Research dating back to 1984. What we study today?

Why and how

Why we started with the gut

At the outset, we built on findings about differences in the gut microbiota as well as on the frequent occurrence of gastrointestinal complaints observed in some children with ASD. From research on prebiotics we also knew that they can serve as a substrate for selected gut microorganisms [3].

That is why we chose the prebiotic intervention POWERbiom+®, which combines galactooligosaccharides (GOS) and components of whole gold kiwi. GOS are well-researched prebiotics and in the available literature are associated, among other things, with support for bifidobacteria [4]. Gold kiwi adds fibre, polyphenols and the enzyme actinidin to the formulation.

Our research question, however, was not aimed only at digestion. If there is a functional two-way connection between the gut and the brain, we wanted to know whether changes observed in the gastrointestinal area would be accompanied by changes in sleep, behaviour, eating habits or overall quality of life.

And it is exactly this question that we decided to follow systematically under real-world conditions of use.

How we looked for the answer

The study participants were children aged 3 to 18 with a diagnosed ASD.

Before supplementation began, parents completed an entry questionnaire capturing the child’s baseline state. Only then did the 60-day POWERbiom+® supplementation begin.

As primary outcomes we monitored the occurrence and intensity of constipation, diarrhoea, abdominal pain, bloating and reflux, together with stool consistency and frequency.

As secondary outcomes we assessed irritability, anxiety, attention, hyperactivity, sensory hypersensitivity, social engagement, sleep quality, eating habits and the child’s parent-rated quality of life.

A first interim assessment after 14 days also let us see whether some changes appear during the first weeks of supplementation.

The final analysis included complete data from 64 children.

Next: Results →

Results

What did the results show us?

The results showed changes in several monitored areas. Digestive parameters were the main theme of the study, but we also recorded interesting developments in other areas of the child’s everyday functioning.

Gastrointestinal complaints

The total score of the five monitored gastrointestinal symptoms – constipation, diarrhoea, abdominal pain, bloating and reflux – decreased over the monitored period.

In the parents’ subjective rating of the overall severity of GI complaints, the average score after 60 days was more than 60% lower than at baseline. The difference was statistically significant for both the combined GI score and the VAS rating.

Stool consistency and frequency

We also recorded a development in the nature of bowel movements.

The proportion of children with physiological stool consistency on the Bristol scale rose from 64.06% to 81.25%.

When we assessed stool consistency and bowel movement frequency together, the proportion of children meeting both physiological criteria rose from 54.69% to 73.44%.

Constipation

We observed a notable result in the group of children with moderate to severe constipation. At the start of the study, 12 children fell into this category.

After 60 days, 10 of them – that is, 83% – moved into the category of mild or no constipation.

In the whole group, the proportion of children with no constipation or only mild constipation rose from 81% to 97%.

Behaviour and sleep

We also recorded an interesting development outside the digestive tract. The composite score for behaviour and sleep covered nine areas – irritability, anxiety, attention, hyperactivity, sensory hypersensitivity, social engagement, falling asleep, waking up and overall sleep quality.

After 60 days, the average score was more than 36% lower than at baseline.

Eating and quality of life

During the study we also recorded a change in the rating of eating habits – food selectivity and appetite.

The parent-rated quality of life of the child rose on a scale of 0 to 10 by 1.25 points on average.

Results in the data

Results of the 60-day open-label single-arm interventional POWERbiom+® study in children with ASD · N = 64

Next: What next →

What next

What the results showed us – and where they take us next

When we look at the results as a whole, it is not only the individual numbers that are interesting.

Over 60 days we observed the development of gastrointestinal complaints and the nature of the stool, together with changes in parent-rated areas of behaviour, sleep, eating and quality of life. It is precisely this broader picture that is one of the most interesting outputs of the project for us.

We interpret the results, however, in the context of the study design. It was an open-label single-arm interventional study without a placebo control group, and much of the assessment was based on parents’ observations. The project did not include an analysis of the composition of the gut microbiome or of biological biomarkers, and follow-up lasted 60 days. We therefore do not see these results as a final answer to the question of the microbiome–gut–brain axis. We see them as a systematically obtained observation that shows us which connections are worth exploring in more detail.

