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− | <nav style='font-family:Lato;font-size:22px;background-color:# | + | <nav style='font-family:Lato;font-size:22px;background-color:#1C77C3;-webkit-transform: translateZ(0);z-index: 100;position: fixed;box-shadow: 0px 0px 10px black;'> |
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− | <a href="https:// | + | <a href="https://2016.igem.org/Team:TAS_Taipei/project"><h4 class="dropdown-toggle disabled" data-toggle="dropdown"><b>PROJECT</b></h4></a> |
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− | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/ | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Description#background">Background</a></h5> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Achievements">Achievements</a></h5> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Collaborations">Collaborations</a></h5> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Parts">Parts</a></h5> |
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<div style='height:140px;'> | <div style='height:140px;'> | ||
<img src="https://static.igem.org/mediawiki/2015/5/5b/Tas_icon_project.png"> | <img src="https://static.igem.org/mediawiki/2015/5/5b/Tas_icon_project.png"> | ||
− | <h4><b>Cataracts</b> - the leading cause of blindness. Find out how we can | + | <h4><b>Cataracts</b> - the leading cause of blindness. Find out how we can noninvasively <b>treat</b> and <b>prevent</b> cataract formation.</b></h4> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Experiments#lensmodel">Lens Cataract Model</a></h5> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Experiments#construct">Prevention and Treatment Constructs </a></h5> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Experiments#prototype">Delivery Prototype</a></h5> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Notebook">Notebook</a></h5> |
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<div style='height:140px;'> | <div style='height:140px;'> | ||
<img src="https://static.igem.org/mediawiki/2015/f/f8/Tas_icon_wetlab.png"> | <img src="https://static.igem.org/mediawiki/2015/f/f8/Tas_icon_wetlab.png"> | ||
− | <h4>We | + | <h4>We don't just come up with great ideas. We show they work. Follow along our discovery of exciting science!</h4> |
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</li> | </li> | ||
<li class="dropdown"> | <li class="dropdown"> | ||
− | <a href="https:// | + | <a href="https://2016.igem.org/Team:TAS_Taipei/Modeling"><h4 class='dropdown-toggle disabled' data-toggle="dropdown"><b>MODEL</b></h4></a> |
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− | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/ | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Model#crystallin">Cataract Damage</a></h5> |
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− | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/ | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Model#gsr25hc">GSR/25HC Pathway</a></h5> |
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− | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/ | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Model#nanoparticle">Nanoparticle Degradation</a></h5> |
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− | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/ | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Model#eyedrop">Eyedrop Model</a></h5> |
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− | <h4><b>Computational Biology</b> provides us models that we cannot easily test. Click to | + | <h4><b>Computational Biology</b> provides us models that we cannot easily test. Click to find out the results of our modeling, and if you want, the math behind it!</h4> |
</div> | </div> | ||
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− | <a href="https:// | + | <a href="https://2016.igem.org/Team:TAS_Taipei/Practices"><h4 class='dropdown-toggle disabled' data-toggle="dropdown"><b>HUMAN PRACTICE</b></h4></a> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Human_Practices#research">Research</a></h5> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Human_Practices#outreach">Outreach</a></h5> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Human_Practices#impact">Impact</a></h5> |
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− | <a href="https:// | + | <a href="https://2016.igem.org/Team:TAS_Taipei/Safety"><h4 class='dropdown-toggle disabled' data-toggle="dropdown"><b>BIOSAFETY</b></h4></a> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Safety#researcher_safety">Researcher Safety</a></h5> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Safety#environmental_safety">Environmental Safety</a></h5> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Safety#biobrick_safety">Biobrick Safety</a></h5> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Safety#local_safety">Local Safety</a></h5> |
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− | <a href="https:// | + | <a href="https://2016.igem.org/Team:TAS_Taipei/Team"><h4 class='dropdown-toggle disabled' data-toggle="dropdown"><b>TEAM</b></h4></a> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Team#members">Members</a></h5> |
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− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/team#tas">About</a></h5> |
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<div> | <div> | ||
− | <h5><a href="https:// | + | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Attributions">Attributions</a></h5> |
+ | </div> | ||
+ | <div> | ||
+ | <h5><a href="https://2016.igem.org/Team:TAS_Taipei/Standard_Pages">Wiki Standard Pages</a></h5> | ||
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<div style='height:140px;'> | <div style='height:140px;'> | ||
<img src="https://static.igem.org/mediawiki/2015/3/34/Tas_icon_team.png"> | <img src="https://static.igem.org/mediawiki/2015/3/34/Tas_icon_team.png"> | ||
− | <h4>Behind every tough iGem project lies a | + | <h4>Behind every tough iGem project lies a hard-working yet cheerful group of students. <b>Meet the team!</b></h4> |
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<h2 style="font-family:'Lato';letter-spacing:10px;color: white; font-size: 60px; margin-top: 0; margin-bottom: 0;"><b> | <h2 style="font-family:'Lato';letter-spacing:10px;color: white; font-size: 60px; margin-top: 0; margin-bottom: 0;"><b> | ||
− | + | C◎UNTERACTS</b></h2> | |
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<li><a href="#overview">Overview</a></li> | <li><a href="#overview">Overview</a></li> | ||
+ | <li><a href="#crystallin">Crystallin</a></li> | ||
<li><a href="#gsr25hc">GSR/25HC</a></li> | <li><a href="#gsr25hc">GSR/25HC</a></li> | ||
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<li><a href="#nanoparticle">Nanoparticles</a></li> | <li><a href="#nanoparticle">Nanoparticles</a></li> | ||
− | <li><a href="# | + | <li><a href="#eyedrop">Eyedrop</a></li> |
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<h1 id='overview'>Modeling</h1> | <h1 id='overview'>Modeling</h1> | ||
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− | <p> | + | |
− | <p> | + | <div class="row"> |
− | < | + | <div class="col-sm-6" > |
− | + | <div class="col-sm-12"> | |
− | + | <h3> Abstract </h4> | |
− | + | <p>We answer two questions: How much GSR to maintain in the lens, and how to maintain that amount? We find the amount of GSR needed in the lens (Model 2) to limit crystallin damage so the resulting cataract is less than LOCS 2.5 (Model 1). Then, we find the optimal design of eyedrops (Model 4) and nanoparticles that will maintain this amount of GSR in the lens (Model 3). These models allow our team to understand the impact of adding GSR-loaded nanoparticles into the lens, and to design a full treatment plan on how to prevent and treat cataracts. </p> | |
+ | </div> | ||
+ | </div> | ||
+ | <div class="col-sm-6" > | ||
+ | <div class="col-sm-12"> | ||
+ | <h3>Achievements</h4> | ||
+ | <ul style="font-size:15px"> | ||
+ | <li>Designed a simple calculator to find amount of GSR or 25HC eyedrops needed for a patient's LOCS score.</li> | ||
+ | <li>Bridged the gap between the medical, biological, and chemical measurement of crystallin damage.</li> | ||
+ | <li>Predicted impact of adding GSR and 25HC on the amount of crystallin damage in the lens.</li> | ||
+ | <li>Created Nanoparticle Customizer for doctors to find a full treatment plan.</li> | ||
+ | <li>Generalized our Customizer to allow other iGEM teams who wish to use nanoparticle drug delivery </li> | ||
