@@ -44,11 +44,11 @@ Preventative care is two-fold, relying on early detection and educating patients
## RCA
Furthering last year's approach with accessibility, affordability, and accuracy, we continued utilizing and improving RCA to assess CAD risk groups while also addressing underrepresented demographics of people. Improvements to last year’s RCA mechanism include testing the biosensor’s specificity, comparing reporter mechanisms, and optimizing reaction times. In addition to RCA modifications, Lambert iGEM pursued a new wet-lab initiative called exponential rolling circle amplification (eRCA). eRCA provides a more sensitive reading of microRNAs with a faster reading time. The emphasis on additional accuracy through eRCA and affordability to end users enables CADlock to be accessible to a wide range of demographics, not limited to race, sex, and economic status (See Wetlab: eRCA).
Furthering last year's approach with accessibility, affordability, and accuracy, we continued utilizing and improving RCA to assess CAD risk groups while also addressing underrepresented demographics of people. Improvements to last year’s RCA mechanism include testing the biosensor’s specificity, comparing reporter mechanisms, and optimizing reaction times. In addition to RCA modifications, Lambert iGEM pursued a new wet-lab initiative called exponential rolling circle amplification (eRCA). eRCA provides a more sensitive reading of microRNAs with a faster reading time. The emphasis on additional accuracy through eRCA and affordability to end users enables CADlock to be accessible to a wide range of demographics, not limited to race, sex, and economic status [See Wetlab: eRCA](https://2023.igem.wiki/lambert-ga/rca/).
## Capillary
Lambert iGEM explored optimizing the reaction time and resources by running the reaction in capillary tubes instead of PCR tubes for DNA amplification and imaging. This quicker response time allows for more patients to be tested in a point-of-care setting. This increase in healthcare accessibility and thus patient retention time allows for an emphasis to be placed on proactive care. MicroRNAs (miRNAs) can easily diffuse through the capillary tube due to the tube's small diameter. This results in high amounts of DNA in isolated locations (see Fig. 4). When exposed to a specific wavelength of light, individual specks of DNA can be seen by the naked eye. DNA can then be counted without the use of complex fluorescence quantification devices such as fluorometers or plate readers, which makes the test more accessible and lowers the technology barrier. Overall, capillary amplification is a reliable, efficient, and user-friendly assay. (See Wetlab: Capillary)
Lambert iGEM explored optimizing the reaction time and resources by running the reaction in capillary tubes instead of PCR tubes for DNA amplification and imaging. This quicker response time allows for more patients to be tested in a point-of-care setting. This increase in healthcare accessibility and thus patient retention time allows for an emphasis to be placed on proactive care. MicroRNAs (miRNAs) can easily diffuse through the capillary tube due to the tube's small diameter. This results in high amounts of DNA in isolated locations (see Fig. 4). When exposed to a specific wavelength of light, individual specks of DNA can be seen by the naked eye. DNA can then be counted without the use of complex fluorescence quantification devices such as fluorometers or plate readers, which makes the test more accessible and lowers the technology barrier. Overall, capillary amplification is a reliable, efficient, and user-friendly assay. [See Wetlab: Capillary](https://2023.igem.wiki/lambert-ga/rca/).
In addition to frugal technologies, Lambert iGEM aimed to decrease the cost of Rolling Circle Amplification (RCA) reagents by producing the enzymes required through protein purification. Phi29 DNA polymerase, priced at $251.00 for 1,250 units from New England Biolabs, not only covers the majority of the cost of RCA reagents but also possesses invaluable qualities such as strand displacement and processive synthesis properties that are necessary for RCA (New England Biolabs, n.d). By producing phi29 DNA polymerase through protein purification, combined with the accuracy of exponential RCA, Lambert iGEM aims to increase the affordability and in turn accessibility, both of which are contributing factors to inclusive care. This allows for more tests because the proteins are much more affordable, increasing the preventative aspect of healthcare. (See Wetlab: Protein Purification)
In addition to frugal technologies, Lambert iGEM aimed to decrease the cost of Rolling Circle Amplification (RCA) reagents by producing the enzymes required through protein purification. Phi29 DNA polymerase, priced at $251.00 for 1,250 units from New England Biolabs, not only covers the majority of the cost of RCA reagents but also possesses invaluable qualities such as strand displacement and processive synthesis properties that are necessary for RCA (New England Biolabs, n.d). By producing phi29 DNA polymerase through protein purification, combined with the accuracy of exponential RCA, Lambert iGEM aims to increase the affordability and in turn accessibility, both of which are contributing factors to inclusive care. This allows for more tests because the proteins are much more affordable, increasing the preventative aspect of healthcare. [See Wetlab: Protein Purification](https://2023.igem.wiki/lambert-ga/protein-purification/).
