AIMS:The present study aims to evaluate the impact of Lactiplantibacillus plantarum LSC3, previously isolated from the stool of an Algerian breastfed infant, on the development of allergic responses to milk proteins in BALB/c mice. METHODS AND RESULTS:BALB/c mice were given orally Lactiplantibacillus plantarum LSC3 and sensitized to β-lactoglobulin according to a food allergy protocol. The results show that oral administration of L. plantarum LSC3 induced a significant decrease in the serum levels of anti β-lactoglobulin IgG and IgE. In addition, oral administration of L. plantarum LSC3 also appears to significantly decrease anaphylactic response ex vivo by reducing short circuit current (Isc) and epithelial conductance (G). Moreover, L. plantarum LSC3 attenuated clinical manifestations induced by the in vivo β-Lg challenge test and protected intestinal morphology from β-Lg-induced tissue alterations. CONCLUSION:These findings reveal that L. plantarum LSC3 exerts immunomodulatory properties, probably through the modulation of systemic and intestinal immune responses.

The gut–brain axis (GBA) has been proposed as a modulator of early neurodevelopment, with growing evidence showing that the gut microbiota influences central nervous system (CNS) functions during infancy and childhood. Postbiotic — non-viable microbial products, metabolites, and structural components — represent a promising therapeutic strategy for modulating neuroimmune interactions and supporting brain health. However, it remains challenging to directly assess their effects on the human paediatric brain.This methodological paper describes an efficient protocol to produce and apply gut microbiota-derived postbiotics to ex vivo human paediatric cortical slices. This approach provides a valuable experimental platform to further investigate the direct effects of microbiota-derived factors on human brain circuits, particularly in the context of neurodevelopment and neuroinflammation. Paediatric cortical tissue obtained from patients with drug-resistant epilepsy provides a valuable ex vivo model for assessing postbiotic effects in pathological contexts. Challenges and future directions in standardizing postbiotic-based interventions for neurodevelopmental and neuroimmune disorders are also discussed.

Probiotics promote host health and have shown potential as both prophylactics and adjuvants for the treatment of several human diseases. Therefore, medical interest in these microorganisms has increased rapidly. Recently, probiotics along with postbiotics, i.e., their inanimated counterparts, have been explored for their capacity to treat cancer, a vast group of diseases characterized by uncontrolled cell proliferation, which represents a major global health burden and is the second leading cause of death worldwide. The present review collects and discusses recent preclinical studies addressing the anticancer action of probiotics and postbiotics from lactic acid bacteria and bifidobacteria, and describes the molecular mechanisms by which they hamper some major hallmarks of tumor cells, i.e., through stimulation of apoptosis, blockade of proliferation and the cell cycle, inhibition of metastasis, metabolic interference, activation of the host anti-cancer immune response. The medical use of probiotics should rely on an extensive understanding of the molecular and cellular mechanisms underlying their anticancer activities. Accordingly, this review aims to provide insights into the advancements and limitations of the current research in this field, which is strongly needed to provide solid rationales for the prospective clinical use of probiotics in cancer.

The maternal gut microbiome plays a pivotal role in shaping pregnancy outcomes. Probiotics are known to support maternal immune function, reduce infection risk, modulate inflammation, and contribute to the prevention of gestational complications such as preeclampsia and diabetes. These effects are largely mediated through alteration in the maternal gut microbiota and the production of microbial metabolites, particularly short-chain fatty acids, which exert direct regulatory effects on the host epigenome.In recent years, increasing attention has focused on the use of probiotics during pregnancy, not only for their benefits on maternal immunity and metabolism but also for their potential to influence the fetal epigenome and, specifically, brain development. Fetal brain epigenetic programming is characterized by a remarkable degree of plasticity and is highly responsive to environmental factors, such as maternal diet, lifestyle, stress, gut microbic status and nutritional supplementation. Understanding the complex relationship between probiotic-induced changes in maternal microbiota and fetal epigenetic modifications, such as DNA methylation, histone modifications, and non-coding RNA regulation, is essential for clarifying how gestational supplementation may shape neurodevelopmental patterns in offspring. This review examines the targeting of pregnancy-associated dysbiosis with probiotics supplementation as a potential and hypothesis-generating strategy that may contribute to favourable epigenetic programming and is associated with neurodevelopmental trajectories relevant to later-life mental health.

