Iron oxide nanoparticles carried by probiotics for iron absorption: a systematic review.

BACKGROUND: One-third of the world's population has anemia, contributing to higher morbidity and death and impaired neurological development. Conventional anemia treatment raises concerns about iron bioavailability and gastrointestinal (GI) adverse effects. This research aims to establish how iron oxide nanoparticles (IONPs) interact with probiotic cells and how they affect iron absorption, bioavailability, and microbiota variation. METHODS: Pointing to the study of the literature and developing a review and critical synthesis, a robust search methodology was utilized by the authors. The literature search was performed in the PubMed, Scopus, and Web of Science databases. Information was collected between January 2017 and June 2022 using the PRISMA (Preferred Reporting Items for Systematic Review and Meta-Analysis) protocols for systematic reviews and meta-analyses. We identified 122 compatible research articles. RESULTS: The research profile of the selected scientific articles revealed the efficacy of IONPs treatment carried by probiotics versus conventional treatment. Therefore, the authors employed content assessment on four topics to synthesize previous studies. The key subjects of the reviewed reports are the characteristics of the IONPs synthesis method, the evaluation of cell absorption and cytotoxicity of IONPs, and the transport of IONPs with probiotics in treating anemia. CONCLUSIONS: To ensure a sufficient iron level in the enterocyte, probiotics with the capacity to attach to the gut wall transport IONPs into the enterocyte, where the maghemite nanoparticles are released.

Cardiometabolic Risk: Characteristics of the Intestinal Microbiome and the Role of Polyphenols.

Cardiometabolic diseases like hypertension, type 2 diabetes mellitus, atherosclerosis, and obesity have been associated with changes in the gut microbiota structure, or dysbiosis. The beneficial effect of polyphenols on reducing the incidence of this chronic disease has been confirmed by numerous studies. Polyphenols are primarily known for their anti-inflammatory and antioxidant properties, but they can also modify the gut microbiota. According to recent research, polyphenols positively influence the gut microbiota, which regulates metabolic responses and reduces systemic inflammation. This review emphasizes the prebiotic role of polyphenols and their impact on specific gut microbiota components in patients at cardiometabolic risk. It also analyzes the most recent research on the positive effects of polyphenols on cardiometabolic health. While numerous in vitro and in vivo studies have shown the interaction involving polyphenols and gut microbiota, additional clinical investigations are required to assess this effect in people.

Vaccinium Species (Ericaceae): Phytochemistry and Biological Properties of Medicinal Plants.

The Vaccinium L. (Ericaceae) genus consists of a globally widespread and diverse genus of around 4250 species, of which the most valuable is the Vaccinioidae subfamily. The current review focuses on the distribution, history, bioactive compounds, and health-related effects of three species: cranberry, blueberry, and huckleberry. Several studies highlight that the consumption of Vaccinium spp. presents numerous beneficial health-related outcomes, including antioxidant, antimicrobial, anti-inflammatory, and protective effects against diabetes, obesity, cancer, neurodegenerative diseases and cardiovascular disorders. These plants' prevalence and commercial value have enhanced in the past several years; thus, the generated by-products have also increased. Consequently, the identified phenolic compounds found in the discarded leaves of these plants are also presented, and their impact on health and economic value is discussed. The main bioactive compounds identified in this genus belong to anthocyanins (cyanidin, malvidin, and delphinidin), flavonoids (quercetin, isoquercetin, and astragalin), phenolic acids (gallic, p-Coumaric, cinnamic, syringic, ferulic, and caffeic acids), and iridoids.

Bioactive Potential of Elderberry (Sambucus nigra L.): Antioxidant, Antimicrobial Activity, Bioaccessibility and Prebiotic Potential.