Where is our research heading?

The ASD project took us from a very practical question about digestive complaints to a broader question:

Why did other parent-rated areas change during the same period?

The next step is therefore not just to repeat the same follow-up in a larger group. We want to get closer to the microbiome–gut–brain axis itself.

In further research, we would like to link subjective and clinical observations with more objective data – for example, analysis of the gut microbiome, its metabolic activity and selected biological markers. Such a design would allow us to investigate not only:

“Which areas change during the intervention?”

but above all:

“Are these changes accompanied by measurable changes in the gut environment and its biological activity?”

Our ambition

For us, the ASD project is not just an evaluation of a single formulation. We started with a very practical question – digestive complaints, which often occur in children with ASD.

Systematic follow-up, however, also gave us a view of other areas and opened new questions about how the gut environment, gastrointestinal complaints, sleep, behaviour, eating and the child’s everyday functioning may be related.

That is exactly where we want to take our research, because we do not want to turn the current results into proof that a change in the gut environment caused changes in behaviour or sleep.

From observation to mechanism. From individual results to a better understanding of how they relate to one another.

Next: Sources →

Sources

Sources and scientific literature

1. Leader G, Abberton C, Cunningham S, Gilmartin K, Grudzien M, Higgins E, Joshi L, Whelan S, Mannion A. Gastrointestinal Symptoms in Autism Spectrum Disorder: A Systematic Review. Nutrients. 2022;14(7):1471. doi:10.3390/nu14071471.
2. Lagod PP, Naser SA. The Role of Short-Chain Fatty Acids and Altered Microbiota Composition in Autism Spectrum Disorder: A Comprehensive Literature Review. Int J Mol Sci. 2023;24(24):17432. doi:10.3390/ijms242417432.
3. Gibson GR, Hutkins R, Sanders ME, Prescott SL, Reimer RA, Salminen SJ, Scott K, Stanton C, Swanson KS, Cani PD, Verbeke K, Reid G. Expert consensus document: The International Scientific Association for Probiotics and Prebiotics (ISAPP) consensus statement on the definition and scope of prebiotics. Nat Rev Gastroenterol Hepatol. 2017;14(8):491–502. doi:10.1038/nrgastro.2017.75.
4. Silk DBA, Davis A, Vulevic J, Tzortzis G, Gibson GR. Clinical trial: the effects of a trans-galactooligosaccharide prebiotic on faecal microbiota and symptoms in irritable bowel syndrome. Aliment Pharmacol Ther. 2009;29(5):508–518. doi:10.1111/j.1365-2036.2008.03911.x.

Next project →

Why and how

Five areas. One follow-up period.

Over the three months, we did not look only at digestion.

The gut microbiome does not function as an isolated system. It communicates with other parts of the body through nervous, immune, endocrine and metabolic pathways. That is also why current research pays attention to, for example, the gut–brain axis or the gut–skin axis.

In our consumer study, we did not measure these biological mechanisms directly.

During the same period, however, we wanted to capture a broader picture of how the individual subjectively rated areas develop with regular use of the product in everyday life.

Why we did not look only at digestion

Mikrobióm+® contains the citrus flavonoids hesperidin and naringin.

Previous experimental findings pointed to their interaction with the gut microbiota and to changes associated with the production of short-chain fatty acids – SCFA, which are among the metabolites produced by the activity of gut microorganisms [1].

Experimental data also showed that supplementation with Mikrobióm+® led in vitro to an increase in bacteria of the genus Roseburia, some species of which are among the producers of butyrate [2].

These findings created the broader scientific context for our project.

The consumer study did not measure these biological mechanisms directly. It did, however, allow us to systematically follow how the individual subjectively rated areas developed over the same three-month period.

How we ran the study

Participants took Mikrobióm+® daily for three months.

The daily dose was 500 mg of Mikrobióm+®, corresponding to 400 mg of hesperidin and 25 mg of naringin. The daily dose was divided into two capsules.

We collected data through online questionnaires at three time points:

before the start of supplementation → after 1 month → after 3 months.

At each time point, participants rated digestive complaints, psychological wellbeing, energy level, sleep quality and skin condition. The final assessment also included questions about adherence to supplementation and overall satisfaction with the product.

