Team:Aboa/Human Practices

Team Aboa 2021

Human Practices

From the very beginning, we wanted to create a project that would be responsible and good for the world as iGEM encourages all teams to do. We started our Human Practices efforts following the questions stated at the iGEM medal criteria page, and had a discussion with our team about all the environmental, social, moral, scientific and other types of values and principles we needed to consider before starting the project. You can find the main points of our discussion below.

Being responsible to us means that we...

  • are not releasing any GMO in the environment.
  • obey the rules and laws of the community around us, whether it is as small as the department we’re working in or as big as the whole EU.
  • are not manipulating our lab results.
  • don’t take credit for other people's work.
  • appreciate and are grateful for every human being taking part in this project.
  • do our best in making our project accessible for everyone.

We also named five core values for our team and made a poster about them which you can see attached below.

The aim of this poster was to remind us of our core values all the way through our project.

A poster with a picture of team Aboa in the middle and five core values written in green balls around it.

Honesty means to us first and foremost not manipulating our lab results but it also has another dimension in the meaning of being honest to each other in our team. Honesty, whether it’s being brutally honest or genuinely optimistic, creates trust between people in the long run, and in a year long project such as iGEM, trust is inevitable between the members of a team for anything at all to work sufficiently.

One of the clearest principles we wanted to obey was one of being biosafe. We made sure from the very beginning that all of the work in the lab would be done in a safe and responsible way so that all GMO used would be disposed of in the right way. This way no GMO would get out in the wild and cause any harm to other organisms or nature in general. Also, we swore by the fact that not one of our lab members would get injured during experiments in the lab.

Another principle that was clear to us was integrity. We wanted to obey the rules of the department we got the lab from and we also wanted to proceed in a way of following the rules and laws of the society around us, meaning the University of Turku, the city of Turku, the nation of Finland and even the EU. This also stemmed from the fact that our PI informed and advised us very clearly of the general rules of the laboratory we got to use.

Respect is a value we need to bring up as one of our core values. We have respect for other teams, competing now and also in the previous competitions, and we give credit to other teams for their work. We don’t put somebody else’s ideas under our own name. We also share respect for each other within our team. We let everybody participate in their own way and we give all a chance to try and develop at their own pace.

Last but not least we mention good sportsmanship as our core value because we understand iGEM is a competition and when taking part in it, we’re following the principles of fair play. We know how to be good winners and losers and we will win and lose as a team, not pointing fingers to anyone or anywhere. We are capable of digesting the end result in a mature way and we can be genuinely happy for other teams’ achievements.

Our project is good to the world because...

We’ve chosen to study a molecule quite unknown to the public and thus we’re shedding light on a bigger problem than just our iGEM project. By doing this, we’re also bringing awareness to the condition of our beloved Baltic Sea and at the same time we’re offering a way to help fix this problematic situation.

In addition, we’re contributing to a better world by producing valuable information about the degradation of diclofenac that our local wastewater treatment plant could possibly utilize in improving the efficiency of their process. Moreover, the enzymes we are working with are not specific to diclofenac, but degrade a variety of other compounds too. Other researchers could hence benefit from our project by refining it to target the issue of pharmaceutical waste in bodies of water on a broader scale, or alternatively, combine these enzymes with others in a bioreactor or biofilter to make it more applicable for their local needs.

Our project is also good to the world as we’re showing an example not only to other areas of Finland but also the rest of the world how we can better the situation of our environment by using the tools of synthetic biology. This way solving our local problem could also have a huge global impact.

The responsible conduct of research

We also wanted to mention separately that our project follows the guidelines of ‘The responsible conduct of research’ (Finnish Advisory Board of Research Integrity, 2012):

  • The research follows the principles that are endorsed by the research community, that is, integrity, meticulousness, and accuracy in conducting research, and in recording, presenting, and evaluating the research results.
  • The methods applied for data acquisition as well as for research and evaluation, conform to scientific criteria and are ethically sustainable. When publishing the research results, the results are communicated in an open and responsible fashion that is intrinsic to the dissemination of scientific knowledge.
  • The researcher takes due account of the work and achievements of other researchers by respecting their work, citing their publications appropriately, and by giving their achievements the credit and weight they deserve in carrying out the researcher’s own research and publishing its results.
  • The researcher complies with the standards set for scientific knowledge in planning and conducting the research, in reporting the research results and in recording the data obtained during the research.
  • The necessary research permits have been acquired and the preliminary ethical review that is required for certain fields of research has been conducted.
  • Before beginning the research or recruiting the researchers, all parties within the research project or team (the employer, the principal investigator, and the team members) agree on the researchers’ rights, responsibilities, and obligations, principles concerning authorship, and questions concerning archiving and accessing data. These agreements may be further specified during the course of the research.
  • Sources of financing, conflicts of interest or other commitments relevant to the conduct of research are announced to all members of the research project and reported when publishing the research results.
  • Researchers refrain from all research-related evaluation and decision-making situations, when there is reason to suspect a conflict of interest.
  • The research organisation adheres to good personnel and financial administration practices and takes into account the data protection legislation.

