Difference between revisions of "Team:UofC Calgary/Safety"
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| − | <h1> Safety Considerations in the Lab<h1> | + | <h1>Safety Considerations in the Lab<h1> |
| − | <h2> How we prepared for lab work</h2> | + | <h2><u>How we prepared for lab work</u></h2> |
| − | <p class="c3"><span class="c1">All Principal Investigators, mentors, and undergraduate researchers were required to complete lab safety training and safety courses developed by the University of Calgary's<a href="http://www.ucalgary.ca/safety/">Environment Health and Safety (EHS)</a> services prior to working in the lab. These mandatory safety training courses included courses on occupational health and safety, laboratory safety, hazard assessment, incident reporting and investigation, spill response, biosafety, bloodborne pathogens, and an updated versions of the WHMIS course. The courses cover biological containment protocols, handling of hazardous materials such as liquid nitrogen, and disposal of waste, as well as standard safety and laboratory practices. All required us to take a test following each course, which certified safe lab work under the EHS Guidelines. All team members, advisors, and mentors received credit for each course and training program listed, and supervisors were present in the lab at all times to oversee undergraduate work.</span> | + | <p class="c3"><span class="c1">All Principal Investigators, mentors, and undergraduate researchers were required to complete lab safety training and safety courses developed by the University of Calgary's <a href="http://www.ucalgary.ca/safety/">Environment Health and Safety (EHS)</a> services prior to working in the lab. These mandatory safety training courses included courses on occupational health and safety, laboratory safety, hazard assessment, incident reporting and investigation, spill response, biosafety, bloodborne pathogens, and an updated versions of the WHMIS course. The courses cover biological containment protocols, handling of hazardous materials such as liquid nitrogen, and disposal of waste, as well as standard safety and laboratory practices. All required us to take a test following each course, which certified safe lab work under the EHS Guidelines. All team members, advisors, and mentors received credit for each course and training program listed, and supervisors were present in the lab at all times to oversee undergraduate work.</span> |
</p> | </p> | ||
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<p class="c3 c11"><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 418.67px; height: 217.33px;"><img alt="" src="https://static.igem.org/mediawiki/2016/4/4a/T--UofC_Calgary--Syed00.jpg" style="width: 418.67px; height: 217.33px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></span> | <p class="c3 c11"><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 418.67px; height: 217.33px;"><img alt="" src="https://static.igem.org/mediawiki/2016/4/4a/T--UofC_Calgary--Syed00.jpg" style="width: 418.67px; height: 217.33px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></span> | ||
</p> | </p> | ||
| − | < | + | <h2><span><u>Structure of the Patch</u></span></h2> |
<li class="c0"><span>The circular design of the packet containing media and <span class="c8 c1">B. subtilis</span> spores encourages equal distribution of liquid pressure around the circumference to prevent the patch from breaking open at the seams. The packets are made of a flexible polyethylene material, which is advantageous for distributing external forces against the patch, such as accidental bumping or squishing. This distribution of pressure prevents the material from rupturing due to accumulated forces along seams or corners.</span> | <li class="c0"><span>The circular design of the packet containing media and <span class="c8 c1">B. subtilis</span> spores encourages equal distribution of liquid pressure around the circumference to prevent the patch from breaking open at the seams. The packets are made of a flexible polyethylene material, which is advantageous for distributing external forces against the patch, such as accidental bumping or squishing. This distribution of pressure prevents the material from rupturing due to accumulated forces along seams or corners.</span> | ||
</li> | </li> | ||
<li class="c0"><span>The fact that our patch is non-invasive and only present in the external part of the body promotes safety better than an implant.</span> | <li class="c0"><span>The fact that our patch is non-invasive and only present in the external part of the body promotes safety better than an implant.</span> | ||
</li> | </li> | ||
| − | < | + | <h2><span><u>Choosing Patch Materials</u></span></h2> |
<li class="c0"><span>All materials used in the patch are chemically and biologically compatible, which decreases the risk of the user's immune response.</span> | <li class="c0"><span>All materials used in the patch are chemically and biologically compatible, which decreases the risk of the user's immune response.</span> | ||
</li> | </li> | ||
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<li class="c0"><span>Release liner: The release liner is disposed after being peeled.</span> | <li class="c0"><span>Release liner: The release liner is disposed after being peeled.</span> | ||
</li> | </li> | ||
| − | < | + | <h2><span><u>Considering Human Use</u></span></h2> |