In total, 97 questionnaire sets were collected. The final analysis included only participants who completed all three assessments within the set time frame. The final analysed group therefore consisted of 56 participants.

Next: Results →

Results

What did the results show us?

One of the most interesting results of the project was that the individual areas did not develop in the same way. For several parameters, statistically significant differences from baseline appeared after the first month.

In some cases they then stayed stable, in others development continued over the following two months, and for some of the monitored indicators a statistically significant change was not confirmed.

It is precisely the dynamics of the results over time, not just a “before” and “after” comparison, that is one of the most interesting outputs of the study for us.

Digestion

Already after one month, we recorded statistically significant changes in digestive symptom scores compared with baseline. These differences were still present at the three-month assessment.

With bloating, however, the development continued – a further statistically significant change was recorded between the first and third month as well.

Psychological wellbeing and energy

Development was not the same for all parameters, however. For emotional wellbeing the difference was significant after the first month, whereas in the comparison of baseline with the third month the result no longer reached the set threshold of statistical significance.

Such a result is an important part of the overall picture – not all monitored parameters develop in a linear way.

Sleep

Sleep was one of the areas where we observed continuing development throughout the three-month period.

The difference was already present after the first month, and further development was also recorded between the first and third month.

Skin

The individual skin parameters did not develop in the same way either. Hydration, itching and the overall subjective rating of skin quality had already changed after the first month.

With elasticity, development continued over the following months, while for redness a statistically significant change was not confirmed.

Results of the 3-month Mikrobióm+® consumer study · final analysed sample N = 56

Next: What next →

What next

What three months showed us – and where they take us next

The results did not show us one universal pattern. And that is exactly what makes them interesting from a research point of view.

Some differences appeared during the first month and then stayed stable. For other parameters, development continued over the following two months. And for some of the monitored indicators, a statistically significant change was not confirmed.

Three months of follow-up thus allowed us to see more than a simple “before” and “after” comparison. They showed us the dynamics of the individual areas over time. And at the same time they opened another question. A consumer study tells us what participants subjectively observed during regular use. It does not yet tell us why these changes appeared or what biological mechanisms accompanied them.

From real-world use to a deeper question

One of the areas that interest us is the gut microbial environment itself and its metabolic activity. Gut microorganisms produce a great many biologically active substances – microbial metabolites. These include short-chain fatty acids, for example butyrate and propionate.

These substances are one of the mechanisms through which the gut microbiome can communicate with the host organism. In this consumer study, we did not measure the microbiome or its metabolites directly.

We therefore do not see the results as proof of a specific biological mechanism, but as a starting point for the next question: Which of the observed changes can we link to objective biological data in the future?

Where is our research heading?

For us, the M+ project shows a natural transition from real-world use of a product to more precise biological investigation.

Further research should help to better understand, above all, the individual variability of response to supplementation. We are interested in what happens to the composition and functional activity of the gut microbiome during supplementation, how its metabolic activity changes, and whether these data can be linked with what participants subjectively observe.

That is how we want to move from the question:

“What do people observe during regular use?”

to a more precise question:

“Why might individual people respond differently to the same intervention?”

This gives further M+ research its own identity – not just a search for a mechanism, but an understanding of the differences between individual responses.

Our ambition

The M+ project captures why our own research does not end with launching a product on the market.

Scientific findings give us a basis for developing a formulation. Real-world use data then show us what participants observed, but also how individual areas developed over time and how differently individual parameters may respond.

That is where we want to continue. Our ambition is not just to repeat the consumer follow-up with a larger number of people. We want to gradually understand why the response to the same intervention need not be the same for everyone.

M+ began with three months in real life. The next question is what mechanisms lie behind the differences between individual responses.

Next: Sources →

Sources

Sources and scientific literature

1. Stevens Y, Van Rymenant E, Grootaert C, Van Camp J, Possemiers S, Masclee A, Jonkers D. The Intestinal Fate of Citrus Flavanones and Their Effects on Gastrointestinal Health. Nutrients. 2019;11(7):1464. doi:10.3390/nu11071464.
2. Sost MM, Ahles S, Verhoeven J, Verbruggen S, Stevens Y, Venema K. A Citrus Fruit Extract High in Polyphenols Beneficially Modulates the Gut Microbiota of Healthy Human Volunteers in a Validated In Vitro Model of the Colon. Nutrients. 2021;13(11):3915. doi:10.3390/nu13113915.

