Team:Thessaly/Human Practices
Overview
In order for a scientific project to be impactful, its design and development need to be executed in relation to the society’s needs and the constraints it poses.
. Our Human Practices methodology consisted of multiple
frameworks which allowed us to effectively assess the status of the problem, interview experts,
stakeholders and regulatory agencies.
Human Practices Methodology
AMALTHEA is a holistic and sustainable approach, aiming to tackle gut dysbiosis both in humans
and animals. Its aim is to offer a proposal for evaluating the state of the gut microflora,
transmitting and storing the evaluation data and alleviating the dysbiosis’ derived symptoms in
a data-driven manner. AMALTHEA’s purpose is to detect, evaluate and ameliorate GI-related
disturbances, giving us results that are reflected in many areas including animal welfare and
human mental health.
Last year, the COCID-19 pandemic made it impossible to conduct proper Human Practices. We understood from revising Amalthea 2020 that the information received was scarce, inconsequential and did very little to broaden our project’s perspective and societal impact.
Through our Integrated Human Practices this year, we emphasized on clarifying:
Figure 1: Engaging with stakeholders.
By receiving feedback and correcting our solution to respond perfectly to our problem can we propose a solution good and responsible for the world.
Figure 2: Integrating stakeholder's feedback.
Figure 3: Our strategy for facing ethical consideration.
Inability to absorb nutrients can lead to IBDs, causing a variety of symptoms, such as diarrhea, anemia, fever, high heart rate and many more, that further aggravate the problem. IBDs are a multifactorial family of interrelated diseases that affect the human gut, which affects 1 in 1000 people worldwide in 2020 (Crohn's & Colitis Foundation,2020).
An aspect of malnutrition-induced immunological deficiency is prevalent in up to 70% of patients with active disease and up to 38% of patients in remission. IBDs have been associated with increased hospitalizations and poor clinical outcome (Balestrieri et al., 2020), as well as being connected to changes in gut microbiota (Lim et al., 2018). In IBD patients, malnutritional deficits are mostly manifested with vitamin A, B, D & K, zinc, and iron deficiencies (Jayawardena et al., 2020)(iGEM Thessaly, 2020).
From the beginning we wanted to tackle the rising problem of gastrointestinal disorders by its root. It is not enough to act upon the diseases when their progression level is high and the condition becomes chronic. Taking into consideration that only around 3% of European Health Budgets are spent on prevention (European Commission, 2017) , our emphasis was based on early detection and ensuring disease prevention in all levels. To tap upon the issue, we turned to those that are on the first line of dealing with the rise of diet-related non-communicable diseases.
Name:Potamianos Spuridon
Affiliation: Professor of Gastroenterology at the Thessaly University School of Medicine
“What did we learn from him?”
Dr. Potamianos informed us that in order to properly diagnose IBD on a patient include their clinical image, microbiological data, endoscopy data and histological pictures. Thus, making diagnosis in late stages extremely difficult and pointless. Also, he laid out that current treatment of IBDs are aminosalicylates, corticosteroids and immunosuppressants which affect in an extreme way the gut health and balance. Finally, he pointed out how much their dietary habits are altered to very simple dietary fibers due to their inability to digest.
“How did we act on his feedback?”
We integrated Dr. Potamianos’ feedback by altering the main direction of the project: We no longer aimed for the diagnosis of gut-related disorders, such as IBD, but for the functional evaluation of the gut microbiome, a more feasible and realistic goal for the device we are designing. The experts Dr. Potamianos proposed were later included in the formation of a multidisciplinary team that through our application could communicate with the patients and give advice upon the user’s personalised data.
Name: Antigoni Chalkia
Affiliation: Clinical Dietitian with MSc. Toxicology
“What did we learn from her?”
Ms. Chalkia informed us that many patients with heavy symptoms visit a dietitian before going to a gastroenterologist, with 50% of all dietitian patients suffering from a GI disorder. In those cases the established examinations take place, such as colonoscopy, and the doctor can then make their diagnosis. In cases, though with very mild or no symptoms, a non-invasive technique would be ideal. Ms. Chalkia was very positive towards the evaluation of the gut microflora through a capsule and proposed the idea of utilising this capsule to collect epidemiological data.
“How did we act on her feedback?”
Since it’s been understood that there’s no specific dietary pattern for patients and that the complexity of the late onset of GI disorders leads us to take further steps for prevention.
Name:Kotsopoulou Theodora
Affiliation: Gastroenterologist
“What did we learn from her?”
Dr. Kotsopoulou laid out the difficulty of identifying and clarifying if someone suffers from Inflammatory Bowel disorders. She also pointed out the severity of pharmaceuticals that a patient might receive while their condition is chronic and not reversible. Also, she has observed a rising tendency of mild gastrointestinal disorders that cannot easily be differentiated.
“How did we act on her feedback?”
We continued our work on both the aforementioned aspects, being more confident on our course of action, and conducted further research on gastrointestinal disorders and checked the stability of the gut microbiome in different conditions.
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Name:Papada Efstathia
Affiliation: Clinical Dietitian, MSc, Ph.D
“What did we learn from her?”
Dr. Papada told us that although every patient is different, a dietary pattern that fits most of them and at the same time is affordable (especially for IBD patients who make a lot of health related expenses), is the Mediterranean diet. She noted that patients are characterized by their symptoms: flare and recession of symptoms. When in flare, patients suffer immensely while a nutrient malabsorption is observed.
“How did we act on her feedback?”
She urged us to check on how dietary patterns can be implemented in a meaningful way on our application, to assist treatment efforts by nutrition.
Name:Konstantinos Gerasimidis
Affiliation: Professor of Clinical Nutrition, University of Glasgow
“What did we learn from him?”
People with increased risk of developing IBD (10 and 20% of patients with IBD having a first-degree relative who also has IBD. The concordance rates for monozygotic twins to be about 60% for CD and 20% for UC) do not present with gut dysbiosis. This is why we shouldn’t restrain ourselves only to IBDs. Very hot topics related with gut dysbiosis: Cancer and Cardiovascular diseases. pH values in the gut can be altered in pathological conditions, such as Crohn’s disease which in turn can lead to microbial disturbances.
“How did we act on his feedback?”
We should see the bigger picture in regards to gastrointestinal disorders and focus on examining how gut conditions in pathologies can affect our detection system
Figure 4.a: Our project’s evolution through our human practices work
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Reforming our approach
Through our discussions with stakeholders, we aimed to reaffirm our goals and at which stage of disease progression our project fits in. We understood that the complexity of Inflammatory Bowel Disease at late stages does not allow efficient medical action, making disease prevention essential, in very early stages of pathogeny. Also, as Mr. Potamianos and Ms. Kotsopoulou suggested, not only are the pharmaceuticals used to tackle IBDs extremely heavy with many side-effects but at the same time there is no non-invasive form to accurately evaluate gut’s physiology.
This fact enhances the limited diagnosis of IBDs in younger ages but also does not help minimize the social stigma and taboo surrounding those disorders.
One other key factor that we integrated was that IBD pathology is only a symptom of the widely spread diet-related, non-communicable diseases. Experts urged us to unravel the missing link between all gastrointestinal disorders, which is their starting point and how we can effectively combat them.
We believe that AMALTHEA will be a strong tool in those experts’ arsenal.
The Rising Threat of Diet-Related NCDs
In Europe, at present, more than half of the adult population is overweight or obese and, children and population groups of lower socio-economic status are the most severely affected. Non-communicable diseases (NCDs) such as cardiovascular diseases, cancer, obesity and diabetes account for 70% of all deaths in the European Union (EU) and their costs represent 70-80% of European healthcare costs (European Commission, 2020). Currently, one in three people worldwide is malnourished while 1.9 billion adults are overweight and 462 million adults are underweight (Fluitman, 2017).
Unsustainable and unhealthy diets with increased demand for livestock products, and calorie-dense, nutrient poor and (hyper) processed food are a leading NCD risk factor and a driver of obesity rates.
NCDs are largely preventable through effective interventions that tackle shared risk factors such as unhealthy diets, physical inactivity and abuse of alcohol.
A better understanding of the mechanisms driving dietary choices will offer opportunities to reduce risk factors for NCDs and facilitate consumers to adhere to a long-lasting, healthy, pleasurable and nutritional (European Commission, 2020).
Figure 4.b: Our project’s evolution through our human practices work
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The Missing Link
“The Microbiome is the next big thing!” Helen Papadimitriou, IBD Specialist
The gastrointestinal (GI) tract harbors trillions of microorganisms (CCFA, 2014). The gut microbiome plays a critical role in the integrity of the intestinal barrier, nutrient absorption, metabolic homeostasis, and immune maturity, particularly in the early stages of life (Robertson, 2020). Dietary components can alter the intestinal microenvironment favoring some bacteria over others. That can affect the microbe–microbe interactions involved in the fermentation of indigestible nutrients, thus the modulation of energy extraction (Xiao, 2019) (Fluitman, 2017).
Figure 4.c: Our project’s evolution through our human practices work
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Our engagement with stakeholders unveiled that every gastrointestinal issue begins from disturbances in the gut microbial community. Those can be characterized as shifts in the abundance of certain strains over others or the decline of their metabolic capabilities (Morgan, 2012) (FAO, 2019).
This phenomenon is known as Gut Dysbiosis
“The next big question is to reveal the relation between dysbiosis and enteropathies” Xenoulis Panagiotis, Associate Professor of Companion Animal Medicine
Figure 4.d: Our project’s evolution through our human practices work
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Since the design of our first year’s effort aimed on the microbiome, we wanted to affirm our expectations and move forward in evolving our system. So, we turned to a wide variety of experts working on microbiome research to receive feedback on how to approach this vast, unknown network of microorganisms and if our system indeed can identify gut dysbiosis
Name:Kotsopoulou Theodora
Affiliation: Gastroenterologist
“What did we learn from her?”
Dr. Kotsopoulou informed us that a lot of research efforts and modernized approaches in gastrointestinal disorders aim on the gut microbiome, thus making it an exceptional new target for diagnostics and therapeutics. However, due to the evasiveness of gi disorders and their detection, a non-invasive method is optimal. She also informed us that that more and more patients come with mild symptoms, however most of them are stigmatized by the issue their facing so their is no real communication between patients and doctors.
“How did we act on her feedback?”
We continued our work on both the aforementioned aspects, being more confident on our course of action, and conducted further research on how to bridge the gap between doctors and patients
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Name:Antigoni Chalkia
Affiliation:Clinical Dietitian with MSc. Toxicology
“What did we learn from her?”
Ms. Chalkia informed us that many patients with heavy symptoms visit a dietitian before going to a gastroenterologist, with 50% of all dietitian patients suffering from a GI disorder. In those cases the established examinations take place, such as colonoscopy, and the doctor can then make their diagnosis. In cases, though with very mild or no symptoms, a non-invasive technique would be ideal. Ms. Chalkia was very positive towards the evaluation of the gut microflora through a capsule and proposed the idea of utilising this capsule to collect epidemiological data. As for the application, she agreed that it is an easy way for a patient to communicate with an expert and that generally speaking dietitians would also find it handy. She also gave us the idea of providing patients with a personalised dietary pattern based on the information collected from the capsule and the notes the user is going to make on the interactive interface. Furthermore, the application would be an easy way for patients to describe to the dietitian details about their clinical picture that are difficult to communicate in person, such as the morphology of their feces.
“How did we act on her feedback?”
Given the prevalence of GI disorders in dietitian’s patients and the positive feedback about the integration of the capsule into their practice, we decided to include dietitians in our multidisciplinary team of experts that our application is going to connect with patients. Ms. Chalkia’s proposal for the utilisation of the capsule for research purposes was our inspiration that later, through our partnership with team Aalto-Helsinki, was formulated into a core aspect of our application. We also endorsed the idea of dietitians providing personalised dietary advice in an asynchronous manner through the application’s interface. Finally, we integrated a stool chart for a fast and easy description of the patient’s stools.
Name:Potamianos Spyridon
Affiliation:Professor of Gastroenterology at the Thessaly University School of Medicine
“What did we learn from him?”
Dr. Potamianos confirmed that SCFAs are a universal biomarker that can be used in the evaluation of the microbiome’s state, but not for diagnosing any kind of gastrointestinal diseases. Regarding the involvement of different experts in gastroenterology patients, he proposed gastroenterologists, dietitians, psychologists and psychiatrists.
“How did we act on his feedback?”
We integrated Dr. Potamianos’ feedback by altering the main direction of the project: We no longer aimed for the diagnosis of gut-related disorders, such as IBD, but for the functional evaluation of the gut microbiome, a more feasible and realistic goal for the device we are designing. The experts Dr. Potamianos proposed, were later included in the formation of a multidisciplinary team that through our application could communicate with the patients and give advice upon the user’s personalised data.
Name:Helen Papadimitriou
Affiliation: IBDs Specialist at the General Hospital of Larissa
As last year, she affirmed that SCFAs are still not a widely used biomarker,even though it's linked with many different gastrointestinal issues. One thing she noted was to be alert about the materials surrounding our detection system and not cause any sort of further inflammation or allergic reaction.
Name:Brian Coombes
Affiliation: Professor and University Scholar Chair, Department of Biochemistry & Biomedical Sciences
Through our interactions in "New Frontiers in Gut Research, Dr Coombes highlighted the role of the microbiome in IBDs: There's an observed enrichment and abundance of certain strains that alter the metabolic capabilities of the gut microbiome. As he suggested in our webinar: "The next holy grail about the microbiome is to target specifically harmful bacteria. Finally on factor that also urged us to took into consideration is that immunosupresants deal with the problem but not the root of it, thus making prevention a key factor in gi disorders.
Name:Panagiotis Xenoulis
Affiliation: Associate Professor, Companion Animal Medicine, University of Thessaly
Dysbiosis shows low levels of SCFAs but we don’t have data supporting if there is a correlation that links dysbiosis with the lowering of SCFAs or if it’s a phenomenon that occurs after dysbiosis is present. He informed us thet in order to evaluate the percentage of SCFAs it is essential that we conduct: 1)Q-PCR on eight different bacterial strains 2)Τargeted Μetabolomics urine and fecal samples
Ιf we wish to check about gut dysbiosis all we have to do is a trial of administering certain feed (therapeutic test) and then check how the gut responds responds
Ιf we have a response to antibiotics this means we had dysbiosis (but sometimes the the usage in a high rate of antibiotics can have reversed results)
Ιf not this means that we must move forward with an endoscopy
He concluded our meeting saying that “The key point is to understand which came first? The gut dysbiosis or the enteropathy”
After revising our last year's design based on the measurements of SCFAs (acetate, propionate, and butyrate) levels, metabolites produced by the fermentation of indigestible carbohydrates (Müller et al., 2019) (iGEM Thessaly, 2020), we received positive feedback to proceed forward:
Figure 4.e: Our project’s evolution through our human practices work
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A synthetic biology solution is an appropriate solution to harness the immense potential in the gut microbiome’s role in pathology. All around the world, there are perspectives which range from being always supportive of synthetic biology, to those who believe synthetic biology is a technology of last resort.
Based on our last year's effort, to accomplish the communication between gut flora and the host organism, we designed a bio-electronic interface to enable real-time monitoring of metabolite deficiencies on the patient’s smartphone.
Our final goal is the development of a micro-bio-electronic ingestible device that will support physicians to provide help to their patients based on their identified needs.
Figure 4.f: Our project’s evolution through our human practices work
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We also modified our detection system and ensured a safe and useful application environment, where data can be received and examined by experts. Synthetic biology allows us to meet the stakeholder needs, elaborate on their call and provide a complete and holistic solution to the problem at hand.
Revising our detection system method
To further develop our detection system we turned to experts to verify if a capsule containing genetically modified bacterial strains is indeed necessary for a proper evaluation of the gut microbiome. As Professor Chatzigeorgiou told us, liquid biopsy is a promising method to ensure non-invasiveness on the patient’s body, but is this achieved by an ingestible capsule? Is there a way to avoid a controversial way of administration or is it crucial to walk this path to receive the most accurate image of the gut flora’s health?