+ | <li>Analyzed sensitivity of prototype, and suggested insights into optimal manufacturing and clinical use of our prototype.</li> | ||
+ | <li>Used Experimental data to develop Models 1 and 3.</li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <h3> Outline </h3> | ||
+ | |||
+ | |||
+ | <h2>Introduction</h2> | ||
+ | |||
+ | <h3> Why Model? </h3> | ||
+ | <p>In the lab, biologists are often unable to test everything experimentally. For example, in our cataracts project, cataract prevention occurs in the long-term, from 20-50 years. Obviously, although short experiments can provide us an idea of what prevention may look like, the power of computational biology allows us to model into the future. As a result, our modeling has been crucial in developing a prototype.</p> | ||
+ | |||
+ | <h3> Focus </h3> | ||
+ | <p>Most iGEM teams perform modeling on gene expression, which we accomplish in model 5. However, as our construct is not directly placed into the eyes, how our synthesized protein impacts the eyes after it is seperately transported is much more interesting. As a result, we spent the majority of our models on understanding the impacts on the eye.</p> | ||
+ | |||
+ | <h3>Guiding Questions </h3> | ||
+ | <ol> | ||
+ | <li>How much GSR do we want inside the lens?</li> | ||
+ | <li>How do we use nanoparticles to control the amount of GSR in the lens?</li> | ||
+ | <li>How do we synthesize GSR, package into NP, and send it into the eye?</li> | ||
+ | </ol> | ||
+ | |||
</div> | </div> | ||
+ | </div> | ||
− | + | ||
− | + | ||
− | + | <div class = "row"> | |
− | + | <div class="col-sm-12"> | |
− | + | <h2 id = 'crystallin'>Model 1: Crystallin Damage</h2> | |
− | + | <h3>Abstract</h3> | |
+ | <p>In our experiments, absorbance measurements are meaningless without understanding how severe a cataract that absorbance measurement means. We use literature research to relate LOCS, the physician's scale of cataract severity) to absorbance, which is how we quantified crystallin damage in experiments. We use experimental data to understand how crystallin damage can be quantified by measuring absorbance. With this model, we can calculate how much crystallin damage we have to limit to reduce LOCS to an acceptable level.</p> | ||
+ | |||
+ | <h3> Purpose </h3> | ||
+ | <p> How much do we need to limit crystallin damage so surgery is not needed? </p> | ||
+ | |||
+ | <h3>Measurement of Cataract Severity</h3> | ||
+ | <div class="row"> | ||
+ | <div class="col-sm-12"> | ||
+ | <p> | ||
+ | There are four ways of measuring cataract severity, each used for a different purpose. | ||
+ | <ol> | ||
+ | <li><b>Lens Optical Cataract Scale (LOCS):</b> Physicians use this scale, from 0 – 6, to grade the severity of cataracts.</li> | ||
+ | <li><b>Opacity (%):</b> This is the physical, quantitative property of the LOC scale.</li> | ||
+ | <li><b>Absorbance at 397.5 nm:</b> This is the experimental method, used by our team in the lab (c.d.).</li> | ||
+ | <li><b>Crystallin Damage: </b>This is a chemical definition. We quantify cataract severity as a function of how much oxidizing agents there are, as well as how long crystalline is exposed to oxidizing agents. We define 1 crystallin damage unit as the damage done to human crystallin when exposed to 1 M hydrogen peroxide, the main oxidizing agent, for 1 hour.</li> | ||
+ | </ol> | ||
+ | |||
+ | </p> | ||
+ | |||
+ | <</div> | ||
+ | </div> | ||
+ | <p>We use the unit of crystallin damage to connect cataract severity with the amount of GSR we add (in Model 2). We want to lower c.d. below so that the resulting cataract is of LOCS 2.5. For the rest of the model, our task is simple: relate each point of the LOCS scale to c.d., in order to connect to Model 2.</p> | ||
+ | |||
+ | <h3>LOCS Equivalence to Absorbance: Literature Research</h3> | ||
+ | <p>Past studies have done numerous studies on how absorbance measurements can be converted to the LOC scale that physicians end up using. With the results of ________ and ________, we construct the first three columns in Table 2.</p> | ||
+ | |||
+ | <h3>Absorbance Equivalence to Crystallin Damage: Experimental Data</h3> | ||
+ | <div class="row"> | ||
+ | <div class="col-sm-7"> | ||
+ | <div class="col-sm-12" > | ||
+ | <p> | ||
+ | We use experimental data from our team’s Cataract Lens Model (link). In each trial, they added H2O2 to crystallin, and measured the resulting absorbance. The data used are shown in Table 1. We can calculate the theoretical c.d., and graph absorbance vs. crystallin damage in Figure 2. | ||
+ | </p> | ||
+ | <p>With this relation in Figure 2, we calculate the equivalent crystallin damage to each LOCS rating and its equivalent absorbance.</p> | ||
+ | <h3>Error Analysis</h3> | ||
+ | <p>It may be surprising that only around 1 M-h is required to induce moderately severe cataracts. Remember that this is done in the absence of antioxidation systems (GSR) and at an extremely high oxidizing concentration of H2O2 (1M of H2O2). In the lens, H2O2 has a much lower concentration, so severe cataracts are induced over months to years.</p> | ||
+ | <p>There are some limitations of the model that arise from our assumptions. We assume that fish and human lens contain similar crystallin proteins that are degraded in the similar manner (Assumption 4). In addition, we made a rough adjustment of data based our diluting procedure. For better results to create a human cataract model, experiments will need to be done on human lens, even better if done in vitro, without any dilutions.</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="col-sm-5" style="margin:0px"> | ||
+ | <div class="col-sm-12"> | ||
+ | <p> | ||
+ | <table class="table table-bordered" style='width: 80%;margin-left:0%;'> | ||
+ | <caption style='caption-side:top;'><b>Table 2: Results of Model 1 – Equivalent values for LOCS, Opacity, Absorbance, and Crystallin Damage.</caption> | ||
+ | <thead> | ||
+ | <tr> | ||
+ | <th>LOCS</th> | ||
+ | <th>Opacity (%)</th> | ||
+ | <th>Absorbance (@397.5 nm)</th> | ||
+ | <th>Crystallin Damage (M-h)</th> | ||
+ | </tr> | ||
+ | </thead> | ||
+ | <tr> | ||
+ | <th>0.0</th> | ||
+ | <th>0.00</th> | ||
+ | <th>0.0000</th> | ||
+ | <th>0.0000</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>0.5</th> | ||
+ | <th>3.24</th> | ||
+ | <th>0.0143</th> | ||
+ | <th>0.1327</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>1.0</th> | ||
+ | <th>6.65</th> | ||
+ | <th>0.0299</th> | ||
+ | <th>0.2774</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>1.5</th> | ||
+ | <th>10.81</th> | ||
+ | <th>0.0497</th> | ||
+ | <th>0.4610</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>2.0</th> | ||
+ | <th>15.88</th> | ||
+ | <th>0.0751</th> | ||
+ | <th>0.6966</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>2.5</th> | ||
+ | <th>21.95</th> | ||
+ | <th>0.1076</th> | ||
+ | <th>0.9981</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>3.0</th> | ||
+ | <th>29.07</th> | ||
+ | <th>0.1492</th> | ||
+ | <th>1.3840</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>4.0</th> | ||
+ | <th>46.37</th> | ||
+ | <th>0.2706</th> | ||
+ | <th>2.5101</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <th>5.0</th> | ||
+ | <th>66.05</th> | ||
+ | <th>0.4691</th> | ||
+ | <th>4.3514</th> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <button class="accordion">Background, Method, Results, Discussion</button> | ||
+ | <div class="panel"> | ||
+ | |||
+ | <div class="accordionmenu1" class ="col-sm-12" > | ||
+ | <ul class="nav nav-tabs"> | ||
+ | <li class="active"><a data-toggle="tab" href="#cryshome">Background</a></li> | ||
+ | <li><a data-toggle="tab" href="#crysmenu1">Assumptions</a></li> | ||
+ | <li><a data-toggle="tab" href="#crysmenu2">Procedure</a></li> | ||
+ | <li><a data-toggle="tab" href="#crysmenu3">Results</a></li> | ||
+ | <li><a data-toggle="tab" href="#crysmenu4">Discussion</a></li> | ||
+ | </ul> | ||
+ | |||
+ | <div class="tab-content"> | ||
+ | <div id="cryshome" class="tab-pane fade in active"> | ||
+ | <h3>Background</h3> | ||
+ | <div class="row"> | ||
+ | <div class="col-sm-8"> | ||
+ | <div class="col-sm-12" style="border:1px solid black"> | ||
+ | <p>There are four ways to measure cataract severity (how blurred the lens is): | ||
+ | <ol> | ||
+ | <li>Lens Optical Cataract Scale III (LOCS) - a scale from 0-6 used by physicians.</li> | ||
+ | <li>Opacity (%) - used to calculate the LOC scale</li> | ||
+ | <li>Absorbance at 397.5 nm - measurable in the lab. </li> | ||