References


Integrated Human Practices

Integrated Human Practices is about demonstrating how our team responded to research, reflections and engagement made during different phases of our project. We consulted different experts at several phases during the year in order to gain more insight about crucial topics. Read further to know who contributed to our project and what was the effect they had on our work.

Biologist

What did we get?

In order to get more information about diclofenac and the effects it may have on the ecosystem of the Baltic Sea, we contacted multiple different biologists and experts from that field. We were really lucky to get Jenni Prokkola to share her knowledge with us. She had previously done research on the effects of diclofenac in fishes.

We addressed some questions to Prokkola. Here are some of the questions and answers briefly summarized (more information can be read from here.):

Has diclofenac some special features in its harmful effects when compared to other pharmaceutical waste?

  • One significant feature is its amount in the bodies of water. Lots of diclofenac is flown down the sink because of its use in pain relieving gels.
  • In addition, diclofenac can accumulate in fishes so that the amount in their tissues can be considerably high even though the exposure was low.

Can you tell more about the specific mechanisms by which diclofenac does harm to the ecosystem?

  • Diclofenac can cause stress to liver metabolism because the liver is in charge of degrading pharmaceuticals.
  • Metabolic enzymes, such as LDH (lactate dehydrogenase), CYP450 1A (cytochrome P450 1A) and GPX (glutathione peroxidase) can react to these events and diclofenac, in turn, can also disturb these responses.
  • Even at low doses, diclofenac has an effect on the fishes’ ability to adapt to hypoxia.

Can you tell whether diclofenac can cause some behaviour disorders in fishes?

  • I have read about these disorders regarding other pharmaceutical waste but not in diclofenac.
  • Nevertheless, it is possible because the behaviour is strictly regulated and physiological mechanisms can interact with it.

In addition, she recommended a few important articles from this topic which could be helpful to us - And they definitely were!

How did this affect our project?

Based on the knowledge we got from Prokkola, we were even more convinced that diclofenac really is a big problem both globally and locally and that it does affect the wildlife of the Baltic Sea. We became more sure that we had chosen the right problem to tackle and got valuable information about the specific effects of diclofenac in fish.

Turun Seudun Puhdistamo Oy

What did we get?

When we had decided that we were going to start working with the wastewater purification process, we contacted the experts at our local wastewater treatment plant. We were aiming to receive answers to specific questions we had posed beforehand. The meeting was scheduled for the end of June and we got to meet experts Jarkko Laanti and Jouko Tuomi from Puhdistamo Oy.

Here are listed our questions and the answers we received.

How much is there diclofenac in the wastewater and how much of it can be removed?

  • At the moment, the efficiency to remove diclofenac is only 27 %. This results in 90 kg of diclofenac ending up in the Baltic Sea every year only from the people of Turku. It’s really expensive to develop new techniques to identify and remove different pharmaceuticals from wastewater so new innovations are very much needed in this field.

Does pH or temperature change between different phases of wwtp?

  • The pH of the wastewater coming into the plant is approximately 7 to 7,5 and it stays the same almost through the whole process. At the lowest it can be 6,5. On the other hand, the temperature of the water changes according to the season from approximately 12 to 25 degrees Celsius. However, in the winter, a lot of meltwater comes into the plant and lowers the temperature of the water up to 8 degrees.

Where could our proposed implementation be added in the wwtp?

  • Your proposed implementation could be added either right before or just after aeration. If it was added earlier in the process, the conditions would be so harsh that the laccases would get denatured. After aeration, there’s only secondary clarification phase and sand filtration phase, so the laccases would still have time to detoxify diclofenac before the water leaves the wastewater treatment process.

After the meeting they gave us the contact information for Hannele Fredriksson who later helped us get samples of the wastewater.

How did this affect our project?

We became even more convinced that diclofenac is a huge problem also locally here in Turku. We got valuable information from the deficiencies of current wastewater purification processes and realized that a diclofenac-degrading system should be added indeed in the wastewater treatment plants.

In addition, we already at this point got important requirements for both our solution and our proposed implementation. Based on these discussions, we had to choose laccases that can be active in pH 7 and at varying temperatures (about 10 to 25 degrees Celsius). Also our final product, in other words, proposed implementation should function in these conditions.

PerkinElmer

What did we get?

We contacted experts at PerkinElmer Finland Oy in the beginning of July. Our aim was to discuss further collaboration actions regarding measurements to be made for the samples we got from our local WWTP. We wanted to find out how much of diclofenac is actually currently in the wastewater around the aeration part where our sample was collected and where our bioreactor would be added. We were unable to measure this ourselves because we didn’t have the equipment needed for this sort of measurement.

How did this affect our project?

We agreed with the experts on bringing our filtered and frozen wastewater samples to PerkinElmer’s lab for them to execute the LC-MS analysis. Our plan was to use this information as a reference data in our own measurements. This collaboration was however put on hold for a while since the experts went out to have their holiday. Unfortunately, our plans in the lab evolved during this time so we never got back to them.

Hannele Fredriksson

What did we get?

