Human Practices

Research is to see what everybody else has seen, and to think what nobody else has thought.
Albert Szent-Györgyi

Overview

Our project, "STAND UP", is not solely the endeavor of us, the iGEM Tokyo Tech 2023 members. We have maintained a close relationship with society throughout all stages of the project - Design, Build, Test, Learn - and have progressed the project, sometimes facing setbacks along the way. The reason why we conduct Human Practices is encapsulated in the words: "Human Practices is the study of how your work affects the world, and how the world affects your work." It involves deeply considering and implementing how we perceive demands from society, integrating thoughts into the project, and how the project can subsequently influence society to refine our project. This is Human Practices in iGEM. Through the reciprocal interaction between society and the project, we have observed what everyone in society is seeing and considered projects that nobody has thought about before. The narrative of &quots;tanding up&quot against Dengue fever originated from the essence of research spoken by Szent-Györgyi and the philosophy of iGEM's Human Practices.

Project Overview

Dengue fever is an infectious disease transmitted by mosquitoes in tropical regions. It is one of the Neglected Tropical Diseases and has not received much attention from the international community thus far. Although vaccine development has advanced recently, many are still suffering. Reducing the number of dengue fever infections contributes to the well-being of people worldwide and is necessary from perspectives like disparity reduction. We initiated our project with the ultimate goal of &quots;reducing the number of dengue fever infections&quots; .

Next, to achieve this goal, through interviews, we pondered which challenges should be addressed and identified three specific objective candidates: (1) Develop and disseminate a vaccine, (2) Conduct vector control, and (3) Engage in awareness reform (for more details, please see ).

Our team discussed how to realize these objectives and decided to prepare for implementing the awareness reform. For vaccine development/dissemination and vector control, we further specified and considered &quots;developing methods for surveillance&quots; and &quots;producing desmethylglycitein using E. coli&quots; (more details in ).

Regarding &quots;producing desmethylglycitein using E. coli,&quots; we explored its feasibility through meetings with synthetic biology experts at Tokyo Tech but decided to abandon the approach, judging that the local use of DMG, i.e., its impact on the ultimate goal, was minimal (please see for more).

For &quots;developing methods for surveillance,&quots; through handovers from the previous fiscal year and meetings with professors at Tokyo Tech, we explored its feasibility and decided to adopt CFNT and 3D-PCR, determining that the demands from society and our impact were sufficiently met (please refer to for more). With this, we aimed to &quots;reduce the number of dengue fever infections&quots; by focusing on &quots;developing methods for surveillance&quots; and &quots;local awareness reform.&quots;

To assess whether our objective was appropriate, we approached potential end-users: government/administration, testing companies, and local people. This activity was helpful in further improving our objectives (see ).

For the objective &quots;developing methods for surveillance,&quots; we progressed while closely communicating the technical contents of two test methods with scientists. We documented the specific content of the interviews (for more details, see ).

We also aimed at &quots;local awareness reform.&quots; For this, we conducted literature reviews and discussions with educational experts, produced booklets for behavior improvement, and developed and implemented educational evaluation indicators to enhance knowledge and attitudes (more details in ).

Lastly, we devised a plan for future social implementation as a concrete result of our HP, discussing with entrepreneurs and experts in social implementation to ensure the project's feasibility is as high as possible (please refer to for more).

sample

Identifying Problems

In order to identify what is necessary to reduce the number of dengue fever infections, we needed to start by understanding the current situation of dengue fever. To do this, we interviewed two individuals and one organization who are familiar with the local situation of dengue fever and identified the issues.

Tomohiko Takasaki image
Tomohiko Takasaki
Adviser, BML, Inc.

Summary

  • Reducing the number of dengue fever infections is most crucially dependent on the proliferation of vaccines.
  • For the effective utilization of vaccines, it is essential to have testing methods that can identify infection history.
  • A testing method is needed that is affordable, requires no machinery, and can clarify which serotype of dengue virus was the cause of infection.
  • Vector control is also important, however, insecticides using organophosphates have shown to be less effective.
Kunihiro Kaihatsu
CEO, VisGene, Ltd.

Summary

  • Promoting vaccines is of utmost importance.
  • It is necessary to increase users through end-user needs assessments.
  • An approach that takes national characteristics into consideration is essential.
Kunihiro Kaihatsu image
J-Trops image
J-Trops
Japanese Society of Tropical Medicine Students’ Branch

Summary

  • In Kenya, people do not go to the hospital even if they contract dengue.
  • There is a need to raise awareness for definitive diagnosis.

Conclusion

From these three interviews, we set three goals necessary to truly reduce the number of dengue fever infections. The first is the development and dissemination of vaccines. Vaccines are the most effective and important measure against dengue fever, and the widespread use of currently available vaccines is needed. The second is controlling mosquitoes. Methods for control include insecticides and repellents for adult mosquitoes, and hormone treatments for larvae. The third is changing the awareness of residents in infected areas. It's necessary to foster basic hygiene awareness among citizens about what dengue fever is, what is necessary to protect oneself from dengue fever, and what is needed to prevent mosquito breeding.