## Estrogen
While researching CAD risk factors and statistics, we came across literature that informed us about disparities among the sexes concerning early CAD diagnosis. We first conferred with Dr. Mindy Gentry, a women’s cardiologist, to get her opinion on the sex differences in cardiovascular health. She confirmed our insights about estrogen being a factor in increased susceptibility to CAD in women. To further our scientific research, we spoke to Dr. DeLisa Fairweather, a director of translational research in the Department of Cardiovascular Medicine at Mayo Clinic's Florida campus. She educated us on estrogen and its role as a CAD suppressor, and explained how women after menopause are more likely to develop heart disease because estrogen levels drastically decrease after menopause, causing women to be more likely to develop inflammation. Dr. Fairweather detailed that women who have inflammation in the heart are more likely to develop autoimmune disease and CAD. Additionally, she also guided us in the further development of our inclusivity by discussing how the hormone affects men as well. She told us that since men have a low estrogen level that also remains stable, they would be an ideal demographic to test for consistency in results, unlike women who have fluctuating levels of the hormone. Under the guidance of Dr. Fairweather, we tested miRNA-20b using Rolling Circle Amplification. In the presence of estrogen, CAD suppressors are activated and miRNA-20b is produced. miR-20b causes a negative feedback loop and suppresses estrogen, which will in turn inhibit the CAD suppressors. Dr. Fairweather explained how in men, a high level of miR-20b means that they are likely to develop CAD, however, in premenopausal women with inflammation symptoms, miR-20b protects them from developing CAD. She explained how if there is a high level of 20b, it will inhibit the estrogen from causing a catalyst between the developing autoimmune disease and CAD. We tested miR-20b with the RCA mechanism and ran a 1% gel to validate the presence of our product (see Fig. 5). Ultimately, increasing the factors that can be tested to signify CAD correlation encompasses a wider yet specified group, allowing for quality proactive care to be implemented. (See Wetlab: Inclusivity)
While researching CAD risk factors and statistics, we came across literature that informed us about disparities among the sexes concerning early CAD diagnosis. We first conferred with Dr. Mindy Gentry, a women’s cardiologist, to get her opinion on the sex differences in cardiovascular health. She confirmed our insights about estrogen being a factor in increased susceptibility to CAD in women. To further our scientific research, we spoke to Dr. DeLisa Fairweather, a director of translational research in the Department of Cardiovascular Medicine at Mayo Clinic's Florida campus. She educated us on estrogen and its role as a CAD suppressor, and explained how women after menopause are more likely to develop heart disease because estrogen levels drastically decrease after menopause, causing women to be more likely to develop inflammation. Dr. Fairweather detailed that women who have inflammation in the heart are more likely to develop autoimmune disease and CAD. Additionally, she also guided us in the further development of our inclusivity by discussing how the hormone affects men as well. She told us that since men have a low estrogen level that also remains stable, they would be an ideal demographic to test for consistency in results, unlike women who have fluctuating levels of the hormone. Under the guidance of Dr. Fairweather, we tested miRNA-20b using Rolling Circle Amplification. In the presence of estrogen, CAD suppressors are activated and miRNA-20b is produced. miR-20b causes a negative feedback loop and suppresses estrogen, which will in turn inhibit the CAD suppressors. Dr. Fairweather explained how in men, a high level of miR-20b means that they are likely to develop CAD, however, in premenopausal women with inflammation symptoms, miR-20b protects them from developing CAD. She explained how if there is a high level of 20b, it will inhibit the estrogen from causing a catalyst between the developing autoimmune disease and CAD. We tested miR-20b with the RCA mechanism and ran a 1% gel to validate the presence of our product (see Fig. 5). Ultimately, increasing the factors that can be tested to signify CAD correlation encompasses a wider yet specified group, allowing for quality proactive care to be implemented. [See Wetlab: Inclusivity](https://2023.igem.wiki/lambert-ga/rca/).
@@ -73,7 +73,7 @@ While researching CAD risk factors and statistics, we came across literature tha
## CADmir
Lambert iGEM developed CADmir as a centralized microRNA (miRNA) database to enhance researchers’ access to functional information about miRNAs more efficiently. (see Fig. 6) Currently, valuable insights on miRNAs are dispersed throughout academic literature, posing a challenge for researchers seeking detailed information about specific miRNAs. CADmir aims to address these issues by offering a cohesive and easily accessible platform that organizes all of the current scientific literature about miRNAs, facilitating quick and efficient searches. Lambert iGEM hopes that with CADmir, more researchers will explore miRNA research and incorporate miRNAs into more projects without requiring substantial amounts of time dedicated to reading through papers. CADmir aims to advance the field of miRNA research by improving information accessibility and reducing the time barrier associated with obtaining high-quality scientific information on these topics.(See Wetlab: Estrogen).