Recent studies have highlighted a close relationship between intestinal dysbiosis and paediatric neurological disorders, such as autism spectrum disorders (ASD), attention deficit hyperactivity disorder (ADHD), epilepsy, cerebral palsy (CP). In the last decades, the relevance of a strong connection between the intestinal microbiota and the brain, i.e., the gut-brain axis, has been broadly ascertained. Through this association, gut microbes contribute to shape development, physiology, and cognitive functions of the brain. The bidirectional communication between gut microbiota and brain involves a complex set of molecular signals that influence synaptic plasticity, learning, memory, mood, and the brain's excitatory-inhibitory balance. Microbial metabolites play a fundamental role in modulating brain activity. Emerging scientific evidence suggests that neurological and psychiatric conditions, including ASD, ADHD, anxiety, and depression, may not simply stem from brain malfunctions, but rather from imbalances in the gut microbiota, which contribute to disrupt brain function and development. In this context, psychobiotics, i.e., a specific group of probiotic microorganisms, have garnered interest for their potential to modulate brain functions. Several pre-clinical studies on psychobiotics have shown promising results in managing paediatric neurological disorders, including ASD and ADHD. However, there is only a limited number of clinical studies to support the use of psychobiotics as therapeutics. This review summarizes and discusses the link between gut microbiota and neurological disorders in children, and the current status of studies investigating the potential application of psychobiotic lactic acid bacteria (LAB) and bifidobacteria for treating these paediatric disorders.

Lactic acid bacteria (LAB) are food-grade microorganisms able to produce and release bioactive compounds of interest to human health. Some LAB strains can synthesize vitamin B2, i.e., riboflavin, a micronutrient essential for cellular metabolism. In this work, six Lactiplantibacillus plantarum isolated from fruits of the Mediterranean area were exposed to the selective pressure of roseoflavin in order to select spontaneous riboflavin-overproducing phenotypes. The best strains, as determined by the level of riboflavin produced, were characterized for some basic probiotic features, including antibacterial activity, production of organic acids, antibiotic resistance, and survival under digestive stresses using an in vitro gut model. The strain L. plantarum Lp 187_B2, which produced the highest riboflavin level (6 mg/L), exhibited good resistance to gastro-intestinal stress and a relevant capacity to antagonize undesired bacteria, was selected for additional investigations to assess its capacity to protect intestinal homeostasis. When used as a postbiotic, Lp 187_B2 significantly increased trans-epithelial electrical resistance (TEER) in Caco-2 cell monolayers, an in vitro model of the intestinal barrier. Moreover, in a Caco-2/THP-1 co-culture system, mimicking the inflamed bowel, Lp 187_B2 postbiotics significantly inhibited the release of TNF-α by macrophages, thus pointing to gut-barrier strengthening and potential anti-inflammatory properties. Though a validation in vivo is required, our preliminary results indicate that L. plantarum Lp 187_B2 could be successfully applied as both probiotic and postbiotic formulations for improving human health.

Neurodegenerative disorders are the main cause of cognitive and physical disabilities, affect millions of people worldwide, and their incidence is on the rise. Emerging evidence pinpoints a disturbance of the communication of the gut–brain axis, and in particular to gut microbial dysbiosis, as one of the contributors to the pathogenesis of these diseases. In fact, dysbiosis has been associated with neuro-inflammatory processes, hyperactivation of the neuronal immune system, impaired cognitive functions, aging, depression, sleeping disorders, and anxiety. With the rapid advance in metagenomics, metabolomics, and big data analysis, together with a multidisciplinary approach, a new horizon has just emerged in the fields of translational neurodegenerative disease. In fact, recent studies focusing on taxonomic profiling and leaky gut in the pathogenesis of neurodegenerative disorders are not only shedding light on an overlooked field but are also creating opportunities for biomarker discovery and development of new therapeutic and adjuvant strategies to treat these disorders. Lactiplantibacillus plantarum (LBP) strains are emerging as promising psychobiotics for the treatment of these diseases. In fact, LBP strains are able to promote eubiosis, increase the enrichment of bacteria producing beneficial metabolites such as short-chain fatty acids, boost the production of neurotransmitters, and support the homeostasis of the gut–brain axis. In this review, we summarize the current knowledge on the role of the gut microbiota in the pathogenesis of neurodegenerative disorders with a particular focus on the benefits of LBP strains in Alzheimer’s disease, Parkinson’s disease, amyotrophic lateral sclerosis, autism, anxiety, and depression.