Enbodying Targets

In the section, we recognized that in order to alleviate the suffering caused by dengue fever, it was necessary to promote dengue fever vaccines, implement effective vector control, and change the awareness of residents in epidemic areas. Following this recognition, our team discussed how these goals could be achieved.

Our team was divided into three groups: the 'Vaccine Development and Dissemination Group, ' the 'Vector Control Group,' and the 'Awareness Change Group.' Based on research papers, each group conducted literature reviews and brainstorming to achieve their respective goals. As a result, the 'Vaccine Dissemination Group' and the 'Vector Control Group' each selected the most influential papers for achieving their goals: 'Best Practices in Dengue Surveillance: A Report from the Asia-Pacific and Americas Dengue Prevention Boards (author: Mark E Beatty, 2020)'[] and 'Molecular action of larvicidal flavonoids on ecdysteroidogenic glutathione S-transferase Noppera-bo in Aedes aegypti (authors: Kazue Inaba, Kana Ebihara, 2022),'[] respectively. Additionally, the 'Awareness Change Group' also found several important papers towards achieving their goal, which will be detailed in the section. Afterward, we conducted interviews with two experts related to these papers.

Ryusuke Niwa
Professor, Life Science Center for Survival Dynamics, Tsukuba Advanced Research Alliance

Summary

  • Desmethylglycitein has a low environmental impact, but large-scale production is essential for practical use.
  • Production of desmethylglycitein in E. coli is not impossible.
  • Large-scale synthesis of desmethylglycitein using E. coli could have a significant impact on the market.
Ryusuke Niwa image
Richard T Mahoney image
Richard T Mahoney
PhD・CEO at RTM Associates

Summary

  • The national healthcare system benefits the most from surveillance data.
  • The data can be used for effective vaccine allocation and monitoring vaccine impact.
  • The data is also useful for infectious disease response planning and requests for funding to the legislature.

Conclusion

We understood that in order to reduce the number of dengue fever infections, it was necessary to specifically explore two methods as concrete approaches to achieve the essential goals of vaccine development and dissemination, mosquito control, and changing the awareness of residents in infected areas. These methods are the production of Desmethylicaritin using E. coli and the development of techniques for surveillance.

Feasibility Study -desmethylglycitein-

In the section , regarding the proposal of synthesizing desmethylglycitein using E. coli, we consulted with experts knowledgeable in substance production using microorganisms to discuss its feasibility.

Tomoaki Matsuura
Professor, Earth-Life Science Institute, Tokyo Institute of Technology

Summary

  • Producing desmethylglycitein in E. coli is challenging, and it might be better to use yeast as a model organism.
  • Desmethylglycitein may not dissolve easily in water, necessitating a review of implementation methods.
  • The yield of the final product is likely to decrease significantly compared to the input.
  • Synthesizing the final product might take a minimum of one year, or even more than five years.
Tomoaki Matsuura image
Toshiaki Fukui image
Toshiaki Fukui
Professor, School of Life Science and Technology, Tokyo Institute of Technology

Summary

  • Designing just a part of the synthesis route for desmethylglycitein might take more than a year.
  • Using yeast is likely to be more effective.
  • Producing the final product might require 2-3 years.

Conclusion

The information obtained from the interview regarding DMG (Desmethylglycitein) is as follows: the synthesis of a small amount of DMG could take a minimum of one year and up to five years. Additionally, there is a possibility that the promising group of enzymes for synthesizing DMG from its precursor, daizein, may not show activity in E. coli, in which case using yeast might be a better option. Given our experimental setup's difficulty in using yeast and the significant amount of time required for production, we concluded that synthesizing DMG should not be adopted for our project.

Feasibility Study -Survaillance-

We conducted interviews with the goal of exploring why surveillance is not currently being conducted and what kind of testing is demanded by society to promote surveillance in the future. Additionally, we aimed to determine the methods by which surveillance testing should be conducted.

Review: Dr. Takasaki and Mr. Kaihatsu
Please see 'Identifying Problems' section.