We turned to several stakeholders to see alternatives on our detection system administration method and confirm or alter our approach.
Name:Chatzigeorgiou Artemis
Affiliation: Professor of Bioinformatics, Department of Electrical and Computer Engineering, University of Thessaly.
She underlined the common ground between our project and contemporary trends of biosciences. Excited with our idea, she emphasized the importance of the microbiome and its relation with its host as a very promising field of study in a very broad spectrum of applications, such as soil microbiome in agriculture, mouth microbiome in human well being etc. She also stated the urgent need for non-invasiveness in modern medical practices, which has already started manifesting itself through the development of methods such as liquid biopsy. Finally, she recommended Dr. Klapa as a very skilled person who could help us with the experimental and modeling parts of our project.
Name:Dimitrios Fatouros
Affiliation:Professor at Pharmaceutical Technology at the department of pharmacy, Aristotle University of Thessaloniki
“What did we learn from him?”
We started our meeting by sharing our thoughts about delivering the SCFASs through a buccal or sublingual patch. However, professor Fatouros told us that SCFAs cannot be absorbed sublingually or buccally unless they undergo a certain processing and be administered as UFA-somes through microneedles on the surface of the patch. When describing our capsule’s design, we asked the Professor whether regular pharmacists would be able to load the bacterial cultures in the tank on their own. He replied that something like that is not possible due to the lack of expertise and proper equipment. In order to work with systems like ours, the pharmacy should fulfil the criteria of Good Manufacturing Practice, something that doesn’t happen even to most university laboratories in Greece. After that, we discussed our plan for sight specificity in our measurements. Professor Fatouros, commented that for something like that to happen we would probably need a coating for the capsule that consists of a pH sensitive polymer that dissolves in the pH of the colon (~7,8). The coating can cover only the semipermeable membrane in order to both start the measurements at the right time point and protect the membrane from dissolving in the stomach’s acidic conditions.
“How did we act on his feedback?”
Firstly, we rejected the idea of sublingual/buccal administration of SCFAs because of the difficulties in the manufacturing process, thus moving on with our second plan, the oral administration. Secondly, we rejected the idea of involving regular pharmacists in the loading and assembly of the capsule, starting to investigate the idea of finding a sole industry that would possess both the required equipment and expertise to take on the manufacturing process from start to finish. Finally, we took the Professor’s advice on the capsule’s coating and started looking for pH sensitive materials that would fit our needs.
Name:Maria Klapa
Affiliation: Head of Metabolic Engineering and Systems Biology Lab (MESBL), FORTH/ICE-HT
She instucted us to revise our administration method by considering other microbial communitites from which we can evaluate the physiology of the host such as the oral or vaginal microbiome. This was an effort to avoid the controversial administration method and ensure maximum safety that won't affect the physiology of the host. However due to the real-time measuring method of our detection system, Dr. Klapa urged us to maximise our safety mechanisms and eliminate any chance of contamination.
Name:Konstantinos Gerasimidis
Affiliation: Professor of Clinical Nutrition, University of Glascow
Our conversation with Dr. Gerasimidis was mostly about the capsule and its capacity to detect and measure the SCFAs concentration in the gut, and partly about our probiotic’s design. He told us that the fact that our device is able to conduct real-time measurements at the site of the SCFAs production, is extremely important, and maybe its biggest advantage, because the data are not impaired. The traditional SCFA-measuring trials are based on stool samples, working with only 5% of the SCFAs produced by the gut. In order to overcome the randomness in the system in pathological conditions, clinical trials with large samples of patients should be conducted. This way we could theoretically standardise the expected pH values.
Developing our application system
In 2020, the approach in designing a medical app was contradictory. According to the 2019 opinion of the Expert Panel on Effective Ways of Investing in Health (EXPH), digital technologies offer new opportunities for delivering more personalized, effective and efficient healthcare. EU policies have consistently emphasized the importance of digital solutions such as eHealth and have accentuated positive aspects of digital innovations. However, there are various concerns on data management and their safety. Doctors and clinical nutritionists last year suggested that it would be dangerous for patients to have a subjective view of their condition from just an app, that can’t accurately diagnose the disease and take into account various factors. (iGEM Thessaly, 2020).
In order to decide whether we should move forward for a digital health system and how to respond to the rising tendencies and needs for digitalization, we had to turn to everyone that may benefit from an application network for the gut microbiome’s data to be evaluated.
Name:Efstathia Papada
Affiliation: Clinical Dietitian, MSc, Ph.D
She was positive towards the idea of integrating a section in our application, in which the user is going to take notes about their dietary habits and the foods that trigger or alleviate their symptoms. This practice can assist the process of nutritional screening, and prevent the manifestation of nutritional deficiencies. On the topic of multidisciplinarity, Dr. Papada commented that generally speaking, although it would be very beneficial for the patient, almost no dietitian sends their patient to mental health experts. Thus, a platform through which this interconnection is facilitated, would be very helpful in the dietitians’ practice. We closed our conversation, with Dr. Papada pointing out two issues: The time window between successive incidences of yoghurt consumption should be defined and, the medical confidentiality between the patient and the experts (gastroenterologist, psychologist and dietitian) should be ensured.
“How did we act on her feedback?”
Dietitians are going to have access to the application as experts in order to consult the users on the selection of the best fitting dietary pattern, while combining their work with a mental health experts’. This way, we achieve better health outcomes for the patient. Medical confidentiality is ensured by allowing each expert to view only the information about the user’s health status, that are relevant to their profession, while being prohibited from viewing the other experts’ consults unless the patient grants them access.
Name:Kotsopoulou Theodora
Affiliation:Gastroenterologist
Concerning the mobile application, she confirmed the importance of communication between different experts, in order to achieve the best outcomes in terms of treatment strategies.
Name:Antigoni Chalkia
Affiliation: Clinical Dietitian with MSc. Toxicology
As for the application, she agreed that it is an easy way for a patient to communicate with an expert and that generally speaking dietitians would also find it handy. She also gave us the idea of providing patients with a personalised dietary pattern based on the information collected from the capsule and the notes the user is going to make on the interactive interface. Furthermore, the application would be an easy way for patients to describe to the dietitian details about their clinical picture that are difficult to communicate in person, such as the morphology of their feces.
Name:Terrovitis Manolis
Affiliation: Postdoctoral Fellow, Institute of Information Systems of the Athena Research Center
Our discussion with Dr. Terrovitis did not focus on a particular subject, yet it provided many useful pieces of information concerning the development of e-AMALTHEA. First of all, he noted the significance of data anonymization and encryption, and how they are achieved with technologies such as blockchain. He made remarks on data safety/privacy, by explaining to us that the user’s data can only be shared with a third person for medical/research purposes only if the former consents, that we have to ensure that our cloud service follows the GDPR regulations for Europe (if developed to follow only certain regulations that are not in effect in Europe that might cause a problem) and that a medical expert is obliged to keep a user’s data up to 10 years from their last visit. He also acknowledged that a Bluetooth transmission can be safe, in the sense that its range is relatively small, so it minimizes the chances of the system being attacked by someone within radius.
Throughout consultations with stakeholders, we’ve implemented their advice and evolved our project in the following ways:
After receiving this feedback, we started ideating a software tool, a mobile application, which would link AMALTHEA’s complex measurement processes with the convenience a mobile device provides. This was the beginning of e-AMALTHEA. At the same time, we began reviewing literature on the subject of how we can alleviate gut dysbiosis.
Priority: Alleviating Gut Dysbiosis
After our discussions with stakeholders, we began to search through literature to understand what a good solution might be to treat gut dysbiosis. Our amelioration system includes metabolic engineering of the gut microbiome and engineered strains that replenish the quantities of missing SCFAs, acting as a cell-factory. Though our design was affirmed last year by Adamantia Kyriakou, Associate Professor in Microbiology at Department of Nutrition and Dietetics at Harokopio University of Athens, we wanted to communicate our idea to more experts to ensure that our approach is optimal.
Name:Konstantinos Gerasimidis
Affiliation: Professor of Clinical Nutrition, University of Glasgow
“What did we learn from him?”
What is also considered a big advantage in our design, this time regarding the genetically engineered probiotic, is its ability to degrade cellulose, thus overcoming the necessity of cross feeders: direct production of SCFAs from cellulose in one single cell, makes the cell independent from the host’s microbiome (which may or may not have the capacity to produce the precursors of SCFAs). However, he expressed his concerns about the pH-sensitive coating of the capsule, upon which the whole site-specificity design was built. pH values can be altered in pathological conditions, such as Crohn’s disease, thus affecting the expected pattern of SCFA concentrations’ fluctuations in the final diagram. Moreover, overproduction of SCFAs in the gut can lead to Irritable Bowel Syndrome symptoms. Finally, he wondered how we overcome the problem of potential development of resistance to cumate.
“How did we act on his feedback?”
In order to overcome the randomness in the system in pathological conditions, clinical trials with large samples of patients should be conducted. This way we could theoretically standardise the expected pH values. A similar approach could be followed also to determine the exact levels of SCFAs, above which symptoms of Irritable Bowel Syndrome occur. Finally, an experiment in which we determine the time window until the cells accumulate enough mutations to become cumate resistant, is vital in order to give proper instructions for the consumer.
Name:Konstantinos Kormas
Affiliation:Professor of Microbial Ecology and Aquatic Environment, Department of Agriculture, Ichthyology & Aquatic Environment
“What did we learn from him?”
Right now, research and therapeutic approaches regarding both humans and fish, focus more on manipulating the existing gut microbiome than adding new microorganisms to it. However, the colonisation of the gut by our genetically engineered probiotic is not prohibitive, because we designed good biosafety systems. Professor Kormas also commented that our overall approach was complete and did not leave any red flags uncovered. He advised us to be careful with the fermentation products of the SCFA producing bacteria, such as H2, CO2, H2S, CH4, because many of them can become toxic. Finally he noted that although SCFAs are already used as a supplement in fish farming, an imbalance in Fe utilisation by the different microbial populations is usually observed.
“How did we act on his feedback?”
We intend to investigate further our probiotic’s tolerance to H2 and the problem of antagonism for Fe utilisation, and consider coadministration of methanogenic bacteria (professor Kormas informed us that they are already tested in fish) in order to detoxify the potentially dangerous byproducts. Furthermore, the adaptation of our bioelectronic capsule, and especially the probiotic, for use in fish farming could be a future vision in the animal welfare aspect of our project.
By interviewing experts, as well as attending global events showcasing similar topics that we could be inspired from, we realised that our approach is relevant and responsive to society’s needs.
However, emerging evidence demonstrates that environmental factors, including diet, may play an important role in pathogenesis. This highlights the need to provide guidance to physicians and patients regarding which foods may be harmful, beneficial, or safe to consume.
We delved deeper into the role of nutrition on a person’s gut health,and evaluated how the gut responds to different forms of nutrition. Our aim was to answer two important questions: Is our synthetic biology solution enough and can it be in line with Greek dietary tradition?
Name:Yiorgos Antonopoulos
Affiliation: Farmer, Owner of Antonopoulos Farms
While the gut microbiome is not closely related to farmland, the relation between what we eat and what we metabolise is interlinked with our inside world. We asked Mr Antonopoulos how can we promote well being and ensure health for all. His answer was simple: learn to hear the world around you and see the unseen. When food is highly industrialized, it is logical to face growing dietary related issues. Only by listening to our inside and "trusting our gut" can we achieve ultimate health.
Name:Katerina Moutou
Affiliation: Associate Professor of Vertabrate Biology, Head of the Laboratory of Genetics, Comparative and Evolutionary Biology, University of Thessaly.
Our Primary Investigator instructed us throughout our project's evolution. When talking about nutrition, she pinpointed on the importance of having proper food with limited additives. She also highlighted that human beings and their microbial communities are not the same organism:they co-exist througout our body, provide nutrients and protection for us while in turn we also provide a safe environment to grow. That's why personalisation is of great importance: We have co-evolved with our microbes and we must listen to their dietary needs, which spring from locally produced food. She urged us to promote through our application a personalized dietarty pattern, in accordance with our birthplace food to avoid risk factors that lead to NCDs
Name:Potamianos Spyridon
Affiliation:Professor of Gastroenterology at the Thessaly University School of Medicine
Dr Potamianos, as said before, gi disorders minimize the nutritional options of the patients health. When nutrients that can be absorbed are not enough to sustain the body, it is logical to face issues that become chronic. We fall into a loop that we cannot improve diet to tackle the rising disorder and the patients condition worsens.
It became evident to us that producing SCFAs solely to enhance metabolic capabilities does not restore the loss of microbial diversity. One other key factor that we took into consideration is that due to the dynamic relationship between microbial populations, it is impossible to alter the microbiome’s composition in a long-term way simply by administering our strain.
Thus, our team has implemented a two-way solution to stop dysbiosis on its root:
Treatment Through Our Food
Hippocrates’ notion “Let food be thy medicine and medicine be thy food” may be an ancient way of saying that dietary intervention is essential to maintain intestinal micro-ecological homeostasis and then impact host physiology and health (Xiao, 2019).
Our nutritional tradition not only has been beneficial for our well-being and development but also contributed to our co-evolution with the gut microbiome. The most common element across the Greek population is the Mediterranean diet which particularly influences microbial plant polysaccharide degradation potential, SCFAs production and pectin metabolism. One of our main questions that arose was how we can implement proper nutritional guidance and at what magnitude can we actually recoup the gut microbiome’s physiology.
Name:Konstantinos Gerasimdis
Affiliation: Professor of Clinical Nutrition, University of Glasgow
Diets that can enhance our microbial community are based on cellulose and are of plant origin. the Mediterranian Diet can be a good template that experts can base on and move forward on adjusting the needs of individual patients.
Name:Efstathia Papada
Affiliation: Clinical Dietitian, MSc, Ph.D
Dr. Papada told us that although every patient is different, a dietary pattern that fits most of them and at the same time is affordable (especially for IBD patients who make a lot of health related expenses), is the Mediterranean diet.
According to our stakeholders' call, we aim to tackle low levels of SCFAs in an integrated manner, improving human health via dietary modification and enhancing the microbiome with a synthetic bacterial strain.
Now that we have successfully grasped the framework in which we operate, we moved towards action, responding to the feedback received in our experimental design, both in a dry and wet lab level.
Figure 4.g: Our project’s evolution through our human practices work
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To ensure our solution was aligned with stakeholder values, we consulted both wet and dry lab experts.
We sought out interdisciplinary experts to meet the absence of substantive work on chemical kinetics modeling and metabolic modeling, which bordered on physics and mathematics respectively.
Our Dry Lab work emphasized in three major goals:
- Designing and implementing an in silico model that supports the wet lab design by providing a mathematical and computational simulation of the underlying genetic mechanisms
Name:Michail Stamatakis
Affiliation: Associate Professor in Chemical Engineering Department of Chemical Engineering University College London
Our meeting with Dr Stamatakis proved to be very useful for the progress of our modeling, considering that he has authored the paper from which we extracted most of our ODEs. Firstly, he emphasized the fluid nature of modeling problems such as ours, indicatively remarking that the parameters we use can vary up to two orders of magnitude, and even that different parts of literature can suggest different values. He also proposed which assumptions we should make, such as extracellular IPTG stability and that all species follow Mass Action Kinetics. By assuming extracellular IPTG stability we simplified our model without sacrificing any information, while Mass Action Kinetics turned out to be the only method compatible with a stochastic simulation. Most importantly, he confirmed that the aforementioned equations are stiff and analyzed the importance of the kind of solvers we would use on them, helping us understand and choose solvers wisely.
