Team:UC Davis

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Latest revision as of 18:07, 21 October 2009

Homepage_UCdavis

            It is estimated that about one in every 133 Americans suffer from an autoimmune disorder called Celiac Disease (14, 11). Celiac Disease is a condition in which the small intestines of an affected individual cannot properly digest gliadin, a protein present in gluten. When consumed, gliadin induces immune system responses, the side-effects of which are, but not limited to: abdominal pain, chronic diarrhea, and vomiting (14). Currently, the only accepted treatment for this disorder is adhering to a gluten-free diet (14, 12). This can be very difficult as great varieties of food contain gluten. This inspired us to imagine an alternative treatment based on a probiotic organism that can survive and take residence in the stomach, where it would secrete an enzyme that degrades gliadin.
           Then the design elements of this organism should satisfy at least two main criteria. First, it should be able to survive only in the stomach; second, it should secrete an enzyme that degrades gliadin. We envision that these parts would not only be useful for our project, but also to other future projects.
       We decided to focus our summer project on building the first of the two key elements required for our system:
                1. Produce an inducible secretion system
                2. Construct a biological pH sensor limiting this secretion system to the stomach

UC Davis iGEM team is sponsored by:

We want to thank Marc Facciotti for his helpful advice.
Special thanks to Suzy Fenton,Wayne Fenton, Jeanne Joe-Fenton and Wayne Fenton Jr.
Also we thank Charles H. Halsted for reviewing our material regarding celiac disease.