Next project →

Research path

The next layer of personalisation: from composition to function

Sequence analysis can give us information about the composition of the microbial community. But the gut environment can also be studied from another angle – through the substances that are formed, transformed or reflect its functional activity there.

This is the layer the new project explores. It is research into a complementary type of biological data that could expand our options for decision-making in the future. But before an analytical result can be used as a basis for further research, we first need to know exactly what the instrument measures, which substances it can capture, whether it can only detect them or also quantify them, and with what accuracy. The project is in this analytical validation phase today.

Next: What we are working on today →

What we are working on today

What are we working on today?

The current phase of the project is analytical. It is not an attempt to replace the way personalised probiotics are made today, nor to decide which existing product is “best” for a particular person. We are investigating a new type of data that could in future complement our knowledge of the gut environment.

One of iProbio’s tasks is to compile a list of metabolites relevant to further research. We then need to verify which of them the new analytical approach can actually detect and quantify, and how the results agree with a calibrated analytical methodology.

Only the results of this phase will show whether the analytical approach is accurate, reproducible and practically usable enough for further experimental steps.

Why are we interested in metabolites?

The composition of the gut microbial community is important information, but it is only one view of the gut environment. Another research layer is its functional activity – that is, what substances are formed and transformed in this environment.

One way to investigate this activity is through metabolites. So for our project it is not only decisive whether the instrument produces an analytical signal. We need to know which biologically relevant substances it can capture reliably and whether it can also quantify them.

If such an approach can be validated, metabolic data could in later phases form one of the layers of a sample’s functional profile.

From one sample to a system

Successfully measuring one sample does not yet mean we can choose an intervention based on the result. For that we need a methodology capable of working with a larger number of samples in the same, reproducible way, and then to verify experimentally whether differently characterised profiles also differ in their response. The planned direction of development therefore proceeds through:

Analytical validation → Functional characterisation → Profile database → In vitro responses → Clinical verification

Linking measurement with response data is the essence of the project. We do not want to automatically derive a recommendation from a single analytical result. We first want to verify whether there is a sufficiently stable and reproducible relationship between a measurable profile and the response at all.

Next: What next →

What next

How does this project fit into iProbio’s approaches?

This is not a ranking from “less” to “more” personalised. The individual approaches address a different question and can complement one another.

These approaches do not compete with each other. They answer different questions – from a targeted formulation through individual preparation to research into whether decision-making could in future be extended with additional functional biological data.

From analytical measurement to a decision-making system

If the individual research steps can be successfully linked, the result should not be just a new way of laboratory measurement. The long-term ambition is to investigate whether it is possible to combine:

Analytical measurement → Functional profile → Experimental response data → Data-supported choice from defined interventions

Such a system would not replace individual personalisation. It would be a new research tool for situations in which we work with a pre-prepared set of interventions and want to better understand for whom each option may make more sense.

Our ambition

At iProbio we already work with both a personalised and a targeted approach today. This project explores how to take their scientific foundation further – from information about who lives in the gut environment to the question of what is functionally happening in this environment and how a different profile may respond to the same intervention.

We are starting with analytical validation because new data has value only if we can say exactly what we measure and with what reliability. Only then does it make sense to explore profiles, responses and their possible link.

Our long-term ambition is to gradually expand the amount of high-quality biological data we can rely on in development and more targeted decision-making. Because the more deeply we understand the gut environment and its response, the more precise the questions we can ask in research.

Next project →

Connections

Allergies as part of a bigger picture

Allergic diseases are a broad group of conditions in which the immune system reacts to substances in the environment that are harmless to most people. Their manifestations can vary and affect the airways, the digestive tract and the skin.

In food allergies, an important question is immune tolerance – the body’s ability to respond appropriately to common components of food. Research looks at the relationships between the gut microbiota, the mucosal barrier and the immune response [4].

