Clinical research with probiotics as an indicator of global valorization since the year 2000.

Probiotics are becoming increasingly popular due to their potential health benefits. With this rise in popularity and demand as indicated by ever-growing market prospects, it seems evident that innovation and valorization are on the rise as well. However, an increasing body of literature shows that innovation is stagnating, which may be detrimental to the exploitation of the benefits of probiotics, for example the development of alternative therapies to manage the increasing prevalence of metabolic and autoimmune disorders. To this end, this study investigated global clinical trials that have been executed since the year 2000 as a first indicator of the status of probiotic valorization. The cumulative number of clinical trials has indeed increased significantly from 0 at the start of the century up to 2,517 registered trials in 2023. However, in Asia, Europe, and North America, the continents with the highest numbers of clinical trials, stagnating or declining trends have been found. In these locations, most clinical trials were funded by non-industry sponsors and targeting probiotic supplements or undefined products. Considering the overall stagnation in clinical trials and viewing these trends in the context of developments in local markets and regulations, the global valorization of probiotics appears to slow down. This could impact the transition from academic research to the development of products that are beneficial and accessible for consumers, either to maintain a healthy lifestyle or to treat medical conditions.

Reviewing the state of the art of probiotics as clinical modalities for brain-gut-microbiota axis associated disorders.

The rise in prevalence of mental and neurological disorders is causing a high burden on society, however adequate interventions are not always available. The brain-gut-microbiota axis (BGMA) may provide a new angle for the development of clinical modalities. Due to the intricate bi-directional signaling between the brain and the gut, it may be helpful to look into interventions that target the gut, such as probiotics. Therefore, this review aimed to investigate the state of the art of probiotics and their potential as clinical modalities for BGMA-associated indications by gaining insight into patents and clinical trials that have been applied for and executed since 1999. A total of 565 patents and 390 clinical trials were found, focusing on probiotic applications for 83 indications. Since the start of the 21st century, the highest numbers of patents and clinical trials were related to primary neuropsychological, affective (depression, anxiety) and cognitive disorders, neurodegenerative and/or inflammatory brain disorders (Alzheimer's disease, Parkinson's disease, amongst others), and gastrointestinal disorders (irritable bowel syndrome). The locations where the most patents and clinical trials were registered included China, the United States, and Iran. From 1999 to ~2013 a slight growth could be seen in the numbers of patents and clinical trials, followed by an almost exponential growth from ~2013 onwards. Overall, the developments of the state of the art were in accordance with previous research, however it appeared that clinical trials showed a slightly slower growth compared to patents, which may have implications for the future implementation of probiotics as clinical modalities for BGMA-associated indications.

Innovations in the veterinary intestinal health field: A patent landscape analysis.

In recent years it has become evident that a healthy intestinal microbiome is beneficial for the overall health of an individual. A healthy microbiome is diverse, increasing stability and resilience and strengthening the immune system. In addition, healthy intestinal metabolisms have a beneficial effect on many physiological processes such as the brain function. Looking from the One Health perspective, which recognizes that health of humans is closely connected to the health of animals and environment, it is inherently beneficial to stimulate the health of animals for the well-being of humans. However, the intensive administration of antibiotics to livestock for prevention and cure of disease, and even stimulation of growth, disrupts a healthy microbiome. With the rapid increase of emerging zoonotic diseases, alternatives to the use of antimicrobial compounds are urgently necessary. This research analyses the development of alternatives for antibiotic use contributing to veterinary intestinal health through an in-depth patent analysis of inventions for fodder additives. In the period 1999-2020, 1269 unique patent families describing the use of probiotics, enzymes and prebiotics for swine, poultry and ruminants were identified. Innovation trends, geography, key applicants, and classification of patents were analysed. Asian industrial applicants applied for the majority of patents comprising the largest share of patents for probiotics and enzymes in combination with fodder for swine. Followed by North American and European industrial applications, applying for patents for probiotics in combination with fodder for poultry, swine, and ruminants. Overall, our results do not show a clear increase in innovations, suggesting that innovations in the use of probiotics and enzymes in animal feed appear to be stalling. While in the near future a combination of the use of antibiotics and alternatives is most likely to be implemented, the use of probiotics stands a good chance of replacing antibiotics in animal husbandry and limiting the adverse effects of antibiotic abuse.

The Gut Microbiota: Master of Puppets Connecting the Epidemiology of Infectious, Autoimmune, and Metabolic Disease.