Name:Maria Klapa
Affiliation: Head of Metabolic Engineering and Systems Biology Lab (MESBL), FORTH/ICE-HT
In our second meeting, Dr Klapa expanded on our modeling work, which she considered really impressive. The most important and interesting piece of advice she gave us was in reference to the switch from GFP to Tyrosine in the lac operon model: if we wanted to replace GFP with Tyrosine, we had to be very careful, given that the two species do not follow the same kinetics. When talking about GFP, she introduced the very useful and on-point terms “imaging” and “semi-quantitative”, to emphasize the crudity of GFP as a reporter gene in comparison with Tyrosine, which can possibly follow a more standard kinetics. In addition, she advised us to conceptually separate endogenous from exogenous species in our model, so that it visualizes the wet lab’s experiments more accurately, even if an in silico simulation may not examine such information per se.
We can conduct experiments in order to achieve a pseudo stable state at our bacterial population inside the capsule.
In order to assess the saturation and toxic effects of SCFAs, we can design a metabolic network in which the rate of bacterial growth is correlated with the concentration of the SCFAs in the microenvironment.
The signal’s accuracy at the final system, depends on the exact amount of tyrosine produced by the capsule’s bacteria as a response to the SCFAs’ concentration. However, tyrosine is an amino acid that exists naturally in the gut, thus interfering with the output’s production. In vitro experiments can determine the ratio of external (gut) tyrosine/internally produced tyrosine (from the capsule’s bacteria) that our bacteria utilise when being induced. We feed the bacteria with a known ratio of unlabeled glucose/UC13 isotope, and then we measure the quantity of the radiolabeled tyrosine that was produced. Each kind of glucose resembles the gut and capsule derived substrates and this test can help us determine the capacity of bacteria to utilise each of them. Thus, we can correlate these results with the conditions that take place during the measurements in the gut, and fine-tune our system’s output.
- Developing an application system that is responsible and safe for the end user and consulting experts to ensure data safety
Name:George Kakarotzas
Affiliation:Associate professor of Software Technology with an Emphasis on Architectures' components
The meeting with Dr Kakarontzas challenged the concept we had in mind at the early stages of the development of our application. He questioned the security of the method we were to use to store the data, and also made us wonder if it was necessary to involve experts in the whole evaluation process. His ideas caused a brainstorm in our team, the fruit of which was the decision that we didn’t want to give up our initial concept, but just adapt to Dr Kakarontzas’s concerns: we improved our application’s safety by using a hybrid (cloud and conventional) database, in which all information about a user is encrypted, and we supported the meaningfulness of an expert’s involvement by basing our opinion more solidly on literature, and, in fact, we considered allowing experts of different specialties to participate in a patient’s evaluation – after all, it proved to be a very sensible tactic to achieve a holistic approach. Furthermore, he advised us on the tools we would find useful in the coding of e-AMALTHEA.
Name:Vlachos Vasilis
Affiliation:Assistant Professor at the Department of Economics, University of Thessaly
Dr Vassilis Vlachos is active in research fields related to cyber security. Therefore, he advised us on security issues, and started our conversation by saying that we did not have the luxury for experimental versions, we had to apply well established methods to secure the users’ data. What really drew our attention was his comment on data leakage policies: to ensure good practices towards our users, we should establish a standard policy which will be followed in case of leakage, which, among others, must include informing the authorities and the victim within 72 hours. He also noted how important data anonymization is and suggested that we use the least information possible to identify users; minimum information required leads to optimal user anonymization and better overall safety. Finally, he advised us not to include certain modern technologies such as cloud only because we were “carried away” by their popularity, but stoically assess the advantages and disadvantages of the methods we will use.
Name:Petros Spachos
Affiliation:Associate Professor at the School of Engineering, University of Guelph
Dr Spachos helped us see our hardware system from a more realistic point of view and gave us insights on how medical IoT (Internet of Things) systems work. He started by analyzing the characteristics that really make a technology “novel” nowadays. Then, he listed the network protocols that are suitable for a system like ours and the ones that are not: he explained that the use of a mobile device narrows down the protocols we can use for data transmission. Thus, he rejected a protocol that would operate on the medical band, and suggested Bluetooth and Wi-Fi connections instead. We chose the former, because we didn’t want the communication between our transmitter and receiver to require a router in between. Lastly, he proposed that we use a method of compression on our data before we transmit them, giving the example of ECG (Electrocardiography) signals, which only transmit their spike (peak) values
- Perfecting our detection system, by providing clear schematics for the capsule dimensions, architecture and specifying the reasoning behind our choices.
Name:Alexandros Katsaounis
Affiliation: Professor at Chemical Technology & Applied Physical Chemistry section, Department of Chemical Engineering, University of Patras
“What did we learn from him?”
We should look into materials that the electrode is covered with that are specific for L-DOPA. In our case we probably need an electrode that works as an effective conductor for the released electrones (e.g. a golden electrode) as the reaction of L-DOPA → L-DOPA quinone + e is a spontaneous one. Professor Katsaounis also pointed out the importance of taking into consideration the medium’s pH when choosing the right electrode: electrodes are often specific for a certain range of pH values, and their mechanical stability also depends on this factor, leading to the electrode’s corrosion in case we don’t make the appropriate adjustments.
“How did we act on his feedback?”
Based on the feedback gained by Dr. Katsaounis, we certified the design of our electrodes sensor and rethought the connection we had established, from the sensor to the processor, for optimal data transfer without noise. When testing the electrodes in our system we are going to produce a calibration curve to correlate SCFAs’ concentration with ΔV, in order to standardise our system.
Name:Paraskevas Kitsos
Affiliation:Associate professor at Department of Electrical and Computer Engineering
Dr Kitsos is a professor at the university of Peloponnese and his research interests focus on integrated circuit design for security systems, cryptographic algorithms and signal processing systems, secure hardware design, Hardware Trojans in integrated circuits and hardware design for embedded systems. While searching for ways to manually encrypt the data transfer from the capsule to a mobile device, we came across one of Dr. Kitsos’s papers which broke down the cryptography of the Bluetooth communication protocol and how that is implemented in hardware. Albeit it was on early Bluetooth versions, it really helped us understand the functionality of its mechanisms. After meeting with him and presenting to him our designs and ideas, he was very intrigued by our project. We mainly talked about our choice of hardware and its possibilities and extensions. We figured out that the data transfer in Bluetooth is encrypted by default with AES/CCM, which are block ciphers algorithms. The main problem we pinpointed was how to implement the authentication between the two devices so that we can eliminate a “man in the middle '' interference. Then, we discussed possible implementations and he gave us some directions on how to continue further.
Name:Dermenoudis Stergios
Affiliation: Biologist and Researcher at Dr Logothetidis’s lab, solid state physics section, Physics Department, Aristotle University of Thessaloniki
Dr Dermenoudis informed us about the main metrics that are involved in choosing the right coating material or membrane for a construct like ours. He emphasized that the membrane materials (and especially semipermeable ones) are mainly classified with respect to the size of their pores, and also that the technique with which they are manufactured plays a major role in their future function and use.
Name:George Papadopoulos
Affiliation: Professor at Department of Chemical Engineering, National Technical University of Athens (NTUA)
We came in contact with Dr Papadopoulos in order to get an insight about the membrane that we were to use as an interface of the biosensor of the capsule. After a brief internet meeting, we came to two conclusions: firstly, biopolymers are a better choice for our application if we can make them work and, secondly, he unfortunately could not help us to the extent that he expected. Nevertheless, he redirected us to Dr. Tsiourvas that specializes in fatty acid specific membranes.
Developing CERAS, our synbiotic supplement
With the advancements in synthetic biology, probiotic engineering offers more flexibility and fine-tuning capacity than others (Aggarwal, 2021). However, we wanted to 1) ensure long term sustainability of our strain to produce large quantities of SCFAs, 2) try to provide our supplement through a non-pharmaceutical way and thus be more friendly to the consumer and 3) ensure that we do not disrupt the microbial community’s natural function.
Name:Persephone Giannouli
Affiliation:Assistant Professor of Technology and Food Quality Control of Plant Origin
“What did we learn from him?”
Professor Giannouli characterised the idea of encapsulating the bacteria as innovative, but expressed some concerns about the effect the alginate may cause on the yoghurt’s pH. She proposed stir yoghurt, a yoghurt that coagulates with stirring, as a good candidate for adding the alginate beads and keeping them intact until consumption. Something that she suggested we mention on our product, is the proposed daily doses. In this case we need to take into consideration not only the substances that are contained solely in our product, but also those that are consumed daily from other sources, too, thus inflicting a cumulative impact. Finally, she mentioned that although there are special and stricter regulations around foods for children (1-2 years old), at the same time there are no warnings on products that should not be consumed by kids.
“How did we act on his feedback?”
We decided to compose a guide that addresses the consumers, giving them detailed instructions about the responsible use of the product, hence ensuring both the safety of the consumer and environmental containment. These instructions include the population groups that should not consume the product, excluding children below the age of five. Finally, the yoghurt’s distribution was decided to take place only from pharmacies to better control the buying public. Future vision: Future experiments are needed to determine the viability of our probiotic in the yoghurt as well as the effect of alginate on the yoghurt’s pH.
Name:Alexandra Meziti
Affiliation:Postdoctoral Researcher, Department of Nutritional Sciences and Dietetics of International Hellenic Universit
“What did we learn from her?”
After being briefed on our project, Dr. Meziti agreed with our approach of preventing the manifestation of gastrointestinal disorders by evaluatiating the gut microbiome and alleviating the dysbiosis. She was also positive towards the colonisation strategy and the in situ production of the desired metabolite, since most modern probiotics lack the long term effect, by dying shortly after the administration. She expressed two main concerns: the first one was the possibility of the SCFA production negatively affecting the metabolic pathways of other bacterial species in the gut. The second one was the competition of our probiotic strain with natural bacteria populations, for food availability.
“How did we act on his feedback?”
We developed a system that gives our bacteria the ability to utilise the cellulose of food, a polymer that naturally goes to waste since gut microbiota do not have the means to degrade it. In this way, we managed to increase the carbon flux to overcome the metabolic strain of SCFA production while achieving to eliminate the competition between our strain and the natural ones over nutrition sources. Future vision: In future experiments and bibliographic research, we intend to elucidate the metabolic relationships between different bacterial strains and predict, with relative precision, the impact of our probiotic’s colonisation to the natural microbial populations.
Name:Nestoras Vlachos
Affiliation: CEO of Epsilon Health
“What did we learn from him?”
Mr. Vlachos proposed that we get in contact with researchers and retrieve data concerning the SCFA concentrations in healthy and pathological conditions and also conduct research about the legislation concerning the administration of GMOs. He introduced us to the list of Generally Recognised and Safe (GRAS) microorganisms that are approved for use as probiotics. Finally, after hearing our idea about administering our probiotic through a yoghurt, he underlined the importance of the conditions in a production line; we should be careful and add our probiotic at a stage that is not going to reduce the probiotic’s viability or the quality of the yoghurt itself.
“How did we act on his feedback?”
We checked the legislation around the administration of GMOs in humans, and realised that our product can not be implemented in Europe. Thus our long term plan, is to promote it to the United States’ market. After that we checked the GRAS list and chose the strain E.coli Nissle 1917 as the best candidate to use as a chassis organism. We also started collecting data about the yoghurt’s production line and arranged new meetings with experts that specialise on food technology.
Name:Kormas Konstantinos
Affiliation:Professor of Microbial Ecology and Aquatic Environment, Department of Agriculture, Ichthyology & Aquatic Environment
“What did we learn from him?”
Right now, research and therapeutic approaches regarding both humans and fish, focus more on manipulating the existing gut microbiome than adding new microorganisms to it. However, the colonisation of the gut by our genetically engineered probiotic is not prohibitive, because we designed good biosafety systems. Professor Kormas also commented that our overall approach was complete and did not leave any red flags uncovered. He advised us to be careful with the fermentation products of the SCFA producing bacteria, such as H2, CO2, H2S, CH4, because many of them can become toxic. Finally he noted that although SCFAs are already used as a supplement in fish farming, an imbalance in Fe utilisation by the different microbial populations is usually observed.
“How did we act on his feedback?”
We intend to investigate further our probiotic’s tolerance to H2 and the problem of antagonism for Fe utilisation, and consider coadministration of methanogenic bacteria (professor Kormas informed us that they are already tested in fish) in order to detoxify the potentially dangerous byproducts. Furthermore, the adaptation of our bioelectronic capsule, and especially the probiotic, for use in fish farming could be a future vision in the animal welfare aspect of our project.
Name:Tassou Chrysoula
Affiliation:Director of Research at the Institute of Agricultural Product Technology (ITPA) of Elliniko Of the Agricultural Organization "DEMETRA"
We met with the team of Dr Tassou and the institue ITPA to discuss our probiotic strains administration method. She suggested to utilise a strain that is well characterized and able to survive fermentation conditions. She suggested the Lactobacillus rabuosis can be used as a reference strain that can withstand low pH levels and the fermentation procedures. However, if it is to achieve a medium of one milion bacterial strains per gr,, our probiotic strain must be administared after the fermentation culture. Finally, she suggested micro-encapsulation of our probiotic strain within prebiotic fibers to withstand the extreme, acidic conditions of the gut.
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Name:Sandrine Uttenweiler-Joseph and Fabien Letisse
Affiliation:Researcher at Biosystems and Process Engineering Laboratory (LISBP)
Fabien and Sandrine are the PIs of INSA-UPS int Toulouse France. They were the ones who provided our constructs for the production of SCFAs and they were interested in learning who the development of our project was going.The gave us credible feedback and the urged us to took into consideration the gut's environment and factors that may halt the production of acetate as well as to consider what may happen when our engineered bacteria die if the DNA released can have any impact on the rest of the existing strains.
Name:Efstathios Giaouris
Affiliation:Associate Professor of Food Microbiology, Department of Food Science and Nutrition of the University of the Aegean
Professor Giaouris presented us the two main methods to produce a probiotic dairy supplement:A probiotic strain added in the fermentation culture or added afterwards. Judging by our expiremental design, Dr Giaouris suggested that our course of action would be determined by how our strain can affect the fermentation procedure and by how the acidic conditions of a yoghurt can affect it. Regardless of our choices, his suggestion was to move forward with administering our probiotic at the end of the fermentation procedure.
Taking into consideration our stakeholders ideas and suggestions, we propose to ameliorate gut dysbiosis with CERAS, a living bio-therapeutic synbiotic supplement, that can act as a living, SCFA producing factory for the gut flora. An engineered probiotic strain encapsulated in prebiotic fibers that produces SCFAs at a pinpoint location in the user’s gut. In a personalized manner, we alleviate gut dysbiosis, ensuring continuing stability on the gut’s microbiome health.
Figure 4.h: Our project’s evolution through our human practices work
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Ensuring Maximum Safety
Since our multi-stage approach to tackle diet related NCDs has a synthetic biology core and taking into consideration that the Greek society is not yet familiarised with concepts of Synthetic Biology, we’ve set as a high priority to provide a safe and responsible solution to the world
That’s why, we tried to answer important safety considerations , safeguarding our project from our certain red flags.
We focused on:
To framework and verify our experimental design, we turned to safety experts to receive feedback and formulate a responsible safety strategy.
Name:Tessa Alexinian
Affiliation:Safety & Security Program Officer
To ensure that our solution is safe and responsible, we've summarized our safety experimental design and decided to contact with the iGEM Safety Commitee. We spoke with Tessa and asked questions surrounding wet lab design, data management and serving our values throughout our activities. Fortunately, Tessa was quite content with our approaches and had a fruitfull convesation surrounding safety issues.
Name:Mathiopoulos Konstantinos
Affiliation:Professor of Molecular Biology, Department of Biochemistry and Biotechnology, University of Thessaly
“What did we learn from him?”
We presented him our whole project and received very positive general feedback. Then, we elaborated more on the biosafety aspect, especially at our probiotic, discussed his concerns. In professor Matthiopoulos’ opinion, the safest way to approach this aspect is by modifying the whole translational machine of our bacteria thus developing a completely orthogonal system.