Microglia are macrophage cells residing in the brain, where they exert a key role in neuronal protection. Through the gut–brain axis, metabolites produced by gut commensal microbes can influence brain functions, including microglial activity. The nuclear factor erythroid 2-related factor 2 (NRF2) is a key regulator of the oxidative stress response in microglia, controlling the expression of cytoprotective genes. Lactobacilli-derived cell-free supernatants (CFSs) are postbiotics that have shown antioxidant and immunomodulatory effects in several in vitro and in vivo studies. This study aimed to explore the effects of lactobacilli CFSs on modulating microglial responses against oxidative stress and inflammation. HMC3 microglia were exposed to lipopolysaccaride (LPS), as an inflammatory trigger, before and after administration of CFSs from three human gut probiotic species. The NRF2 nuclear protein activation and the expression of NRF2-controlled antioxidant genes were investigated by immunoassay and quantitative RT-PCR, respectively. Furthermore, the level of pro- and anti-inflammatory cytokines was evaluated by immunoassay. All CFSs induced a significant increase of NRF2 nuclear activity in basal conditions and upon inflammation. The transcription of antioxidant genes, namely heme oxygenase 1, superoxide dismutase (SOD), glutathione-S transferase, glutathione peroxidase, and catalase also increased, especially after inflammatory stimulus. Besides, higher SOD1 activity was detected relative to inflamed microglia. In addition, CFSs pre-treatment of microglia attenuated pro-inflammatory TNF-α levels while increasing anti-inflammatory IL-10 levels. These findings confirmed that gut microorganisms’ metabolites can play a relevant role in adjuvating the microglia cellular response against neuroinflammation and oxidative stress, which are known to cause neurodegenerative diseases. Gut-brain crosstalk: molecular point of view. Metabolites contained in the supernatant derived from Lactobacilli can cross the gut barrier and reach the central nervous system, where they are taken up by microglial cells. They induce the activation of the NRF2 pathway and the production of inflammatory mediators. This interaction attenuates two important events: oxidation (with high levels of NRF2) and inflammation (with high levels of IL-10 and low levels of TNF-α).

The food industry constantly seeks new starter cultures with superior characteristics to enhance the sensory and overall quality of final products. Starting from a collection of Algerian dairy (goat and camel) lactic acid bacteria, this work focused on the exploration of the technological and probiotic potential of Weissella cibaria (VR81 and LVT1) and Lactiplantibacillus plantarum R12 strains isolated from raw camel milk and fermented milk, respectively. These bioactive strains were selected for their high performance among ten other LAB strains and were used as starter cultures to develop a novel and nutritionally enhanced dairy-like plant-based yogurt using quinoa (Chenopodium quinoa Willd) as a raw matrix. The strains were evaluated for their antagonistic effects against Listeria innocua, Listeria ivanovii, Staphylococcus aureus, Escherichia coli, Salmonella enterica, and Pseudomonas aeruginosa, resilience to acidic and osmotic challenges, and tolerance to gastrointestinal mimicking conditions (i.e., pepsin and bile salt). Their aggregation and adhesion profiles were also analyzed. Furthermore, L. plantarum and W. cibaria were tested in single and co-culture for the fermentation and biocontrol of quinoa. The strains exhibited probiotic properties, including a high potential for biocontrol applications, specifically against L. innocua and P. aeruginosa (20 mm diameter zone with the neutralized cell-free supernatant), which disappeared after protease treatment, suggesting that bioactive peptides might be responsible for the observed antimicrobial effect. Additionally, they demonstrated resilience to acidic (pH 2) and osmotic challenges (1M sucrose), tolerance to gastro-intestinal conditions, as well as good aggregation and adhesion profile. Furthermore, the strains were able to produce metabolites of interest, such as exopolysaccharide (yielding up to 4.7 mg/mL) and riboflavin, reaching considerable production levels of 2.5 mg/L upon roseoflavin selection. The application of W. cibaria and L. plantarum as primary starters (both in single and co-culture) for fermenting quinoa resulted in effective acidification of the matrix (ΔpH of 2.03 units) and high-quality beverage production. in vivo challenge tests against L. innocua showed the complete inhibition of this pathogen when L. plantarum was included in the starter, either alone or in combination with W. cibaria. Both species also inhibited Staphylococcus and filamentous fungi. Moreover, the co-culture of mutant strains of L. plantarum R12d and W. cibaria VR81d produced riboflavin levels of 175.41 µg/100 g in fermented quinoa, underscoring their potential as starters for the fermentation, biopreservation, and biofortification of quinoa while also displaying promising probiotic characteristics.