| − | <li class="c0"><span>Using a patch gives | + | <li class="c0"><span>Using a patch gives the user the freedom to take medication even with an upset stomach, as the drug will not need to go through the digestive system.</span> |
</li> | </li> | ||
<li class="c0"><span>Accidental breakage of the patch while attached to skin may cause infections if not disinfected properly.</span> | <li class="c0"><span>Accidental breakage of the patch while attached to skin may cause infections if not disinfected properly.</span> | ||
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<li class="c0"><span>Patch may cause severe harm if swallowed.</span> | <li class="c0"><span>Patch may cause severe harm if swallowed.</span> | ||
</li> | </li> | ||
| − | <li class="c0"><span>Potential skin irritation may occur. | + | <li class="c0"><span>Potential skin irritation may occur. However, our initial mouse trials have shown no signs of inflammation.</span> |
</li> | </li> | ||
| − | <li class="c0"><span>The safety of the radioprotective patch was assessed by conducting </span><span class="c8 c1">in vivo</span><span class="c1"> mouse trails to test the patch, safety of the bacteria and | + | <li class="c0"><span>The safety of the radioprotective patch was assessed by conducting </span><span class="c8 c1">in vivo</span><span class="c1"> mouse trails to test the patch, safety of the bacteria and our mBBI peptide. <b>Our mouse trial protocol was followed and approved by the <a href="http://www.ucalgary.ca/research/researchers/ethics-compliance/animal-use-protocols">Health Sciences Animal Care Committee (HSACC)</a> who review the ethics concerning animal care and testing at the University of Calgary.</b></span> |
</li> | </li> | ||
<li class="c0"><span class="c1">Documentation of ethics approval from the animal studies is available upon request as it contains confidential information that can not be made publicly available.</span> | <li class="c0"><span class="c1">Documentation of ethics approval from the animal studies is available upon request as it contains confidential information that can not be made publicly available.</span> | ||
</li> | </li> | ||
| − | < | + | <h2><span><u>Containment</u></span></h2> |
| − | <li class="c0"><span class="c1">Contamination issues may be a problem in space if the bacteria were to escape. To protect against the event that our radioprotective patch causes contamination in space or on earth, | + | <li class="c0"><span class="c1">Contamination issues may be a problem in space if the bacteria were to escape. To protect against the event that our radioprotective patch causes contamination in space or on earth, our patch has several engineering controls in place, such as the protective physical barriers of the patch that would prevent the outbreak of bacteria. The backing layer and the size-controlling membrane prevents the bacteria from escaping outside of the patch and being released onto the skin, respectively.</span> |
| − | + | <li class="c0"><span>Although our initial diffusion assays have shown that bacteria can diffuse through our semipermeable membrane, we have begun testing with other smaller filters including peptide dialysis membranes and 0.2 micron size filters. We have also shown success with membranes used to filter-sterilize liquids.</span> | |
| − | + | ||
| − | + | ||
| − | <li class="c0"><span>Although our initial diffusion assays have shown that bacteria can diffuse through our semipermeable membrane, we have begun testing with other smaller filters including peptide dialysis membranes and 0.2 micron size filters.</span> | + | |
</li> | </li> | ||
<li class="c0"><span>See pages 15 - 16 of the <a href="https://drive.google.com/file/d/0B25REnfhyiNWM0hsNGtjRl9GWkk/view?usp=sharing" style="padding-right:0px";>Device Manual</a> to tackle contamination issues.</span> | <li class="c0"><span>See pages 15 - 16 of the <a href="https://drive.google.com/file/d/0B25REnfhyiNWM0hsNGtjRl9GWkk/view?usp=sharing" style="padding-right:0px";>Device Manual</a> to tackle contamination issues.</span> | ||
</li> | </li> | ||
| − | < | + | <h2><span><u>Future Considerations for Patch Design</u></span></h2> |
<p class="c3"><span class="c1">If we can determine a better membrane that prevents the diffusion of the bacteria, we can use a two layer semi permeable system where the first layer prevents the diffusion of the bacteria and a second layer which further filters BBI for diffusion.</span> | <p class="c3"><span class="c1">If we can determine a better membrane that prevents the diffusion of the bacteria, we can use a two layer semi permeable system where the first layer prevents the diffusion of the bacteria and a second layer which further filters BBI for diffusion.</span> | ||
</p> | </p> | ||
| − | < | + | <h2><span><u>Safe disposal:</u></span></h2> |
<li class="c0"><span>See pages 11 and 12 of the <a href="https://drive.google.com/file/d/0B25REnfhyiNWM0hsNGtjRl9GWkk/view?usp=sharing" style="padding-right:0px";>Device Manual</a> for disposal information.</span> | <li class="c0"><span>See pages 11 and 12 of the <a href="https://drive.google.com/file/d/0B25REnfhyiNWM0hsNGtjRl9GWkk/view?usp=sharing" style="padding-right:0px";>Device Manual</a> for disposal information.</span> | ||
</li> | </li> | ||
<p class="c2"><span></span> | <p class="c2"><span></span> | ||
</p> | </p> | ||