“How did we act on his feedback?”
We explained to him that due to restrictions of time, it is not possible for us to execute such an ambitious plan. For that reason, so we concluded that, realistically, our best option is to combine at least two kill switches in each bacterial category -probiotic bacteria and bacteria in the detection system of the capsule- to ensure both safety for the patient and environmental containment.
Name:Vourtchis Dionisis
Affiliation:Representative of EBSA in Greece
“What did we learn from him?”
We made a presentation of our biosafety plan and discussed its details. Mr. Vourtsis insisted on the importance of environmental containment and the potential dangers that derive from the process of loading the bacteria in the tank.
“How did we act on his feedback?”
The environmental containment aspect is covered by two specific kill switches, one in each one of our products -engineered probiotic and metabolite detection bacteria in the capsule- that are responsible for the bacteria’s elimination once they exit the human body.
Name:Dimitrios Karpouzas
Affiliation:Professor of Enviromental Microbiology & Biotechnology, University of Thessaly
Our last year’s kill switch, for the environmental containment via the cold-inducible kill switch was rejected from Prof. Dimitrios Karpouzas, in view of the extensive heat waves. He told us that the cold-inducible switch won’t be applicable to regions with higher temperatures. We replaced it with a knockout strain for the dapA so the bacteria can’t survive without the supplementation of the cell wall component dapA. This switch is an extra layer of environmental containment. This way, the bacteria will die from starvation inside the capsule after its rejection, eliminating a possibility of environmental contamination.
Name:Karras Panagiotis
Study:Departement of Biochemistry and Biotechnology, University of Thessaly
The capsule, after its defecation, will be flushed through the toilet. The capsule cannot be collected after usage, so we had to predict and prevent possible environmental contamination from capsule accumulation. So, we contacted Dr.Panagiotis Karas, an expert on toxic waste management. He highlighted that the capsule, if not collected, will eventually break, and both the batteries and the microchip will be released. He proposed that the capsules should be collected from the trash piles after the regular separation of these types of waste. The capsules should be disassembled, and the batteries and the microchip should be distributed to appropriate recycling companies.
After expert interviews and literature research, we finalized our safety plan and addressed this issue holistically for AMALTHEA, which is summarized as follows.
AMALTHEA’s original design aimed for the functional evaluation of the gut microbiome and the amelioration of gastrointestinal symptoms related to dysbiosis. Through our bibliographic research we came across a considerable amount of literature concerning the role that gut microbes play in our general well-being, including both our physical and mental health. Our starting point was the inadequately investigated, often non-communicable and always extremely costly issue of comorbidity of enteric and mental disorders. Aiming to shed light on this perspective and due to our partnership with team Aalto-Helsinki, we elaborated on the subject of mental health.
Our response to see if we could act upon this well-documented issue was to engage with experts in the field of mental health experts.
Name: Paraskevi Angelopoulou
Affiliation: Doctoral researcher at Aristotle University of Thessaloniki
“What did we learn from her?”
As Ms. Angelopoulou informed us, in Greece, no Medical school, except for one, has any subject related to the patient’s psychology or the way in which a doctor should make unpleasant announcements. Stigma, depression and anger are very common in chronic patients making the communication with the doctor difficult. This in turn, often leads to a negative attitude on behalf of the doctors which can lead to medical errors, mistakes in drug prescriptions and bad communication. Another problem that exists almost everywhere, and especially in Greece, is the lack of multidisciplinarity in the treatment of gut-related disorders, which translated mostly as absence of mental health experts from the development of treatment strategies for gastroenterology patients. Our conversation about the application was also very fruitful. Ms. Angelopoulou was very positive towards our idea and commented that an application like ours can increase the patient’s accessibility to experts without stigmatising them. We shared our idea of organising emotional support groups in which users that share the same or similar problem, communicate and support each other. However, she was sceptical about this idea, since teams like these need to be coordinated by an expert who poses the appropriate boundaries. Her general proposals for new features at the app were the following: connecting the users with experts in an easy and fast way, make the interface friendly and use questions that focus at the present time.
“How did we act on his feedback?”
The information about the lack of gastroenterologist’s training on mental health issues, inspired us to design a booklet that addresses them and offers basic information about the comorbidity of gut and mental disorders as well as the importance of taking into consideration the patient’s mental state when engaging with one. Moreover, we built upon the fact that the formation of multidisciplinary teams of experts is necessary to achieve higher quality of treatment in gastroenterology patients, adding psychologists/psychiatrists as experts that can use the application to evaluate the patients results and provide advice. The questionnaires we were thinking of integrating in the app were articulated in a way that focuses the user’s mind in their somatic symptoms and feelings more at the present time. Regarding the application, we started off by rejecting our initial idea to assemble support groups from strangers without the ceaseless presence of a psychologist. After that we tried to develop a friendly interface that would seem welcoming and appealing for the users, and finally we articulated the questions we were thinking of integrating in the app in a way that focuses the user’s mind in their somatic symptoms and feelings more at the present time. Future vision: We plan to develop a map on which psychologists and psychiatrists near the location of the user will be marked with a pin. This way someone with the need for help can click on the pin and arrange an appointment in an easy and fast manner.
Name: Agorastos Agorastou
Affiliation: Assisistan Professor of Psychiatry, Aristotle University of Thessaloniki
“What did we learn from him?”
80% of chronic diseases are related to stress and thus constitute a big demographic group. Depression, one of the most predominant stress related disorders, poses the No1 cause of disability, while 60% of patients with depression underestimate the effect of stress on their condition. The capsule’s non-invasiveness is a big advantage. Patients, especially those with depression will be positive to the idea of using the capsule in order to assess their gut microbiome’s state and possibly improve their condition. Regarding the probiotic, the average psychiatrist does not make suggestions for dietary adaptations to their patients in order to alleviate their disorder’s symptoms. One possible cause for this phenomenon is the lack of guidelines regarding the prescription of probiotics to psychiatric patients. However, doctors are open to the idea of integrating probiotics to their treatment practices. Finally, something that we need to take into consideration is that gastroenterologists prescribe psychiatric drugs that empirically work for the alleviation of gut-related symptoms, disregarding their effects on the patients mental state
“How did we act on his feedback?”
We decided to include psychiatric patients that suffer from diseases that fall into the category of stress related disorders such as posttraumatic stress disorder (PTSD), Panic, Generalized anxiety and Alcohol abuse, and focus especially on depression due to both its high prevalence and severity. We intend to approach psychiatrists in the future as an appealing target group that can promote AMALTHEA (capsule) and CERAS (yoghurt with SCFA producing probiotic) to a new and considerably broad audience. We are going to focus on better understanding the psychiatrists’ background knowledge about diet and design a strategy about informing them in a constructive way, especially on the probiotic prescription. Last but not least, we are going to turn to gastroenterologists, talk about the exact cases of psychiatric drugs’ prescription and inform them about the destabilising effects of those drugs on the patients’ mental health.
Name: Paraskevi Angelopoulou
Study: Doctoral researcher at Aristotle University of Thessaloniki
“What did we learn from her?”
We started off our meeting by talking about the initial version of our booklet for gastroenterologists. Ms. Angelopoulou, noted that it felt close to the reader but it probably would be more useful, at its current form, for gastroenterologists’ patients. Moreover, she proposed adding some information that would make the link between mental and gut disorders clearer. After that, we moved on to discussing the application in more detail, compared to our first meeting. We concluded that a feature that organises the data collected from the weekly filling of the questionnaire would be organised in 6-month diagrams that could be relatively easy to be reassessed by mental health experts. She proposed that we add some questions relative to the user’s medical history that would be answered when the user first opened the application, along with functionality, socialization and mood related questions that would be asked on a weekly basis. The Initial questions together with the medical history of the user, can be used to personalise the application, while the integration of a mindfulness exercise could close the questionnaires’ filling so that the user leaves with a feeling of relaxation.
“How did we act on his feedback?”
We took Ms. Angelopoulou comments under consideration and developed two new booklets, one for gastroenterologists and one for patients, keeping in the last one the friendly layout the first version had, while designing a completely different one for the gastroenterologists. Regarding the application, we integrated a small number of questions that will be answered the first time someone opens the application, as well as three distinct sections of questions related to the functionality, the socialization and the mental state of the user. Future vision: The Initial questions will be combined with the medical history of the user, in order to personalise the application; certain answers to selected questions will slightly change the apps’ interface to better suit the psychological needs of the user. Moreover, a mindfulness exercise will accompany the questionnaires, promoting relaxation and “psychological rehabilitation” for the user. Activities like this, also function as a tool to connect an experience such as filling a questionnaire, with emotions that attract the user to repeat the experience and hence, be responsible towards the task. Regarding the 6-month collections of data, we think that the best way to present the information is through a somatic symptoms versus time, and a mental disorders’ symptoms versus time diagram.
Name:Agorastos Agorastou
Affiliation: Assisistan Professor of Psychiatry, Aristotle University of Thessaloniki
“What did we learn from him?”
We received extensive feedback on the first booklet we designed, with the goal of briefing gastroenterologists about the prevalence of mental health disorders in their patients as well as the comorbidity of gut and mental disorders. The main concerns dr.Agorastos expressed, are: 1) The images used in the booklet should be very carefully selected, in order not to enhance, but alleviate the stigma and discrimination against individuals with mental health disorders. 2) The phraseology should also be chosen very carefully. For example, he proposed changing the title: “What is a mental disorder?” into “How to you conceive the concept of mental health?”. 3) He encouraged us to design a booklet for gastroenterology patients, in order to inform them on basic concepts concerning their mental health.
“How did we act on his feedback?”
We worked on every aspect of the booklet that was pointed out as concerning and composed two new versions of the material, one for gastroenterologists and one for their patients.
Eventually we decided to investigate this issue more thoroughly, starting by asking two basic questions:
During these conversations, and in spite of many specialists having very divergent backgrounds, we repeatedly received a prompting towards the consideration of mental health in the evaluation of the general impact that gut microbiota imbalances inflict on the human body.
Microbiota imbalances reported by the capsule can lead psychiatrists to the conclusion that tackling the problem with diet modifications might be a partial solution. The fact that the measurements are going to be taken in a non-invasive manner, plays a major role in the inclination of patients to take the capsule. Studies have shown that targeting the microbiota by probiotic supplementation could alleviate psychological distress including an index of depression (Raypole, C., 2019). At this point a patient warned by the doctor, can alter their diet either using our application, or directly consuming our probiotic, as it is a product personalized and safe for general use at the same time. The probiotic can be proposed by the doctor even without the patient using the capsule, as it is a product that can be used only as a prevention measure.
Figure 4.i: Our project’s evolution through our human practices work
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When we approached Professor Konstantinos Kormas, Professor of Microbial Ecology and Aquatic Environment at the University of Thessaly, to consult him regarding how the gut conditions affect our synbiotic’s survivability, we were introduced by Dr Kormas in an area of research which is the fish gut microbiome. The similarities in morphology of the gut microbiome between humans and fish, as well as the common strategies used to tackle general gastrointestinal pathologies raised our interest to review the literature and implement our project to the animal gut microbiome.
At a similar time period, our work was to be presented at the 9th Panhellenic Veterinary Students Conference that was organised by I.V.S.A.- International Veterinary Students ’Association. The aim of this action was to inform the student community about how synthetic biology could contribute to safeguarding animal health and One health.
Applying the One Health approach to the microbiome allows for consideration of both pathogenic and non-pathogenic microbial transfer between humans, animals, and the environment (Trinh et al, 2018). Different animal species house unique microbiomes, often of equal or greater complexity compared to the human microbiome. As in humans, animal microbiomes influence the health of livestock, pets, disease vectors, and foundational species that uphold ecosystems. Only by diving into and understanding such interactions, will we be able to generate innovative interventions to prevent and manage a variety of human and animal health conditions.
Those two events were instrumental to the final iteration of our project:
As always, we turned to relevant researchers and production companies to assess for whom our project would be helpful.
Name: Konstantinos Kormas
Affiliation: Professor of Microbial Ecology and Aquatic Environment, Department of Agriculture, Ichthyology & Aquatic Environment
Dr Kormas, being an expert on the fish gut microbiome, he inspired us to work and evaluate how can our system could be implemented on animal populations
Name:Panagiotis Xenoulis
Affiliation: Associate Professor, Companion Animal Medicine, University of Thessaly
Dr Xenoulis main research interest is the study of the domesticated animal gut microbiome, specifically on cats. We were shocked to find out the percentages of animals hospitilized with gi disorders as well as the interest of the scientific community and public in solving those issues
Name: Institute of Animal Health
Affiliation: DEMETRA
During our visit to the Research Institute of Animal Science, we discussed our project on its whole and we concluded that it is importan to from it move away from the constant usage of antibiotics and move to a more preventative approach, by monitoring in real-time the microbial conditions in the gut of production animals and through probiotic administration strengthen their immunity and performance. The use of antibiotics as growth factors was very extensive in recent years. Although there were efforts to reduce antibiotics in animal production, a lot of farmers continue to use them. Also, they expressed their interest of using our project as a research toolkit to receive valuable data on the microbial communitites on livestock.
Through our discussion, we understood that different approaches must be implemented for different animal species and purposes.
Our implementation in this manner helps us to fulfill the aim of the project to include a One Health context.
Figure 4.j: Our project’s evolution through our human practices work
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Having ideated and designed a solution that was developed from the advice and insight of wet and dry lab experts, and born from the values of our stakeholders, our team looked towards proposed implementation.
General Implementation
Detection System
Given the complexity of the capsule (especially the engineered bacteria’s sensitivity), the whole device should be assembled strictly by pharmaceutical companies that possess both the required equipment and the expertise. At this point, we need to underline the fact that the most delicate part of our device is the biological one, and for the aforementioned reasons a pharmaceutical company can be assigned the task of preparing and loading the bacterial cultures in the capsule’s tank. On the other hand, the electronic parts will not stand in the way of the whole process, due to their ease of assembly and their affordability; thus, no third-party manufacturer is required in the overall system’s mass production. This way, we ensure environmental containment and safety for the manufacturers themselves and ,at the same time, we reduce the possibility of error, which rises with respect to the number of intermediaries. Right after manufacturing, the capsule is distributed under strict conditions, (especially temperature ones) to clinics and pharmacies.
e-Amalthea
As in all components of AMALTHEA, we considered data security very seriously in our application’s development process, too. We protect our user’s data on many levels: first and foremost, e-AMALTHEA strictly follows the GDPR regulations, and, additionally, we connect it with a hybrid database that combines the advantages of a cloud and a conventional database. Knowing the importance of data anonymization, we also ensured that the data is encrypted with standard algorithms the cloud database provides, so that no user can be linked or correlated to their sensitive information such as SSN, TIN, medical results etc.
CERAS
We add our probiotic in the yoghurt in the late stages of its production, separately from the bacteria that conduct its fermentation. After achieving the desired composition and bacterial load, the yoghurt gets sealed and stored at low temperatures so that we preserve the probiotic’s viability and capacity for the SCFAs’ production.
Name:Epsilon Health
Affiliation: Private Sector
To effectively launch our probiotic supplement into the market, we reached out to Epsilon Health, one of the key probiotic-producing companies in Greece. Nestora Vlachos, the company’s CEO instructed us to evaluate current legislation on the development of engineered probiotic strains as well as ensuring the strain’s safety for market release.
As discussed, there are already hybrid dairy products with probiotics but not many of them administer their probiotic strain outside the fermentation culture due to survivability issues and product's taste alteration. However, due to our integrated alginate bids design, Mr Athanasoula was interested in the potential this project can have if implemented after fermenatation. Also, the 2 month period of lifespan of a yoghurt can be achieved, thus making our project financially viable.