Upon dietary administration, probiotic microorganisms can reach as live cells the human gut, where they interact with the microbiota and host cells, thereby exerting a beneficial impact on host functions, mainly through immune-modulatory activities. Recently, attention has been drawn by postbiotics, i.e. non-viable probiotic microbes, including their metabolic products, which possess biological activities that benefit the host. Lactiplantibacillus plantarum is a bacterial species that comprises recognised probiotic strains. In this study, we investigated in vitro the probiotic (and postbiotic) potential of seven L. plantarum strains, including five newly isolated from plant-related niches. The strains were shown to possess some basic probiotic attributes, including tolerance to the gastrointestinal environment, adhesion to the intestinal epithelium and safety. Besides, their cell-free culture supernatants modulated cytokine patterns in human macrophages in vitro, promoting TNF-α gene transcription and secretion, while attenuating the transcriptional activation and secretion of both TNF-α and IL-8 in response to a pro-inflammatory signal, and enhancing the production of IL-10. Some strains induced a high IL-10/IL-12 ratio that may correlate to an anti-inflammatory capacity in vivo. Overall, the investigated strains are good probiotic candidates, whose postbiotic fraction exhibits immunomodulatory properties that need further in vivo studies. The main novelty of this work consists in the polyphasic characterisation of candidate beneficial L. plantarum strains obtained from relatively atypical plant-associated niches, by an approach that explores both probiotic and postbiotic potentials, in particular studying the effect of microbial culture-conditioned media on cytokine pattern, analysed at both transcriptional and secretion level in human macrophages.

Luteolin (3′,4′,5,7-tetrahydroxyflavone), a member of the flavonoid family derived from plants and fruits, shows a wide range of biomedical applications. In fact, due to its anti-inflammatory, antioxidant and immunomodulatory activities, Asian medicine has been using luteolin for centuries to treat several human diseases, including arthritis, rheumatism, hypertension, neurodegenerative disorders and various infections. Of note, luteolin displays many anti-cancer/anti-metastatic properties. Thus, the purpose of this review consists in highlighting the relevant mechanisms by which luteolin inhibits tumor progression in metastasis, i.e., affecting epithelial-mesenchymal transition (EMT), repressing angiogenesis and lysis of extracellular matrix (ECM), as well as inducing apoptosis.

The continued exploration of the diversity of lactic acid bacteria in little-studied ecological niches represents a fundamental activity to understand the diffusion and biotechnological significance of this heterogeneous class of prokaryotes. In this study, Lactiplantibacillus plantarum (Lpb. plantarum) strains were isolated from Tunisian vegetable sources, including fermented olive and fermented pepper, and from dead locust intestines, which were subsequently evaluated for their antimicrobial activity against foodborne pathogenic bacteria, including Escherichia coli O157:H7 CECT 4267 and Listeria monocytogenes CECT 4031, as well as against some fungi, including Penicillium expansum, Aspergilus niger, and Botrytis cinerea. In addition, their resistance to oro-gastro-intestinal transit, aggregation capabilities, biofilm production capacity, adhesion to human enterocyte-like cells, and cytotoxicity to colorectal adenocarcinoma cell line were determined. Further, adhesion to tomatoes and the biocontrol potential of this model food matrix were analyzed. It was found that all the strains were able to inhibit the indicator growth, mostly through organic acid production. Furthermore, these strains showed promising probiotic traits, including in vitro tolerance to oro-gastrointestinal conditions, and adhesion to abiotic surfaces and Caco-2 cells. Moreover, all tested Lpb. plantarum strains were able to adhere to tomatoes with similar rates (4.0–6.0 LogCFU/g tomato). The co-culture of LAB strains with pathogens on tomatoes showed that Lpb. plantarum could be a good candidate to control pathogen growth. Nonetheless, further studies are needed to guarantee their use as probiotic strains for biocontrol on food matrices.