+ | <li>Crystallin Damage - used to quantify how much crystallin has been reacted with hydrogen peroxide to create insoluble, damaged crystallin. The following definition of crystallin damage is used:</li> | ||
+ | </ol> | ||
+ | \[c.d.(t) = \int_{0}^{\infty} [H_2O_2]_t dt\] | ||
+ | </p> | ||
+ | |||
+ | <p> In other words, 1 unit of crystallin damage, in M-h, is equal to the damage caused by 1 molar concentration of hydrogen peroxide reacting crystallin in the eyes for 1 hour.</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="col-sm-4" style="background-color:lightpink;margin:0px"> | ||
+ | <div class="col-sm-12" style="border:1px solid black"> | ||
+ | <h3>LOCS Scale</h3><br> <br> <br><br> <br> <br> <br><br><br><br> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | </div> | ||
+ | |||
+ | <div id="crysmenu1" class="tab-pane fade"> | ||
+ | <h3>Assumptions</h3> | ||
+ | <div class="row"> | ||
+ | <div class="col-sm-12"> | ||
+ | <div class="col-sm-12"> | ||
+ | <p> | ||
+ | <ol> | ||
+ | <li>Definition of crystallin damage: Crystallin damage is proportional to the concentration of hydrogen peroxide, and the time of exposure. This is a valid assumption, supported by the fact that the reaction between cysteine (molecules on crystallin) and hydrogen peroxide is linear. </li> | ||
+ | <li>We assume that the amount of crystallin is far greater than the amount oxidized. Our product is meant for long-term cataract prevention and minor treatment, and is not suggested for patients with extremely severe cataracts. </li> | ||
+ | <li>When the experiments diluted the cataract lens protein, the amount of crystallin is diluted. However, the final absorbance of degraded crystallin is also diluted, so we assume any errors in absorbance is canceled out.</li> | ||
+ | <li>We assume that fish and human lens contain similar crystallin proteins.</li> | ||
+ | |||
+ | |||
+ | </ol> | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | </div> | ||
+ | <div id="crysmenu2" class="tab-pane fade"> | ||
+ | <h3>Procedure</h3> | ||
+ | |||
+ | <div class="row"> | ||
+ | <div class="col-sm-8"> | ||
+ | <div class="col-sm-12" style="border:1px solid black"> | ||
+ | <p> | ||
+ | <ol> | ||
+ | <li>In the first part, we find how the absorbance measurements in the lab are related to the severity of the cataracts. Through literature data, we can relate LOCS to the opacity of the lens. Then, via physical calculations, we can relate the opacity of the lens to the absorbance of the lens at 400 nm.</li> | ||
+ | <li>Then, we use our team’s experimental data in the cataract model. For each trial, the concentration of H2O2 and the length of exposure are given, so we can calculate the theoretical crystallin damage using the definition above and the assumptions we made. In each trial we also measured the absorbance, so we have a relation between crystallin damage and absorbance. </li> | ||
+ | <li>However, we need to make a minor adjustment, because absorbance is affected by dilution. When the fish lens was isolated, they were placed in Tris buffer and diluted. We calculate the ratio of volumes from diluted volume to the Tris buffer, and multiply each absorbance measurement by this value.</li> | ||
+ | </ol> | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="col-sm-4" style="background-color:lightpink;margin:0px"> | ||
+ | <div class="col-sm-12" style="border:1px solid black"> | ||
+ | <h3>LOCS Scale</h3><br> <br> <br><br> <br> <br> <br><br><br><br> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | <div id="crysmenu3" class="tab-pane fade"> | ||
+ | <h3>Results</h3> | ||
+ | |||
+ | <table class="table table-bordered" style='width: 70%;margin-left:15%;'> | ||
+ | <caption style='caption-side:top;'><b>Table 1: Results of Model 1 - Equivalent values for LOCS, Opacity, Absorbance, and Crystallin Damage. </b> </caption> | ||
<tbody> | <tbody> | ||
<tr> | <tr> | ||
− | <td> | + | <td>LOCS</td> |
− | <td>0. | + | <td>0.0</td> |
− | <td> | + | <td>0.5</td> |
− | <td> | + | <td>1.0</td> |
− | + | <td>1.5</td> | |
− | + | <td>2.0</td> | |
− | <td> | + | <td>2.5</td> |
− | + | <td>3.0</td> | |
− | <td> | + | <td>4.0</td> |
− | <td> | + | <td>5.0</td> |
− | <td> | + | |
− | + | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
− | <td> | + | <td>Degree</td> |
− | <td> | + | <td>None</td> |
− | <td> | + | <td colspan="2">Trace</td> |
− | <td> | + | <td colspan="3">Mild</td> |
− | <td> | + | <td colspan="2">Moderate</td> |
− | <td> | + | <td>Severe</td> |
− | <td> | + | </tr> |
− | <td> | + | <tr> |
− | <td> | + | <td>Opacity (%)</td> |
− | <td> | + | <td>0</td> |
− | <td> | + | <td>3.24</td> |
− | <td>4</td> | + | <td>6.65</td> |
+ | <td>10.81</td> | ||
+ | <td>15.88</td> | ||
+ | <td>21.95</td> | ||
+ | <td>29.07</td> | ||
+ | <td>46.37</td> | ||
+ | <td>66.05</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Absorbance (a.u.)</td> | ||
+ | <td>0.0000</td> | ||
+ | <td>0.0143</td> | ||
+ | <td>0.0299</td> | ||
+ | <td>0.0497</td> | ||
+ | <td>0.0751</td> | ||
+ | <td>0.1076</td> | ||
+ | <td>0.1492</td> | ||
+ | <td>0.2706</td> | ||
+ | <td>0.4691</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Crystallin Damage (c.d.)</td> | ||
+ | <td>0.0000</td> | ||
+ | <td>0.1327</td> | ||
+ | <td>0.2774</td> | ||
+ | <td>0.4610</td> | ||
+ | <td>0.6966</td> | ||
+ | <td>0.9981</td> | ||
+ | <td>1.3840</td> | ||
+ | <td>2.5101</td> | ||
+ | <td>4.3514</td> | ||
</tr> | </tr> | ||
</tbody> | </tbody> | ||
− | + | </table> | |
− | + | <br><br><br> | |
− | + | <table class="table table-bordered" style='width: 70%;margin-left:15%;'> | |
− | + | <caption style='caption-side:top;'><b>Table 2: Experimental Data used for Model 1 from Cataract Lens Model (TAS) - Absorbance vs. Crystallin Damage </b></caption> | |
− | + | <tbody> | |
− | + | <thead> | |
− | + | <td>Trial</td> | |
− | + | <td>H2O2 Concentration (M)</td> | |
− | + | <td>Exposure Time (h)</td> | |
− | </div> | + | <td>Crystallin Damage (c.d.)</td> |
+ | <td>Measured Absorbance (abs @400 nm)</td> | ||
+ | </thead> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>0.100</td> | ||
+ | <td>24.0</td> | ||
+ | <td>2.40</td> | ||
+ | <td>0.105</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td>0.100</td> | ||
+ | <td>46.5</td> | ||
+ | <td>4.65</td> | ||
+ | <td>0.451</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td>0.100</td> | ||
+ | <td>72.0</td> | ||
+ | <td>7.20</td> | ||
+ | <td>0.0.695</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td>0.100</td> | ||
+ | <td>20.0</td> | ||
+ | <td>2.00</td> | ||
+ | <td>0.089</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>5</td> | ||
+ | <td>0.100</td> | ||
+ | <td>42.0</td> | ||
+ | <td>4.20</td> | ||
+ | <td>0.392</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>6</td> | ||
+ | <td>0.100</td> | ||
+ | <td>15.0</td> | ||
+ | <td>1.50</td> | ||
+ | <td>0.093</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>7</td> | ||
+ | <td>0.100</td> | ||
+ | <td>42.0</td> | ||
+ | <td>0.340</td> | ||
+ | <td>0.340</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>8</td> | ||
+ | <td>0.100</td> | ||
+ | <td>67.0</td> | ||
+ | <td>6.70</td> | ||
+ | <td>0.563</td> | ||
+ | </tr> | ||
+ | |||
+ | </tbody> | ||
+ | </table> | ||
+ | |||
+ | </div> | ||
− | + | <div id="crysmenu4" class="tab-pane fade"> | |
− | + | <h3>Discussion</h3> | |
− | + | <h4>Model Result</h4> | |
− | + | <div class="row"> | |
− | + | <div class="col-sm-6"> | |
− | + | <div class="col-sm-12" > | |
− | + | <p> | |
− | + | The model successfully relates LOCS, opacity of lens, absorbance measurements, and the equivalent crystallin damage of the lens. The purpose of relating LOCS to crystallin damage, is that in Model 2, we will use chemical kinetics to determine how adding GSR to the lens will decrease the amount of crystallin damage. Exactly how much crystallin damage we need to decrease is determined by the desired LOCS. For example, if we want to have a LOCS rating of less than 2.5, then we must lower crystalline damage to only 0.9981 M-h. | |
− | + | </p> | |
− | + | </div> | |
− | + | </div> | |
+ | |||
+ | <div class="col-sm-6" style="background-color:lightpink;margin:0px"> | ||
+ | <div class="col-sm-12"> | ||
+ | <p> | ||
+ | <br><br><br><br><br> | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | <h4>Model Adjustment</h4> | ||
+ | <div class="row"> | ||
+ | <div class="col-sm-6" style="background-color:lightpink;margin:0px"> | ||
+ | <div class="col-sm-12" > | ||
+ | <p> | ||
+ | <br><br><br><br><br> | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="col-sm-6" > | ||