Summary

The essential requirements for the test kit are as follows:

  • Affordable
  • No machinery needed and user-friendly
  • Capable of determining the serotype
Review: Dr. Takasaki and Mr. Kaihatsu image
Discussion image
Discussion
iGEM Tokyo Tech

Summary

  • Professor Suzuki's neutralization test is effective.
  • A test kit using the isothermal amplification method also holds potential.
Ryosuke Suzuki
Chief, Laboratory of Hepatitis Viruses II, National Institute of Infectious Diseases

Summary

  • Professor Suzuki's testing method reduces the testing time to 1/3 compared to existing methods.
  • This can lead to savings in reagent costs and labor expenses.
Ryosuke Suzuki image
Masayuki Yamamura / Ken Komiya image
Masayuki Yamamura / Ken Komiya
School of Computing, Tokyo Institute of Technology

Summary

  • Professor Komiya's isothermal amplification technique has not been confirmed for practical use at the research stage.
  • This technique requires skilled technicians.
Yoshihisa Matsumoto
Laboratory for Zero-Carbon of EnergyInstitute of Innovative Research,Tokyo Institute of Technology

Summary

  • 3D-PCR has roughly the same sensitivity as traditional PCR, but it is high-throughput and low-cost.
Yoshihisa Matsumoto image

Conclusion

Through interviews with three experts, we reached the following conclusions. The improved system based on Dr. Suzuki's lineage for the PRNT method is effective for surveillance and highly feasible. Isothermal amplification in dengue fever diagnosis is promising for its simplicity and effectiveness in surveillance, but its practicality is not yet assured, and there are issues with our technique. The 3D-PCR developed by Dr. Matsumoto has significant potential for widespread surveillance use due to its high efficiency and cost-effectiveness in the current state of PCR, making it highly feasible. Therefore, we decided to construct STAND UP using the CFNT based on Dr. Suzuki's system and Dr. Matsumoto's 3D-PCR. For specific technical details, please refer to the Design section.

Communication with Endusers

The impact of our project extends to various organizations and individuals worldwide. As stated in the Description, our goal is to gather and publish data, fostering increased interest among all stakeholders in dengue fever. However, the end users defined are governments that use the data we provide to make decisions on vaccine introduction, assess the effectiveness of vector control, and determine which areas to focus vector control efforts. Another end user is the testing company that will directly use the diagnostic system we developed. We developed the diagnostic method while understanding what diagnostic approach is demanded by society. In this phase, we conducted interviews to re-verify that our product is indeed in high demand by society.

Tokyo Metropolitan Government image
Tokyo Metropolitan Government

Summary

  • Tokyo's vector-borne disease prevention measures consist of preventing mosquito breeding, early detection of vector mosquitos, and preventing the spread of infection when a case arises.
  • In Japan, the importance of surveillance is not emphasized.
How to Confront Unknown Viruses! Pandemic Research Working Group
Pandemic Research Working Group

Summary

  • In the case of COVID-19, large-scale surveillance proved beneficial for policy decision-making.
How to Confront Unknown Viruses! Pandemic Research Working Group image

Conclusion

From the efforts of Tokyo Metropolitan Government in tackling dengue fever and the discussions in the "Stand Up to Unknown Viruses! Pandemic Research Working Group," we became convinced that there is a significant need for surveillance and the data derived from it in regions where the dengue virus is prevalent. Furthermore, from an ethical and social perspective concerning personal information protection, we understood that data should be anonymized before accumulation, rather than accumulating data and then anonymizing it.

H.U. Group Holdings, Inc. image
H.U. Group Holdings, Inc.

Summary

  • The two testing methods can significantly improve the cost and efficiency of dengue fever diagnosis.
  • They can be used as companion diagnostics, and their implementation can have a direct impact on local medical treatments, especially concerning drug administration.
Reaw / Search
Chulalongkorn University

Summary

  • Mainly PCR tests and antigen tests are used for COVID-19 testing.
  • The cost of testing is sometimes paid by institutions like universities, depending on the case.
  • In Thailand, preventive care is not very common, and culture and mindset have a big impact.
  • Regular health check-ups are performed mainly for the elderly, but young people do not use them much.
  • A proposal has been made to conduct new testing methods for free.
  • Free testing alone is not enough incentive, and more motivation is needed to take the test.
Reaw / Search image
Tahanachot image
Tahanachot
Chulalongkorn University

Summary

  • Antigen testing at home is common, but PCR tests can only be done at hospitals.
  • PCR tests have different types and price ranges, presenting some barriers to access.
  • In Thailand, cultural influences on infectious disease testing and prevention are significant.
  • Health check-ups and hospital tests tend to be conducted in specific situations (such as international travel, annual health checks, etc.)
  • National hospitals are crowded, doctors are overworked, and time has become a bottleneck for testing.
  • There is awareness about dengue fever, but proactive measures and testing are hard to conduct.
  • When initial symptoms appear, other infectious diseases like influenza are usually suspected first.
  • It is believed that testing methods led by the country or government provide a sense of reassurance.

Conclusion

From interviews with the testing companies and people living in dengue fever epidemic areas, who are our anticipated end users, we received several pieces of feedback for our project. First, from the testing companies, we understood that our developed testing methods and STAND UP are indeed in demand by society. Additionally, considering ethical issues, we realized the necessity of distributing pamphlets after testing results to provide knowledge for prevention as a part of fulfilling our duty to explain. Furthermore, people living in epidemic areas might not take the test regardless of the cost, and this is largely influenced by cultural attitudes towards healthcare. Therefore, it is essential to first align with their culture.