The probiotic’s lifespan is approximately two months, a period long enough for the product’s distribution and its stay on the pharmacy’s shelf. The CERAS’ package, except for the yoghurt itself, also includes the proper amount of cumate for the colonisation termination and a detailed guideline for the product’s safe consumption.
In Vitro, Pre-Clinical and Clinical Trials
Moving beyond our experimental design and our project’s ideation, the first step that every scientific project must circumvent is to test our project’s function through multilear trials. Since the gut microbiome and its bacterial formation varies from person to person, we were skeptical on how we can lay out a proper framework to conduct clinical trials to meet the microbiome’s delicate nature.
Once again, we turned to experts to pinpoint a plausible course of action to cover any ethical, safety and fidelity concerns on our core project aspects.
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Name: Adele Costabile
Affiliation: Associate Professor (Reader) in Nutrition at Roehampton University
Dr. Costabile is an expert on the pathology of the gut microbiome, especially on colon cancer. We were introduced to her by Dr. Meziti to receive feedback on how can we standirize our methods and assess our systems fidelity. She instructed us to conduct an in vitro gut microbiome environment. This means that we simulate the bacterial strains, metabolite, pH conditions and temperature in 72-h setting. Moreover, to achieve microbiome objectivity to be sure about the testing limits, she suggested a dietary protocol that can prepare a gut microbiome to our tes.
Name: Nikolaos Kostomitsopoulos
Study: Vice-President of the Federation of European Laboratory Animal Science Associations (FELASA)
Nikolaos Kostomitsopoulos was our go-to choice to ensure and include every propable outcome in our implementation designs. He instructed us that our pre-clinical trials should be conducted to a rodent and non-rodent animals. So, for our capsule system to be tested judging by our systems dimensions and morphology of the animal GI tract, he suggested to move to dog and a pig for our animal selections. He also supported us with all the necessary documentation (listed below) to conduct our pre-clinical trials.
After our interview, we managed to lay out a course of action to properly conduct in vitro, preclinical clinical and trials
The endpoint of Synthetic Biology is industrialisation. Although as a iGEM project we serve the fundamental scientific discipline such as synthetic genomics, engineering and molecular biology, it is also important to integrate it to the market. Thus, we aimed to build bridges with the industry and relevant stakeholders, such as pharmaceutical and probiotic companies to properly access our place in the market and strategize our next moves for introducing AMALTHEA to the industry sector.
Name:EFSA
Affiliation: European Organisation
In order to assess the legislative landscape of the European Union, we tried contacting the European Food and Safety Administration. We've send our request to meet with representatives of EFSA and receive feedback about our dairy products safety and risk assesment, gmo marketing analysis and set-backs as well as on what does the future hold on implementing a synthetic biology project in our world. Unfortunately, due to the European Union's strict COVID restrictions, with most health experts fighting on the frontlines, our meeting was postponed and we await further reply.
Project Phase 1 Reflection
To properly evolve previous work, we must first understand it, examine its intention, and aim and make
proper adjustments to move forward. In order to do so, it was necessary to consult our last year team’s
effort, evaluate the project’s direction and fill-in the missing links.
Last year, the COCID-19 pandemic made it impossible to conduct proper Human Practices. We understood from revising Amalthea 2020 that the information received was scarce, inconsequential and did very little to broaden our project’s perspective and societal impact.
Through our Integrated Human Practices this year, we emphasized on clarifying:
- The effectiveness of our approach solely to Inflammatory Bowel Disorders and the project’s evolution to the broader spectrum of gastrointestinal disorders.
- Whether to invest on a cell-free, salivary calprotectin detection system or some iteration to our modular system or not.
- Maximize our efforts to provide safe data transmission and management as well as providing expert feedback on everyday users.
- Highlight nutrition’s catalytic role in disease progression and severity.
Integration
Our Human Practices work throughout this year aims to understand the mutualistic relationship between
people and science. This means asking the right questions, having sustained and meaningful conversations
with stakeholders and field experts while implementing knowledge received in every developmental step of
our journey.
Figure 1: Engaging with stakeholders.
By receiving feedback and correcting our solution to respond perfectly to our problem can we propose a solution good and responsible for the world.
Figure 2: Integrating stakeholder's feedback.
Ethics and Safety
When approaching delicate issues, we must be vigilant and mindful of the impact our project may have. So, we followed a framework in which we must pursue and clear out any doubts and scepticism around our project, trying to consolidate any probable outcome and reframe our approach.
Figure 3: Our strategy for facing ethical consideration.
Integrated Human Practices Framework
Surveying the Problem
Overview Of The Problem
Gastrointestinal diseases, especially inflammatory bowel diseases (IBDs),
Crohn’s disease (CD,) and Ulcerative Colitis (UC), are chronic inflammatory conditions,
which have been increasing in incidents, prevalence, and severity, in recent years(Elmaliklis et al., 2019).
Inability to absorb nutrients can lead to IBDs, causing a variety of symptoms, such as diarrhea, anemia, fever, high heart rate and many more, that further aggravate the problem. IBDs are a multifactorial family of interrelated diseases that affect the human gut, which affects 1 in 1000 people worldwide in 2020 (Crohn's & Colitis Foundation,2020).
An aspect of malnutrition-induced immunological deficiency is prevalent in up to 70% of patients with active disease and up to 38% of patients in remission. IBDs have been associated with increased hospitalizations and poor clinical outcome (Balestrieri et al., 2020), as well as being connected to changes in gut microbiota (Lim et al., 2018). In IBD patients, malnutritional deficits are mostly manifested with vitamin A, B, D & K, zinc, and iron deficiencies (Jayawardena et al., 2020)(iGEM Thessaly, 2020).
Reflecting First Responders
“Based on statistics, 1 in 10.000 in Greece suffer from IBDs. But, those are the ones that came forth. Everyone else, suffers in silence”
Kostas Demertzis, HELLESCC
From the beginning we wanted to tackle the rising problem of gastrointestinal disorders by its root. It is not enough to act upon the diseases when their progression level is high and the condition becomes chronic. Taking into consideration that only around 3% of European Health Budgets are spent on prevention (European Commission, 2017) , our emphasis was based on early detection and ensuring disease prevention in all levels. To tap upon the issue, we turned to those that are on the first line of dealing with the rise of diet-related non-communicable diseases.
Potamianos Spyridon
University of Thessaly
University of Thessaly
Name:Potamianos Spuridon
Affiliation: Professor of Gastroenterology at the Thessaly University School of Medicine
Dr. Potamianos informed us that in order to properly diagnose IBD on a patient include their clinical image, microbiological data, endoscopy data and histological pictures. Thus, making diagnosis in late stages extremely difficult and pointless. Also, he laid out that current treatment of IBDs are aminosalicylates, corticosteroids and immunosuppressants which affect in an extreme way the gut health and balance. Finally, he pointed out how much their dietary habits are altered to very simple dietary fibers due to their inability to digest.
“How did we act on his feedback?”
We integrated Dr. Potamianos’ feedback by altering the main direction of the project: We no longer aimed for the diagnosis of gut-related disorders, such as IBD, but for the functional evaluation of the gut microbiome, a more feasible and realistic goal for the device we are designing. The experts Dr. Potamianos proposed were later included in the formation of a multidisciplinary team that through our application could communicate with the patients and give advice upon the user’s personalised data.
Antigoni Chalkia
Dietician
Dietician
Name: Antigoni Chalkia
Affiliation: Clinical Dietitian with MSc. Toxicology
Ms. Chalkia informed us that many patients with heavy symptoms visit a dietitian before going to a gastroenterologist, with 50% of all dietitian patients suffering from a GI disorder. In those cases the established examinations take place, such as colonoscopy, and the doctor can then make their diagnosis. In cases, though with very mild or no symptoms, a non-invasive technique would be ideal. Ms. Chalkia was very positive towards the evaluation of the gut microflora through a capsule and proposed the idea of utilising this capsule to collect epidemiological data.
“How did we act on her feedback?”
Since it’s been understood that there’s no specific dietary pattern for patients and that the complexity of the late onset of GI disorders leads us to take further steps for prevention.
Kotsopoulou Theodora
Gastroenterologist
Gastroenterologist
Name:Kotsopoulou Theodora
Affiliation: Gastroenterologist
Dr. Kotsopoulou laid out the difficulty of identifying and clarifying if someone suffers from Inflammatory Bowel disorders. She also pointed out the severity of pharmaceuticals that a patient might receive while their condition is chronic and not reversible. Also, she has observed a rising tendency of mild gastrointestinal disorders that cannot easily be differentiated.
“How did we act on her feedback?”
We continued our work on both the aforementioned aspects, being more confident on our course of action, and conducted further research on gastrointestinal disorders and checked the stability of the gut microbiome in different conditions.
Papada Efstathia
Clinical Dietitian, MSc, Ph.D
Clinical Dietitian, MSc, Ph.D
Name:Papada Efstathia
Affiliation: Clinical Dietitian, MSc, Ph.D
Dr. Papada told us that although every patient is different, a dietary pattern that fits most of them and at the same time is affordable (especially for IBD patients who make a lot of health related expenses), is the Mediterranean diet. She noted that patients are characterized by their symptoms: flare and recession of symptoms. When in flare, patients suffer immensely while a nutrient malabsorption is observed.
“How did we act on her feedback?”
She urged us to check on how dietary patterns can be implemented in a meaningful way on our application, to assist treatment efforts by nutrition.
Konstantinos Gerasimidis
University of Glasgow
University of Glasgow
Name:Konstantinos Gerasimidis
Affiliation: Professor of Clinical Nutrition, University of Glasgow
People with increased risk of developing IBD (10 and 20% of patients with IBD having a first-degree relative who also has IBD. The concordance rates for monozygotic twins to be about 60% for CD and 20% for UC) do not present with gut dysbiosis. This is why we shouldn’t restrain ourselves only to IBDs. Very hot topics related with gut dysbiosis: Cancer and Cardiovascular diseases. pH values in the gut can be altered in pathological conditions, such as Crohn’s disease which in turn can lead to microbial disturbances.
“How did we act on his feedback?”
We should see the bigger picture in regards to gastrointestinal disorders and focus on examining how gut conditions in pathologies can affect our detection system
Figure 4.a: Our project’s evolution through our human practices work
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Reforming our approach
Through our discussions with stakeholders, we aimed to reaffirm our goals and at which stage of disease progression our project fits in. We understood that the complexity of Inflammatory Bowel Disease at late stages does not allow efficient medical action, making disease prevention essential, in very early stages of pathogeny. Also, as Mr. Potamianos and Ms. Kotsopoulou suggested, not only are the pharmaceuticals used to tackle IBDs extremely heavy with many side-effects but at the same time there is no non-invasive form to accurately evaluate gut’s physiology.
This fact enhances the limited diagnosis of IBDs in younger ages but also does not help minimize the social stigma and taboo surrounding those disorders.
One other key factor that we integrated was that IBD pathology is only a symptom of the widely spread diet-related, non-communicable diseases. Experts urged us to unravel the missing link between all gastrointestinal disorders, which is their starting point and how we can effectively combat them.
We believe that AMALTHEA will be a strong tool in those experts’ arsenal.
Approaching The Public Perception Of The Microbiome
In Europe, at present, more than half of the adult population is overweight or obese and, children and population groups of lower socio-economic status are the most severely affected. Non-communicable diseases (NCDs) such as cardiovascular diseases, cancer, obesity and diabetes account for 70% of all deaths in the European Union (EU) and their costs represent 70-80% of European healthcare costs (European Commission, 2020). Currently, one in three people worldwide is malnourished while 1.9 billion adults are overweight and 462 million adults are underweight (Fluitman, 2017).
Unsustainable and unhealthy diets with increased demand for livestock products, and calorie-dense, nutrient poor and (hyper) processed food are a leading NCD risk factor and a driver of obesity rates.
NCDs are largely preventable through effective interventions that tackle shared risk factors such as unhealthy diets, physical inactivity and abuse of alcohol.
A better understanding of the mechanisms driving dietary choices will offer opportunities to reduce risk factors for NCDs and facilitate consumers to adhere to a long-lasting, healthy, pleasurable and nutritional (European Commission, 2020).
Figure 4.b: Our project’s evolution through our human practices work
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The Missing Link
“The Microbiome is the next big thing!” Helen Papadimitriou, IBD Specialist
The gastrointestinal (GI) tract harbors trillions of microorganisms (CCFA, 2014). The gut microbiome plays a critical role in the integrity of the intestinal barrier, nutrient absorption, metabolic homeostasis, and immune maturity, particularly in the early stages of life (Robertson, 2020). Dietary components can alter the intestinal microenvironment favoring some bacteria over others. That can affect the microbe–microbe interactions involved in the fermentation of indigestible nutrients, thus the modulation of energy extraction (Xiao, 2019) (Fluitman, 2017).
Figure 4.c: Our project’s evolution through our human practices work
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Our engagement with stakeholders unveiled that every gastrointestinal issue begins from disturbances in the gut microbial community. Those can be characterized as shifts in the abundance of certain strains over others or the decline of their metabolic capabilities (Morgan, 2012) (FAO, 2019).
This phenomenon is known as Gut Dysbiosis
“The next big question is to reveal the relation between dysbiosis and enteropathies” Xenoulis Panagiotis, Associate Professor of Companion Animal Medicine
Figure 4.d: Our project’s evolution through our human practices work
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Since the design of our first year’s effort aimed on the microbiome, we wanted to affirm our expectations and move forward in evolving our system. So, we turned to a wide variety of experts working on microbiome research to receive feedback on how to approach this vast, unknown network of microorganisms and if our system indeed can identify gut dysbiosis
Kotsopoulou Theodora
Gastroenterologist
Gastroenterologist
Name:Kotsopoulou Theodora
Affiliation: Gastroenterologist
Dr. Kotsopoulou informed us that a lot of research efforts and modernized approaches in gastrointestinal disorders aim on the gut microbiome, thus making it an exceptional new target for diagnostics and therapeutics. However, due to the evasiveness of gi disorders and their detection, a non-invasive method is optimal. She also informed us that that more and more patients come with mild symptoms, however most of them are stigmatized by the issue their facing so their is no real communication between patients and doctors.
“How did we act on her feedback?”
We continued our work on both the aforementioned aspects, being more confident on our course of action, and conducted further research on how to bridge the gap between doctors and patients
Antigoni Chalkia
Dietician
Dietician
Name:Antigoni Chalkia
Affiliation:Clinical Dietitian with MSc. Toxicology
Ms. Chalkia informed us that many patients with heavy symptoms visit a dietitian before going to a gastroenterologist, with 50% of all dietitian patients suffering from a GI disorder. In those cases the established examinations take place, such as colonoscopy, and the doctor can then make their diagnosis. In cases, though with very mild or no symptoms, a non-invasive technique would be ideal. Ms. Chalkia was very positive towards the evaluation of the gut microflora through a capsule and proposed the idea of utilising this capsule to collect epidemiological data. As for the application, she agreed that it is an easy way for a patient to communicate with an expert and that generally speaking dietitians would also find it handy. She also gave us the idea of providing patients with a personalised dietary pattern based on the information collected from the capsule and the notes the user is going to make on the interactive interface. Furthermore, the application would be an easy way for patients to describe to the dietitian details about their clinical picture that are difficult to communicate in person, such as the morphology of their feces.
“How did we act on her feedback?”
Given the prevalence of GI disorders in dietitian’s patients and the positive feedback about the integration of the capsule into their practice, we decided to include dietitians in our multidisciplinary team of experts that our application is going to connect with patients. Ms. Chalkia’s proposal for the utilisation of the capsule for research purposes was our inspiration that later, through our partnership with team Aalto-Helsinki, was formulated into a core aspect of our application. We also endorsed the idea of dietitians providing personalised dietary advice in an asynchronous manner through the application’s interface. Finally, we integrated a stool chart for a fast and easy description of the patient’s stools.