| − | < | + | <h1><span>Safety Considerations of Biobrick Parts</span></h1> |
| − | <li class="c0"><span class="c1">Public safety: | + | <li class="c0"><span class="c1">Public safety: threonine auxotrophy was employed where integrated cassettes containing flanking regions of homology for the </span><span class="c1 c8">B. subtilis thrC</span><span class="c1"> gene were used. This knocked out threonine synthase in our chassis, making it unlikely to survive outside of conditions where theronine is supplied.</span> |
</li> | </li> | ||
| − | <li class="c0"><span class="c1">Environmental safety: It’s possible our </span><span class="c8 c1"> | + | <li class="c0"><span class="c1">Environmental safety: It’s possible that our </span><span class="c8 c1">B. subtilis</span><span class="c1"> transformed with mBBI could outcompete native microbes and potentially disrupt existing ecologies; however, the integrated mBBI genetic construct would not make a significant difference in the bacteria’s survivability outside the lab. Given the nature of our project and safeguards we have implemented, none of our BioBricks confer unsafe risks to individuals nor the environment.</span> |
</li> | </li> | ||
| − | < | + | <h1>Future Considerations</h1> |
| − | <p class="c3"><span class="c1">We would engineer inducible kill switches that could eradicate the bacteria if need be | + | <p class="c3"><span class="c1">We would engineer inducible kill switches that could eradicate the bacteria if need be. Additionally, integrating mBBI in various essential genes required for amino acid synthesis could provide more opportunities for auxotrophy, increasing the safety of using </span><span class="c8 c1">B. subtilis </span><span class="c1">in the device.</span> |
</p> | </p> | ||
<p class="c2"><span class="c1"></span> | <p class="c2"><span class="c1"></span> | ||
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<p class="c2"><span></span> | <p class="c2"><span></span> | ||
</p> | </p> | ||
| − | < | + | <h1><span>Safety Forms</span></h1> |
<li class="c0"><span class="c1"><a href="https://2016.igem.org/Safety/About_Our_Lab">About Our Lab</a></span> | <li class="c0"><span class="c1"><a href="https://2016.igem.org/Safety/About_Our_Lab">About Our Lab</a></span> | ||
</li> | </li> | ||
Revision as of 23:55, 16 October 2016
Safety
Safety Considerations in the Lab
How we prepared for lab work
How we prepared for lab work
All Principal Investigators, mentors, and undergraduate researchers were required to complete lab safety training and safety courses developed by the University of Calgary's Environment Health and Safety (EHS) services prior to working in the lab. These mandatory safety training courses included courses on occupational health and safety, laboratory safety, hazard assessment, incident reporting and investigation, spill response, biosafety, bloodborne pathogens, and an updated versions of the WHMIS course. The courses cover biological containment protocols, handling of hazardous materials such as liquid nitrogen, and disposal of waste, as well as standard safety and laboratory practices. All required us to take a test following each course, which certified safe lab work under the EHS Guidelines. All team members, advisors, and mentors received credit for each course and training program listed, and supervisors were present in the lab at all times to oversee undergraduate work.
The University of Calgary has a university-wide Biosafety Committee, whose guidelines for safe biological laboratory practices were adhered to throughout the project. The team’s lab benches and experimental plans were assessed and deemed safe to proceed with by this Biosafety Committee. The Univerity's Environment Health and Safety (EHS) services provided additional training for individuals working with radiation and irradiated cells.
Our project utilized Bacillus subtilis and a commonly used lab-strain of Escherichia coli, TOP10. Both are non-pathogenic and non-infectious, and are classified as Biosafety Level 1 organisms (BSL-1). Therefore, these organisms posed no significant risk to researchers. Since the BSL-1 cells (E. coli and B. subtilis) have GRAS labelling, the main cloning component of out project did not require ethics approval by review boards. Some team members worked with HCT116 and 1BR3 primary cell lines, which are human colon carcinoma and human skin fibroblast cell lines and are classified as Biosafety Level 2 (BSL-2).The cell lines were received from completely anonymous donors. We handled these cell lines at containment level 2 in accordance with the Bloodborne Pathogens Standard and Biosafety Committee guidelines.
Safety Considerations for the Device
Structure of the Patch
Choosing Patch Materials
Considering Human Use
Containment
Future Considerations for Patch Design
If we can determine a better membrane that prevents the diffusion of the bacteria, we can use a two layer semi permeable system where the first layer prevents the diffusion of the bacteria and a second layer which further filters BBI for diffusion.
Safe disposal:
Safety Considerations of Biobrick Parts
Future Considerations
We would engineer inducible kill switches that could eradicate the bacteria if need be. Additionally, integrating mBBI in various essential genes required for amino acid synthesis could provide more opportunities for auxotrophy, increasing the safety of using B. subtilis in the device.
Safety Forms