Technology

Ryosuke Suzuki
Chief, Laboratory of Hepatitis Viruses II, National Institute of Infectious Diseases

Summary

  • The genes, dC and dE, that make up SRIP have clear roles, and without these genes, SRIP does not mature.
  • We gained insight into how much dilution should be done and where to set the positive standard when testing with CFNT.
Ryosuke Suzuki image
Mami Matsuda image
Mami Matsuda
Researcher, Laboratory of Hepatitis Viruses II, National Institute of Infectious Diseases

Summary

  • For SRIP, the gene introduced must be just right in length; not too long, not too short. Genes between 333bp and 375bp need to be set as reporter genes.
  • Considering cell damage and complexity of the operation, lipofection, not electroporation, is suitable. Additionally, using PEI instead of lipofection reagents was suggested. This is also noted on the Engineering Success page.
Vinayak
IISER Pune2_India 2022

Summary

  • The C10 antibody has been reported to have the ability to neutralize all dengue viruses.
  • We have revised the expression infection history we originally used to can identify the amount of neutralizing antibodies against the dengue virus.
Vinayak image

Conclusion

The above feedback had a significant impact on our project. For specific details, please refer to the Engineering Success page.

Improving KAP

In the section, we realized that changing the awareness of people in dengue-infected areas is a necessary strategy to reduce the number of dengue infections. Indeed, multiple surveys have pointed out the need to improve some or all aspects of dengue fever's knowledge, attitudes, and practices (KAP) in epidemic areas worldwide[]. In STAND UP, we conduct surveillance, but to obtain cooperation for this or to use surveillance as a catalyst for reducing the number of infections, it is desirable to pair it with improving dengue fever's KAP. This is because, unlike vaccines and treatments, surveillance does not have a direct effect but produces an indirect effect. Below, we will introduce discussions and practices aimed at enhancing activities to improve dengue fever's KAP.

Decision on Direction

From our previous interviews and literature reviews, we identified the following issues with Knowledge, Attitudes, and Practices (KAP): ① There is variation in the level and stages of KAP across different countries and regions, ② The factors causing this variation include accessibility to information and government response, and ③ The stage of practices is often low in achievement in every country and region. From this, we realized the need to focus particularly on the practices stage of KAP and to think of strategies to improve knowledge and attitudes for some individuals. Regarding the emphasis on the practices stage, we discussed creating guidelines for specific strategies and the possibility of organizing events to motivate actual engagement. Locally, since information about dengue is often obtained through passive mass media such as TV and radio, we concluded that preparing guidelines for future prevention campaigns through these media would be more effective. Therefore, we decided to work on creating pamphlets. Regarding knowledge and attitudes, we found that there is a significant lack of knowledge about mosquito control measures, which are the vectors of dengue fever, compared to knowledge about the disease itself[]. Therefore, to encourage people to engage in activities that prevent mosquito breeding, it is important to first supplement their knowledge. Additionally, in areas where the dengue epidemic is still not widespread, general knowledge about dengue fever is still insufficient[]. To improve this knowledge and attitudes, we must not just transmit information unidirectionally, but engage in bidirectional interactions with the people, expanding their knowledge and improving attitudes. We decided to explore these methods through practical experience and accumulate know-how in bidirectional communication, in organic collaboration with Education.

Creation of the Pamphlet (Dengue Defence Book; DDB)

To create a guideline booklet that could be used in practice, we first needed to refer to credible and accurate information. Therefore, we decided to refer to general information published by the Tokyo Metropolitan Government, the administrative district of Japan's capital, and the Ministry of Health, Labour and Welfare, which is responsible for health administration in Japan. Additionally, we referred to a pamphlet created by iGEM Pune2_india, who conducted a dengue fever project last year, summarizing methods to suppress the outbreak of dengue fever. The booklet is composed of four pages. The first page contains basic information about dengue fever, the second page describes methods to prevent the breeding of mosquitoes, the third page offers methods to avoid mosquito bites, and the fourth page includes an introduction to us, the creators, and credits. Our focus was on the content of the second and third pages. On the second page, we used illustrations to explain methods to suppress the breeding of mosquito larvae, known as wrigglers. On the first page, we had already used diagrams to explain that dengue fever is transmitted by mosquitoes, so we introduced ways to prevent their breeding. We specifically illustrated places like buckets and old tires where water accumulates and does not drain, as these are common breeding sites for wrigglers, making it easier for readers to reference and practice. The third page explained methods to avoid getting bitten by mosquitoes. We discussed reducing mosquito adult habitats such as bushes and grass, minimizing skin exposure, and installing mosquito nets on windows, all illustrated for clarity. These methods are recommended not only by Japanese administrative agencies but also by organizations like the CDC[], and we used specific illustrations to encourage practical implementation. The completed booklet was translated into five languages: Japanese, English, Thai, Indonesian, Hindi, and Malay, and posted on our website.