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-<title>UCDavis_preliminary project description</title>
+<title>Homepage_UCdavis</title>
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-<body>
+<body style="background-color: rgb(255, 255, 255); color: rgb(0, 0, 0);"
-<div style="text-align: left;">
+alink="#ee0000" link="#0000ee" vlink="#551a8b">
-<div style="text-align: center;">
+<div style="text-align: center;"><span
-<div style="text-align: left;"><b><span
+style="color: rgb(255, 255, 153);">&nbsp;
+</span><b style="color: rgb(255, 255, 153);"><span
 style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><span
-style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span><b><span
+style="color: rgb(255, 255, 153);"><img src="" alt=""
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><big><big><span
+style="width: 151px; height: 161px;"></span><b
-style="font-weight: bold;"></span></big></big><br>
+style="color: rgb(255, 255, 153);"><span
-<big><big><span style="font-weight: bold;"></span></big></big></div>
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><b
-<big><big><span style="font-weight: bold;">Preliminary project
+style="color: rgb(255, 255, 153);"><span
-description:</span></big></big><small><br>
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><a
-</small></div>
+href="https://2009.igem.org/Team:UC_Davis/About_Us"><b
-<small><b><span
+style="color: rgb(255, 255, 153);"><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><br>
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b></a><b
-<b><span
+style="color: rgb(255, 255, 153);"><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><span
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><a
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">1
+href="https://2009.igem.org/Team:UC_Davis/About_Us"><b
-in 133 Americans are suffering
+style="color: rgb(255, 255, 153);"><span
-from a form of autoimmune disorder called Celiac Disease. Celiac
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b></a><b
-Disease is a
+style="color: rgb(255, 255, 153);"><span
-condition where the small intestines of an affected individual cannot
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><b
-digest
+style="color: rgb(255, 255, 153);"><span
-gliadin, a protein present in gluten. If consumed, the body kick starts
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big>&nbsp;
-its
+</big></big></big></big></span></b><b style="color: rgb(255, 255, 153);"><span
-immune system response. The effects of the immune response include, but
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
-are not
+href="https://2009.igem.org/Team:UC_Davis/About_Us"><img alt=""
-limited to abdominal pain and vomiting.</span><span
+src="https://static.igem.org/mediawiki/2009/d/d1/UCDAVIS_PIC4.png"
-style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span><span
+style="border: 0px solid ; width: 92px; height: 46px;"></a> </big></big></big></big></span></b><b
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span><br>
+style="color: rgb(255, 255, 153);"><span
-<span style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span><br>
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
-<span style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">One
+href="https://2009.igem.org/Team:UC_Davis/Project"><img alt=""
-of our teammate's friends is
+src="https://static.igem.org/mediawiki/2009/b/b9/UCDAVIS_PIC8.png"
-currently suffering from this illness, which
+style="border: 0px solid ; width: 92px; height: 46px;"></a> </big></big></big></big></span></b><b
-prompted us to look into Celiac Disease as our main focus. What we
+style="color: rgb(255, 255, 153);"><span
-learned that currently, no cure has been found and the only treatment
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
-is a gluton free diet.
+href="https://2009.igem.org/Team:UC_Davis/Notebook"><img alt=""
-This treatment is very difficult as an overwhelming majority of food
+src="https://static.igem.org/mediawiki/2009/2/2f/UCDAVIS_PIC5.png"
-contain
+style="border: 0px solid ; width: 92px; height: 46px;"></a> </big></big></big></big></span></b><b
-gluten and gliadin.</span><span
+style="color: rgb(255, 255, 153);"><span
-style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></small>
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><b
-</div>
+style="color: rgb(255, 255, 153);"><span
-<div><small>
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><a
-</small></div>
+href="https://2009.igem.org/Team:UC_Davis/Parts"><img alt=""
+src="https://static.igem.org/mediawiki/2009/a/a6/UCDAVIS_PIC6.png"
+style="border: 0px solid ; width: 92px; height: 46px;"></a>&nbsp;<a
+href="https://2009.igem.org/Team:UC_Davis/Contacts_References"><img
+alt="" src="https://static.igem.org/mediawiki/2009/1/1d/UCDAVIS_PIC7.png"
+style="border: 0px solid ; width: 92px; height: 46px;"></a></big></big></big></big></span></b><b
+style="color: rgb(0, 0, 0);"><span
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big><big><big><big><small>
+<img style="width: 191px; height: 147px;" alt="" src=""><br>
+</small></big></big></big></big></span></b></div>
+<hr style="width: 100%; height: 2px;"><b style="color: rgb(0, 0, 0);"><span
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><big><span
+style="color: rgb(0, 0, 0);"></span></big><b
+style="color: rgb(0, 0, 0);"><span
+style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><span
+style="font-family: Symbol;"></span><span
+style="font-family: Arial Narrow; font-weight: bold;"></span></span></b><span
+style="color: rgb(0, 0, 0); font-family: Arial Narrow; font-weight: bold;"><span
+style="font-size: 13pt; line-height: 115%;"></span></span>
 <p class="MsoNormal"
-style="margin-bottom: 0.0001pt; line-height: normal; color: rgb(0, 0, 0); background-color: rgb(255, 255, 255); text-align: left;"><small><span
+style="margin-bottom: 0.0001pt; line-height: normal;"><b><span
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">&nbsp;</span><span
+style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;">&nbsp;&nbsp;&nbsp;
-style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span><br>
+&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;<big> </big></span></b><span
-<span style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">This
+style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black; font-weight: bold;"><big>&nbsp;
-year the University of
+</big><big>It
-California,Davis entered the 2009 iGEM competition with a focus on
+is estimated that about one in
-creating
+every 133 Americans suffer from an autoimmune disorder called Celiac
-a new
+Disease
-way to treat Celiac. Our project has been split into two tasks. The
+(14, 11). Celiac Disease is a condition in which the small intestines of
-first is to
+an
-produce an inducible secretion system; the second is to construct a
+affected individual cannot properly digest gliadin, a protein present
-biological
+in gluten.
-pH sensor to compartmentalize this secretion system in the stomach.
+When
-Moreover, this
+consumed, gliadin induces immune system responses, the
-pH system will act as a kill switch if this system were to ever leave
+side-effects of which are,
+but not limited to: abdominal pain, chronic diarrhea, and vomiting (14).
+Currently, the only accepted treatment for this disorder is adhering to
+a
+gluten-free diet (14, 12). This can be very difficult as great
+varieties of