In inhalant allergies, attention is also paid to the relationships between the microbiota, mucosal immunity and allergic inflammation. Links with both the gut and the local microbial environment are being studied [3, 5].

One specific area we want to explore in more detail is atopic eczema. It is a chronic inflammatory skin disease in which a combination of genetic, immunological and environmental factors plays a part, and impairment of the skin barrier also has a significant role. Among its most prominent features is itching, which can significantly interfere with sleep and quality of life [1, 2].

It is precisely the combination of a broader allergology perspective with a closer look at atopic eczema that opens further questions about the relationships between the gut environment, the immune system and the various manifestations of allergic diseases.

Next: Research questions →

Research questions

From the gut microbiome to immune tolerance

The gut microbiome is a complex community of microorganisms that is in constant contact with the host organism. Through microbial metabolites and interactions with the gut barrier, it may also be linked to processes of immune regulation.

Immune tolerance is one of the important areas of research into allergic diseases. The immune system must be able to respond to a potential danger, while at the same time tolerating the many common substances the body encounters every day.

In food allergies, researchers study the relationships between the gut microbiota, the mucosal barrier and the immune response. The importance of individual microorganisms is not necessarily the same in every circumstance. Their effect may depend on diet, age, the host organism and other environmental factors [4].

In inhalant allergies, links between the gut and mucosal microbiota and allergic inflammation are being studied. The research brings interesting observations, but the exact role of the gut microbiome in individual allergic diseases remains a subject of further study [3, 5].

So this is not a simple relationship in which the specific composition of the gut microbiome directly determines whether an allergy develops. Allergic diseases are the result of a complex interaction of genetic, immunological and environmental factors.

Atopic eczema: when the skin enters the picture too

In atopic eczema, another important element is added to the immune mechanisms – the skin barrier.

The skin is one of the main interfaces between the organism and the external environment. In atopic eczema its barrier function is impaired and, together with complex immune dysregulation, contributes to the development of the typical skin symptoms [1].

Atopic eczema is therefore an interesting model for studying how barrier functions, immune mechanisms and the microbial environment can come together.

It can also occur together with other allergic diseases. That does not mean, however, that there is one universal mechanism or an inevitable sequence linking them. It is the very complexity of these relationships that makes it worthwhile to study them in a broader biological context.

Why the gut–skin axis interests us

The skin and the gut are anatomically distant from each other, yet both organs are major interfaces between the organism and the external environment, and both work closely with the immune system.

This is why current research is looking at a concept known as the gut–skin axis [1].

That does not mean, however, that there is one simple path by which the gut “controls” the skin. It is a complex network of relationships in which the composition and activity of the gut microbiota, the integrity of the gut barrier, microbial metabolites and immune mechanisms may all play a role [1].

In the context of ALLERGIC+®, this concept interests us above all as one of the areas of broader research into allergic diseases. It allows us to look at atopic eczema not only through the visible signs on the skin, but also through the biological connections that may link the gut, the immune system and the skin barrier.

Next: What next →

What next

Where do we want to take the research?

The project is part of our broader research interest in the gut microbiome and the possibilities of its targeted modulation. We want to develop knowledge about the relationships between the gut environment, the immune system and allergic diseases. We are interested in the biological connections that may link the microbiota, barrier functions and immune regulation.

Within this broader view, we pay particular attention to atopic eczema, which makes it possible to explore another important part of this system – the skin and its barrier function.

Our aim is to contribute gradually to a better understanding of this area and to build a scientific foundation for further research questions.

Our ambition

For us, the ALLERGIC+® project is not just research into the skin. It is another branch of our scientific interest in the gut microbiome and its relationship with the immune system.

We want to develop knowledge that links microbiology, immunology, allergology and dermatology. We are interested in what we can gain by not looking at individual organs and diseases in isolation, but in a broader biological context.

The gut, the immune system and the barrier organs are parts of one complex organism. Their mutual relationships are scientific questions to which we want to look for answers step by step.