Usually, the search for new candidate probiotics starts from strain isolation, followed by genotypic and phenotypic characterisations. For the best candidates, the final selection criteria, i.e., an efficient biomass production and the survival to stressful conservation processes, may often represent a bottleneck. The aim of this study is to reverse this classic bottom-up approach, thereby evaluating the in vitro probiotic properties of microbes that are already commercialized and employed in the dairy sector. The major advantage of reversing the traditional scheme is to deal with strains that are already suitable for the scale-up at the industrial level. In this work, four lactobacilli strains were analysed, belonging to the species of Lactiplantibacillus plantarum (strains PLA and PLA2) and Lacticaseibacillus rhamnosus (strains PAR4 and RHM). Both L. plantarum strains showed the best survival under simulated oro-gastrointestinal stress; PLA and PAR4 had the strongest inhibitory activity against all the tested harmful bacteria, with the latter strain showing also the highest percentage of Caco-2 adhesion; RHM was the best biofilm producer on abiotic surface. Finally, cell-free surnatants from all the strain cultures exhibited anti-inflammatory action on THP-1 macrophages. For all the studied strains, it is possible to claim beneficial functional properties other than the technological ones for which they are already marketed. The possible use of the four strains in a mixture could represent a strategy to diversify and maximize their beneficial potential. Nonetheless, future studies are necessary to validate in vivo the observed beneficial properties and to evaluate any effect of the vehicle product on the probiotic aptitude.

Fabry disease (FD) is an X-linked lysosomal disease due to a deficiency in the activity of the lysosomal-galactosidase A (GalA), a key enzyme in the glycosphingolipid degradation pathway. FD is a complex disease with a poor genotype–phenotype correlation. In the early stages, FD could involve the peripheral nervous system (acroparesthesias and dysautonomia) and the ski (angiokeratoma), but later kidney, heart or central nervous system impairment may significantly decrease life expectancy. The advent of omics technologies offers the possibility of a global, integrated and systemic approach well-suited for the exploration of this complex disease. In this narrative review, we will focus on the main metabolomic studies, which have underscored the importance of detecting biomarkers for a diagnostic and prognostic purpose in FD. These investigations are potentially useful to explain the wide clinical, biochemical and molecular heterogeneity found in FD patients. Moreover, the quantitative mass spectrometry methods developed to evaluate concentrations of these biomarkers in urine and plasma will be described. Finally, the complex metabolic biomarker profile depicted in FD patients will be reported, which varies according to gender, types of mutations, and therapeutic treatment.

Honey is a valuable reservoir of lactic acid bacteria (LAB) and, particularly, of fructophilic LAB (FLAB), a relatively novel subgroup of LAB whose functional potential for human and food application has yet to be explored. In this study, FLAB and LAB strains have been isolated from honeys of different floral origins and selected for their broad antimicrobial activity against typical foodborne pathogenic bacteria and spoilage filamentous fungi. The best candidates, two strains belonging to the species Lactiplantibacillus plantarum and Fructobacillus fructosus , were submitted to partial characterisation of their cell free supernatants (CFS) in order to identify the secreted metabolites with antimicrobial activity. Besides, these strains were examined to assess some major functional features, including in vitro tolerance to the oro-gastrointestinal conditions, potential cytotoxicity against HT-29 cells, adhesion to human enterocyte-like cells and capability to stimulate macrophages. Moreover, when the tested strains were applied on table grapes artificially contaminated with pathogenic bacteria or filamentous fungi, they showed a good ability to antagonise the growth of undesired microbes, as well as to survive on the fruit surface at a concentration that is recommended to develop a probiotic effect. In conclusion, both LAB and FLAB honey-isolated strains characterised in this work exhibit functional properties that validate their potential use as biocontrol agents and for the design of novel functional foods. We reported antimicrobial activity, cytotoxic evaluation, probiotic properties and direct food application of a F. fructosus strain, improving the knowledge of this species, in particular, and on FLAB, more generally.