+ | <div class="col-sm-12"> | ||
+ | <p> | ||
+ | When determining the relationship between absorbance and crystallin, in Figure 1 the best fit line has a x – intercept that is nonzero. However, when converting each absorbance rating to equivalent crystallin damage in Table 2, we ignore the constant term. When doing the experiments, the fish lens may have contained GSH that is still active, so the fact that the crystallin is exposed to H2O2, the degradation reaction does not happen until all GSH is depleted, and crystallin damage begins to form. We subtract around 1 unit of crystallin damage from all values. | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | <h4>Error Analysis</h4> | ||
+ | <div class="row"> | ||
+ | <div class="col-sm-12"> | ||
+ | <div class="col-sm-12" > | ||
+ | <p> | ||
+ | It may be surprising that only around 1 M-h is required to induce moderately severe cataracts. Remember that this is done in the absence of antioxidation systems (GSR) and at an extremely high oxidizing concentration of H2O2 (1M of H2O2). In the lens, H2O2 has a much lower concentration, so severe cataracts are induced over months to years. | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | </div> | ||
+ | |||
+ | <div class="row"> | ||
+ | <div class="col-sm-12"> | ||
+ | <div class="col-sm-12" > | ||
+ | <p> | ||
+ | There are some limitations of the model that arise from our assumptions. We assume that fish and human lens contain similar crystallin proteins that are degraded in the similar manner (Assumption 4). Also, to simplify the experiments, the lens were diluted in Tris buffer. Because of this dilution, the actual crystallin damage is much lower, but so is the actual absorbance. We assume that the decrease in crystallin damage and absorbance is the same, so no adjustments need to be made for the relation between crystallin damage and absorbance (Assumption 3). For better results to create a human cataract model, experiments will need to be done on human lens, even better if done in vitro, without any dilutions. | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | </div> | ||
+ | |||
+ | </div> | ||
− | + | </div> | |
− | + | </div> | |
− | <h2 id =" | + | |
− | + | ||
− | <caption style='caption-side:top;'><b>Table | + | |
− | < | + | </div> <!-- End Menu --> |
+ | |||
+ | |||
+ | <h3> Conclusion</h3> | ||
+ | <div class="row"> | ||
+ | <div class="col-sm-6" style="background-color:lightpink;margin:0px"> | ||
+ | <div class="col-sm-12" > | ||
+ | <p> | ||
+ | <br><br><br><br><br> | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="col-sm-6"> | ||
+ | <div class="col-sm-12"> | ||
+ | <p> | ||
+ | For surgery to not be needed, the LOCS value has to be below 2.5. This is equivalent to 21.95% in light opacity or 0.1076 abs units. Based on the results of our experiments, this is equivalent to 0.9981 units of crystallin damage, the damage done to crystallin if exposed to 0.9981 M of H2O2 for 1 hr. | ||
+ | For future models, this value 0.9981 units of c.d. will be called the crystallin damage threshold for LOCS 2.5. | ||
+ | |||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | </div> <!-- Container --> | ||
+ | </div> <!-- Container --> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <div class = "row"> | ||
+ | <div class="col-sm-12"> | ||
+ | <h2 id = 'gsr25hc'>Model 2: GSR/25HC Chemical Pathway</h2> | ||
+ | |||
+ | <div class="row"> | ||
+ | <div class="col-sm-6" > | ||
+ | <h3> Abstract </h3> | ||
+ | <p> The key question: <b>How much GSR to add?</b> Now that we know how much we need to limit crystallin damage, we use systems of ordinary differential equation to model the GSR Pathway. We calculate the necessary GSR concentration to be maintained over 50 years so that the resulting cataract is below LOCS 2.5./p> | ||
+ | </p> | ||
+ | </div> | ||
+ | <div class="col-sm-6" > | ||
+ | <h3> Purpose </h3> | ||
+ | <p> How much GSR do we need to maintain in the lens so that the crystallin damage recorded over 50 years is below the threshold for LOCS 2.5? </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | <h3> Chemical Kinetics Model: Differential Equations </h3> | ||
+ | <div class="row"> | ||
+ | <div class="col-sm-12" > | ||
+ | <p> By the Law of Mass Action, Michaelis-Menten Enzyme kinetics, Ping-pong mechanism, and the Law of Passive Diffusion, we build a system of 10 differential equations based on 6 chemical reactions. All parameters, constants, and initial conditions are based off literature data. Estimates made are also shown with assumptions and reasoning. The details are shown in the collapsible for interested readers. </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="row"> | ||
+ | <div class="col-sm-6" > | ||
+ | <h3>Blackbox Approach: Testing GSR Impact </h3> | ||
+ | <p> Image</p> | ||
+ | </div> | ||
+ | <div class="col-sm-6" > | ||
+ | <h3> </h3> | ||
+ | <p> We will vary the input, Initial GSR concentration, from 0 to 100 uM, holding all other variables constant, and numerically solve for the amount of hydrogen peroxide over time. With this graph, we can find the amount of crystallin damage accumulated over 20 to 50 years if different levels of GSR is maintained. </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <button class="accordion">Background, Method, Results, Discussion</button> | ||
+ | <div class="panel"> | ||
+ | |||
+ | <div class="accordionmenu1" class ="col-sm-12" > | ||
+ | <ul class="nav nav-tabs"> | ||
+ | <li class="active"><a data-toggle="tab" href="#gsrhome">Background</a></li> | ||
+ | <li><a data-toggle="tab" href="#gsrmenu1">Method</a></li> | ||
+ | <li><a data-toggle="tab" href="#gsrmenu2">Results Part 1</a></li> | ||
+ | <li><a data-toggle="tab" href="#gsrmenu2">Results Part 2</a></li> | ||
+ | <li><a data-toggle="tab" href="#gsrmenu3">Discussion</a></li> | ||
+ | </ul> | ||
+ | |||
+ | <div class="tab-content"> | ||
+ | <div id="gsrhome" class="tab-pane fade in active"> | ||
+ | <h4>Background</h4> | ||
+ | |||
+ | <p> The following 6 reactions describe the antioxidant system inside the cortex and nucleus.</p> | ||
+ | |||
+ | $$GP_{xr}+[H_2O_2]_{in}+H^+ \xrightarrow[]{k_1} GP_{xo}+H_2O ...(1)$$ | ||
+ | $$GP_{xo}+GSH+H^+ \xrightarrow[]{k_2} [GS-GP_x]+H_2O ...(2)$$ | ||
+ | $$[GS-GP_x] + GSH \xrightarrow[]{k_3} GP_{xr}+GSSG+H^+ ...(3)$$ | ||
+ | $$NADPH \xrightarrow[GSR]{k4, K_{4M}} NADP^+ ...(4)$$ | ||
+ | $$GSSG \xrightarrow[GSR']{k_5, K_{5M}} 2GSH ...(5)$$ | ||
+ | $$[H_2O_2]_{out} \xrightarrow[]{k_5} [H_2O_2]_{in} ...(6)$$ | ||
+ | |||
+ | <p> Each reaction will be discussed in detail, and we will derive rate equations.</p> | ||
+ | |||
+ | <p><b>Reaction 1:</b>As hydrogen ions are numerous are negligible in the reaction, we will ignore it. By the <b>law of mass</b> action, the rate of this reaction is: $$r_1=k_1[GP_{xr}][H_2O_2]_{in}$$</p> | ||
+ | |||
+ | <p><b>Reaction 2:</b> By the <b>law of mass</b> action, the rate of this reaction is: $$r_2=k_2[GP_{xo}][GSH]$$</p> | ||
+ | |||
+ | <p><b>Reaction 3:</b> By the <b>law of mass</b> action, the rate of this reaction is: $$r_3=k_3[GS-GP_x][GSH]$$</p> | ||
+ | |||
+ | <p><b>Summary of Reaction 1-3:</b>In these reactions, hydrogen peroxide is reduced to water. GSH is consumed to recycle the enzyme GPx back into reduced form, to neutralize more hydrogen peroxide.</p> | ||
+ | <br> | ||
+ | |||
+ | <p><b>Reaction 4:</b> By<b>Michaelis-Menten kinetics </b>and the <b>Ping-Pong mechanism</b>, the rate of this reaction, with rate constant k4, and Michaelis-Menten constant Km4, is: $$r_4=k_4\frac{[NADPH]}{K_{4M}+[NADPH]}$$</p> | ||
+ | |||
+ | <p><b>Reaction 5:</b> By<b>Michaelis-Menten kinetics </b>and the <b>Ping-Pong mechanism</b>, the rate of this reaction, with rate constant k5, and Michaelis-Menten constant Km5, is: $$r_5=k_5\frac{[GSSG]}{K_{4M}+[GSSG]}$$</p> | ||
+ | |||
+ | <p><b>Summary of Reaction 4-5</b>: In these, GSSG is reduced back to form GSH, using the enzyme GSR. This is necessary for antioxidation to continue as reaction 1 is constantly using GSH, converting them to GSSG.</p> | ||
+ | <br> | ||
+ | |||
+ | <p><b>Reaction 6:</b> By the <b>Law of Passive Diffusion</b>, the rate of diffusion into the cortex and lens is: is: $$r_6=k_6([H_2O_2]_{out}-[H_2O_2]_{in})$$</p> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | <div id="gsrmenu1" class="tab-pane fade"> | ||
+ | <h4>Differential Equations</h4> | ||
