Difference between revisions of "Team:NTU-LIHPAO-Taiwan/Modeling/Conclusion"

 
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<ul class="subs">
 
<li><a href="https://2015.igem.org/Team:NTU-LIHPAO-Taiwan/Description">Description</a></li>
 
<li><a href="https://2015.igem.org/Team:NTU-LIHPAO-Taiwan/Description">Description</a></li>
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<li><a href="https://2015.igem.org/Team:NTU-LIHPAO-Taiwan/Basic_Part">Basic Parts</a></li>
 
<li><a href="https://2015.igem.org/Team:NTU-LIHPAO-Taiwan/Basic_Part">Basic Parts</a></li>
 
<li><a href="https://2015.igem.org/Team:NTU-LIHPAO-Taiwan/Composite_Part">Composite Parts</a></li>
 
<li><a href="https://2015.igem.org/Team:NTU-LIHPAO-Taiwan/Composite_Part">Composite Parts</a></li>
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<li><a href="https://2015.igem.org/Team:NTU-LIHPAO-Taiwan/Part_Collection">Part Collection</a></li>
 
 
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<ul class="main-Content">
 
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<span class="title">Prototype</span>
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<span class="title">Modeling Summary</span>
 
<ul class="sub-Content">
 
<ul class="sub-Content">
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<li><a href="#First">Prototype</a></li>
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<li><a href="#First1">Hypothesis</a></li>
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</ul>
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<li><a href="#First2">Conclusion</a></li>
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<li>
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<span class="title">Main Part</span>
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<ul class="sub-Content">
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<li><a href="#Second1">Part:BBa_K1841001</a></li>
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<li><a href="#Second2">Part:BBa_K1841004</a></li>
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<li><a href="#Second3">Part:BBa_K1841007</a></li>
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</li>
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<li>
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<span class="title">Test Part</span>
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<ul class="sub-Content">
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<li><a href="#Third1">RFP</a></li>
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<li><a href="#Third2">His-PYY</a></li>
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<li><a href="#Third3">Signal peptide-PYY-His</a></li>
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<li><a href="#Third4">Y2R</a></li>
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<span class="title">Reference</span>
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<li><a href="#Fourth">Reference</a></li>
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<div class="ContentHolder">
 
<div class="ContentHolder">
 
<div class="Text1">Modeling Summary</div>
 
<div class="Text1">Modeling Summary</div>
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<div class="Text2" id="First">Hypothesis</div>
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<div class="Text2" id="First1">Hypothesis</div>
 
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<div class="Text3">
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After orally ingested, certain amounts of modified probiotic <i>L. casei</i> can survive under acidic condition when passing through the stomach. Owning to gastric acid, bile acid and digestive enzymes, the presence of these bacteria in the intestinal tract only last for a limited time, and then are washed out in faeces.<a href="#Reference1">[1]</a> During the process of transition and temporary colonization, CPP-PYY complexes are produced and exert the following effect.
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<ol class="part2">
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        <li>Total oral ingestion of 10<sup>9</sup> <i>L. casei</i> on the first day.</li>
 +
        <li>The varying ratio (%) of bacteria number to total ingested number attached on the intestine against time were according to the mouse experiment.</li>
 +
        <li>Incorporate suicide mechanism: 4 days for bacteria survival.</li>
 +
        <li>TAT penetrates through villi by an uncharacterized pinocytosis/endocytosis related mechanism, which is also receptor-independent.</li>
 +
        <li>CPP-PYY complex production rate per <i>L. casei</i> : 10<sup>-10</sup> μg/min.</li>
 +
        <li>One-fourth of the products secreted from <i>L. casei</i> penetrate through the small intestine epithelial cells, without being digested by the intestine fluid.</li>
 +
        <li>Effective permeability of TAT-insulin conjugates from literature as reference for TAT-PYY complex : P<sub>eff</sub> = 1.62×10<sup>-5</sup> (cm/s).</li>
 +
        <li>Mean total mucosal surface of the small intestine interior averages 32 m<sup>2</sup>.</li>
 +
        <li><i>L. casei</i> cell size range = 0.7-1.1 x 2.0-4.0 μm. One-tenth of the bacteria height multiplied the mucosa surface area for the donor chamber volume.</li>
 +
        <li>Fick’s Law of Binary Diffusion applied for CPP-PYY complex adsorption into bloodstream. Interstitial fluid diffusion coefficient of bovine serum albumin in tissues was taken for reference : D<sub>AB</sub> = 5.8 x 10<sup>-7</sup> cm<sup>2</sup>/s.</li>
 +
        <li>The distance from epithelial cells to capillaries is 4 μm.</li>
 +
        <li>The diffusion is rapid enough to adsorb all the products in every minute.</li>
 +
        <li>All the calculation was based on reaching steady-state condition in the interval of every minute discretely.</li>
 +
        <li>Thrombin cleavage efficiency of Arg-Ser(R-S) site in CPP-PYY complex was referred to Arg-Thr(R-T) site in salmon calcitonin.</li>
 +
        <li>Nearly 100 % of the complexes would be cleaved during 30 minutes due to the fluid motion in circulation.</li>
 +
        <li>PYY concentration of 100 pg/ml has the power of appetite suppression without drug resistance.</li>
 +
        <li>Average total blood volume of 5 liter in human body.</li>
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</ol>
 
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<div class="Text2" id="First">Conclusion</div>
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<div class="Text2" id="First2">Conclusion</div>
 
<div class="Text3">
 
<div class="Text3">
 
Ideally, the optimal concentration of PYY is 0.1 μg/L for appetite suppression, giving little side effect. Here we performed the simulation with 10<sup>9</sup> of total <i>L. casei</i> intake, and 10<sup>-10</sup> μg/min of CPP-PYY complex production rate per <i>L. casei</i>. The results is shown below.
 