Infectious, autoimmune, and metabolic diseases put an enormous pressure on both quality of life and the economy. For all three disease types, it is known that the quality of the gut microbiota composition is correlated to both onset and progression of disease. Hence, maintaining eubiosis and preventing gradual irreversible loss of beneficial microbes within the gut microbial ecosystem is of utmost importance. As such, the epidemiological trends of these disease types may serve as proxies for the integrity of the human gut microbiota. Here, we present incidence data covering the last decades for prototypical infectious diseases (tuberculosis and measles), autoimmune disorders (type-1 diabetes and multiple sclerosis), and the prevalence of metabolic syndrome. Our findings reveal that vaccination efforts correlate with relatively low levels of archetypal infectious disease incidence. However, autoimmune and metabolic disorders are, together with the usage of antibiotics, steeply on the rise. These findings suggest that the status of the gut microbiota is persistently deteriorating, as reflected by the proxies. As such, the epidemiological trends shown here may serve as a starting point for a mechanistic understanding of the interplay between these different disease types that can be used for future prevention and mitigation strategies like targeted stimulation and suppletion of microorganisms by means of, e.g., fermented foods, prebiotics and probiotics.

Probiotics for the Management of Infectious Diseases: Reviewing the State of the Art.

This review aims to provide insight into the potential of probiotics as a clinical modality targeted at infectious diseases by creating a comprehensive overview of the state of the art of research and development efforts as shown by patents and clinical trials of the past 20 years. Data were retrieved from patent and clinical trial databases to reflect the long- and short-term developments of probiotics research. The data were analyzed to extract information on the total number of patents and trials for each indication, application date and location, and applicant/sponsor type. A total of 80 infectious diseases were investigated, precipitating in 789 patents and 602 clinical trials for 67 indications studied as targets of probiotics. An increasing trend was seen for the number of patents and clinical trials that were applied for since 1999 with the highest number of patents and clinical trials targeted to digestive tract, respiratory, and urogenital indications. Overall, research demonstrated a substantial interest in probiotics targeting infectious diseases, which was in line with reported unmet needs and global probiotics sales estimates. However, the declining rate of translation from patents to clinical trials indicates that there are some barriers obstructing the research process.

Nurturing by nutrition: On the future of gut microbiota management strategies for autoimmune disease.

The incidence of autoimmune disease continues to rise, which urges for new prevention and treatment modalities. The composition of the gut microbiota is associated with both susceptibility and progression of disease. Nutrition significantly shapes the gut microbial composition, and poses as such a modality for both prevention and treatment/adjuvant therapy. At very young age, nutritional intervention targeting the gut microbiota is still possible within a one-size-fits all regime, accompanied by a relatively high effect size. As ageing results in higher interindividual variation induced by cumulative exposome factors, a more personalized approach is needed, having a higher effect size than that of current nutritional intervention. As such, supplementation of microbial consortia consisting of keystone taxa and microbial guilds that are involved in the pathophysiology seem a promising direction to lower the burden of autoimmune disease.

On the Verge of a Catastrophic Collapse? The Need for a Multi-Ecosystem Approach to Microbiome Studies.

While the COVID-19 pandemic has led to increased focus on pathogenic microbes that cross the animal-human species barrier, calls to include non-pathogenic interactions in our perspective on public health are gaining traction in the academic community. Over generations, the diversity of the human gut microbiota is being challenged by external perturbations and reduced acquisition of symbiotic species throughout life. When such reduced diversity concerns not only the microbial species, but also the higher taxonomic levels and even the guild level, adequate compensation for possible losses may be lacking. Shifts from a high-abundance to a low-abundance state, known as a tipping point, may result in simultaneous shifts in covarying taxa and ultimately to a catastrophic collapse in which the ecosystem abruptly and possibly irreversibly shifts to an alternative state. Here, we propose that co-occurrence patterns within and between microbial communities across human, animal, soil, water, and other environmental domains should be studied in light of such critical transitions. Improved mechanistic understanding of factors that shape structure and function is needed to understand whether interventions can sustainably remodel disease-prone microbiota compositions to robust and resilient healthy microbiota. Prerequisites for a rational approach are a better understanding of the microbial interaction network, both within and inter-domain, as well as the identification of early warning signs for a catastrophic collapse, warranting a timely response for intervention. We should not forget that mutualism and pathogenicity are two sides of the same coin. Building upon the planetary health concept, we argue that microbiome research should include system level approaches to conserve ecosystem resilience. HIGHLIGHTS 1. Non-pathogenic interactions between ecosystems play a key role in maintaining health. 2. The human gut microbiome may be on the verge of a catastrophic collapse. 3. Research should identify keystone taxa and guilds that interconnect different domains. 4. We should not forget that mutualism and pathogenicity are two sides of the same coin.

Towards a rational design of faecal transplant analogues.

Faecal transplants (microbiota transfer) have shown to be promising therapies having a wide range of therapeutic applications. However, current safety considerations hamper further valorisation. As such, well designed faecal transplant analogues provide an interesting alternative to minimize possible safety aspects. However, to date little knowledge on how to rationally design such analogues exists. Here, we show by applying first order basic graph theory that such analogues dedicated to restoring a specific physiological functionality (a microbial guild) should consist of 5-6 species to maximize stability, efficiency, and minimize safety issues and production costs.