+ | <p>We have six reaction rates derived from above. Now, we will form differential equations, where every time a species is used as a reactant, the reaction rate will be subtracted from the species’ derivative, while each time it is formed as a product, the reaction rate will be added to the species’ derivative. We will go through each species in detail:</p> | ||
+ | <p>Substituting the rate of each reaction, we get the following system of differential equations. </p> | ||
+ | <p> | ||
+ | $$\frac{d[GP_{xr}]}{dt}=k_3[GS-GP_x][GSH]-k_1[GP_{xr}][H_2O_2]_{in}$$ | ||
+ | |||
+ | $$\frac{d[H_2O_2]}{dt}=k_6[[H_2O_2]_{out}-{H_2O_2]_{in}]-k_1[GP_{xr}][H_2O_2]_{in}$$ | ||
+ | |||
+ | $$\frac{d[H_2O_2]}{dt}=k_1([H_2O_2]_{out} - [H_2O_2]_{in})-k_1[GP_{xr}][H_2O_2]_{in}$$ | ||
+ | |||
+ | $$\frac{d[GP_{x0}]}{dt}=k_1[GP_{xr}][H_2O_2]_{in}-k_2[GP_{xo}][GSH]$$ | ||
+ | |||
+ | $$\frac{d[H_2O]}{dt}=k_1[GP_{xr}][H_2O_2]_{in}+k_2[GP_{xo}][GSH]$$ | ||
+ | |||
+ | $$\frac{d[GSH]}{dt}=2k_5[GSR']\frac{[GSSG]}{K_{5M}+[GSSG]}-k_2[GP_{xo}][GSH]-k_3[GS-GP_x][GSH]$$ | ||
+ | |||
+ | $$\frac{d[GS-GP_x]}{dt}=k_2[GP_{xo}][GSH]-k_3[GS-GP_x][GSH]$$ | ||
+ | |||
+ | $$\frac{d[GSSG]}{dt}=k_3[GS-GP_x][GSH]-k_5[GSR']\frac{[GSSG]}{K_{5M}+[GSSG]}$$ | ||
+ | |||
+ | $$\frac{d[NADPH]}{dt}=-k_4[GSR]\frac{[NADPH]}{K_{4M}+[NADPH]}$$ | ||
+ | |||
+ | $$\frac{d[GSR]}{dt}=k_5[GSR']\frac{[GSSG]}{K_{5M}+[GSSG]}-k_4[GSR]\frac{[NADPH]}{K_{4M}+[NADPH]}$$ | ||
+ | |||
+ | $$\frac{d[GSR']}{dt}=k_4[GSR]\frac{[NADPH]}{K_{4M}+[NADPH]}-k_5[GSR']\frac{[GSSG]}{K_{5M}+[GSSG]}$$ | ||
+ | |||
+ | </p> | ||
+ | |||
+ | |||
+ | </div> | ||
+ | <div id="gsrmenu2" class="tab-pane fade"> | ||
+ | <h4>Part 1 Results</h4> | ||
+ | |||
+ | <table class="table table-bordered" style='width: 70%;margin-left:15%;'> | ||
+ | <caption style='caption-side:top;'><b>Table 1: Data obtained from XXX<b> relating each value of the LOCS scale, to opacity values. </caption> | ||
+ | <tbody> | ||
<tr> | <tr> | ||
− | < | + | <td>LOCS</td> |
− | < | + | <td>0.0</td> |
− | < | + | <td>0.5</td> |
− | < | + | <td>1.0</td> |
+ | <td>2.0</td> | ||
+ | <td>3.0</td> | ||
+ | <td>4.0</td> | ||
+ | <td>5.0</td> | ||
+ | <td>6.0</td> | ||
</tr> | </tr> | ||
− | |||
− | |||
<tr> | <tr> | ||
− | <td> | + | <td>Degree</td> |
− | <td> | + | <td>None</td> |
− | <td> | + | <td></td> |
− | <td> | + | <td>Trace</td> |
+ | <td>Mild</td> | ||
+ | <td>Surgery Suggested</td> | ||
+ | <td>Moderate</td> | ||
+ | <td>Severe</td> | ||
+ | <td>Very Severe</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
− | <td> | + | <td>Opacity (%)</td> |
− | <td> | + | <td>0.34</td> |
− | <td>0. | + | <td>4.24</td> |
− | <td>0. | + | <td>5.80</td> |
+ | <td>18.88</td> | ||
+ | <td>23.60</td> | ||
+ | <td>49.14</td> | ||
+ | <td>65.61</td> | ||
+ | <td>90+</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Absorbance (a.u.)</td> | ||
+ | <td>0.001</td> | ||
+ | <td>0.019</td> | ||
+ | <td>0.026</td> | ||
+ | <td>0.091</td> | ||
+ | <td>0.117</td> | ||
+ | <td>0.294</td> | ||
+ | <td>0.464</td> | ||
+ | <td>1.3+</td> | ||
</tr> | </tr> | ||
</tbody> | </tbody> | ||
</table> | </table> | ||
− | + | <p> We wish to remain below clinically significant levels, so we will reach attempt to lower the LOCS rating of a cataract to below grade 2.5, which means we want to control GSR such that the crystallin damage results in less than 0.108 a.u. at absorbance at 397.5 nm. </p> | |
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | + | ||
− | < | + | </div> |
− | + | <div id="gsrmenu3" class="tab-pane fade"> | |
− | + | ||
− | + | </div> | |
− | + | ||
− | + | <div id="gsrmenu4" class="tab-pane fade"> | |
− | + | <h4>Menu 3</h4> | |
− | + | <p>Eaque ipsa quae ab illo inventore veritatis et quasi architecto beatae vitae dicta sunt explicabo.</p> | |
− | <caption style='caption-side:top;'><b>Table | + | </div> |
− | < | + | |
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | </div> <!-- End Menu --> | ||
+ | |||
+ | <h3></h3> | ||
+ | <div class="row"> | ||
+ | <div class="col-sm-6"> | ||
+ | <div class="col-sm-12" > | ||
+ | <p> | ||
+ | <br><br><br><br><br> | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <div class="col-sm-6" > | ||
+ | <div class="col-sm-12"> | ||
+ | <p> | ||
+ | When determining the relationship between absorbance and crystallin, in Figure 1 the best fit line has a x – intercept that is nonzero. However, when converting each absorbance rating to equivalent crystallin damage in Table 2, we ignore the constant term. When doing the experiments, the fish lens may have contained GSH that is still active, so the fact that the crystallin is exposed to H2O2, the degradation reaction does not happen until all GSH is depleted, and crystallin damage begins to form. We subtract around 1 unit of crystallin damage from all values. | ||
+ | </p> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <h4> Conclusion</h4> | ||
+ | <p> Conclusion</p> | ||
+ | |||
+ | |||
+ | |||
+ | </div> <!-- Container --> | ||
+ | </div> <!-- Container --> | ||
+ | |||
+ | |||
+ | |||
+ | <div class = "row"> | ||
+ | <div class="col-sm-12"> | ||
+ | <h2 id = 'nanoparticle'>Model 3: Nanoparticles</h2> | ||
+ | <h4> Abstract </h4> | ||
+ | <p> Abstract </p> | ||
+ | |||
+ | <h4> Purpose </h4> | ||
+ | <p> Purpose </p> | ||
+ | |||
+ | |||
+ | |||
+ | <button class="accordion">Background, Method, Results, Discussion</button> | ||
+ | <div class="panel"> | ||
+ | |||
+ | <div class="accordionmenu1" class ="col-sm-12" > | ||
+ | <ul class="nav nav-tabs"> | ||
+ | <li class="active"><a data-toggle="tab" href="#nphome">Background</a></li> | ||
+ | <li><a data-toggle="tab" href="#npmenu1">Method</a></li> | ||
+ | <li><a data-toggle="tab" href="#npmenu2">Results Part 1</a></li> | ||
+ | <li><a data-toggle="tab" href="#npmenu2">Results Part 2</a></li> | ||
+ | <li><a data-toggle="tab" href="#npmenu3">Discussion</a></li> | ||
+ | </ul> | ||
+ | |||
+ | <div class="tab-content"> | ||
+ | <div id="home" class="tab-pane fade in active"> | ||
+ | <h4>Background</h4> | ||
+ | |||
+ | <p>There are three quantifiers of how severe cataract formation is, two are measurable, one is not. | ||
+ | <ol> | ||
+ | <li>Absorbance @ 397.5 nm, which is measured with lab equipment.</li> | ||
+ | <li>LOCS scale, subjectively measured by physicians on a scale from 0 - 6. | ||
+ | </li> | ||
+ | <li>Crystallin Damage, which we define as the following (for any time $t$) </li> | ||
+ | </ol> | ||
+ | |||
+ | </p> | ||
+ | |||
+ | \[c.d.(t) = \int_{0}^{\infty} [H_2O_2]_t dt\] | ||
+ | |||
+ | <p> In other words, 1 unit of crystallin damage, in M-h, is equal to the damage caused by 1 molar concentration of hydrogen peroxide reacting crystallin in the eyes for 1 hour.</p> | ||
+ | |||
+ | <p> In making this definition, we assume that crystallin damage is directly proportional to the amount of time crystallin is exposed to hydrogen peroxide. Hydrogen peroxide causes damage by forming disulfide bridges within cysteine molecules on crystallin. This changes the structure of crystallin, causing misfolding and cataract damage. Our linear assumption is valid because the rate for this reaction is first order with respect to hydrogen peroxide concentration. </p> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | <div id="npmenu1" class="tab-pane fade"> | ||
+ | <h4>Method</h4> | ||
+ | <p>We will relate the three by doing the following: | ||
+ | <ol> | ||
+ | <li>Relate LOCS scale to opacity via literature research. </li> | ||
+ | <li>Relate opacity to light transmittance via literature research. </li> | ||
+ | <li>Relate light transmittance to absorbance via physical calculations. </li> | ||
+ | <li>Relate absorbance to crystallin damage via experimental data. </li> | ||
+ | |||
+ | |||
+ | </ol> | ||
+ | </p> | ||
+ | |||
+ | |||
+ | </div> | ||
+ | <div id="npmenu2" class="tab-pane fade"> | ||
+ | <h4>Part 1 Results</h4> | ||
+ | |||
+ | <table class="table table-bordered" style='width: 70%;margin-left:15%;'> | ||
+ | <caption style='caption-side:top;'><b>Table 1: Data obtained from XXX<b> relating each value of the LOCS scale, to opacity values. </caption> | ||
+ | <tbody> | ||
<tr> | <tr> | ||
− | < | + | <td>LOCS</td> |
− | < | + | <td>0.0</td> |
− | < | + | <td>0.5</td> |
+ | <td>1.0</td> | ||
+ | <td>2.0</td> | ||
+ | <td>3.0</td> | ||
+ | <td>4.0</td> | ||
+ | <td>5.0</td> | ||
+ | <td>6.0</td> | ||
</tr> | </tr> | ||
− | </ | + | <tr> |
+ | <td>Degree</td> | ||
+ | <td>None</td> | ||
+ | <td></td> | ||
+ | <td>Trace</td> | ||
+ | <td>Mild</td> | ||
+ | <td>Surgery Suggested</td> | ||
+ | <td>Moderate</td> | ||
+ | <td>Severe</td> | ||
+ | <td>Very Severe</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Opacity (%)</td> | ||