Engagement with Education

There is a need to improve knowledge and attitudes towards dengue fever in future infected areas. We have been conducting activities in Education to improve knowledge and attitudes about synthetic biology (for more details, see Education ). In this, we explored effective event methodologies. Educational activities to improve knowledge and attitudes are one of the important issues in the world of pedagogy and are conducted daily in schools. This holds true whether it's for dengue fever or synthetic biology – the essence is the same. We interviewed experts in educational technology and high school biology teachers to explore the necessary conditions for successful events.

Hiroko Sano image
Hiroko Sano
Koishikawa Secondary School

Summary

  • the importance of designing classes to sustain student interest
  • implementing systems for group discussions and feedback to enhance knowledge and attitudes.
Saori Okada
Associate Professor, Institute for Liberal Arts, Tokyo Institute of Technology

Summary

  • gained valuable insights into guiding and deepening participants' understanding through the clarification of evaluation criteria and the introduction of specific examples
Saori Okada image

Educational indicators

Based on the above meetings, in our outreach efforts, we have gained confidence that by refining the format of the classes and adapting the content and educational materials, we can enhance the interest and engagement of the participants. Consequently, we recognized the need to consider indicators for evaluating this. After conducting discussions within our team, we decided to base our evaluation on the concept of "active learning" and "deep learning through active participation," which is a part of Japan's educational system. We are currently examining various evaluation methods being implemented in Japan and considering the development of an iGEM-specific evaluation method.[.][..]This will serve as a specific educational material evaluation method that other iGEM teams can also utilize!

The indicators are as follows:

Teaching Materials and Slides
  • Innovate to ensure participants can recognize the issues they should learn about here
    • Make the title easily understandable
    • Introduce the theme at the beginning
  • Design to broaden the participants' interest
    • Address gaps in the current educational curriculum
    • Connect with other educational materials
  • Were there strategies to promote practical actions by participants?
    • Did it touch upon both scientific theory and its societal implementation?
    • Did it provide opportunities for participants to contemplate their future activities?
Event Format
  • Promote mutual learning among participants
    • Ensure time for group work activities
    • Provide opportunities for presenting the results of the work
  • Establish a system for organizers to learn.
    • Ensure participants have room for spontaneous activities.
    • Secure time for participants and organizers to engage both inside and outside the class.
  • Innovate to promote participants' proactive actions.
    • Address both scientific theories and their social implementation.
    • Provide an opportunity for participants to contemplate their future activities

Specific Practices - Three Examples in Education -

Aiming to meet evaluation criteria, specific activities were carried out in the field of Education (for more details, please refer to the Education page). Among them, here are three examples that had a particularly significant impact.

Chiba Prefectural Funabashi High School

Students from Chiba Prefectural Funabashi High School visited TokyoTech twice, where we conducted classes on synthetic biology and infectious diseases. We believe it is important to educate high school students, who will likely be responsible for the future development of science and technology. Therefore, we thought it essential to implement what we consider effective educational practices, while also using this as an opportunity to learn and develop better teaching methods.

One of our concerns is that the Japanese educational curriculum does not sufficiently cover topics related to synthetic biology, such as genetic manipulation. Hence, when educating high school students, we made an effort to link the content to the biology curriculum they are already studying in high school, ensuring that what they learned was continuous and connected to their existing knowledge. Indeed, based on this approach, the first class we conducted received comments like "It was easy to understand and interesting, including the content we learned at school." The surveys on interest in synthetic biology and infectious diseases also showed positive feedback, with over 80% of respondents expressing increased interest. Furthermore, we received comments about how this education could be applied to food production and medical fields using synthetic biology, as well as deepened understanding of testing methods. From this, we can say that this educational initiative provided an opportunity for students to think about applications of synthetic biology.

However, we also received comments from students who were not particularly interested. Indeed, the survey results showed that we were unable to capture the interest of 10-20% of the students. Therefore, we shifted our focus from the structure of the lessons and the content of the slides to aspects we had not paid much attention to before, such as the way we spoke and the frequency of questions, focusing more on the method of communication with the participants. We believed that even if we used good slides and had a well-structured class, it would not be effective if the delivery was not engaging. So, in our second class, we made efforts to increase the opportunities for students to ask questions, actively engaged them in discussion, varied our tone of voice to make the class more appealing, and made the class more interactive. As a result, even though we used the same slides and had the same class structure as the first time, over 90% of the participants reported an increased interest in synthetic biology and infectious diseases. Furthermore, we received more specific questions than before, such as "How are bacteria used in synthetic biology chosen?" and "What is the structure of vaccines that respond to complex viruses?", suggesting a higher level of understanding among the participants.

From these experiences, we learned that when conducting education, it is important to consider the attributes and backgrounds of the participants and link the education to these aspects. Making the educational content as interactive and engaging as possible to capture the interest of participants is crucial. We believe these elements are important in all situations and can be applied to other opportunities.