Potamianos Spyridon
University of Thessaly
University of Thessaly
Name:Potamianos Spyridon
Affiliation:Professor of Gastroenterology at the Thessaly University School of Medicine
Dr. Potamianos confirmed that SCFAs are a universal biomarker that can be used in the evaluation of the microbiome’s state, but not for diagnosing any kind of gastrointestinal diseases. Regarding the involvement of different experts in gastroenterology patients, he proposed gastroenterologists, dietitians, psychologists and psychiatrists.
“How did we act on his feedback?”
We integrated Dr. Potamianos’ feedback by altering the main direction of the project: We no longer aimed for the diagnosis of gut-related disorders, such as IBD, but for the functional evaluation of the gut microbiome, a more feasible and realistic goal for the device we are designing. The experts Dr. Potamianos proposed, were later included in the formation of a multidisciplinary team that through our application could communicate with the patients and give advice upon the user’s personalised data.
Helen Papadimitriou
IBDs Specialist
IBDs Specialist
Name:Helen Papadimitriou
Affiliation: IBDs Specialist at the General Hospital of Larissa
Brian Coombes
Department of Biochemistry & Biomedical Sciences
Department of Biochemistry & Biomedical Sciences
Name:Brian Coombes
Affiliation: Professor and University Scholar Chair, Department of Biochemistry & Biomedical Sciences
Panagiotis Xenoulis
Associate Professor, Companion Animal Medicine
Associate Professor, Companion Animal Medicine
Name:Panagiotis Xenoulis
Affiliation: Associate Professor, Companion Animal Medicine, University of Thessaly
Ιf we wish to check about gut dysbiosis all we have to do is a trial of administering certain feed (therapeutic test) and then check how the gut responds responds
Ιf we have a response to antibiotics this means we had dysbiosis (but sometimes the the usage in a high rate of antibiotics can have reversed results)
Ιf not this means that we must move forward with an endoscopy
He concluded our meeting saying that “The key point is to understand which came first? The gut dysbiosis or the enteropathy”
After revising our last year's design based on the measurements of SCFAs (acetate, propionate, and butyrate) levels, metabolites produced by the fermentation of indigestible carbohydrates (Müller et al., 2019) (iGEM Thessaly, 2020), we received positive feedback to proceed forward:
Utilize SCFAs as an indicator for the functional evaluation of the gut microbiome.
Figure 4.e: Our project’s evolution through our human practices work
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AMALTHEA: Incorporating Synthetic Biology
Based on our last year's effort, to accomplish the communication between gut flora and the host organism, we designed a bio-electronic interface to enable real-time monitoring of metabolite deficiencies on the patient’s smartphone.
Our final goal is the development of a micro-bio-electronic ingestible device that will support physicians to provide help to their patients based on their identified needs.
Figure 4.f: Our project’s evolution through our human practices work
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We also modified our detection system and ensured a safe and useful application environment, where data can be received and examined by experts. Synthetic biology allows us to meet the stakeholder needs, elaborate on their call and provide a complete and holistic solution to the problem at hand.
Illustrate an Effective Solution Defining Our Purpose
To further develop our detection system we turned to experts to verify if a capsule containing genetically modified bacterial strains is indeed necessary for a proper evaluation of the gut microbiome. As Professor Chatzigeorgiou told us, liquid biopsy is a promising method to ensure non-invasiveness on the patient’s body, but is this achieved by an ingestible capsule? Is there a way to avoid a controversial way of administration or is it crucial to walk this path to receive the most accurate image of the gut flora’s health?
We turned to several stakeholders to see alternatives on our detection system administration method and confirm or alter our approach.
Chatzigeorgiou Artemis
University of Thessaly
University of Thessaly
Name:Chatzigeorgiou Artemis
Affiliation: Professor of Bioinformatics, Department of Electrical and Computer Engineering, University of Thessaly.
Dimitrios Fatouros
Aristotle University of Thessaloniki
Aristotle University of Thessaloniki
Name:Dimitrios Fatouros
Affiliation:Professor at Pharmaceutical Technology at the department of pharmacy, Aristotle University of Thessaloniki
We started our meeting by sharing our thoughts about delivering the SCFASs through a buccal or sublingual patch. However, professor Fatouros told us that SCFAs cannot be absorbed sublingually or buccally unless they undergo a certain processing and be administered as UFA-somes through microneedles on the surface of the patch. When describing our capsule’s design, we asked the Professor whether regular pharmacists would be able to load the bacterial cultures in the tank on their own. He replied that something like that is not possible due to the lack of expertise and proper equipment. In order to work with systems like ours, the pharmacy should fulfil the criteria of Good Manufacturing Practice, something that doesn’t happen even to most university laboratories in Greece. After that, we discussed our plan for sight specificity in our measurements. Professor Fatouros, commented that for something like that to happen we would probably need a coating for the capsule that consists of a pH sensitive polymer that dissolves in the pH of the colon (~7,8). The coating can cover only the semipermeable membrane in order to both start the measurements at the right time point and protect the membrane from dissolving in the stomach’s acidic conditions.
“How did we act on his feedback?”
Firstly, we rejected the idea of sublingual/buccal administration of SCFAs because of the difficulties in the manufacturing process, thus moving on with our second plan, the oral administration. Secondly, we rejected the idea of involving regular pharmacists in the loading and assembly of the capsule, starting to investigate the idea of finding a sole industry that would possess both the required equipment and expertise to take on the manufacturing process from start to finish. Finally, we took the Professor’s advice on the capsule’s coating and started looking for pH sensitive materials that would fit our needs.
Maria Klapa
FORTH/ICE-HT
FORTH/ICE-HT
Name:Maria Klapa
Affiliation: Head of Metabolic Engineering and Systems Biology Lab (MESBL), FORTH/ICE-HT
Konstantinos Gerasimidis
University of Glascow
University of Glascow
Name:Konstantinos Gerasimidis
Affiliation: Professor of Clinical Nutrition, University of Glascow
Developing our application system
In 2020, the approach in designing a medical app was contradictory. According to the 2019 opinion of the Expert Panel on Effective Ways of Investing in Health (EXPH), digital technologies offer new opportunities for delivering more personalized, effective and efficient healthcare. EU policies have consistently emphasized the importance of digital solutions such as eHealth and have accentuated positive aspects of digital innovations. However, there are various concerns on data management and their safety. Doctors and clinical nutritionists last year suggested that it would be dangerous for patients to have a subjective view of their condition from just an app, that can’t accurately diagnose the disease and take into account various factors. (iGEM Thessaly, 2020).
In order to decide whether we should move forward for a digital health system and how to respond to the rising tendencies and needs for digitalization, we had to turn to everyone that may benefit from an application network for the gut microbiome’s data to be evaluated.
Efstathia Papada
Clinical Dietitian
Clinical Dietitian
Name:Efstathia Papada
Affiliation: Clinical Dietitian, MSc, Ph.D
“How did we act on her feedback?”
Dietitians are going to have access to the application as experts in order to consult the users on the selection of the best fitting dietary pattern, while combining their work with a mental health experts’. This way, we achieve better health outcomes for the patient. Medical confidentiality is ensured by allowing each expert to view only the information about the user’s health status, that are relevant to their profession, while being prohibited from viewing the other experts’ consults unless the patient grants them access.
Kotsopoulou Theodora
Gastroenterologist
Gastroenterologist
Name:Kotsopoulou Theodora
Affiliation:Gastroenterologist
Antigoni Chalkia
Dietician
Dietician
Name:Antigoni Chalkia
Affiliation: Clinical Dietitian with MSc. Toxicology
Terrovitis Manolis
Athena Research Center
Athena Research Center
Name:Terrovitis Manolis
Affiliation: Postdoctoral Fellow, Institute of Information Systems of the Athena Research Center
- To avoid contamination and to safely receive data, we moved to wireless data transport, but it was instructed by experts that data safety and management should be taken very closely into consideration.
- It is very important for the patient to not receive any feedback in an automated way about their results, since the evaluation of the microbial community is not something that can be generalised and must receive expert feedback.
- As mentioned, self diagnosis must be evaded at all costs since it leads patients to wrong choices, self isolation and increases the distance between people and the next generation’s medical care. Thus, experts must be present and give detailed advice regarding the capsule’s result
- Although acknowledging the condition and the metabolic physiology of the gut flora can give significant insights on the user's gut health, when we identify a disturbance, we must act upon them.
After receiving this feedback, we started ideating a software tool, a mobile application, which would link AMALTHEA’s complex measurement processes with the convenience a mobile device provides. This was the beginning of e-AMALTHEA. At the same time, we began reviewing literature on the subject of how we can alleviate gut dysbiosis.
Priority: Alleviating Gut Dysbiosis
After our discussions with stakeholders, we began to search through literature to understand what a good solution might be to treat gut dysbiosis. Our amelioration system includes metabolic engineering of the gut microbiome and engineered strains that replenish the quantities of missing SCFAs, acting as a cell-factory. Though our design was affirmed last year by Adamantia Kyriakou, Associate Professor in Microbiology at Department of Nutrition and Dietetics at Harokopio University of Athens, we wanted to communicate our idea to more experts to ensure that our approach is optimal.
Konstantinos Gerasimidis
University of Glascow
University of Glascow
Name:Konstantinos Gerasimidis
Affiliation: Professor of Clinical Nutrition, University of Glasgow
What is also considered a big advantage in our design, this time regarding the genetically engineered probiotic, is its ability to degrade cellulose, thus overcoming the necessity of cross feeders: direct production of SCFAs from cellulose in one single cell, makes the cell independent from the host’s microbiome (which may or may not have the capacity to produce the precursors of SCFAs). However, he expressed his concerns about the pH-sensitive coating of the capsule, upon which the whole site-specificity design was built. pH values can be altered in pathological conditions, such as Crohn’s disease, thus affecting the expected pattern of SCFA concentrations’ fluctuations in the final diagram. Moreover, overproduction of SCFAs in the gut can lead to Irritable Bowel Syndrome symptoms. Finally, he wondered how we overcome the problem of potential development of resistance to cumate.
“How did we act on his feedback?”
In order to overcome the randomness in the system in pathological conditions, clinical trials with large samples of patients should be conducted. This way we could theoretically standardise the expected pH values. A similar approach could be followed also to determine the exact levels of SCFAs, above which symptoms of Irritable Bowel Syndrome occur. Finally, an experiment in which we determine the time window until the cells accumulate enough mutations to become cumate resistant, is vital in order to give proper instructions for the consumer.
Konstantinos Kormas
University of Thessaly
University of Thessaly
Name:Konstantinos Kormas
Affiliation:Professor of Microbial Ecology and Aquatic Environment, Department of Agriculture, Ichthyology & Aquatic Environment
Right now, research and therapeutic approaches regarding both humans and fish, focus more on manipulating the existing gut microbiome than adding new microorganisms to it. However, the colonisation of the gut by our genetically engineered probiotic is not prohibitive, because we designed good biosafety systems. Professor Kormas also commented that our overall approach was complete and did not leave any red flags uncovered. He advised us to be careful with the fermentation products of the SCFA producing bacteria, such as H2, CO2, H2S, CH4, because many of them can become toxic. Finally he noted that although SCFAs are already used as a supplement in fish farming, an imbalance in Fe utilisation by the different microbial populations is usually observed.
“How did we act on his feedback?”
We intend to investigate further our probiotic’s tolerance to H2 and the problem of antagonism for Fe utilisation, and consider coadministration of methanogenic bacteria (professor Kormas informed us that they are already tested in fish) in order to detoxify the potentially dangerous byproducts. Furthermore, the adaptation of our bioelectronic capsule, and especially the probiotic, for use in fish farming could be a future vision in the animal welfare aspect of our project.
However, emerging evidence demonstrates that environmental factors, including diet, may play an important role in pathogenesis. This highlights the need to provide guidance to physicians and patients regarding which foods may be harmful, beneficial, or safe to consume.
We delved deeper into the role of nutrition on a person’s gut health,and evaluated how the gut responds to different forms of nutrition. Our aim was to answer two important questions: Is our synthetic biology solution enough and can it be in line with Greek dietary tradition?
Yiorgos Antonopoulos
Farmer
Farmer
Name:Yiorgos Antonopoulos
Affiliation: Farmer, Owner of Antonopoulos Farms
Katerina Moutou
University of Thessaly
University of Thessaly
Name:Katerina Moutou
Affiliation: Associate Professor of Vertabrate Biology, Head of the Laboratory of Genetics, Comparative and Evolutionary Biology, University of Thessaly.
Potamianos Spyridon
University of Thessaly
University of Thessaly
Name:Potamianos Spyridon
Affiliation:Professor of Gastroenterology at the Thessaly University School of Medicine
It became evident to us that producing SCFAs solely to enhance metabolic capabilities does not restore the loss of microbial diversity. One other key factor that we took into consideration is that due to the dynamic relationship between microbial populations, it is impossible to alter the microbiome’s composition in a long-term way simply by administering our strain.
Thus, our team has implemented a two-way solution to stop dysbiosis on its root:
Nutrition assessment and personalized nutrition, accompanied with a living bio-therapeutic synbiotic supplement.
Treatment Through Our Food
“The best gift you can give someone is a good, traditional plate”
Antonia Trichopoulou, President of the Hellenic Health Foundation
Antonia Trichopoulou, President of the Hellenic Health Foundation
Hippocrates’ notion “Let food be thy medicine and medicine be thy food” may be an ancient way of saying that dietary intervention is essential to maintain intestinal micro-ecological homeostasis and then impact host physiology and health (Xiao, 2019).
Our nutritional tradition not only has been beneficial for our well-being and development but also contributed to our co-evolution with the gut microbiome. The most common element across the Greek population is the Mediterranean diet which particularly influences microbial plant polysaccharide degradation potential, SCFAs production and pectin metabolism. One of our main questions that arose was how we can implement proper nutritional guidance and at what magnitude can we actually recoup the gut microbiome’s physiology.
Gerasimidis Konstantinos
University of Glascow
University of Glascow
Name:Konstantinos Gerasimdis
Affiliation: Professor of Clinical Nutrition, University of Glasgow
Efstathia Papada
Clinical Dietitian, MSc, Ph.D
Clinical Dietitian, MSc, Ph.D
Name:Efstathia Papada
Affiliation: Clinical Dietitian, MSc, Ph.D
According to our stakeholders' call, we aim to tackle low levels of SCFAs in an integrated manner, improving human health via dietary modification and enhancing the microbiome with a synthetic bacterial strain.
Now that we have successfully grasped the framework in which we operate, we moved towards action, responding to the feedback received in our experimental design, both in a dry and wet lab level.
Figure 4.g: Our project’s evolution through our human practices work
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Rationalize and Formulate Our Solution
Integrating Advice: Dry Lab Experts
Our Dry Lab work emphasized in three major goals:
- Designing and implementing an in silico model that supports the wet lab design by providing a mathematical and computational simulation of the underlying genetic mechanisms
Michail Stamatakis
University College London
University College London
Name:Michail Stamatakis
Affiliation: Associate Professor in Chemical Engineering Department of Chemical Engineering University College London
Maria Klapa
FORTH/ICE-HT
FORTH/ICE-HT
Name:Maria Klapa
Affiliation: Head of Metabolic Engineering and Systems Biology Lab (MESBL), FORTH/ICE-HT
- Developing an application system that is responsible and safe for the end user and consulting experts to ensure data safety
George Kakarotzas
University of Thessaly
University of Thessaly
Name:George Kakarotzas
Affiliation:Associate professor of Software Technology with an Emphasis on Architectures' components
Vlachos Vasilis
Economist
Economist
Name:Vlachos Vasilis
Affiliation:Assistant Professor at the Department of Economics, University of Thessaly
Petros Spachos
Software Engineer
Software Engineer
Name:Petros Spachos
Affiliation:Associate Professor at the School of Engineering, University of Guelph
- Perfecting our detection system, by providing clear schematics for the capsule dimensions, architecture and specifying the reasoning behind our choices.