Next: Sources →

Sources

Sources and scientific literature

1. Sadowsky RL, Sulejmani P, Lio PA. Atopic Dermatitis: Beyond the Skin and Into the Gut. J Clin Med. 2023;12(17):5534. doi:10.3390/jcm12175534.
2. Jeon C, Yan D, Nakamura M, Sekhon S, Bhutani T, Berger T, Liao W. Frequency and Management of Sleep Disturbance in Adults with Atopic Dermatitis: A Systematic Review. Dermatol Ther (Heidelb). 2017;7(3):349–364. doi:10.1007/s13555-017-0192-3.
3. Koidl L, Untersmayr E. An update on the clinical implications of the microbiome in the development of allergy diseases. Allergo J Int. 2024;33:263–268. doi:10.1007/s40629-024-00300-0.
4. Delaroque C, Desai MS. Context-dependent roles of the gut microbiome in food allergy tolerance versus sensitization. Gut Microbes. 2025;17(1):2590830. doi:10.1080/19490976.2025.2590830.
5. Hu Y, Zhang R, Li J, Wang H, Wang M, Ren Q, Fang Y, Tian L. Association Between Gut and Nasal Microbiota and Allergic Rhinitis: A Systematic Review. J Asthma Allergy. 2024;17:633–651. doi:10.2147/JAA.S472632

Next project →

What we will monitor

We are not interested only in whether a person falls ill

The main question of the upcoming project is simple:

What will the course of upper respiratory tract infections look like during 12 weeks of supplementation?

The number of infections alone, however, is only one part of the picture. Two people can go through the same number of infections over the same period and yet their course can be completely different. One episode may be short and mild. Another may last longer, run a more pronounced course or require a visit to the doctor.

That is why we want to monitor four basic dimensions of respiratory infections: how often they appear, how long they last, how severe they are and how long the participant stays free of infection between episodes. It is their combination that can give us a more accurate picture of the whole respiratory season than the simple answer: “fell ill / didn’t fall ill”.

Next: Why this way →

Why this way

Why we want to run the research during the respiratory season

Respiratory infections do not occur evenly throughout the year. It is the autumn and winter period that brings natural conditions for observing repeated episodes of upper respiratory tract infections, so we plan to run the project during the respiratory season, placing the follow-up in the period when the likelihood of URTI is naturally higher.

From published findings to our own data

The individual components of iMunobiom+® do not enter the project without scientific context. In developing the formulation we build on published findings about its individual components, and the task of our own study is not to repeat existing work.

We want to follow the finished formulation as a whole under respiratory-season conditions and obtain our own systematic data on the course of infection episodes. Such a first step will let us find out which parameters are the most informative, how well they can be recorded in practice and which of them are worth using as the basis of a more robust clinical design.

Why we are interested in infection-free time too

With respiratory infections, attention naturally focuses on the period when a person is ill. From a research point of view, however, the period between individual episodes is interesting too.

If infections recur during follow-up, we want to know not only their number and duration, but also how much time passes from the end of one episode to the start of the next. That is why we included infection-free time among the main monitored parameters. This parameter can complement the classic view of disease frequency and help characterise a more complex picture of the whole season, not just individual days of illness.

Next: What next →

What next

Where do we want to take the research?

We see the planned iMunobiom+® study as the first phase of our own research into the finished formula during the respiratory season. Its purpose will be to provide the first systematically collected data and help us refine the design of further clinical research.

The data we collect may show:

• which endpoints best capture the course of infections,
• which data are practically easy to collect,
• what variability we can expect,
• and which questions deserve controlled verification.

In the next phase, we want to supplement clinical and subjective observations with objective laboratory biomarkers. Such a step could create a basis for a future randomised controlled clinical study – an RCT.

Our ambition

For us, the iMunobiom+® project is about more than counting colds. We want to understand the course of an entire respiratory season. How often an infection occurs. How long it lasts. How it progresses. And how much time a person spends without another episode.

The first study may not give us a definitive answer to the question of the formulation’s effect. It is meant, however, to create a data foundation on which we can build higher-quality further clinical research. Our ambition is to move gradually from simple tracking of infection episodes to a more robust model that can link the course of the respiratory season, objective biological data and a controlled clinical design.

One respiratory season may not give us all the answers. But it can show us how to ask the next question more precisely.