School Festival(Koudai Festival)

The Koudai Festival is an annual school festival held at TokyoTech. It attracts various people, including those interested in TokyoTech and those wishing to pursue further studies there. We conducted lessons and presentations about genetic manipulation at this event. Although many visitors were already interested in TokyoTech, our exhibit attracted a diverse crowd of all ages and genders, which we saw as an excellent opportunity to explore and practice methods of sharing knowledge with a wide range of people. Consequently, we decided to conduct surveys and collect comments from the participants after the lessons.

We had planned this because we believed that while people are increasingly hearing the term "genetic manipulation," they seldom have the opportunity to learn about it comprehensively. As a result, we received comments like, "Although I often hear about it, having a comprehensive explanation deepened my understanding," confirming that this indeed met the listeners' needs and was an important factor in spreading knowledge. Additionally, we received numerous suggestions regarding the pace of speech and the use of language. Specifically, comments like, "Please use simpler language" and "Please explain more slowly." From this, we learned to speak slowly and use simple language when addressing a diverse audience. These insights have become indicators in shaping our future educational events at TokyoTech.

iGEM Internship

We planned and held an iGEM Internship targeted at high school students, believing that it was necessary to permeate the values of synthetic biology and increase the recognition of iGEM among students. The purpose of this internship was to conduct a hackathon-style activity where Tokyo Institute of Technology's iGEM members and high school students collaborated to develop new infectious disease-related iGEM projects. The internship was designed as a two-day program, with participants divided into two main groups: high school students and college students. On Day 1, each group selected a project theme, with the high school group choosing Crimean-Congo hemorrhagic fever and the college group opting for dengue fever. Day 2 involved collecting materials and refining ideas based on the themes decided on the first day, followed by presentations of the completed projects to the entire group. Notably, the high school team proposed a diagnostic kit for Crimean-Congo hemorrhagic fever based on sheep feces that emits fluorescence in a positive reaction. This proposal was particularly well-received due to its uniqueness and rationality. After the presentations, an active Q&A session further deepened the understanding of the projects. The internship concluded with a round-table discussion to address the high school students' questions about their future educational paths and careers. Throughout the internship, there was a valuable exchange of ideas and collaboration across different academic years, making it a meaningful experience for all participants.

Conclusion

To improve Knowledge, Attitudes, and Practices (KAP) regarding dengue fever, we undertook the creation of a booklet and the development of indicators for a consciousness-raising event. In creating the booklet, we focused on vector control, which is crucial in suppressing mosquito breeding but is currently not widely practiced. We explained strategies to prevent mosquito bites using diagrams. Additionally, translating the booklet into multiple languages is expected to enhance its effectiveness. Regarding the creation of indicators for the event, we integrated it with our educational activities and developed evaluation indicators that could also be used by other iGEM teams, along with their implementation. These two initiatives are organically connected and, through local activities, can significantly contribute to the prevention of the spread of dengue fever.

Implimentation

The purpose of this chapter is to examine the project from an entrepreneurial perspective and to emphasize the social necessity of STAND UP. Up until now, we have discussed how STAND UP has evolved, but we had not considered the direction for its future implementation in society. Through the following interviews, we were able to discern the necessary direction for social implementation. As a result, we were able to clearly define the specific direction for the social implementation of STAND UP.

Takumi Ohashi
Associate Professor, School of Environment and Society, Tokyo Institute of Technology

Summary

  • For the social implementation of the project, it is crucial to have passionate individuals who persistently conduct proof of concept.
  • It is necessary to gather overarching data about potential users, compare it with others, and demonstrate differentiation and strengths.
Takumi Ohashi image
Nao Hosokawa image
Nao Hosokawa
INDEST, Tokyo Institute of Technology

Summary

  • The two testing methods that make up STAND UP, 3D-PCR and CFNT, are in demand in society.
  • The data obtained through the testing methods we are developing have shown the potential to contribute to pharmaceutical companies in determining the production volume of medical resources.
  • We confirmed that STAND UP holds universal value towards social implementation.
Tomohiko Kaihatsu
CEO, VisGene, Ltd.

Summary

  • In the development of dengue fever testing methods, specificity is required to be 100% in terms of distinguishing between other viruses and the dengue virus, as well as identifying the serotype.
  • Whether the sensitivity is not inferior compared to other testing methods becomes one of the criteria.
  • We were able to learn about the specific process of bringing a testing method to society from our competitor, Mr. Kaihatsu.
Tomohiko Kaihatsu image

Implimentation Plan

Based on the insights obtained from the interviews, we have conceived a specific implementation plan for STAND UP:

  • Dengue fever is an extremely important issue that needs to be resolved, as mentioned in SDG 3.3, "End the epidemics of AIDS, tuberculosis, malaria and neglected tropical diseases and combat hepatitis, water-borne diseases and other communicable diseases." Currently, vaccines are under development, and there are no effective treatments, leaving prevention from mosquito bites and symptomatic treatment as the only options.
  • Support for these measures and research for future vaccine introduction are necessary for solving dengue fever. Public institutions like WHO and local health authorities should respond to these needs, but there is a limitation in areas where dengue infection data is publicly available.
  • We propose a low-cost, high-throughput, and highly sensitive method to facilitate the collection of dengue infection data. We aim to accumulate testing data for 1.4 billion people living in endemic areas, which include current outbreak status obtained through 3D-PCR and past infection history of individuals obtained through anonymized CFNT.

