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<!doctype html>
<html lang="en">
<head>
<meta charset="utf-8">
<meta name="viewport" content="width=device-width, initial-scale=1.0, maximum-scale=1.0, user-scalable=no">
<title>Clostridioides difficile</title>
<link rel="stylesheet" href="reveal.js/dist/reset.css">
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<link rel="stylesheet" href="resources/style.css">
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</head>
<body>
<div class="reveal">
<div class="slides">
<section data-background="#222" data-background-video="resources/title_vid.webm" data-background-size="contain"
data-background-video-loop="1">
<h2 style="color:darkorange; text-shadow: 1px 1px 1px black">
Motility of<br><em>Clostridioides difficile</em>
</h2>
<!---<h2 style="font-family: 'Geneva', sans-serif; color: #55f; text-shadow: 1px 1px 1px black"><div class="umgsquares"></div>UMG</h2> --->
<div style="margin-top: 18%">
<div style="text-shadow: 1px 1px 1px black">
<p style="text-shadow: 1px 1px 1px black; ">
Dr. Julian Schwanbeck</p>
<p style="text-shadow: 1px 1px 1px black; font-size: .9em; margin-top: 5%">
April 6th 2022<br>Göttingen
</p>
</div>
</div>
</section>
<section>
<p style="font-size: 2em; color: darkorange; margin-top: -5%"><em>Clostridioides difficile</em></p>
<div style="float: left; width: 45%; align-items: center; display: table; justify-content: center; height: 450px; margin-left: 40px">
<ul style="display: table-cell; vertical-align: middle; font-size: .9em">
<li style="margin-bottom: .25em">Formerly <em>Clostridium difficile</em></li>
<li style="margin-bottom: .25em">Firmicutes</li>
<li style="margin-bottom: .25em">Obligate anaerobe</li>
<li style="margin-bottom: .25em">Spore forming</li>
<li style="margin-bottom: .25em">Human pathogen</li>
<li style="margin-bottom: .25em">Nosocomial infection</li>
<li style="color: darkorange">Peritrichous flagella</li>
</ul>
</div>
<div style="float:right; alignment: right; width: 50%">
<img src="resources/cdiff_bright_bg.png" alt style="background:none; border:none; box-shadow:none; width:70%"> <!-- 10px solid transparent; -->
<p style="font-size: .5em;">Picture in collaboration with PD Dr. Hoppert</p>
</div>
<aside class="notes">
Gram Positive
Peritrichous erklären
</aside>
</section>
<section>
<!-- <section>-->
<h3 style="color:darkorange; ">The Pathogen</h3>
<div style="float: left; width: 55%; align-items: center; display: table; justify-content: center; height: 450px; margin-left: 20px"> <!-- -->
<ul style="display: table-cell; vertical-align: middle; font-size: .8em">
<li style="margin-bottom: .25em">Illness is <em>C</em>. <em>difficile</em> infection (CDI)</li>
<li style="margin-bottom: .25em">Healthy microbiome offers protection against <em>C</em>. <em>difficile</em></li>
<!-- <li style="margin-bottom: .25em"><em>C.</em> <em>difficile</em> infection (CDI) risk is strongly increased<br>by a preceding course of antibiotics</li> -->
<li style="margin-bottom: .25em"><em>C</em>. <em>difficile</em> mainly infects the ascending colon</li>
<!-- <li style="margin-bottom: .25em">Three main toxins: <em>tcdA</em>, <em>tcdB</em>, binary toxin CDT</li> -->
<li style="margin-bottom: .25em">Toxins lead to cell lysis of epithelial cells</li>
<li style="margin-bottom: .25em">Spores are found throughout the environment</li>
</ul>
</div>
<div style="float: right; width: 40%; align-items: center; display: table; justify-content: center; height: 450px; margin: auto">
<img src="resources/lower_intestine2.png" alt style="background:none; border:none; box-shadow:none; height: 400px">
</div>
<!-- </section>-->
<!-- <section>-->
<!-- <img src="resources/gru.jpg" alt style="background:none; border:none; box-shadow:none; ">-->
<!-- </section>-->
<aside class="notes">
Spores: lakes, fresh water, rivers (40-80%), soil, manure, private residences, any pets/livestock<br>
fecal oral circle<br>
secondary bile acids deoxycholate and lithocholate<br>