Small heat-shock proteins (sHSP) are ubiquitous ATP-independent chaperones that prevent irreversible aggregation of heat-damaged denaturing proteins. Lactiplantibacillus plantarum is a widespread Gram-positive bacterium with probiotic claims and vast potential for agro-food, biotechnological and biomedical applications. L. plantarum possesses a family of three sHSP, which were previously demonstrated to be involved in its stress tolerance mechanisms. Here, the three L. plantarum sHSP were heterologously expressed, purified and shown to have a chaperone activity in vitro, measuring their capacity to suppress protein aggregation, as assayed spectrophotometrically by light scattering. Their anti-aggregative capacity was found to be differently influenced by pH. Differences were also found relative to their holdase function and their capacity to modulate liposome membrane fluidity, suggesting interplays between them and indicating diversified activities. This is the first study assessing the chaperone action of sHSP from a probiotic model. The different roles of the three sHSP can increase L. plantarum's capabilities to survive the various types of stress characterising the diverse habitats of this highly adaptable species. Reported evidence supports the interest in L. plantarum as one of the model species for bacteria that have three different sHSP-encoding genes in their genomes.

Lactiplantibacillus plantarum (L. plantarum) is a well-studied and versatile species of lactobacilli. It is found in several niches, including human mucosal surfaces, and it is largely employed in the food industry and boasts a millenary tradition of safe use, sharing a long-lasting relationship with humans. L. plantarum is generally recognised as safe and exhibits a strong probiotic character, so that several strains are commercialised as health-promoting supplements and functional food products. For these reasons, L. plantarum represents a valuable model to gain insight into the nature and mechanisms of antimicrobials as key factors underlying the probiotic action of health-promoting microbes. Probiotic antimicrobials can inhibit the growth of pathogens in the gut ensuring the intestinal homeostasis and contributing to the host health. Furthermore, they may be attractive alternatives to conventional antibiotics, holding potential in several biomedical applications. The aim of this review is to investigate the most relevant papers published in the last ten years, bioprospecting the antimicrobial activity of characterised probiotic L. plantarum strains. Specifically, it focuses on the different chemical nature, the action spectra and the mechanisms underlying the bioactivity of their antibacterial and antiviral agents. Emerging trends in postbiotics, some in vivo applications of L. plantarum antimicrobials, including strengths and limitations of their therapeutic potential, are addressed and discussed.

Objective We investigated whether the knock out of small heat shock protein (sHSP) genes ( hsp1 , hsp2 and hsp3 ) impact on probiotic features of Lactiplantibacillus plantarum WCFS1, aiming to find specific microbial effectors involved in microbe-host interplay. Results The probiotic properties of L. plantarum WCFS1 wild type, hsp1 , hsp2 and hsp3 mutant clones were evaluated and compared through in vitro trials. Oro-gastro-intestinal assays pointed to significantly lower survival for hsp1 and hsp2 mutants under stomach-like conditions, and for hsp3 mutant under intestinal stress. Adhesion to human enterocyte-like cells was similar for all clones, though the hsp2 mutant exhibited higher adhesiveness. L. plantarum cells attenuated the transcriptional induction of pro-inflammatory cytokines on lipopolysaccharide-treated human macrophages, with some exception for the hsp1 mutant. Intriguingly, this clone also induced a higher IL10/IL12 ratio, which is assumed to indicate the anti-inflammatory potential of probiotics. Conclusions sHSP genes deletion determined some differences in gut stress resistance, cellular adhesion and immuno-modulation, also implying effects on in vivo interaction with the host. HSP1 might contribute to immunomodulatory mechanisms, though additional experiments are necessary to test this feature.

The dietary consumption of probiotics in the form of pharmaceuticals or functional food can improve human health and contribute to disease prevention. However, the biological activity and health potential of food-delivered probiotics can be severely compromised by the stress conditions encountered by the microorganisms throughout the manufacture process, from probiotic preparation to their inclusion into food, subsequent storage and ingestion. Here, we give an account of the stress factors that can have major negative impacts on probiotic viability and functionality, with a focus on food-related environmental adverse conditions. We also describe some of the mechanisms elicited by the microbial cells to counteract these stresses and summarize a few relevant approaches proposed in literature to develop more robust and competitive probiotics by enhancing their stress tolerance, with the aim to improve the efficacy and health value of probiotic functional food.