These data will be made publicly available worldwide as open-source. There are four anticipated uses:

  • First, it can contribute to the activities of health organizations and governments, promoting accurate epidemic predictions, medical resource distribution, determining and evaluating mosquito vector eradication areas, and appropriate policy implementation for dengue. It can also provide referable data for research and surveys for vaccine introduction and verification of vaccine effectiveness.
  • Second, it can be used by individuals who have undergone testing. Their test results can be made available, encouraging them to actively engage in basic preventive measures against dengue and activities aimed at eliminating mosquito breeding grounds. We have created a guidebook for dealing with dengue to encourage this behavioral change in individuals. We also plan to hold events to raise awareness about the dangers of dengue.
  • Third, it can be utilized in the societies of endemic areas. Local media, businesses, and individuals can measure the spread of dengue in their area and assess the risk of contracting severe dengue. This directly links to infection prevention in that society.
  • Fourth, it can be used by people outside endemic areas. Such extensive surveillance data can be used by data scientists worldwide, making it valuable from an informatics perspective. The open-source publication of dengue data can increase research in informatics with a focus on dengue and create interest in the disease among many people. This is extremely important in shifting the status of dengue from a 'neglected tropical disease' to a disease that naturally garners attention.
  • The 'open-sourcing' of dengue data ultimately reduces the likelihood of individuals contracting dengue and developing severe dengue fever (DHF). This means addressing the economic disparities caused by endemic diseases. In other words, it can contribute to SDG 10.2, &qout;By 2030, empower and promote the social, economic and political inclusion of all, irrespective of age, sex, disability, race, ethnicity, origin, religion or economic or other status."
  • PCR tests and antigen tests are primarily used for COVID-19 testing.
  • The cost of tests is sometimes covered by institutions like universities.
  • In Thailand, preventive care is not very common, and culture and mindset have a significant impact.
  • Regular health check-ups are conducted mainly among the elderly, but not much among the young.
  • It is proposed that the new testing methods be implemented free of charge.
  • Being free alone is not sufficient as an incentive, and further motivation is needed to get people to take the test.