ascending colon, decending ileum<br>
pseudomembranous colitis<br>
Tox A/B<br>
lowest O2 conc. in colon<br>
</aside>
</section>
<section>
<section>
<h3 style="color:darkorange; ">The best treatment is prevention</h3>
<p>Failing that, current treatment options for CDI are:
<ul>
<li>Metronidazole</li>
<li>Vancomycin</li>
<li>Fidaxomicin</li>
<li>Feacal microbial transplant</li>
</ul>
</p>
</section>
<section>
<!-- <h3 style="color:darkorange; ">Introduction: the treatment</h3> -->
<img src="resources/fdx_screen_whiteBG.png" alt style="background:none; border:none; box-shadow:none; ">
</section>
<aside class="notes">
Hospital bed/room cleaning is a huge issue<br>
Aerosolic hydrogen peroxide treatment of hospital rooms
has been tested, but is fraught with health and safety hazards (Steindl et al., 2015).<br>
Vanco/Metro most commen<br>
Vanco: third line Antibiotic (D)<br>
Metro suffers from higher recurrence rates and resistance rates in 027<br>
FDX next to no resistance, though expensive<br>
</aside>
</section>
<section>
<h3 style="color:darkorange; ">Some numbers</h3>
<!-- CDC graph joke -->
<img src="resources/CDC_2019_cdiff_scary_sub.png" alt style="background:none; border:none; box-shadow:none; ">
<p style="font-size: .7em"><a href="https://www.doi.org/10.15620/cdc:82532">Source: CDC: Antibiotic resistance threats in the United States, 2019</a> </p>
</section>
<section>
<section>
<h3 style="color:darkorange; ">Bacterial Motility</h3>
<!-- Gram neg, peritrichous flagella, ..., spore forming -->
<div style="float: left; width: 55%; display: table; font-size: .75em; justify-content: center; height: 450px; margin-left: 20px">
<ul style="display: table-cell; vertical-align: middle; ">
<li style="margin-bottom: .55em">Bacteria face a miniscule Reynolds number</li>
<li style="margin-bottom: .55em">
Bacteria employ motility for a variety of reasons
<ul>
<li style="font-size: .9em; ">To explore for areas to colonize</li>
<li style="font-size: .9em; ">As a means for dispersal into the environment</li>
<li style="font-size: .9em; ">In conjunction with chemotaxis for directed motility</li>
</ul>
</li>
<li style="margin-bottom: .45em">Almost all sequenced <em>C</em>. <em>difficile</em> strains possess
a chemotaxis operon
<ul>
<li style="font-size: .9em; ">But not all have a functioning flagellar operon</li>
</ul>
</li>
<li style="margin-bottom: .55em">Analysis of motility requires observation of single cells
<!-- <ul>
<li style="font-size: .9em; margin-bottom: .45em">For example “run and tumble” in <em>E</em>.
<em>coli</em></li>
</ul> -->
</li>
</ul>
</div>
<div style="float: right; width: 40%; align-items: center; display: table; justify-content: center; height: 450px; margin: auto">
<!-- <img src="resources/chemtaxsys_light.png" alt style="background:none; border:none; box-shadow:none; height: 200px"> -->
<img src="resources/Ecoli_Tumbling.png" alt style="background:none; border:none; box-shadow:none; height: 400px">
</div>
</section>
<section>
<div style="height: 700px; vertical-align: center">
<img data-src="resources/chemtaxsys_whiteBG.png" alt
style="background:none; border:none; box-shadow:none;">
</div>
</section>
<aside class="notes">
low reynolds number: purcell 1976 lecture<br>
viscosity of medium and size of object make the number<br>
10^4 human, 10^2 guppy 10^-4 - 10^-5 bacteria<br>
explore for colonyz.: attachement (flagellum tip FliD)
Directed motility: needs to be faster than osmosis<br>
The main chemotaxis machinery (MCP, CheA, CheY) is needed for signal sensing and transduction<br>
Flagellar motility is a requirement for chemotaxis<br>
</aside>
</section>
<!-- Some published data -->
<section data-background="#222" data-background-video="resources/twitching_background.webm"
data-background-size="contain" data-background-video-loop="1">
<h3 style="color: darkorange; text-shadow: 1px 1px 1px black">Motility in liquid medium</h3>