Next: Sources →

Sources

Sources and scientific literature

1. Villena J, Kitazawa H. The Modulation of Mucosal Antiviral Immunity by Immunobiotics: Could They Offer Any Benefit in the SARS-CoV-2 Pandemic? Front Physiol. 2020;11:699. doi:10.3389/fphys.2020.00699.
2. Auinger A, Riede L, Bothe G, Busch R, Gruenwald J. Yeast (1,3)-(1,6)-beta-glucan helps to maintain the body’s defence against pathogens: a double-blind, randomized, placebo-controlled, multicentric study in healthy subjects. Eur J Nutr. 2013;52(8):1913–1918. doi:10.1007/s00394-013-0492-z.
3. Dharsono T, Rudnicka K, Wilhelm M, Schoen C. Effects of Yeast (1,3)-(1,6)-Beta-Glucan on Severity of Upper Respiratory Tract Infections: A Double-Blind, Randomized, Placebo-Controlled Study in Healthy Subjects. J Am Coll Nutr. 2019;38(1):40–50. doi:10.1080/07315724.2018.1478339.
4. Martens PJ, Gysemans C, Verstuyf A, Mathieu C. Vitamin D’s Effect on Immune Function. Nutrients. 2020;12(5):1248. doi:10.3390/nu12051248.

Next project →

Hypothesis

From a scientific discovery to our own hypothesis

At iProbio we do not see published scientific studies as the end of knowledge. For us they are the beginning of further research.

The discovery of 4-hydroxyindole did not inspire us to look for a way to simply add this substance from outside. We were far more interested in the question of whether it is possible to create conditions that support the metabolic activity of the gut microbiome itself and the natural production of biologically active metabolites. We built our research direction on this hypothesis.

At the same time, we recognise that the biological processes in endometriosis cannot be explained by a single molecule. That is why our research does not focus on 4-HI alone. We are interested in the wider context – the gut microbiome, its metabolic activity, microbial metabolites, inflammatory processes, the integrity of the gut barrier and other mechanisms that may be interconnected. We consider this more complex view important for further study of the relationship between the microbiome and endometriosis. For us, 4-HI is therefore the starting point of a research question, not the sole explanation of the disease.

Next: ENDOM Study →

ENDOM Study

iProbio ENDOM Study - The first step of our research programme

Every high-quality clinical study needs reliable, systematically collected data. That is why we have prepared the iProbio ENDOM Study – a prospective non-interventional observational study, which is the first phase of our own research programme in the field of endometriosis.

The aim is to follow women with clinically confirmed endometriosis during three months of supplementation with the combination of LADYbiom+®, POWERbiom+® and Mikrobióm+®, recording clinical manifestations, quality of life and selected biological parameters.

We plan to enrol enough women so that 50 participants aged 19 to 45 complete the whole follow-up period. The diagnosis of endometriosis must be clinically confirmed, and on entering the study the women must have at least one of the monitored symptoms – dysmenorrhoea, chronic pelvic pain or dyspareunia.

Each participant will be followed for three months of taking the combination of products, will attend three planned clinical visits and a voluntary check-up 30 days after the end of the supplementation period. The follow-up will include validated questionnaires, the participant’s diary and the provision of biological samples.

So we are not interested only in general feedback from patients; we want to start systematically linking what women observe during the supplementation period with clinical and biological data.

What will we be monitoring during the study?

One of the main values of the project for us is linking several layers of information.

Pain

We will mainly monitor the intensity of dysmenorrhoea, chronic pelvic pain and dyspareunia using the NRS numeric rating scale (0 to 10).

Quality of life

The second important perspective will be quality of life. We will assess it using the validated EHP-30 questionnaire, created specifically for women with endometriosis.

Biological context

Alongside the clinical assessment, we also want to monitor selected biological parameters related to the research hypothesis. Specifically, there will be two levels:

• Inflammatory markers in the blood, specifically IL-6 and TNF-alpha as key pro-inflammatory cytokines associated with chronic inflammation in endometriosis, and hsCRP (high-sensitivity C-reactive protein) as a marker of low-grade systemic inflammation. In addition, standard CRP and a full blood count with a leukocyte differential will also be monitored.
• Microbial indole metabolites in a stool sample, including 4-hydroxyindole, which was described in the study by Talwar et al. (2025) as reduced in women with endometriosis. In a mouse model, its administration reduced endometriotic lesions and eased pain. These are preclinical results that have not yet been verified in humans. In addition to stool, we will also monitor 4-hydroxyindole isomers in blood plasma.

