Team:UCL/Demonstrate
<!DOCTYPE html>
DEMONSTRATE
Do our devices work in real life?
OVERVIEW
Talking to Aubrey de Grey and Filipe Cabreiro has stressed the need for us to be able to explore through not just conceptional design but through wet lab, how we can prove that our ideas will work in the real world, in the complex human body. Real world conditions to us is defined as the human body. How will our biobricks interact under real conditions? Will our biobrick survive in the human body? Will they interact or affect other processes in the body? How can we then ensure that the result of this exploration can feed back onto the design of our devices to make them safer and more controlled.
IrrE biobrick for lubricant
IrrE
This construct already exists in the registry, however, we wanted to expand the pre-existing functionality. Here we have shown that this gene goes beyond helping growth in high salt concentration solutions by demonstrating that it actually exists bacterial growth in lubricant.
To achieve this we used lubricant from the market, specifically, Superdrug (Code: 380900), head quartered in Croydon, Surrey, to demonstrate this in order to prove the gene would be functional in real-world conditions.
Our results indicate our proof of concept that under simulated conditions to be a success. This is because E. coli transformed with IrrE had improved growth in LB and lubricant compared to wild type.
We believe this is the first step to demonstrate that a living organism would be capable of living in lubricant.
Our functionality for our GMO lubricant would be two fold.
Initially we envisaged an STI-detecting lubricant which would change colour when in contact with an organism such as syphilis and chlamydia trachomatis.
When talking about ageing most of the diseases and implications that come to mind are quite obvious: loss of sight, hearing or mental capacity, arthritis and
many more. One that doesn’t come to mind very often are Sexually Transmitted Infections, such as chlamydia, syphilis and gonorrhoea. The fact of the matter is that between 2007 and 2011 the number of chlamydia and syphilis increased by 31%and respectively 52% in the American population over the age of 65. 1This may initially come as a surprise but it is a reality nowadays, especially in elderly care facilities.
With people living for longer, medication countering the effects of erectile dysfunction and a lot of free time it can be crudely compared to an American frat house. This might not yet be seen as a problem as people of all ages should be able to enjoy themselves however they want to. Yet the real problems start when there are secondary effects from the STIs, such as arthritis, vision loss and other symptoms. Those will usually be classified as just age-related problems and treated as such, rather than being treated appropriately.
However, it came to our attention the array of medical conditions that require lubricant, especially those associated with the elderly. We saw the potential of the addition of a safe GMO within them to prevent bacterial infections.
In hospitals nowadays it is very common to have a urinary catheter inserted to drain the bladder. Catheters can either be intermittent, where they are only inserted temporarily and removed as soon as the bladder is empty, or indwelling, where they are in place for a prolonged period of time (days or weeks.[i]) The NHSN (National Healthcare Safety Network) had found that in the EU around 17.5% of hospital patients need indwelling catheters: 45-79% of which were in a critical care unit, 17% in the medical ward, 23% in the surgical ward and 9% in rehabilitation[ii].
Around about 73% of the patients receiving these catheters are above the age of 65. Now the problem arises as about 28% of these elderly patients will develop a UTI (urinary tract infection), due to the use of a urinary catheter. 60-80% of patients with indwelling catheters receive antimicrobials to counteract asymptomatic bacteriuria, which can lead to microbial resistance. In practice catheter lubricant is used to try to reduce the risk of infection, pain, friction and general discomfort.[iii]
From the data shown in the graph above it clearly shown that the E.Coli transformed with IrrE grows better in the 40%Lubricant/60%LB solution in direct comparison with the Wild type E.Coli, which hardly grew at all. This data thus suggests that with the IrrE the E.Coli is better adapted to living in, which would allow further studies being conducted on the maximum concentration of Lubricant it can be grown in. Furthermore once that has been determined the pathogen detecting aspect of the concept can be realised.
Thus overall this experiment has allowed us to show that IrrE increases the growth of the E. Coli in the Superdrug Lubricant containing: purified water, glycerine, Carbopol 940, Triethanolamine and Sodium Butyl Paraben. This supports the previous experiments that concluded that IrrE allows for better growth in saline conditions[iv], except this time in Superdrug Lubricant.
For further conceptualisation and experimentation it should be taken into consideration that within a commercialisable product the bacteria will not be under ideal growth conditions and the nutrients will be a limiting factor, especially when considering that the lubricant concentration will have to be a lot higher than in this experiment.
References:
[i] "Urinary Catheterisation - NHS Choices." NHS Choices. Department of Health, n.d. Web. 9 Oct. 2016.
[ii] Nicolle, Lindsay E. "Catheter Associated Urinary Tract Infections." Antimicrob Resist Infect Control Antimicrobial Resistance and Infection Control 3.1 (2014): 23. Web.
[iii] June, 2015 22, and Lakshmi Muthuraman4 March, 2016 10:32 Am. "Selecting Gel Types for Urinary Catheter Insertion." Nursing Times. N.p., n.d. Web. 9 Oct. 2016.
[iv] http://parts.igem.org/Part:BBa_K729005
Lycopene in the gut
We wanted to show that lycopene would have improved growth under oxidative stress. We have achieved this in the lab however we wanted to go beyond this and did so with a collaboration with Dundee schools who had created a gut device. We wanted to ensure our lycopene probiotic would withstand the gut, and following the results, have proven the robustness of the system by in a simulation. The graph below shows that our bacteria are indeed capable of surviving in the gut!