Konstantinos Beltsios
Chemical Engineer
Chemical Engineer
Name:Konstantinos Beltsios
Affiliation:School of Chemical Engineering NTUA Department of Materials Science and Engineering
We contacted Dr Beltsios for his expertise on materials science, and especially polymers. In our discussion he pointed out the inherent instability of semipermeable membranes as constructs, as well as some key points we should pay attention to when looking for the appropriate membrane for our capsule: he claimed that to achieve our idea, we needed a membrane that is able to perform reverse osmosis, and, for that to happen, we also had to construct the capsule’s tank so that there is greater osmotic pressure on the inside than on the outside of the membrane. This, along with further bibliographical research, lead to a more considerate choice for our capsule’s semipermeable membrane.
Affiliation:School of Chemical Engineering NTUA Department of Materials Science and Engineering
Alexandros Katsaounis
Chemical Engineer
Chemical Engineer
Name:Alexandros Katsaounis
Affiliation: Professor at Chemical Technology & Applied Physical Chemistry section, Department of Chemical Engineering, University of Patras
We should look into materials that the electrode is covered with that are specific for L-DOPA. In our case we probably need an electrode that works as an effective conductor for the released electrones (e.g. a golden electrode) as the reaction of L-DOPA → L-DOPA quinone + e is a spontaneous one. Professor Katsaounis also pointed out the importance of taking into consideration the medium’s pH when choosing the right electrode: electrodes are often specific for a certain range of pH values, and their mechanical stability also depends on this factor, leading to the electrode’s corrosion in case we don’t make the appropriate adjustments.
“How did we act on his feedback?”
Based on the feedback gained by Dr. Katsaounis, we certified the design of our electrodes sensor and rethought the connection we had established, from the sensor to the processor, for optimal data transfer without noise. When testing the electrodes in our system we are going to produce a calibration curve to correlate SCFAs’ concentration with ΔV, in order to standardise our system.
Paraskevas Kitsos
University of Peloponnese
University of Peloponnese
Name:Paraskevas Kitsos
Affiliation:Associate professor at Department of Electrical and Computer Engineering
Dermenoudis Stergios
Solid State Physics Section
Solid State Physics Section
Name:Dermenoudis Stergios
Affiliation: Biologist and Researcher at Dr Logothetidis’s lab, solid state physics section, Physics Department, Aristotle University of Thessaloniki
George Papadopoulos
Chemical Engineering
Chemical Engineering
Name:George Papadopoulos
Affiliation: Professor at Department of Chemical Engineering, National Technical University of Athens (NTUA)
Dimitris Tsiourvas
Chemical Engineer
Chemical Engineer
Name:Dimitris Tsiourvas
Affiliation:Department of Physical Chemistry, NCSR “Demokritos”
Dr Tsiourvas, as a very experienced researcher, who was in fact recommended by Dr Papadopoulos, discussed with us some details concerning semipermeable membranes and their characteristics. He was able to pinpoint a particular membrane that might be optimal for the biochemical characteristics of SCFAs.
Affiliation:Department of Physical Chemistry, NCSR “Demokritos”
Integrating Advice: Wet Lab Experts
With the advancements in synthetic biology, probiotic engineering offers more flexibility and fine-tuning capacity than others (Aggarwal, 2021). However, we wanted to 1) ensure long term sustainability of our strain to produce large quantities of SCFAs, 2) try to provide our supplement through a non-pharmaceutical way and thus be more friendly to the consumer and 3) ensure that we do not disrupt the microbial community’s natural function.
Persephone Giannouli
University of Thessaly
University of Thessaly
Name:Persephone Giannouli
Affiliation:Assistant Professor of Technology and Food Quality Control of Plant Origin
Professor Giannouli characterised the idea of encapsulating the bacteria as innovative, but expressed some concerns about the effect the alginate may cause on the yoghurt’s pH. She proposed stir yoghurt, a yoghurt that coagulates with stirring, as a good candidate for adding the alginate beads and keeping them intact until consumption. Something that she suggested we mention on our product, is the proposed daily doses. In this case we need to take into consideration not only the substances that are contained solely in our product, but also those that are consumed daily from other sources, too, thus inflicting a cumulative impact. Finally, she mentioned that although there are special and stricter regulations around foods for children (1-2 years old), at the same time there are no warnings on products that should not be consumed by kids.
“How did we act on his feedback?”
We decided to compose a guide that addresses the consumers, giving them detailed instructions about the responsible use of the product, hence ensuring both the safety of the consumer and environmental containment. These instructions include the population groups that should not consume the product, excluding children below the age of five. Finally, the yoghurt’s distribution was decided to take place only from pharmacies to better control the buying public. Future vision: Future experiments are needed to determine the viability of our probiotic in the yoghurt as well as the effect of alginate on the yoghurt’s pH.
Alexandra Meziti
Microbiome Expert
Microbiome Expert
Name:Alexandra Meziti
Affiliation:Postdoctoral Researcher, Department of Nutritional Sciences and Dietetics of International Hellenic Universit
After being briefed on our project, Dr. Meziti agreed with our approach of preventing the manifestation of gastrointestinal disorders by evaluatiating the gut microbiome and alleviating the dysbiosis. She was also positive towards the colonisation strategy and the in situ production of the desired metabolite, since most modern probiotics lack the long term effect, by dying shortly after the administration. She expressed two main concerns: the first one was the possibility of the SCFA production negatively affecting the metabolic pathways of other bacterial species in the gut. The second one was the competition of our probiotic strain with natural bacteria populations, for food availability.
“How did we act on his feedback?”
We developed a system that gives our bacteria the ability to utilise the cellulose of food, a polymer that naturally goes to waste since gut microbiota do not have the means to degrade it. In this way, we managed to increase the carbon flux to overcome the metabolic strain of SCFA production while achieving to eliminate the competition between our strain and the natural ones over nutrition sources. Future vision: In future experiments and bibliographic research, we intend to elucidate the metabolic relationships between different bacterial strains and predict, with relative precision, the impact of our probiotic’s colonisation to the natural microbial populations.
Nestoras Vlachos
CEO of Epsilon Health
CEO of Epsilon Health
Name:Nestoras Vlachos
Affiliation: CEO of Epsilon Health
Mr. Vlachos proposed that we get in contact with researchers and retrieve data concerning the SCFA concentrations in healthy and pathological conditions and also conduct research about the legislation concerning the administration of GMOs. He introduced us to the list of Generally Recognised and Safe (GRAS) microorganisms that are approved for use as probiotics. Finally, after hearing our idea about administering our probiotic through a yoghurt, he underlined the importance of the conditions in a production line; we should be careful and add our probiotic at a stage that is not going to reduce the probiotic’s viability or the quality of the yoghurt itself.
“How did we act on his feedback?”
We checked the legislation around the administration of GMOs in humans, and realised that our product can not be implemented in Europe. Thus our long term plan, is to promote it to the United States’ market. After that we checked the GRAS list and chose the strain E.coli Nissle 1917 as the best candidate to use as a chassis organism. We also started collecting data about the yoghurt’s production line and arranged new meetings with experts that specialise on food technology.
Kormas Konstantinos
University of Thessaly
University of Thessaly
Name:Kormas Konstantinos
Affiliation:Professor of Microbial Ecology and Aquatic Environment, Department of Agriculture, Ichthyology & Aquatic Environment
Right now, research and therapeutic approaches regarding both humans and fish, focus more on manipulating the existing gut microbiome than adding new microorganisms to it. However, the colonisation of the gut by our genetically engineered probiotic is not prohibitive, because we designed good biosafety systems. Professor Kormas also commented that our overall approach was complete and did not leave any red flags uncovered. He advised us to be careful with the fermentation products of the SCFA producing bacteria, such as H2, CO2, H2S, CH4, because many of them can become toxic. Finally he noted that although SCFAs are already used as a supplement in fish farming, an imbalance in Fe utilisation by the different microbial populations is usually observed.
“How did we act on his feedback?”
We intend to investigate further our probiotic’s tolerance to H2 and the problem of antagonism for Fe utilisation, and consider coadministration of methanogenic bacteria (professor Kormas informed us that they are already tested in fish) in order to detoxify the potentially dangerous byproducts. Furthermore, the adaptation of our bioelectronic capsule, and especially the probiotic, for use in fish farming could be a future vision in the animal welfare aspect of our project.
Tassou Chrysoula
DEMETRA
DEMETRA
Name:Tassou Chrysoula
Affiliation:Director of Research at the Institute of Agricultural Product Technology (ITPA) of Elliniko Of the Agricultural Organization "DEMETRA"
Sandrine Uttenweiler-Joseph and Fabien Letisse
LISBP
LISBP
Affiliation:Researcher at Biosystems and Process Engineering Laboratory (LISBP)
Efstathios Giaouris
Food Microbiologist
Food Microbiologist
Name:Efstathios Giaouris
Affiliation:Associate Professor of Food Microbiology, Department of Food Science and Nutrition of the University of the Aegean
Taking into consideration our stakeholders ideas and suggestions, we propose to ameliorate gut dysbiosis with CERAS, a living bio-therapeutic synbiotic supplement, that can act as a living, SCFA producing factory for the gut flora. An engineered probiotic strain encapsulated in prebiotic fibers that produces SCFAs at a pinpoint location in the user’s gut. In a personalized manner, we alleviate gut dysbiosis, ensuring continuing stability on the gut’s microbiome health.
Figure 4.h: Our project’s evolution through our human practices work
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Ensuring Maximum Safety
Since our multi-stage approach to tackle diet related NCDs has a synthetic biology core and taking into consideration that the Greek society is not yet familiarised with concepts of Synthetic Biology, we’ve set as a high priority to provide a safe and responsible solution to the world
That’s why, we tried to answer important safety considerations , safeguarding our project from our certain red flags.
We focused on:
- - Conduct safe laboratory work
- - The capsule itself provides the physical borders separating the bacteria from the gut. However, a mechanical or biological malfunction must be averted at all costs.
- - Moving to a new digital age, we must ensure that user data is protected and not used in any malicious way.
- - Our synbiotic supplement and later our probiotic strain must not cause any harm to the gut flora or cause immunity to the user’s organism.
- - Ensure environmental sustainability by avoiding contamination and creating super-resistant strains.
To framework and verify our experimental design, we turned to safety experts to receive feedback and formulate a responsible safety strategy.
iGEM Safety Commitee
iGEM HQ
iGEM HQ
Name:Tessa Alexinian
Affiliation:Safety & Security Program Officer
Mathiopoulos Konstantinos
Molecular Biologist
Molecular Biologist
Name:Mathiopoulos Konstantinos
Affiliation:Professor of Molecular Biology, Department of Biochemistry and Biotechnology, University of Thessaly
We presented him our whole project and received very positive general feedback. Then, we elaborated more on the biosafety aspect, especially at our probiotic, discussed his concerns. In professor Matthiopoulos’ opinion, the safest way to approach this aspect is by modifying the whole translational machine of our bacteria thus developing a completely orthogonal system.
“How did we act on his feedback?”
We explained to him that due to restrictions of time, it is not possible for us to execute such an ambitious plan. For that reason, so we concluded that, realistically, our best option is to combine at least two kill switches in each bacterial category -probiotic bacteria and bacteria in the detection system of the capsule- to ensure both safety for the patient and environmental containment.
Vourtchis Dionisis
EBSA - European Biosafety Association
EBSA - European Biosafety Association
Name:Vourtchis Dionisis
Affiliation:Representative of EBSA in Greece
We made a presentation of our biosafety plan and discussed its details. Mr. Vourtsis insisted on the importance of environmental containment and the potential dangers that derive from the process of loading the bacteria in the tank.
“How did we act on his feedback?”
The environmental containment aspect is covered by two specific kill switches, one in each one of our products -engineered probiotic and metabolite detection bacteria in the capsule- that are responsible for the bacteria’s elimination once they exit the human body.
Dimitrios Karpouzas
Environmental Biotechnologist
Environmental Biotechnologist
Name:Dimitrios Karpouzas
Affiliation:Professor of Enviromental Microbiology & Biotechnology, University of Thessaly
Karras Panagiotis
Toxic Waste Management
Toxic Waste Management
Name:Karras Panagiotis
Study:Departement of Biochemistry and Biotechnology, University of Thessaly
Dieter Vanderheuvel
University of Antwerp
University of Antwerp
Name:Dieter Vanderheuvel
Affiliation:Postdoctoral Researcher at the Laboratory of Applied Microbiology and Biotechnology of the University of Antwerp
Dr. Vandenheuvel, Postdoctoral Researcher at the Laboratory of Applied Microbiology and Biotechnology of the University of Antwerp, pointed out that we will have issues implementing the probiotic in food products, because of the E. coli testing. Our E. coli will be present in food but the test cannot differentiate the pathogenic E. coli from ours (FDA, 2021). Our future final probiotic will be a Lactobacili genus as he proposed, in order to make our engineered probiotic suitable to be implemented in a fermented product like yogurt.
Affiliation:Postdoctoral Researcher at the Laboratory of Applied Microbiology and Biotechnology of the University of Antwerp
After expert interviews and literature research, we finalized our safety plan and addressed this issue holistically for AMALTHEA, which is summarized as follows.
- Genome Integration to avoid gene transfer from plasmids. (Thomas & Nielsen, 2005).
- L-arabinose inducible kill switch in the event that the capsule's bacterial tank breaks
- A knockout strain for the dapA so the bacteria can’t survive without the supplementation of the cell wall component dapA in environmental conditions, ensuring environmental containment of the bacteria containing tank in our detection system.
- Cumate: A completely user-controlled system that is activated upon external administration which is distributed along with CERAS, in order to end the probiotic’s colonization.
- Tryptophan System1:The second kill switch, a system based on the amino acid tryptophan, which suppresses cell death. When the bacteria exit the host’s body, the tryptophan that naturally occured in the gut environment is missing, hence allowing a toxin to be produced and the bacteria to die.
Highlighting Mental Health’s Gravity
“An invisible disease, it has major impacts on our mental health”
Anonymous Patient
Anonymous Patient
AMALTHEA’s original design aimed for the functional evaluation of the gut microbiome and the amelioration of gastrointestinal symptoms related to dysbiosis. Through our bibliographic research we came across a considerable amount of literature concerning the role that gut microbes play in our general well-being, including both our physical and mental health. Our starting point was the inadequately investigated, often non-communicable and always extremely costly issue of comorbidity of enteric and mental disorders. Aiming to shed light on this perspective and due to our partnership with team Aalto-Helsinki, we elaborated on the subject of mental health.
Our response to see if we could act upon this well-documented issue was to engage with experts in the field of mental health experts.
Paraskevi Angelopoulou
Health Psychologist
Health Psychologist
Name: Paraskevi Angelopoulou
Affiliation: Doctoral researcher at Aristotle University of Thessaloniki
As Ms. Angelopoulou informed us, in Greece, no Medical school, except for one, has any subject related to the patient’s psychology or the way in which a doctor should make unpleasant announcements. Stigma, depression and anger are very common in chronic patients making the communication with the doctor difficult. This in turn, often leads to a negative attitude on behalf of the doctors which can lead to medical errors, mistakes in drug prescriptions and bad communication. Another problem that exists almost everywhere, and especially in Greece, is the lack of multidisciplinarity in the treatment of gut-related disorders, which translated mostly as absence of mental health experts from the development of treatment strategies for gastroenterology patients. Our conversation about the application was also very fruitful. Ms. Angelopoulou was very positive towards our idea and commented that an application like ours can increase the patient’s accessibility to experts without stigmatising them. We shared our idea of organising emotional support groups in which users that share the same or similar problem, communicate and support each other. However, she was sceptical about this idea, since teams like these need to be coordinated by an expert who poses the appropriate boundaries. Her general proposals for new features at the app were the following: connecting the users with experts in an easy and fast way, make the interface friendly and use questions that focus at the present time.