The mechanistic link between health and gut microbiota diversity.

Although numerous reports link a decreased diversity of the gut microbiota to a declined health status, to date no mechanistic motivation for this exists. Here, we show by applying first principles basic graph theory on small networks that higher diversity within such a network indeed leads to more efficient systems and redundancy. Our results quantitatively support earlier hypothetical considerations on gut microbiota richness with respect to these parameters. Our simulations show that higher species diversity leads to higher resilience within small microbiological ecosystems, like being present in the gut microbiota. This notion should provide an ingredient when developing new interventional strategies within the domain of microbiota management.

The underexposed role of food matrices in probiotic products: Reviewing the relationship between carrier matrices and product parameters.

Probiotic microorganisms are increasingly incorporated into food matrices in order to confer proposed health benefits on the consumer. It is important that the health benefits, sensory properties, shelf-life and probiotic gastrointestinal tract (GIT) survival of these products are carefully balanced as they determine functionality and drive consumer acceptance. The strain-specific effects of probiotic species are imperative in this process but carrier matrices may play a pivotal role as well. This study therefore recapitulates the wealth of knowledge on carrier matrices and their interaction with probiotic strains. The most substantiated carrier matrices, factors that influence probiotic functionality and matrix effects on shelf-life, GIT survival and clinical efficacy are reviewed. Results indicate that carrier matrices have a significant impact on the quality of probiotic products. Matrix components, such as proteins, carbohydrates and flavoring agents are shown to alter probiotic efficacy and viability. In vivo studies furthermore revealed strain-dependent matrix effects on the GIT survival of probiotic bacteria. However, only a limited number of studies have specifically addressed the effects of carrier matrices on the aforementioned product-parameters; most studies seem to focus solely on the strain-specific effects of probiotic microorganisms. This hampers the innovation of probiotic products. More human studies, comparing not only different probiotic strains but different carrier matrices as well, are needed to drive the innovation cycle.

Probing the structure of a rotaxane with two-dimensional infrared spectroscopy.

Femtosecond 2D-IR spectroscopy has been used to study the structure of a [2]rotaxane composed of a benzylic amide macrocycle that is mechanically interlocked onto a succinamide-based thread. Both the macrocycle and the thread contain carbonyl groups, and by determining the coupling between the stretching modes of these groups from the cross-peaks in the 2D-IR spectrum, the structure of the macrocycle-thread system has been probed. Our results demonstrate that 2D-IR spectroscopy can be used to observe structural changes in molecular devices on a picosecond time scale.

Ultrafast dynamics of isolated model photoactive yellow protein chromophores: "Chemical perturbation theory" in the laboratory.

Pump-probe and pump-dump probe experiments have been performed on several isolated model chromophores of the photoactive yellow protein (PYP). The observed transient absorption spectra are discussed in terms of the spectral signatures ascribed to solvation, excited-state twisting, and vibrational relaxation. It is observed that the protonation state has a profound effect on the excited-state lifetime of p-coumaric acid. Pigments with ester groups on the coumaryl tail end and charged phenolic moieties show dynamics that are significantly different from those of other pigments. Here, an unrelaxed ground-state intermediate could be observed in pump-probe signals. A similar intermediate could be identified in the sinapinic acid and in isomerization-locked chromophores by means of pump-dump probe spectroscopy; however, in these compounds it is less pronounced and could be due to ground-state solvation and/or vibrational relaxation. Because of strong protonation-state dependencies and the effect of electron donor groups, it is argued that charge redistribution upon excitation determines the twisting reaction pathway, possibly through interaction with the environment. It is suggested that the same pathway may be responsible for the initiation of the photocycle in native PYP.

Vibrational relaxation of the bending mode of HDO in liquid D2O.

The vibrational relaxation of the bending mode of HDO in liquid D2O has been studied using time-resolved mid-infrared pump-probe spectroscopy. At short delays, the transient spectrum clearly shows the v = 1 --> 2 induced absorption and v = 1 --> 0 bleaching and stimulated emission, whereas at long delays, the transient spectrum is dominated by the spectral changes caused by the temperature increase in the sample after vibrational relaxation. From the decay of the v = 1 --> 2 induced absorption, we obtain an estimate of 390 +/- 50 fs for the vibrational lifetime, in surprisingly good agreement with recent theoretical predictions. In the v = 0 --> 1 frequency region, the decay of the absorption change involves a second, slower component, which suggests that after vibrational relaxation the system is not yet in thermal equilibrium.

Vibrational relaxation of the H2O bending mode in liquid water.

We have studied the vibrational relaxation of the H(2)O bending mode in an H(2)O:HDO:D(2)O isotopic mixture using infrared pump-probe spectroscopy. The transient spectrum and its delay dependence reveal an anharmonic shift of 55+/-10 cm(-1) for the H(2)O bending mode, and a value of 400+/-30 fs for its vibrational lifetime.