+ | <td>0.34</td> | ||
+ | <td>4.24</td> | ||
+ | <td>5.80</td> | ||
+ | <td>18.88</td> | ||
+ | <td>23.60</td> | ||
+ | <td>49.14</td> | ||
+ | <td>65.61</td> | ||
+ | <td>90+</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Absorbance (a.u.)</td> | ||
+ | <td>0.001</td> | ||
+ | <td>0.019</td> | ||
+ | <td>0.026</td> | ||
+ | <td>0.091</td> | ||
+ | <td>0.117</td> | ||
+ | <td>0.294</td> | ||
+ | <td>0.464</td> | ||
+ | <td>1.3+</td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | <p> We wish to remain below clinically significant levels, so we will reach attempt to lower the LOCS rating of a cataract to below grade 2.5, which means we want to control GSR such that the crystallin damage results in less than 0.108 a.u. at absorbance at 397.5 nm. </p> | ||
+ | |||
+ | |||
+ | |||
+ | </div> | ||
+ | <div id="npmenu3" class="tab-pane fade"> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | <div id="npmenu4" class="tab-pane fade"> | ||
+ | <h4>Menu 3</h4> | ||
+ | <p>Eaque ipsa quae ab illo inventore veritatis et quasi architecto beatae vitae dicta sunt explicabo.</p> | ||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | </div> <!-- End Menu --> | ||
+ | |||
+ | |||
+ | <h4> Conclusion</h4> | ||
+ | <p> Conclusion </p> | ||
+ | |||
+ | |||
+ | |||
+ | </div> <!-- Container --> | ||
+ | </div> <!-- Container --> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <div class = "row"> | ||
+ | <div class="col-sm-12"> | ||
+ | <h2 id = 'eyedrop'>Model 4: Eyedrops</h2> | ||
+ | <h4> Abstract </h4> | ||
+ | <p> Abstract </p> | ||
+ | |||
+ | <h4> Purpose </h4> | ||
+ | <p> Purpose </p> | ||
+ | |||
+ | |||
+ | |||
+ | <button class="accordion">Background, Method, Results, Discussion</button> | ||
+ | <div class="panel"> | ||
+ | |||
+ | <div class="accordionmenu1" class ="col-sm-12" > | ||
+ | <ul class="nav nav-tabs"> | ||
+ | <li class="active"><a data-toggle="tab" href="#eyehome">Background</a></li> | ||
+ | <li><a data-toggle="tab" href="#eyemenu1">Method</a></li> | ||
+ | <li><a data-toggle="tab" href="#eyemenu2">Results Part 1</a></li> | ||
+ | <li><a data-toggle="tab" href="#eyemenu2">Results Part 2</a></li> | ||
+ | <li><a data-toggle="tab" href="#eyemenu3">Discussion</a></li> | ||
+ | </ul> | ||
+ | |||
+ | <div class="tab-content"> | ||
+ | <div id="eyehome" class="tab-pane fade in active"> | ||
+ | <h4>Background</h4> | ||
+ | |||
+ | <p>There are three quantifiers of how severe cataract formation is, two are measurable, one is not. | ||
+ | <ol> | ||
+ | <li>Absorbance @ 397.5 nm, which is measured with lab equipment.</li> | ||
+ | <li>LOCS scale, subjectively measured by physicians on a scale from 0 - 6. | ||
+ | </li> | ||
+ | <li>Crystallin Damage, which we define as the following (for any time $t$) </li> | ||
+ | </ol> | ||
+ | |||
+ | </p> | ||
+ | |||
+ | \[c.d.(t) = \int_{0}^{\infty} [H_2O_2]_t dt\] | ||
+ | |||
+ | <p> In other words, 1 unit of crystallin damage, in M-h, is equal to the damage caused by 1 molar concentration of hydrogen peroxide reacting crystallin in the eyes for 1 hour.</p> | ||
+ | |||
+ | <p> In making this definition, we assume that crystallin damage is directly proportional to the amount of time crystallin is exposed to hydrogen peroxide. Hydrogen peroxide causes damage by forming disulfide bridges within cysteine molecules on crystallin. This changes the structure of crystallin, causing misfolding and cataract damage. Our linear assumption is valid because the rate for this reaction is first order with respect to hydrogen peroxide concentration. </p> | ||
+ | |||
+ | </div> | ||
+ | |||
+ | <div id="eyemenu1" class="tab-pane fade"> | ||
+ | <h4>Method</h4> | ||
+ | <p>We will relate the three by doing the following: | ||
+ | <ol> | ||
+ | <li>Relate LOCS scale to opacity via literature research. </li> | ||
+ | <li>Relate opacity to light transmittance via literature research. </li> | ||
+ | <li>Relate light transmittance to absorbance via physical calculations. </li> | ||
+ | <li>Relate absorbance to crystallin damage via experimental data. </li> | ||
+ | |||
+ | |||
+ | </ol> | ||
+ | </p> | ||
+ | |||
+ | |||
+ | </div> | ||
+ | <div id="eyemenu2" class="tab-pane fade"> | ||
+ | <h4>Part 1 Results</h4> | ||
+ | |||
+ | <table class="table table-bordered" style='width: 70%;margin-left:15%;'> | ||
+ | <caption style='caption-side:top;'><b>Table 1: Data obtained from XXX<b> relating each value of the LOCS scale, to opacity values. </caption> | ||
<tbody> | <tbody> | ||
<tr> | <tr> | ||
− | <td> | + | <td>LOCS</td> |
− | <td> | + | <td>0.0</td> |
− | <td>34 | + | <td>0.5</td> |
+ | <td>1.0</td> | ||
+ | <td>2.0</td> | ||
+ | <td>3.0</td> | ||
+ | <td>4.0</td> | ||
+ | <td>5.0</td> | ||
+ | <td>6.0</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Degree</td> | ||
+ | <td>None</td> | ||
+ | <td></td> | ||
+ | <td>Trace</td> | ||
+ | <td>Mild</td> | ||
+ | <td>Surgery Suggested</td> | ||
+ | <td>Moderate</td> | ||
+ | <td>Severe</td> | ||
+ | <td>Very Severe</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Opacity (%)</td> | ||
+ | <td>0.34</td> | ||
+ | <td>4.24</td> | ||
+ | <td>5.80</td> | ||
+ | <td>18.88</td> | ||
+ | <td>23.60</td> | ||
+ | <td>49.14</td> | ||
+ | <td>65.61</td> | ||
+ | <td>90+</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
− | <td> | + | <td>Absorbance (a.u.)</td> |
− | <td> | + | <td>0.001</td> |
− | <td> | + | <td>0.019</td> |
+ | <td>0.026</td> | ||
+ | <td>0.091</td> | ||
+ | <td>0.117</td> | ||
+ | <td>0.294</td> | ||
+ | <td>0.464</td> | ||
+ | <td>1.3+</td> | ||
</tr> | </tr> | ||
</tbody> | </tbody> | ||
</table> | </table> | ||
+ | <p> We wish to remain below clinically significant levels, so we will reach attempt to lower the LOCS rating of a cataract to below grade 2.5, which means we want to control GSR such that the crystallin damage results in less than 0.108 a.u. at absorbance at 397.5 nm. </p> | ||
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− | + | </div> | |
− | + | <div id="eyemenu3" class="tab-pane fade"> | |
+ | |||
+ | </div> | ||
+ | |||
+ | <div id="eyemenu4" class="tab-pane fade"> | ||
+ | <h4>Menu 3</h4> | ||
+ | <p>Eaque ipsa quae ab illo inventore veritatis et quasi architecto beatae vitae dicta sunt explicabo.</p> | ||
+ | </div> | ||
+ | |||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | |||
+ | </div> <!-- End Menu --> | ||
+ | |||
+ | |||
+ | <h4> Conclusion</h4> | ||
+ | <p> Conclusion</p> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | </div> <!-- Container --> | ||
+ | </div> <!-- Container --> | ||
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<div class="col-sm-12"> | <div class="col-sm-12"> | ||
− | <h2 id | + | <h2 id ="X">Conclusion</h2> |
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− | + | <p class="col-sm-12">Yay</p> | |
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− | + | <div class = "row"> | |
− | + | <div class="col-sm-12"> | |
− | + | <h3>Citations</h3> | |
− | + | <br> <br> <br> <br> <br> <br> <br> <br> | |
</div> | </div> | ||
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− | <canvas id="canvas-container" style = "z-index:-1"></canvas> | + | |
− | <script type="text/javascript" src='https://2015.igem.org/Template:TAS_Taipei/js/field?action=raw&ctype=text/javascript'></script> | + | |
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <style type='text/css'> | ||
+ | #bg { position: fixed; top: 0; left: 0; } | ||
+ | .bgwidth { width: 100%; } | ||
+ | .bgheight { height: 100%; } | ||
+ | </style> | ||
+ | |||
+ | <div class="backgroundpic"> | ||
+ | <img src="https://static.igem.org/mediawiki/2016/d/dd/T--TAS_Taipei--TaipeiBackground-reflected3.png" style="z-index:0" width="100%" height="100%" id="bg" alt="" > | ||
+ | </div> | ||
+ | |||
+ | <script> | ||
+ | var nowRadius = 0 | ||
+ | $(function() { | ||
+ | $({blurRadius: 0}).animate({blurRadius: 10}, { | ||
+ | duration: 20000, | ||
+ | easing:'swing', // or "linear" | ||
+ | // use jQuery UI or Easing plugin for more options | ||
+ | step: function() { | ||
+ | console.log(this.blurRadius); | ||
+ | if ($("#bluebutton").hasClass("isOn") ) {return;}; | ||
+ | if($('#redbutton').hasClass("isOn") ) {return;} | ||
+ | $('.backgroundpic').css({ | ||
+ | "-webkit-filter": "blur("+this.blurRadius+"px)", | ||
+ | "filter": "blur("+this.blurRadius+"px)" | ||
+ | |||
+ | }); | ||
+ | nowRadius = this.blurRadius; | ||
+ | var LOCSnum = Math.round(nowRadius*6/9); | ||
+ | if (LOCSnum > 6) LOCSnum = "6+"; | ||
+ | $('#LOCS').text(LOCSnum+""); | ||
