber tappers of the Amazon: searching for caterpillar-bristles toxic components. PLoS Negl Trop Dis 6(2): e1531 10.1371/journal.pntd.0001531
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