+food contain gluten. This inspired us to imagine an alternative
+treatment based on a probiotic organism
+that can
+survive and take residence in the stomach, where it would secrete an
+enzyme that
+degrades gliadin.</big></span><big style="font-weight: bold;"><span
+style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
+&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp; &nbsp;&nbsp; Then the design
+elements of this organism should satisfy at least two main
+criteria.
+First, it
+should be able to survive only in the stomach; second, it should secrete an
+enzyme
+that degrades gliadin<i>.</i> We envision that these parts would
+not
+only be useful for our project, but also to other future projects.</span><span
+style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span><br>
+</big><span style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big
+style="font-weight: bold;">&nbsp;&nbsp;&nbsp;
+&nbsp; &nbsp;
+&nbsp;&nbsp; We decided to focus our summer project on building the
+first of the two key elements
+required for
+our system:<o:p></o:p><br>
+&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp;
+&nbsp;&nbsp;&nbsp; &nbsp;&nbsp;&nbsp; 1. <a
+href="https://2009.igem.org/Team:UC_Davis/secretion">Produce an
+inducible secretion
+system</a><br>
+</big>
+<big style="font-weight: bold;">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+. <a href="https://2009.igem.org/Team:UC_Davis/ph_sensor1">Construct a
+biological pH sensor limiting this secretion system to
 the
-stomach by inducing cell death via pH change. After weeks of reading
+stomach</a></big></span><a
-numerous
+href="https://2009.igem.org/Team:UC_Davis/ph_sensor1"><span
-articles, we are confident that with correct parts and timing, our
+style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><span
-two
+style="text-decoration: underline;"></span></span></a><span
-systems can be built in time to present to the iGEM community.</span><span
+style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><big
-style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span></small></p>
+style="font-weight: bold;"><br>
-<div style="text-align: left;"><small>
+</big></span></p>
-</small></div>
 <p class="MsoNormal"
-style="margin-bottom: 0.0001pt; line-height: normal; color: rgb(0, 0, 0); background-color: rgb(255, 255, 255); text-align: left;"><small><span
+style="margin-bottom: 0.0001pt; line-height: normal;"><span
-style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p>&nbsp;</o:p></span></small><small><span
+style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span><a
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br>
+href="https://2009.igem.org/Team:UC_Davis/ph_sensor1"><b
-We
+style="color: rgb(0, 0, 0);"><span
-have recently moved into the synthesis
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b></a><span
-and assembly stage of our two systems.</span><span
+style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
-style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></small><span
+&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;
-style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p></span><span
+&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;
-style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">Though
+&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;
-this is our first year in the competition, we feel
+&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;
-that we are moving at a good pace, working out kinks along the way.</span><o:p></o:p></p>
+&nbsp; &nbsp; &nbsp; &nbsp; &nbsp;
-<small><span
+&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
-style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></small>
+&nbsp;</span><a href="https://2009.igem.org/Team:UC_Davis/ph_sensor1"><span
-<div style="text-align: left;"></div>
+style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><span
-<small></small>
+style="text-decoration: underline;"></span></span></a><a
+href="https://2009.igem.org/Team:UC_Davis/Project_Summary"><img alt=""
+src="https://static.igem.org/mediawiki/2009/a/ac/UCDAVIS_summary.png"
+style="border: 0px solid ; width: 63px; height: 50px;"></a><span
+style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+<span style="text-decoration: underline;"><br>
+</span> </span><a href="https://2009.igem.org/Team:UC_Davis/ph_sensor1"><b
+style="color: rgb(0, 0, 0);"><span
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b></a><span
+style="font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span><b
+style="color: rgb(0, 0, 0);"><span
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b></p>
+<hr style="width: 100%; height: 2px;">
+<p class="MsoNormal"
+style="margin-bottom: 0.0001pt; line-height: normal; color: rgb(0, 0, 0); text-align: center; background-color: rgb(255, 255, 255);"><b><span
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><b><span
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><b><span
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b><b><span
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"></span></b></p>
+<ul style="background-color: rgb(255, 255, 255); color: rgb(0, 0, 0);">
+</ul>
+<div style="text-align: center;"><big> </big></div>
+<div style="text-align: center;">
 <div style="text-align: left;">
+<div style="text-align: center;"><big><span style="font-weight: bold;"><big>&nbsp;&nbsp;
+&nbsp;&nbsp;<small>
+UC
+Davis iGEM team is sponsored by:</small></big></span></big><br>
+<big><span style="font-weight: bold;"></span></big></div>
+<div style="text-align: center;">&nbsp;&nbsp; &nbsp;&nbsp; <img
+src="https://static.igem.org/mediawiki/2009/8/88/UCDAVIS_Biomedical_engineering.jpg"
+alt="" style="width: 83px; height: 109px;"><img
+style="width: 105px; height: 109px;" alt=""
+src="https://static.igem.org/mediawiki/2009/4/43/UCDAVIS_college_of_engineering.png"><img
+style="width: 135px; height: 99px;" alt=""
+src="https://static.igem.org/mediawiki/2009/8/8e/UCDAVIS_Accutek1.jpg"><img
+style="width: 121px; height: 109px;" alt=""
+src="https://static.igem.org/mediawiki/2009/6/6b/UCDAVIS_mrgen.gif"><img
+style="width: 182px; height: 108px;" alt=""
+src="https://static.igem.org/mediawiki/2009/c/c6/UCDAVIS_genome_center.jpg"><br>
+<font size="-1">We want to thank <span style="font-weight: bold;">Marc</span>
+<span style="font-weight: bold;">Facciotti</span> for his helpful
+advice.<br>
+Special thanks to <span style="font-weight: bold;">Suzy
+Fenton</span>,<span style="font-weight: bold;">Wayne Fenton, Jeanne
+Joe-Fenton </span>and<span style="font-weight: bold;"> Wayne Fenton Jr.</span>
+<br>
+Also we thank <span style="font-size: 10pt; line-height: 115%;"><b>Charles
+H. <span class="il">Halsted </span></b><span class="il">for reviewing
+our material regarding celiac disease.</span></span></font><br>
+<br>
 </div>
-<small></small>
+</div>
-<div style="text-align: left;"></div>
+</div>
-<p
+<ul style="background-color: rgb(255, 255, 255); color: rgb(0, 0, 0);">
-style="color: rgb(0, 0, 0); background-color: rgb(255, 255, 255); text-align: left;"
+<span style="font-weight: bold;"></span>
-class="MsoNormal"><small><o:p>&nbsp;</o:p></small></p>
+</ul>
+<p class="MsoNormal"
+style="margin-bottom: 0.0001pt; line-height: normal; color: rgb(0, 0, 0); text-align: left; background-color: rgb(255, 255, 255);"><b><span
+style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;
+</span></b></p>
+<span style="color: rgb(0, 0, 0);"></span>
 </body>
 </html>