So we do not want to answer only the question:

“Do we observe a change during the supplementation period?”

We are also interested in:

“What is happening in the biological context during the same period?”

Accompanying parameters

The participant’s diary also records daily gastrointestinal symptoms (bloating, intestinal cramps, stool consistency on the Bristol scale), the phase of the menstrual cycle, any concomitant treatment (for example analgesics for breakthrough pain) and adherence to supplementation. This information is important for us so that we can also take into account the influence of other factors that may affect the response to supplementation.

Why a combination of three formulations?

The combination being followed is not built on a single isolated component. We will follow women who take the combination of LADYbiom+®, Mikrobióm+® and POWERbiom+®.

Among other ingredients, LADYbiom+® contains the probiotic strain Lacticaseibacillus rhamnosus CA15, Mikrobióm+® the citrus flavonoids hesperidin and naringin, and POWERbiom+® galactooligosaccharides and components from dried kiwi.

The research rationale for the combination rests on the assumption that the individual components may act on different, interconnected aspects of the microbial environment.

That is why we find it interesting to observe the combination as a whole and its possible synergy, rather than retrospectively attributing the observed result to one isolated component.

Why are we starting with a non-interventional observational study?

The iProbio ENDOM Study is the proof-of-concept phase of our research. Its task is to bring the first standardised data of our own, verify the feasibility of the research concept and help us refine the methodology for the next phase.

We therefore do not see the current project as final proof, but as a systematic first step before a larger controlled clinical validation. Pilot data can show us which parameters are the most informative, what variability we can expect and how to set up the next phase of research as precisely as possible.

Next: What next →

What next

Where do we want to take the research?

After the iProbio ENDOM Study, we want to follow up with the 4HI-SYNBIO project – an upcoming randomised, double-blind and placebo-controlled clinical study.

The concept envisages a larger number of women aged 19 to 45 with clinically confirmed endometriosis and a longer intervention period. Participants would be randomly assigned to a group taking the investigated intervention or placebo. Before and after the intervention, the project envisages more detailed blood and stool analyses, regular ultrasound examinations (TVUS), ongoing assessment using the validated EHP-30 and NRS instruments, and further clinical examinations.

An important difference from the first phase is to be the depth of the biological analyses.

The concept includes quantifying 4-HI and a broader indole panel using LC-MS/MS, analysing the gut microbiome using 16S rRNA sequencing and external biostatistics. Such a design would allow us to link three layers of information:

what the patient observes → what the clinical parameters show → what changes at the level of the microbiome and its metabolites.

We see exactly this depth as the next logical step of the project.

The 4HI-SYNBIO project is currently a concept in preparation. Its final design and scope will be refined in the next phase of project preparation.

Our ambition

For us, the ENDOMETRIOSIS project is not a single isolated study. It is a research programme that arose from a specific scientific discovery and is gradually turning it into our own testable hypothesis.

We started from a published finding about 4-HI. The first step of our own is the iProbio ENDOM Study, which is meant to bring systematic pilot data. If these data allow us to refine the research question further, the next step is to be the controlled clinical validation 4HI-SYNBIO.

Our ambition is therefore not only to observe whether a single symptom changes during supplementation. Over time we want to link the patient’s clinical experience, objective biological data, the gut microbiome and its metabolic activity.

And to find out whether a new scientific question can, step by step, become a clinically testable concept.

From discovery to hypothesis. From hypothesis to our own data. From our own data to clinical validation.

Next: Sources →

Sources

Sources and scientific literature

1. Talwar C, Davuluri GVN, Kamal AHM, Coarfa C, Han SJ, Veeraragavan S, Parsawar K, Putluri N, Hoffman K, Jimenez P, Biest S, Kommagani R. Identification of distinct stool metabolites in women with endometriosis for non-invasive diagnosis and potential for microbiota-based therapies. Med. 2025;6(2):100517. doi:10.1016/j.medj.2024.09.006.

Next project →