<div class="fragment" style="margin: auto; width: 1000px">
<div style="float: left; width: 60%">
<video data-autoplay width="100%" height="auto">
<source data-src="resources/r20_twitch.webm" type="video/webm">
</video>
</div>
<div style="float: right; width: 40%; text-shadow: 1px 1px 1px black">
<ul>
<li>No noticeable<br>displacement</li>
</ul>
<p><br></p>
<ul>
<li>Very high rate<br>of trembling</li>
</ul>
</div>
</div>
<aside class="notes">
[Going forward: some published data]<br>
Explain beginning:<br>
- Motility in capillary assay<br>
- Motility in agarose<br>
- Motility in stich agar
</aside>
</section>
<section data-background-transition="none" data-background-size="20%" data-background-image="resources/pvp.png"
data-background-position="center" data-background-repeat="no-repeat" data-background="#222">
<h3 style="color: darkorange; text-shadow: 1px 1px 1px black">Motility with Polyvinylpyrrolidone</h3>
<p>PVP is a high molecular weight polymer</p>
<video data-autoplay class="fragment" width="auto" height="500px" autoplay loop controls>
<source data-src="resources/motile_liquid.webm">
</video>
<aside class="notes">
1.3 Mega Dalton<br>
MW 360.000 g/mol<br>
Similarity to mucin?<br>
</aside>
</section>
<section data-background="#222" data-background-video="resources/annot_vid.webm" data-background-size="contain"
data-background-video-loop="1" data-preload>
<aside class="notes">
pink: straight motility<br>
Yellow: curvy<br>
blue: back and forth motility<br>
green: inntermediate, turnarounds
</aside>
</section>
<section>
<div style="position: relative; height: 700px; left: -470px; margin-top: 0%">
<img data-src="resources/back_track.png" alt
style="position: absolute; background:none; border:none; box-shadow:none;">
<img data-src="resources/back_track_dist.png" class="fragment" alt
style="position: absolute; background:none; border:none; box-shadow:none;">
</div>
<aside class="notes">
TA: turnarounds [red]<br>
Displ.: displacement<br>
Dist.: total distance<br>
</aside>
</section>
<section>
<!--
<h4 style="color:darkorange; "><em>Clostridioides difficile</em> single cell swimming strategy</h4>
margin-top: 3%
-->
<div style="height: 700px; vertical-align: center">
<img src="resources/Figure_1.png" alt
style="background:none; border:none; box-shadow:none;">
</div>
<aside class="notes">
Origin/rose plot erklären
Mucin erwähnen (kommt später)
</aside>
</section>
<section>
<div style="height: 70%; vertical-align: center; margin-top: 0%">
<img data-src="resources/fig2.png" alt
style="background:none; border:none; box-shadow:none;">
</div>
</section>
<!--- <section>
<div style="height: 700px; vertical-align: center">
<img data-src="resources/cys.png" alt
style="background:none; border:none; box-shadow:none;">
</div>
<aside class="notes">
Langsamer werden erzählen
</aside>
</section>
<section>
<div style="position: relative; height: 720px; left: -612px;">
<img data-src="resources/mot_model1.png" alt
style="position: absolute; background:none; border:none; box-shadow:none;">
<img data-src="resources/mot_model2.png" class="fragment" alt
style="position: absolute; background:none; border:none; box-shadow:none;">
<img data-src="resources/mot_model3.png" class="fragment" alt
style="position: absolute; background:none; border:none; box-shadow:none;">
</div>
</section> -->
<!-- <section>
<h3 style="color:darkorange; ">Future Perspectives and Collaborations</h3>
<ul>
<li>Establish chemotaxis assay on a single cell basis, for example microfluidic based assay
<ul>
<li style="font-size: .9em; margin-bottom: .45em">Test candidate substrates <em>(from metabolomic
studies in the consortium)</em> for their influence in chemotaxis & motility
</li>
</ul>
</li>
<li>Generation of knock-out mutants for chemotaxis genes and test for chemotaxis defects
<ul>
<li style="font-size: .9em; margin-bottom: .45em">Test mutants from the Consortium for defects in
chemotaxis & motility
</li>
</ul>
</li>
<li>
If prior work is promising confirm hypothesis in an animal model