Ideally, the optimal concentration of PYY is 0.1 μg/L for appetite suppression, giving little side effect. Here we performed the simulation with 10<sup>9</sup> of total <i>L. casei</i> intake, and 10<sup>-10</sup> μg/min of CPP-PYY complex production rate per <i>L. casei</i>. The results is shown below.
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<img src="https://static.igem.org/mediawiki/2015/d/de/Pathway_Figure7.png" width="500px"/>
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<div class="Article_PictureText1"><div class="Text_Picture">[Fig.2-1] Final concentration of PYY (μg/L) in the blood vessel</div></div>
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<blue>[Final concentration of PYY in the blood (μg/L)] = 11745 × [Total <i>L. casei</i> intake] × [Distribution ratio of <i>L. casei</i>] × [CPP-PYY complex production rate per <i>L. casei</i> (μg/min)]</blue>
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<b>[Final concentration of PYY in the blood (μg/L)] = 11745 × [Total <i>L. casei</i> intake] × [Distribution ratio of <i>L. casei</i>] × [CPP-PYY complex production rate per <i>L. casei</i> (μg/min)]</b>
 
</div>
 
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<div class="Text3">
 
<div class="Text3">
 
This general formula is based on one-dose response, and continuous drug intake should be further modified by successive accumulation for determining the appetite suppression effect.
 
This general formula is based on one-dose response, and continuous drug intake should be further modified by successive accumulation for determining the appetite suppression effect.
 
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</div>
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<div class="Text3">
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<b>—></b>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<a href="https://2015.igem.org/Team:NTU-LIHPAO-Taiwan/Modeling"><b>[Click on here back to Modeling Introduction]</b></a>
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<div class="Text3">
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<b>—></b>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<a href="https://2015.igem.org/Team:NTU-LIHPAO-Taiwan/Modeling/Conclusion"><b>[Click on here to see our Simulation Strategy and Results]</b></a>
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Latest revision as of 13:17, 18 September 2015

NTU-LIHPAO-Taiwan

Modeling Summary
Hypothesis
  1. Total oral ingestion of 109 L. casei on the first day.
  2. The varying ratio (%) of bacteria number to total ingested number attached on the intestine against time were according to the mouse experiment.
  3. Incorporate suicide mechanism: 4 days for bacteria survival.
  4. TAT penetrates through villi by an uncharacterized pinocytosis/endocytosis related mechanism, which is also receptor-independent.
  5. CPP-PYY complex production rate per L. casei : 10-10 μg/min.
  6. One-fourth of the products secreted from L. casei penetrate through the small intestine epithelial cells, without being digested by the intestine fluid.
  7. Effective permeability of TAT-insulin conjugates from literature as reference for TAT-PYY complex : Peff = 1.62×10-5 (cm/s).
  8. Mean total mucosal surface of the small intestine interior averages 32 m2.
  9. L. casei cell size range = 0.7-1.1 x 2.0-4.0 μm. One-tenth of the bacteria height multiplied the mucosa surface area for the donor chamber volume.
  10. Fick’s Law of Binary Diffusion applied for CPP-PYY complex adsorption into bloodstream. Interstitial fluid diffusion coefficient of bovine serum albumin in tissues was taken for reference : DAB = 5.8 x 10-7 cm2/s.
  11. The distance from epithelial cells to capillaries is 4 μm.
  12. The diffusion is rapid enough to adsorb all the products in every minute.
  13. All the calculation was based on reaching steady-state condition in the interval of every minute discretely.
  14. Thrombin cleavage efficiency of Arg-Ser(R-S) site in CPP-PYY complex was referred to Arg-Thr(R-T) site in salmon calcitonin.
  15. Nearly 100 % of the complexes would be cleaved during 30 minutes due to the fluid motion in circulation.
  16. PYY concentration of 100 pg/ml has the power of appetite suppression without drug resistance.
  17. Average total blood volume of 5 liter in human body.
Conclusion
Ideally, the optimal concentration of PYY is 0.1 μg/L for appetite suppression, giving little side effect. Here we performed the simulation with 109 of total L. casei intake, and 10-10 μg/min of CPP-PYY complex production rate per L. casei. The results is shown below.
[Fig.2-1] Final concentration of PYY (μg/L) in the blood vessel
By building the model for simulation step by step, we summarized the general formula with varies parameters to determine the dose response :
[Final concentration of PYY in the blood (μg/L)] = 11745 × [Total L. casei intake] × [Distribution ratio of L. casei] × [CPP-PYY complex production rate per L. casei (μg/min)]
This general formula is based on one-dose response, and continuous drug intake should be further modified by successive accumulation for determining the appetite suppression effect.
—>     [Click on here back to Modeling Introduction]
—>     [Click on here to see our Simulation Strategy and Results]
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