“How did we act on his feedback?”
The information about the lack of gastroenterologist’s training on mental health issues, inspired us to design a booklet that addresses them and offers basic information about the comorbidity of gut and mental disorders as well as the importance of taking into consideration the patient’s mental state when engaging with one. Moreover, we built upon the fact that the formation of multidisciplinary teams of experts is necessary to achieve higher quality of treatment in gastroenterology patients, adding psychologists/psychiatrists as experts that can use the application to evaluate the patients results and provide advice. The questionnaires we were thinking of integrating in the app were articulated in a way that focuses the user’s mind in their somatic symptoms and feelings more at the present time. Regarding the application, we started off by rejecting our initial idea to assemble support groups from strangers without the ceaseless presence of a psychologist. After that we tried to develop a friendly interface that would seem welcoming and appealing for the users, and finally we articulated the questions we were thinking of integrating in the app in a way that focuses the user’s mind in their somatic symptoms and feelings more at the present time. Future vision: We plan to develop a map on which psychologists and psychiatrists near the location of the user will be marked with a pin. This way someone with the need for help can click on the pin and arrange an appointment in an easy and fast manner.
Agorastos Agorastou
Professor of Psychiatry
Professor of Psychiatry
Name: Agorastos Agorastou
Affiliation: Assisistan Professor of Psychiatry, Aristotle University of Thessaloniki
80% of chronic diseases are related to stress and thus constitute a big demographic group. Depression, one of the most predominant stress related disorders, poses the No1 cause of disability, while 60% of patients with depression underestimate the effect of stress on their condition. The capsule’s non-invasiveness is a big advantage. Patients, especially those with depression will be positive to the idea of using the capsule in order to assess their gut microbiome’s state and possibly improve their condition. Regarding the probiotic, the average psychiatrist does not make suggestions for dietary adaptations to their patients in order to alleviate their disorder’s symptoms. One possible cause for this phenomenon is the lack of guidelines regarding the prescription of probiotics to psychiatric patients. However, doctors are open to the idea of integrating probiotics to their treatment practices. Finally, something that we need to take into consideration is that gastroenterologists prescribe psychiatric drugs that empirically work for the alleviation of gut-related symptoms, disregarding their effects on the patients mental state
“How did we act on his feedback?”
We decided to include psychiatric patients that suffer from diseases that fall into the category of stress related disorders such as posttraumatic stress disorder (PTSD), Panic, Generalized anxiety and Alcohol abuse, and focus especially on depression due to both its high prevalence and severity. We intend to approach psychiatrists in the future as an appealing target group that can promote AMALTHEA (capsule) and CERAS (yoghurt with SCFA producing probiotic) to a new and considerably broad audience. We are going to focus on better understanding the psychiatrists’ background knowledge about diet and design a strategy about informing them in a constructive way, especially on the probiotic prescription. Last but not least, we are going to turn to gastroenterologists, talk about the exact cases of psychiatric drugs’ prescription and inform them about the destabilising effects of those drugs on the patients’ mental health.
Paraskevi Angelopoulou
Health Psychologist (2nd Meeting)
Health Psychologist (2nd Meeting)
Name: Paraskevi Angelopoulou
Study: Doctoral researcher at Aristotle University of Thessaloniki
We started off our meeting by talking about the initial version of our booklet for gastroenterologists. Ms. Angelopoulou, noted that it felt close to the reader but it probably would be more useful, at its current form, for gastroenterologists’ patients. Moreover, she proposed adding some information that would make the link between mental and gut disorders clearer. After that, we moved on to discussing the application in more detail, compared to our first meeting. We concluded that a feature that organises the data collected from the weekly filling of the questionnaire would be organised in 6-month diagrams that could be relatively easy to be reassessed by mental health experts. She proposed that we add some questions relative to the user’s medical history that would be answered when the user first opened the application, along with functionality, socialization and mood related questions that would be asked on a weekly basis. The Initial questions together with the medical history of the user, can be used to personalise the application, while the integration of a mindfulness exercise could close the questionnaires’ filling so that the user leaves with a feeling of relaxation.
“How did we act on his feedback?”
We took Ms. Angelopoulou comments under consideration and developed two new booklets, one for gastroenterologists and one for patients, keeping in the last one the friendly layout the first version had, while designing a completely different one for the gastroenterologists. Regarding the application, we integrated a small number of questions that will be answered the first time someone opens the application, as well as three distinct sections of questions related to the functionality, the socialization and the mental state of the user. Future vision: The Initial questions will be combined with the medical history of the user, in order to personalise the application; certain answers to selected questions will slightly change the apps’ interface to better suit the psychological needs of the user. Moreover, a mindfulness exercise will accompany the questionnaires, promoting relaxation and “psychological rehabilitation” for the user. Activities like this, also function as a tool to connect an experience such as filling a questionnaire, with emotions that attract the user to repeat the experience and hence, be responsible towards the task. Regarding the 6-month collections of data, we think that the best way to present the information is through a somatic symptoms versus time, and a mental disorders’ symptoms versus time diagram.
Agorastos Agorastou
Professor of Psychiatry
Professor of Psychiatry
Name:Agorastos Agorastou
Affiliation: Assisistan Professor of Psychiatry, Aristotle University of Thessaloniki
We received extensive feedback on the first booklet we designed, with the goal of briefing gastroenterologists about the prevalence of mental health disorders in their patients as well as the comorbidity of gut and mental disorders. The main concerns dr.Agorastos expressed, are: 1) The images used in the booklet should be very carefully selected, in order not to enhance, but alleviate the stigma and discrimination against individuals with mental health disorders. 2) The phraseology should also be chosen very carefully. For example, he proposed changing the title: “What is a mental disorder?” into “How to you conceive the concept of mental health?”. 3) He encouraged us to design a booklet for gastroenterology patients, in order to inform them on basic concepts concerning their mental health.
“How did we act on his feedback?”
We worked on every aspect of the booklet that was pointed out as concerning and composed two new versions of the material, one for gastroenterologists and one for their patients.
Eventually we decided to investigate this issue more thoroughly, starting by asking two basic questions:
- - What kind of relationship is established between the gut microbiome and the Central Nervous System (CNS) -especially the brain- and
- - At what degree could this relationship lead to mental health disorders? As the time passed and the points of intersection between our main project and the mental health perspective increased, we decided to take this research a step further and start acting upon related issues that emerge at the individual, as well as at the societal level.
During these conversations, and in spite of many specialists having very divergent backgrounds, we repeatedly received a prompting towards the consideration of mental health in the evaluation of the general impact that gut microbiota imbalances inflict on the human body.
Microbiota imbalances reported by the capsule can lead psychiatrists to the conclusion that tackling the problem with diet modifications might be a partial solution. The fact that the measurements are going to be taken in a non-invasive manner, plays a major role in the inclination of patients to take the capsule. Studies have shown that targeting the microbiota by probiotic supplementation could alleviate psychological distress including an index of depression (Raypole, C., 2019). At this point a patient warned by the doctor, can alter their diet either using our application, or directly consuming our probiotic, as it is a product personalized and safe for general use at the same time. The probiotic can be proposed by the doctor even without the patient using the capsule, as it is a product that can be used only as a prevention measure.
Figure 4.i: Our project’s evolution through our human practices work
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Safeguarding Animal Welfare
At a similar time period, our work was to be presented at the 9th Panhellenic Veterinary Students Conference that was organised by I.V.S.A.- International Veterinary Students ’Association. The aim of this action was to inform the student community about how synthetic biology could contribute to safeguarding animal health and One health.
Applying the One Health approach to the microbiome allows for consideration of both pathogenic and non-pathogenic microbial transfer between humans, animals, and the environment (Trinh et al, 2018). Different animal species house unique microbiomes, often of equal or greater complexity compared to the human microbiome. As in humans, animal microbiomes influence the health of livestock, pets, disease vectors, and foundational species that uphold ecosystems. Only by diving into and understanding such interactions, will we be able to generate innovative interventions to prevent and manage a variety of human and animal health conditions.
Those two events were instrumental to the final iteration of our project:
Implement Amalthea to the Animal Welfare
As always, we turned to relevant researchers and production companies to assess for whom our project would be helpful.
Konstantinos Kormas
University of Thessaly
University of Thessaly
Name: Konstantinos Kormas
Affiliation: Professor of Microbial Ecology and Aquatic Environment, Department of Agriculture, Ichthyology & Aquatic Environment
Panagiotis Xenoulis
Companion Animal Medicine
Companion Animal Medicine
Name:Panagiotis Xenoulis
Affiliation: Associate Professor, Companion Animal Medicine, University of Thessaly
Institute of Animal Research
DEMETRA
DEMETRA
Name: Institute of Animal Health
Affiliation: DEMETRA
Nireus
Fish Feed Production Industry
Fish Feed Production Industry
Name: Nireus
Affiliation: Fish Feed Production Industry
Through our discussions with Nireus, we were interested to learn if any aspects of our project can be implemented to the fishing industry. Even though there was an interest in using SCFAs as a feed additive, probiotic strain administration in fish is commercialy rare due to their aquatic lifestyle. Thus, implemented AMALTHEA in fish was found with skepticism.
Affiliation: Fish Feed Production Industry
Through our discussion, we understood that different approaches must be implemented for different animal species and purposes.
Our implementation in this manner helps us to fulfill the aim of the project to include a One Health context.
Figure 4.j: Our project’s evolution through our human practices work
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Closing the loop
Our Proposed Implementation
Detection System
Given the complexity of the capsule (especially the engineered bacteria’s sensitivity), the whole device should be assembled strictly by pharmaceutical companies that possess both the required equipment and the expertise. At this point, we need to underline the fact that the most delicate part of our device is the biological one, and for the aforementioned reasons a pharmaceutical company can be assigned the task of preparing and loading the bacterial cultures in the capsule’s tank. On the other hand, the electronic parts will not stand in the way of the whole process, due to their ease of assembly and their affordability; thus, no third-party manufacturer is required in the overall system’s mass production. This way, we ensure environmental containment and safety for the manufacturers themselves and ,at the same time, we reduce the possibility of error, which rises with respect to the number of intermediaries. Right after manufacturing, the capsule is distributed under strict conditions, (especially temperature ones) to clinics and pharmacies.
e-Amalthea
As in all components of AMALTHEA, we considered data security very seriously in our application’s development process, too. We protect our user’s data on many levels: first and foremost, e-AMALTHEA strictly follows the GDPR regulations, and, additionally, we connect it with a hybrid database that combines the advantages of a cloud and a conventional database. Knowing the importance of data anonymization, we also ensured that the data is encrypted with standard algorithms the cloud database provides, so that no user can be linked or correlated to their sensitive information such as SSN, TIN, medical results etc.
CERAS
We add our probiotic in the yoghurt in the late stages of its production, separately from the bacteria that conduct its fermentation. After achieving the desired composition and bacterial load, the yoghurt gets sealed and stored at low temperatures so that we preserve the probiotic’s viability and capacity for the SCFAs’ production.
Epsilon Health
Probiotic Producing Industry
Probiotic Producing Industry
Name:Epsilon Health
Affiliation: Private Sector
OLYMPOS
Dairy Company
Dairy Company
Name: OLYMPOS
Affiliation: Private Sector
As our goal is to implement CERAS to a well accepted and traditionally friendly food in Greek Society, we met with TYRAS A.E., the biggest dairy industry in Greece. After we laid out our project idea, we spoke with Athanasios Athanasoulas, Head of Research and Development. He briefly presented to us what it takes financially to launch a probiotic dairy product and discussed the probability of CERAS being produced and deployed.Affiliation: Private Sector
As discussed, there are already hybrid dairy products with probiotics but not many of them administer their probiotic strain outside the fermentation culture due to survivability issues and product's taste alteration. However, due to our integrated alginate bids design, Mr Athanasoula was interested in the potential this project can have if implemented after fermenatation. Also, the 2 month period of lifespan of a yoghurt can be achieved, thus making our project financially viable.
The probiotic’s lifespan is approximately two months, a period long enough for the product’s distribution and its stay on the pharmacy’s shelf. The CERAS’ package, except for the yoghurt itself, also includes the proper amount of cumate for the colonisation termination and a detailed guideline for the product’s safe consumption.
In Vitro, Pre-Clinical and Clinical Trials
Moving beyond our experimental design and our project’s ideation, the first step that every scientific project must circumvent is to test our project’s function through multilear trials. Since the gut microbiome and its bacterial formation varies from person to person, we were skeptical on how we can lay out a proper framework to conduct clinical trials to meet the microbiome’s delicate nature.
Once again, we turned to experts to pinpoint a plausible course of action to cover any ethical, safety and fidelity concerns on our core project aspects.
Adele Costabile
University of Roehampton
University of Roehampton
Affiliation: Associate Professor (Reader) in Nutrition at Roehampton University
Nikolaos Kostomitsopoulos
Vice-President of FELASA
Vice-President of FELASA
Name: Nikolaos Kostomitsopoulos
Study: Vice-President of the Federation of European Laboratory Animal Science Associations (FELASA)
After our interview, we managed to lay out a course of action to properly conduct in vitro, preclinical clinical and trials
- Bioinformatics, modeling and in vitro tests simulating the GI tract conditions regarding microbial composition, pH levels and nutrients, receiving measurements in a 72-h period of time as Prof. Costabile suggested
- Conduct pre-clinical trials on a rodent and non-rodent animal surrounding toxicity, safety and efficacy by providing a systematic review and meta analysis based on Arrive 2.0 guidelines.
- Standardization of our measure techniques and normal levels, by testing how sensitive is our method in sampling test groups of healthy and gi disorders suffering populations.
Consolidation of Society-Science-Industry
The endpoint of Synthetic Biology is industrialisation. Although as a iGEM project we serve the fundamental scientific discipline such as synthetic genomics, engineering and molecular biology, it is also important to integrate it to the market. Thus, we aimed to build bridges with the industry and relevant stakeholders, such as pharmaceutical and probiotic companies to properly access our place in the market and strategize our next moves for introducing AMALTHEA to the industry sector.
EFSA
European Food and Safety Administration
European Food and Safety Administration
Name:EFSA
Affiliation: European Organisation
EnzyQuest
Biotechnology Company
Biotechnology Company
Name:Koutsioulis Dimitris
Affiliation: CEO of EnzyQuest
SUPERFOOD
Probiotic Company
Probiotic Company
Name: SUPERFOODS
Affiliation: R&D Department
Superfood is a fellow probiotic producing company. Our team members got the chance to meet with representatives of the company to discuss laboratory production, industry capabilities in the Greek Area and several concerns over CERAS.
Affiliation: R&D Department
Pfizer Hellas
Pharmaceutical Company
Pharmaceutical Company
Name:Pfizer Hellas
Affiliation: Board of Directors
NOVARTIS
Pharmaceutical Company
Pharmaceutical Company
Name:NOVARTIS
Affiliation: Private Sector
Ministry of Development and Investment
Public Sector
Public Sector
Name: Michalis Dritzas
Study: Head of Deputy Minister of Development and Investments in Research, Technology & Innovation, of Greece
Uphold our Ideals
(1) provides a complete and prevention toolkit for the rising threat of diet-related non communicable diseases (NCDs)
(2) addresses holistically the task at hand, respecting dietary traditions and our society’s beliefs and opinions
(3) is safe for humans, animals and the natural environment
(4) ensures data privacy for users
(5) is financially viable and scalable.
Figure 4.k: Our project’s evolution through our human practices work
.
Future Directions
Also, after discussions with our PIs, we’ve arranged contacting Effimia Panagiotidou, a legal advisor of the Research Committee Technology Transfer Office of the University of Thessaly to guide us further on what is required to move forward with an application for intellectual property on AMALTHEA.
Our goal, our dream after the competition is to move even higher and begin marching on our research career.
Figure 5: Our project aspiration for future development.
References
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