</li>
</ul>
</section> -->
<section>
<section>
<h3 style="color:darkorange; ">What does that imply?</h3>
<div style="height: 700px; vertical-align: center; margin-top: 0%">
<img src="resources/lower_intestine2.png" alt style="background:none; border:none; height: 80%; box-shadow:none; ">
</div>
</section>
<section>
<div style="height: 700px; vertical-align: center">
<img src="resources/suppl_Fig_8.png" alt
style="background:none; border:none; box-shadow:none;">
</div>
</section>
</section>
<section>
<h3 style="color:darkorange; ">Minimal Motility Media</h3>
<p>
Find the minimum set of additives<br>which sustain the best population wide motility
</p>
<div style="float: left; width: 45%; align-items: center; display: table; justify-content: center; height: 450px; margin-left: 40px">
<ul style="font-size: .9em">
<li>Amino acids:
<ul>
<li class="fragment strike" data-fragment-index="1">Tryptophane</li>
<li class="fragment strike" data-fragment-index="1">Methionine</li>
<li class="fragment strike" data-fragment-index="1">Leucine</li>
<li class="fragment strike" data-fragment-index="1">Valine</li>
<li>Proline</li>
<li>Isoleucine</li>
<li>Cysteine</li>
</ul>
</li>
</ul>
</div>
<div style="float: right; width: 45%; align-items: center; display: table; justify-content: center; height: 450px; margin-right: 40px">
<ul style="font-size: .9em">
<li class="fragment strike" data-fragment-index="1">Salts</li>
<li class="fragment strike" data-fragment-index="1">Trace salts</li>
<li class="fragment strike" data-fragment-index="1">Trace elements</li>
<li class="fragment strike" data-fragment-index="1">Iron</li>
<li>Glucose</li>
</ul>
</div>
</section>
<section>
<h3 style="color:darkorange; ">Minimal Motility Media</h3>
<img data-src="resources/220404_violin_plots.png" alt
style="background:none; border:none; box-shadow:none;">
</section>
<section>
<h3 style="color:darkorange; ">Motility over time</h3>
<img data-src="resources/220403_1min_vid_overview.png" alt
style="background:none; border:none; box-shadow:none;">
</section>
<section>
<section>
<div style="position: relative; height: 720px; left: -612px;">
<img src="resources/mot_model1.png" alt style="position: absolute; background:none; border:none; box-shadow:none;">
<img src="resources/mot_model2.png" class="fragment" alt style="position: absolute; background:none; border:none; box-shadow:none;">
<img src="resources/mot_model3.png" class="fragment" alt style="position: absolute; background:none; border:none; box-shadow:none;">
</div>
<aside class="notes">
Cysteine export<br>
</aside>
</section>
<section>
<h3 style="color:darkorange; ">Computer Model</h3>
<div style="height: 500px; vertical-align: center; margin-top: 0%">
<img data-src="resources/210518_wat0.png" alt
style="background:none; border:none; box-shadow:none;">
</div>
</section>
</section>
<section>
<div style="margin: auto; width: 1000px; text-align: center"> <!-- text-shadow: 1px 1px 1px black; -->
<h3 style="color: darkorange">Thank you for your Attention</h3>
</div>
<h4 style="">Special thanks to:</h4> <!-- text-shadow: 1px 1px 1px black; -->
<p style="font-size: .8em">
<b>Friends & Colleagues:</b><br>
Emi,
Pia,
Rosi,
Sabrina,
Rike,
Ruben,
Vincent,
Anastasia,
Tenzin,
Noémie,
Annika,
Cera,
Avi,
Maik,
Ruth,
Hr. Zickenrott
</p>
<p style="font-size: .8em">
<b>Collaborators:</b><br>
Prof. Rolf Daniel,
Dr. Wolfgang Bohne,
Prof. Fabian M. Commichau,
Jerôme Dretzke,
Carina Fischer,
Katrina Funkner,
Nadine Gänstaller,
Prof. Uwe Groß,
PD Dr. Michael Hoppert,
Igor Khassanov,
Anna Klassen,
Kelly Libnik,
Prof. Burkhard Morgenstern,
<b style="color: deeppink;">Ines Oehmig</b>,
Dr. Jörg Overmann,
Prof. Jörg Stülke,
Prof. Andreas E. Zautner
</p>
<div style="font-size:.7em; ">
<p>This Project was funded by the Federal State of Lower Saxony<br>
Niedersächsisches Vorab (VWZN2889/3215/3266)</p>
</div>
</section>
</div>
</div>
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