Reference

  1. Azami NAM, Moi ML, Ami Y, Suzaki Y, Taniguchi S, Tajima S, Saijo M, Takasaki T, Kurane I, Lim CK. Genotype-Dependent Immunogenicity of Dengue Virus Type 2 Asian I and Asian/American Genotypes in Common Marmoset (Callithrix jacchus): Discrepancy in Neutralizing and Infection-Enhancing Antibody Levels between Genotypes. Microorganisms. 2021 Oct 21;9(11):2196. doi: 10.3390/microorganisms9112196. PMID: 34835327; PMCID: PMC8618970.
  2. Sasaki T, Moi ML, Saito K, Isawa H, Takasaki T, Sawabe K.Aedes albopictus Strain and Dengue Virus Serotype in the Dengue Fever Outbreaks in Japan: Implications of Wolbachia Infection. Jpn J Infect Dis. 2022 Mar 24;75(2):140-143. doi: 10.7883/yoken.JJID.2021.376. Epub 2021 Aug 31. PMID: 34470970.
  3. Azami NAM, Moi ML, Salleh SA, Neoh HM, Kamaruddin MA, Jalal NA, Ismail N, Takasaki T, Kurane I, Jamal R. Dengue epidemic in Malaysia: urban versus rural comparison of dengue immunoglobulin G seroprevalence among Malaysian adults aged 35-74 years. Trans R Soc Trop Med Hyg. 2020 Nov 6;114(11):798-811. doi: 10.1093/trstmh/traa056. PMID: 32735681.
  4. Runtuwene LR, Kawashima S, Pijoh VD, Tuda JSB, Hayashida K, Yamagishi J, Sugimoto C, Nishiyama S, Sasaki M, Orba Y, Sawa H, Takasaki T, James AA, Kobayashi T, Eshita Y.The Lethal(2)-Essential-for-Life [L(2)EFL] Gene Family Modulates Dengue Virus Infection in Aedes aegypti. Int J Mol Sci. 2020 Oct 12;21(20):7520. doi: 10.3390/ijms21207520. PMID: 33053895; PMCID: PMC7593908.
  5. Kato F, Nio Y, Yagasaki K, Suzuki R, Hijikata M, Miura T, Miyazaki I, Tajima S, Lim CK, Saijo M, Takasaki T, Hishiki T. Identification of inhibitors of dengue viral replication using replicon cells expressing secretory luciferase. Antiviral Res. 2019 Dec;172:104643. doi: 10.1016/j.antiviral.2019.104643. Epub 2019 Oct 31. PMID: 31678478.
  6. Dengue sanofi.https://www.sanofi.com/en/your-health/vaccines/dengue. Accessed: 2023/12/14
  7. Takeda’s Dengue Vaccine Candidate Provides Continued Protection Against Dengue Fever Through 4.5 Years in Pivotal Clinical TrialDengue Takeda Pharmaceutical Company Limited.https://www.takeda.com/newsroom/newsreleases/2022/takedas-dengue-vaccine-candidate-provides-continued-protection-against-dengue-fever-through-4.5-years-in-pivotal-clinical-trial. Accessed: 2023/12/14
  8. History of smallpox vaccinationWHO.https://www.who.int/news-room/spotlight/history-of-vaccination/history-of-smallpox-vaccination. Accessed: 2023/12/14
  9. History of polio vaccinationWHO.https://www.who.int/news-room/spotlight/history-of-vaccination/history-of-polio-vaccination. Accessed: 2023/12/14
  10. Infectious Disease Testing BusinessVisGene, ltd..https://visgene.com/services/infection/. Accessed: 2023/12/14
  11. Japanese Society of Tropical Medicine Students’ BranchJ-Trops.https://www.j-trops.com/. Accessed: 2023/12/14
  12. Beatty ME, Stone A, Fitzsimons DW, Hanna JN, Lam SK, Vong S, Guzman MG, Mendez-Galvan JF, Halstead SB, Letson GW, Kuritsky J, Mahoney R, Margolis HS; Asia-Pacific and Americas Dengue Prevention Boards Surveillance Working Group. Best practices in dengue surveillance: a report from the Asia-Pacific and Americas Dengue Prevention Boards. PLoS Negl Trop Dis. 2010 Nov 16;4(11):e890. doi: 10.1371/journal.pntd.0000890. PMID: 21103381; PMCID: PMC2982842.
  13. Inaba, K., Ebihara, K., Senda, M. et al. Molecular action of larvicidal flavonoids on ecdysteroidogenic glutathione S-transferase Noppera-bo in Aedes aegypti. BMC Biol 20, 43 (2022). https://doi.org/10.1186/s12915-022-01233-2
  14. Matsuda, M., Yamanaka, A., Yato, K. et al. High-throughput neutralization assay for multiple flaviviruses based on single-round infectious particles using dengue virus type 1 reporter replicon. . Sci Rep 8, 16624 (2018). https://doi.org/10.1038/s41598-018-34865-y
  15. Shell”lockiGEM Montpellier 2022.https://2022.igem.wiki/montpellier/. Accessed: 2023/12/14
  16. Mosquito Prevention MonthTokyo Metropolitan Government Bureau of Public Health and Medical Care. https://www.hokeniryo.metro.tokyo.lg.jp/. Accessed: 2023/12/14
  17. Elson WH, Ortega E, Kreutzberg-Martinez M, et alCross-sectional study of dengue-related knowledge, attitudes and practices in Villa El Salvador, Lima, PeruBMJ Open 2020;10:e037408. doi: 10.1136/bmjopen-2020-037408
  18. Hossain MI, Alam NE, Akter S, Suriea U, Aktar S, Shifat SK, Islam MM, Aziz I, Islam MM, Islam MS, Mohiuddin AKM. Knowledge, awareness and preventive practices of dengue outbreak in Bangladesh: A countrywide study. PLoS One. 2021 Jun 10;16(6):e0252852. doi: 10.1371/journal.pone.0252852. PMID: 34111157; PMCID: PMC8192001.
  19. Shafie, A.A.; Moreira, E.D., Jr.; Di Pasquale, A.; Demuth, D.; Yin, J.Y.S.Knowledge, Attitudes and Practices toward Dengue Fever, Vector Control, and Vaccine Acceptance Among the General Population in Countries from Latin America and Asia Pacific: A Cross-Sectional Study (GEMKAP). Vaccines 2023, 11, 575. https://doi.org/10.3390/vaccines11030575
  20. Chinnakali P, Gurnani N, Upadhyay RP, Parmar K, Suri TM, Yadav K.High Level of Awareness but Poor Practices Regarding Dengue Fever Control: A Cross-sectional Study from North India. N Am J Med Sci. 2012 Jun;4(6):278-82. doi: 10.4103/1947-2714.97210. PMID: 22754880; PMCID: PMC3385365.
  21. Prevent Mosquito BitesCDC. https://www.cdc.gov/mosquitoes/mosquito-bites/prevent-mosquito-bites.html. Accessed: 2023/12/14