+ | if (LOCSnum ==0) document.getElementById('bluebutton').click(); | ||
+ | } | ||
+ | }); | ||
+ | |||
+ | }); | ||
+ | |||
+ | startBlur= function(speed) { | ||
+ | |||
+ | $({blurRadius: nowRadius}).animate({blurRadius: 10}, { | ||
+ | duration: speed, | ||
+ | easing: 'swing', // or "linear" | ||
+ | // use jQuery UI or Easing plugin for more options | ||
+ | step: function() { | ||
+ | console.log(this.blurRadius); | ||
+ | if ($("#bluebutton").hasClass("isOn") ) {return;}; | ||
+ | if($('#redbutton').hasClass("isOn") ) {return;} | ||
+ | |||
+ | $('.backgroundpic').css({ | ||
+ | "-webkit-filter": "blur("+this.blurRadius+"px)", | ||
+ | "filter": "blur("+this.blurRadius+"px)" | ||
+ | |||
+ | }); | ||
+ | |||
+ | nowRadius = this.blurRadius; | ||
+ | var LOCSnum = Math.round(nowRadius*6/9); | ||
+ | if (LOCSnum > 6) LOCSnum = "6+"; | ||
+ | $('#LOCS').text(LOCSnum+""); | ||
+ | |||
+ | } | ||
+ | }); | ||
+ | }; | ||
+ | |||
+ | stopBlur= function(speed) { | ||
+ | |||
+ | $({blurRadius: nowRadius}).animate({blurRadius: 0}, { | ||
+ | duration: speed, | ||
+ | easing: 'swing', // or "linear" | ||
+ | // use jQuery UI or Easing plugin for more options | ||
+ | step: function() { | ||
+ | console.log(this.blurRadius); | ||
+ | if ($("#redbutton").hasClass("isOn") ) {} else{return;}; | ||
+ | $('.backgroundpic').css({ | ||
+ | "-webkit-filter": "blur("+this.blurRadius+"px)", | ||
+ | "filter": "blur("+this.blurRadius+"px)" | ||
+ | |||
+ | }); | ||
+ | nowRadius = this.blurRadius; | ||
+ | var LOCSnum = Math.round(nowRadius*6/9); | ||
+ | if (LOCSnum > 6) LOCSnum = "6+"; | ||
+ | $('#LOCS').text(LOCSnum+""); | ||
+ | |||
+ | } | ||
+ | }); | ||
+ | |||
+ | |||
+ | }; | ||
+ | |||
+ | function chooseBlur() { | ||
+ | if ($("#redbutton").hasClass("isOn") ) | ||
+ | { | ||
+ | stopBlur(3500); | ||
+ | |||
+ | } | ||
+ | |||
+ | else { | ||
+ | if($("#bluebutton").hasClass("isOn") ) {} | ||
+ | else {startBlur(12000);} | ||
+ | } | ||
+ | }; | ||
+ | |||
+ | |||
+ | </script> | ||
+ | |||
+ | <script> | ||
+ | |||
+ | function switchToggleB() { | ||
+ | if ( $("#bluebutton").hasClass("isOn") ) { | ||
+ | $("#bluebutton").removeClass("isOn"); } | ||
+ | else { $("#bluebutton").addClass("isOn"); } | ||
+ | }; | ||
+ | function switchToggleR() { | ||
+ | if ( $("#redbutton").hasClass("isOn") ) { | ||
+ | $("#redbutton").removeClass("isOn"); } | ||
+ | else { $("#redbutton").addClass("isOn"); } | ||
+ | }; | ||
+ | |||
+ | |||
+ | </script> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <canvas id="canvas-container" style = "z-index:-1" hidden></canvas> | ||
+ | <script type="text/javascript" src='https://2015.igem.org/Template:TAS_Taipei/js/field?action=raw&ctype=text/javascript' hidden></script/> | ||
+ | |||
+ | |||
+ | |||
+ | <script type="text/javascript"> | ||
+ | /* Toggle between adding and removing the "active" and "show" classes when the user clicks on one of the "Section" buttons. The "active" class is used to add a background color to the current button when its belonging panel is open. The "show" class is used to open the specific accordion panel */ | ||
+ | var acc = document.getElementsByClassName("accordion"); | ||
+ | var i; | ||
+ | |||
+ | for (i = 0; i < acc.length; i++) { | ||
+ | acc[i].onclick = function(){ | ||
+ | this.classList.toggle("active"); | ||
+ | this.nextElementSibling.classList.toggle("show"); | ||
+ | } | ||
+ | } | ||
+ | </script> | ||
+ | |||
+ | <script type="text/javascript"> | ||
+ | $("#category_navbar a").on('click', function(event) { | ||
+ | // Make sure this.hash has a value before overriding default behavior | ||
+ | if (this.hash !== "") { | ||
+ | // Prevent default anchor click behavior | ||
+ | event.preventDefault(); | ||
+ | |||
+ | // Store hash0 | ||
+ | var hash = this.hash; | ||
+ | |||
+ | // Using jQuery's animate() method to add smooth page scroll | ||
+ | // The optional number (800) specifies the number of milliseconds it takes to scroll to the specified area | ||
+ | $('html, body').animate({ | ||
+ | scrollTop: $(hash).offset().top | ||
+ | }, 800, function(){ | ||
+ | |||
+ | // Add hash (#) to URL when done scrolling (default click behavior) | ||
+ | window.location.hash = hash; | ||
+ | }); | ||
+ | } // End if | ||
+ | }); | ||
+ | |||
+ | |||
+ | </script> | ||
+ | |||
+ | <div id="slideout"> | ||
+ | <div id="slidecontent"> | ||
+ | <h3>Prevention</h3> | ||
+ | <h5>GSR Eyedrop</h5> | ||
+ | <label class="switch"> | ||
+ | <input id="bluebutton" onClick="switchToggleB(); chooseBlur()" type="checkbox"> | ||
+ | <div class="slider round bluecolorbutton"></div> | ||
+ | </label> | ||
+ | <br> | ||
+ | |||
+ | <h3>Treatment</h3> | ||
+ | <h5>25HC Eyedrop</h5> | ||
+ | <label class="switch"> | ||
+ | <input id="redbutton" onClick="switchToggleR(); chooseBlur()" type="checkbox"> | ||
+ | <div class="slider round redcolorbutton"></div> | ||
+ | </label> | ||
+ | <h4><b> LOCS: <span id="LOCS">0</span> </b></h4> | ||
+ | |||
+ | </div> | ||
+ | <div id="clickme"> | ||
+ | <h2 class="vertical-text" style="Serif" > | ||
+ | Eyedrops | ||
+ | </h2> | ||
+ | </div> | ||
+ | </div> | ||
+ | |||
+ | <style type='text/css'> | ||
+ | |||
+ | #slideout { | ||
+ | background: #FFD700; | ||
+ | position: fixed; | ||
+ | height: 300px; | ||
+ | width: 200px; | ||
+ | top: 30%; | ||
+ | right:-150px; | ||
+ | padding-left: 60px; | ||
+ | z-index:30; | ||
+ | border-radius: 25px; | ||
+ | |||
+ | |||
+ | } | ||
+ | |||
+ | #clickme { | ||
+ | position: absolute; | ||
+ | top: 0; left: 0; | ||
+ | height: 300px; | ||
+ | width: 50px; | ||
+ | background: #C0FF3E; | ||
+ | z-index:30; | ||
+ | border-radius: 25px 5px 5px 25px; | ||
+ | } | ||
+ | |||
+ | #slidecontent { | ||
+ | float:left; | ||
+ | } | ||
+ | |||
+ | .vertical-text { | ||
+ | transform: rotate(90deg); | ||
+ | transform-origin: left bottom 0; | ||
+ | float: left; | ||
+ | } | ||
+ | /* The switch - the box around the slider */ | ||
+ | .switch { | ||
+ | position: relative; | ||
+ | display: inline-block; | ||
+ | width: 60px; | ||
+ | height: 34px; | ||
+ | } | ||
+ | |||
+ | /* Hide default HTML checkbox */ | ||
+ | .switch input {display:none;} | ||
+ | |||
+ | /* The slider */ | ||
+ | .slider { | ||
+ | position: absolute; | ||
+ | cursor: pointer; | ||
+ | top: 0; | ||
+ | left: 0; | ||
+ | right: 0; | ||
+ | bottom: 0; | ||
+ | background-color: #ccc; | ||
+ | -webkit-transition: .4s; | ||
+ | transition: .4s; | ||
+ | } | ||
+ | |||
+ | |||
+ | .slider:before { | ||
+ | position: absolute; | ||
+ | content: ""; | ||
+ | height: 26px; | ||
+ | width: 26px; | ||
+ | left: 4px; | ||
+ | bottom: 4px; | ||
+ | background-color: white; | ||
+ | -webkit-transition: .4s; | ||
+ | transition: .4s; | ||
+ | } | ||
+ | |||
+ | input:checked + .slider.bluecolorbutton { | ||
+ | background-color: #2196F3; | ||
+ | } | ||
+ | |||
+ | input:focus + .slider.bluecolorbutton { | ||
+ | box-shadow: 0 0 1px #2196F3; | ||
+ | } | ||
+ | |||
+ | input:checked + .slider.redcolorbutton { | ||
+ | background-color: #FF0000; | ||
+ | } | ||
+ | |||
+ | input:focus + .slider.redcolorbutton { | ||
+ | box-shadow: 0 0 1px #FF0000; | ||
+ | } | ||
+ | |||
+ | |||
+ | |||
+ | input:checked + .slider:before { | ||
+ | -webkit-transform: translateX(26px); | ||
+ | -ms-transform: translateX(26px); | ||
+ | transform: translateX(26px); | ||
+ | } | ||
+ | |||
+ | /* Rounded sliders */ | ||
+ | .slider.round { | ||
+ | border-radius: 34px; | ||
+ | } | ||
+ | |||
+ | .slider.round:before { | ||
+ | border-radius: 50%; | ||
+ | } | ||
+ | </style> | ||
+ | <script> | ||
+ | $(function () { | ||
+ | var rightVal = -150; | ||
+ | $("#clickme").click(function () { | ||
+ | rightVal = (rightVal * -1) - 150; | ||
+ | $(this).parent().animate({right: rightVal + 'px'}, {queue: false, duration: 500}); | ||
+ | }); | ||
+ | }); | ||
+ | |||
+ | |||
+ | </script> | ||
+ | |||
</body> | </body> | ||
</html> | </html> |
Latest revision as of 14:51, 6 October 2016
Modeling
Abstract
We answer two questions: How much GSR to maintain in the lens, and how to maintain that amount? We find the amount of GSR needed in the lens (Model 2) to limit crystallin damage so the resulting cataract is less than LOCS 2.5 (Model 1). Then, we find the optimal design of eyedrops (Model 4) and nanoparticles that will maintain this amount of GSR in the lens (Model 3). These models allow our team to understand the impact of adding GSR-loaded nanoparticles into the lens, and to design a full treatment plan on how to prevent and treat cataracts.