Lambert iGEM developed CADmir as a centralized microRNA (miRNA) database to enhance researchers’ access to functional information about miRNAs more efficiently. (see Fig. 6) Currently, valuable insights on miRNAs are dispersed throughout academic literature, posing a challenge for researchers seeking detailed information about specific miRNAs. CADmir aims to address these issues by offering a cohesive and easily accessible platform that organizes all of the current scientific literature about miRNAs, facilitating quick and efficient searches. Lambert iGEM hopes that with CADmir, more researchers will explore miRNA research and incorporate miRNAs into more projects without requiring substantial amounts of time dedicated to reading through papers. CADmir aims to advance the field of miRNA research by improving information accessibility and reducing the time barrier associated with obtaining high-quality scientific information on these topics. [See Drylab: Software](https://2023.igem.wiki/lambert-ga/software/).
@@ -82,11 +82,11 @@ Lambert iGEM developed CADmir as a centralized microRNA (miRNA) database to enha
## LabPilot
LabPilot was targeted towards underfunded laboratories and independent researchers to increase accessibility to this crucial equipment. The leading retailing liquid handler costs around \$10,000, prohibiting less-privileged laboratories from purchasing liquid handlers and instead manually pipetting (OT-2 Robot, 2023). LabPilot is primarily made with 3D printed parts, making it affordable and easily replicable by any community with a 3D printer at a build cost of \$250 and a pipette cost of \$409 (Eppendorf Research Plus Adjustable Volume, Single Channel Pipette, 2 - 20 uL, Yellow)(See Parts List), and about 6.5% the cost of a commercial device (OT-2 Robot). LabPilot upholds the standard of accuracy found in other liquid handlers by having a significant difference when comparing the volume of liquid pipetted by LabPilot and a human (See Proof of Concept). LabPilot’s intuitive software utilizes critical user experience (UX) principles: a user can quickly transfer liquid from one location to another with just three taps. Additionally, LabPilot’s software conforms to the Web Content Accessibility Guidelines 2.1 standards, ensuring a quality user experience for people with disabilities. (See Hardware: LabPilot)
LabPilot was targeted towards underfunded laboratories and independent researchers to increase accessibility to this crucial equipment. The leading retailing liquid handler costs around \$10,000, prohibiting less-privileged laboratories from purchasing liquid handlers and instead manually pipetting (OT-2 Robot, 2023). LabPilot is primarily made with 3D printed parts, making it affordable and easily replicable by any community with a 3D printer at a build cost of \$250 and a pipette cost of \$409 (Eppendorf Research Plus Adjustable Volume, Single Channel Pipette, 2 - 20 uL, Yellow)(See Parts List), and about 6.5% the cost of a commercial device (OT-2 Robot). LabPilot upholds the standard of accuracy found in other liquid handlers by having a significant difference when comparing the volume of liquid pipetted by LabPilot and a human (See Proof of Concept). LabPilot’s intuitive software utilizes critical user experience (UX) principles: a user can quickly transfer liquid from one location to another with just three taps. Additionally, LabPilot’s software conforms to the Web Content Accessibility Guidelines 2.1 standards, ensuring a quality user experience for people with disabilities. (See Hardware: LabPilot) [See Drylab: Hardware](https://2023.igem.wiki/lambert-ga/hardware/).
## Micro-Q
Many medical diagnostics utilize fluorescence reporter mechanisms with mathematical models to quantify substances. However, commercial fluorometers are often bulky and cost over \$10,000, making these diagnostics inaccessible. Last year, Micro-Q exhibited single-tube green fluorescence quantification for \$16 but was limited to one sample. This year, Micro-Q Plus allows multi-sample, multi-spectral quantification with camera based fluorescence detection enabling automatic generation of fluorescence/protein expression curves. All these capabilities fit in a pocket-sized portable box, increasing accessibility to underfunded labs and the public. (See Measurement: MicroQ)
Many medical diagnostics utilize fluorescence reporter mechanisms with mathematical models to quantify substances. However, commercial fluorometers are often bulky and cost over \$10,000, making these diagnostics inaccessible. Last year, Micro-Q exhibited single-tube green fluorescence quantification for \$16 but was limited to one sample. This year, Micro-Q Plus allows multi-sample, multi-spectral quantification with camera based fluorescence detection enabling automatic generation of fluorescence/protein expression curves. All these capabilities fit in a pocket-sized portable box, increasing accessibility to underfunded labs and the public. (See Measurement: MicroQ)[See Drylab: Measurement](https://2023.igem.wiki/lambert-ga/measurement/).