Hannele is a person who has stayed with us for quite a long time during the project. We first met with her in the beginning of July when we went to visit the wastewater treatment plant in Kakolanmäki. She was kind enough to prepare a guided tour for us and answer our questions about the technical side of the process. During our tour, when we reached the aeration part, she took the samples for us from a six meter deep pool.

After the tour we kept in contact with her because we knew that she would be a very useful person to consult when designing our proposed implementation more thoroughly. In addition to working in the WWTP, she was also a student of energy and environmental technique from the University of Applied Sciences of Turku. For a long time earlier in the project timeline, we were planning to organize an ideation workshop for students in her school to help us further develop our proposed implementation. Unfortunately, this plan didn’t get fulfilled due to problems in scheduling but we kept in touch still.

As we’re writing this on September 24th, our next plan is to organize a meeting where she and her colleagues could meet with our partnership team’s local WWTP’s experts and together we could refine our proposed implementations. We would thus make sure that each of our implementations would be applicable to the other one’s local process. This means that we would take a deeper look into the laws and regulations in addition to the technical side of the implementation. If a meeting can not be organized, then the wanted result could be achieved by sending a similar e-mail questionnaire to both sides.

How did this affect our project?

Based on the discussions with Hannele, we could modify our proposed implementation so that it could be implemented in our local WWTP in reality. In addition, cooperation with Hannele enabled our partnership with the team from Chicago to become true despite the problems we encountered in the lab. To conclude, Hannele had a remarkably big role in the success of our project.

Biochemistry Experts

What did we get?

Biochemistry experts for us entails the researchers and staff at the biochemistry unit at the Department of Life Technologies at our university with whom we discussed project details. From them we received most notably valuable insights on protein purification methods and technical help for our project. We went through our plans first in a meeting with Jarmo Käpylä and later with Anu Salminen who was available when problems arose in the lab. They helped us decide on the exact protocols for enzyme production, cell lysis and protein purification, as well as asked us questions about our enzymes that we had not realized to consider earlier. These were all matters that we had been struggling with in the team and it helped us a lot to get their advice on these issues. Their expertise also solved our SDS-PAGE dilemma since we had been confused about all available options, and were wondering which gel would suit our purposes the best.

How did this affect our project?

Being in contact with biochemistry experts had a significant effect on our project. We would most likely not have been able to produce and extract our enzymes in a nearly as unproblematic fashion and as early on as without their support. Especially Anu made a big impact by offering lab assistance in moments of doubt during enzyme purification steps. She also proposed ideas for getting started with enzyme activity measurements. (Read more)

Legislation

What did we get?

We explored the legislation of the EU regarding genetically modified organisms (GMOs). We learned that the deliberate release of GMOs is really strictly legislated in EU countries. For example, when introducing GMOs into the environment, the step-to-step principles need to be obeyed, meaning that the area and volume must be very carefully gradually expanded - during the long course of time. In addition, we need to be ultimately sure to prevent our cells from spreading into surroundings since they have antibiotic resistance genes that can possibly be transferred to other microbes in the environment.

How did this affect our project?

These legislation issues affected our project so that we focused even more deeply to design different solutions to prevent the spread of cells into the environment. From our safety related solutions you can read more from Proposed Implementation and Safety.

Local Pharmacies

What did we get?

To find out more about people's habits and attitudes towards returning drugs to pharmacies, we interviewed two local pharmacies. We found out that on average about 215 litres of unused pharmaceuticals are returned to the pharmacy every week. Most of the returned pharmaceuticals are prescription tablets. In addition people tend to return injectable drugs as well as cough mixtures. There is still a lot to improve in people’s habits as they don’t regard for example pain gels or inhalators for the treatment of asthma as drugs that need to be returned to pharmacies. In addition to different pharmaceuticals, people also tend to return apparatus and cosmetics there - even though they shouldn’t be taken to the pharmacy.

Another of our local pharmacies told us that they don't have any place to return the unused drugs. Instead, people should simply give the drugs to the personnel. Another pharmacy, in turn, told us that they have an easily reachable place for the customers to return the drugs. There are also clear guidelines on how to return the drugs. Some instructions on how to return drugs are given out to the customers in both pharmacies - but clearly not to all. In addition, at least one of the pharmacies tries to remind people of the returning the drugs in their social media channels. From pharmacies the pharmaceutical waste is transformed to the central factory of hazardous waste. There the waste is burned in high temperatures and treated so that it changes to less harmful compounds.

Based on this survey, we concluded that there is a lot to improve in people’s habits and attitudes towards returning the unused drugs. We asked for an interview from twenty local pharmacies and got answered from only two. Maybe that also tells something about attitudes. It can be possible that for pharmacies it’s even humiliating to admit that they don’t have any visible instructions or reminders to return the unused drugs back to the pharmacy. By conducting this survey, we raised awareness for them to realize that the public is concerned about their environmental effects.

How did this affect our project?

As a part of our project, we wanted to pay attention to the habits of people concerning the return on unused or expired drugs back to pharmacies. After getting information about this, we agreed that there might not be almost any problems at all with pharmaceutical waste if people would only return their drug to pharmacies. Based on the information we got, we decided to add more information about returning drugs to pharmacies in our poster of the Baltic Sea Day.