Achievements
- Designed a simple calculator to find amount of GSR or 25HC eyedrops needed for a patient's LOCS score.
- Bridged the gap between the medical, biological, and chemical measurement of crystallin damage.
- Predicted impact of adding GSR and 25HC on the amount of crystallin damage in the lens.
- Created Nanoparticle Customizer for doctors to find a full treatment plan.
- Generalized our Customizer to allow other iGEM teams who wish to use nanoparticle drug delivery
- Analyzed sensitivity of prototype, and suggested insights into optimal manufacturing and clinical use of our prototype.
- Used Experimental data to develop Models 1 and 3.
Outline
Introduction
Why Model?
In the lab, biologists are often unable to test everything experimentally. For example, in our cataracts project, cataract prevention occurs in the long-term, from 20-50 years. Obviously, although short experiments can provide us an idea of what prevention may look like, the power of computational biology allows us to model into the future. As a result, our modeling has been crucial in developing a prototype.
Focus
Most iGEM teams perform modeling on gene expression, which we accomplish in model 5. However, as our construct is not directly placed into the eyes, how our synthesized protein impacts the eyes after it is seperately transported is much more interesting. As a result, we spent the majority of our models on understanding the impacts on the eye.
Guiding Questions
- How much GSR do we want inside the lens?
- How do we use nanoparticles to control the amount of GSR in the lens?
- How do we synthesize GSR, package into NP, and send it into the eye?
Model 1: Crystallin Damage
Abstract
In our experiments, absorbance measurements are meaningless without understanding how severe a cataract that absorbance measurement means. We use literature research to relate LOCS, the physician's scale of cataract severity) to absorbance, which is how we quantified crystallin damage in experiments. We use experimental data to understand how crystallin damage can be quantified by measuring absorbance. With this model, we can calculate how much crystallin damage we have to limit to reduce LOCS to an acceptable level.
Purpose
How much do we need to limit crystallin damage so surgery is not needed?
Measurement of Cataract Severity
There are four ways of measuring cataract severity, each used for a different purpose.
- Lens Optical Cataract Scale (LOCS): Physicians use this scale, from 0 – 6, to grade the severity of cataracts.
- Opacity (%): This is the physical, quantitative property of the LOC scale.
- Absorbance at 397.5 nm: This is the experimental method, used by our team in the lab (c.d.).
- Crystallin Damage: This is a chemical definition. We quantify cataract severity as a function of how much oxidizing agents there are, as well as how long crystalline is exposed to oxidizing agents. We define 1 crystallin damage unit as the damage done to human crystallin when exposed to 1 M hydrogen peroxide, the main oxidizing agent, for 1 hour.
We use the unit of crystallin damage to connect cataract severity with the amount of GSR we add (in Model 2). We want to lower c.d. below so that the resulting cataract is of LOCS 2.5. For the rest of the model, our task is simple: relate each point of the LOCS scale to c.d., in order to connect to Model 2.
LOCS Equivalence to Absorbance: Literature Research
Past studies have done numerous studies on how absorbance measurements can be converted to the LOC scale that physicians end up using. With the results of ________ and ________, we construct the first three columns in Table 2.
Absorbance Equivalence to Crystallin Damage: Experimental Data
We use experimental data from our team’s Cataract Lens Model (link). In each trial, they added H2O2 to crystallin, and measured the resulting absorbance. The data used are shown in Table 1. We can calculate the theoretical c.d., and graph absorbance vs. crystallin damage in Figure 2.
With this relation in Figure 2, we calculate the equivalent crystallin damage to each LOCS rating and its equivalent absorbance.
Error Analysis
It may be surprising that only around 1 M-h is required to induce moderately severe cataracts. Remember that this is done in the absence of antioxidation systems (GSR) and at an extremely high oxidizing concentration of H2O2 (1M of H2O2). In the lens, H2O2 has a much lower concentration, so severe cataracts are induced over months to years.
There are some limitations of the model that arise from our assumptions. We assume that fish and human lens contain similar crystallin proteins that are degraded in the similar manner (Assumption 4). In addition, we made a rough adjustment of data based our diluting procedure. For better results to create a human cataract model, experiments will need to be done on human lens, even better if done in vitro, without any dilutions.
LOCS | Opacity (%) | Absorbance (@397.5 nm) | Crystallin Damage (M-h) |
---|---|---|---|
0.0 | 0.00 | 0.0000 | 0.0000 |
0.5 | 3.24 | 0.0143 | 0.1327 |
1.0 | 6.65 | 0.0299 | 0.2774 |
1.5 | 10.81 | 0.0497 | 0.4610 |
2.0 | 15.88 | 0.0751 | 0.6966 |
2.5 | 21.95 | 0.1076 | 0.9981 |
3.0 | 29.07 | 0.1492 | 1.3840 |
4.0 | 46.37 | 0.2706 | 2.5101 |
5.0 | 66.05 | 0.4691 | 4.3514 |
Conclusion
For surgery to not be needed, the LOCS value has to be below 2.5. This is equivalent to 21.95% in light opacity or 0.1076 abs units. Based on the results of our experiments, this is equivalent to 0.9981 units of crystallin damage, the damage done to crystallin if exposed to 0.9981 M of H2O2 for 1 hr. For future models, this value 0.9981 units of c.d. will be called the crystallin damage threshold for LOCS 2.5.
Model 2: GSR/25HC Chemical Pathway
Abstract
The key question: How much GSR to add? Now that we know how much we need to limit crystallin damage, we use systems of ordinary differential equation to model the GSR Pathway. We calculate the necessary GSR concentration to be maintained over 50 years so that the resulting cataract is below LOCS 2.5./p>
Purpose
How much GSR do we need to maintain in the lens so that the crystallin damage recorded over 50 years is below the threshold for LOCS 2.5?
Chemical Kinetics Model: Differential Equations
By the Law of Mass Action, Michaelis-Menten Enzyme kinetics, Ping-pong mechanism, and the Law of Passive Diffusion, we build a system of 10 differential equations based on 6 chemical reactions. All parameters, constants, and initial conditions are based off literature data. Estimates made are also shown with assumptions and reasoning. The details are shown in the collapsible for interested readers.
Blackbox Approach: Testing GSR Impact
Image
We will vary the input, Initial GSR concentration, from 0 to 100 uM, holding all other variables constant, and numerically solve for the amount of hydrogen peroxide over time. With this graph, we can find the amount of crystallin damage accumulated over 20 to 50 years if different levels of GSR is maintained.
When determining the relationship between absorbance and crystallin, in Figure 1 the best fit line has a x – intercept that is nonzero. However, when converting each absorbance rating to equivalent crystallin damage in Table 2, we ignore the constant term. When doing the experiments, the fish lens may have contained GSH that is still active, so the fact that the crystallin is exposed to H2O2, the degradation reaction does not happen until all GSH is depleted, and crystallin damage begins to form. We subtract around 1 unit of crystallin damage from all values.
Conclusion
Conclusion
Model 3: Nanoparticles
Abstract
Abstract
Purpose
Purpose
Conclusion
Conclusion
Model 4: Eyedrops
Abstract
Abstract
Purpose
Purpose
Conclusion
Conclusion
Conclusion
Yay
Citations
![](https://static.igem.org/mediawiki/2016/d/dd/T--TAS_Taipei--TaipeiBackground-reflected3.png)