Due to its abundance of physiologically active ingredients, one of the oldest medicinal herbs, elderberry (EB) Sambucus nigra L., is beneficial for both therapeutic and dietary purposes. This study determined the bioaccessibility of the phenolic compounds and the prebiotic potential of the polyphenols from freeze-dried EB powder (FDEBP), along with the antioxidant and antimicrobial activities of this extract. The most significant phenolic compounds in black EB are represented by anthocyanins (41.8%), predominating cyanidin-sambubiosides and cyanidin-glucosides (90.1% of the identified anthocyanins). The FRAP assay obtained the highest antioxidant activity value (185 ± 0.18 µmol Fe2+/g DW). The most sensitive to the antimicrobial activity of the extract was proven to be Staphylococcus aureus, and Pseudomonas aeruginosa had the lowest minimum inhibitory concentration of 1.95 mg/mL. To determine the prebiotic potential of the polyphenols, the cell growth of five probiotic strains (Lactobacillus plantarum, L. casei, L. rhamnosus, L. fermentum and Saccharomyces boulardii) was tested. The influence on cell growth was positive for all five probiotic strains used. Overall, the most significant increase (p < 0.05) was recorded at 1.5% FDEBP, on L. casei with a growth index (GI) of 152.44%, very closely followed by GI at 0.5% and 1% concentrations. The stability of the total phenolic compounds through simulated gastronitestinal digestion was increased (93%), and the bioaccessibility was also elevated (75%).

Evaluation of the Astragalus exscapus L. subsp. transsilvanicus Roots' Chemical Profile, Phenolic Composition and Biological Activities.

Novel and natural molecules for pharmaceutical applications are a contemporary preoccupation in science which prompts research in underexplored environments. Astragalus exscapus ssp. transsilvanicus (Schur) Nyár. (ASTRA) is a plant species endemic to Transylvania, having a very similar root system with that of A. membranaceus (Fisch.) Bunge, known for its health promoting properties. The present study endeavored to perform basic characterization of the ASTRA roots by proximate analysis, to investigate the fatty acid profile of the lipids extracted from the ASTRA roots, to examine the phenolic composition of the root extracts by liquid chromatography, and to evaluate the biological activities through determination of the antioxidant, antimicrobial and cytotoxic capacities of the extracts. The primary compounds found in the ASTRA roots were carbohydrates and lipids, and the fatty acid composition determined by GC-MS showed linoleic acid as preponderant compound with 31.10%, followed by palmitic, oleic and α-linolenic acids with 17.30%, 15.61% and 14.21%, respectively. The methanol extract of the ASTRA roots presented highest phenolic content, Astragaloside IV being the predominant compound with 425.32 ± 0.06 µg/g DW. The antimicrobial assay showed remarkable antimicrobial potential of the extract at a concentration of 0.356 and 0.703 mg ASTRA root powder (DW)/mL, highlighting its efficacy to inhibit S. aureus and S. epidermidis bacterial strains. Furthermore, the cell proliferation assessment showed the noteworthy proficiency of the treatment in inhibiting the proliferation of B16F10 melanoma cells.

In Vitro Bioaccessibility of Bioactive Compounds from Rosehip-Enriched Corn Extrudates.

The rosehip (Rosa canina L.) fruit has gained researchers' attention due to its rich chemical composition in vitamin C, phenols, carotenoids, and high antioxidant activity; meanwhile, polymers such as pea protein are generally recognized as exhibiting a protection role against the extrusion process. Corn snacks extrudates obtained by replacing corn flour with 10% R. canina powder (R) and 10% R. canina with pea protein (RPP) were evaluated for the physicochemical, textural, optical, and nutritional characteristics. A sample manufactured without R. canina powder was used as a control. Hardness, crispiness, chewiness, and solubility index (WSI) of the final extrudates were improved by addition of R. canina and pea protein powder (PP); meanwhile, b* (yellow/blue coordinate), C (chroma), and h* (tone) optical parameters were significantly different from the control sample (p < 0.05). Extrusion highlighted a negative impact on total phenols, carotenoids, vitamin C, and antioxidant activity extrudates, while PP exhibited a good protection against the extrusion process. In vitro digestion increased the bioaccessibility of vitamin C, folate, antioxidant activity, total phenols, and total carotenoids mainly on RPP extrudates.

Carotenoid Recovery from Tomato Processing By-Products through Green Chemistry.

The recovery of bioactive compounds from agro-industry-derived by-products sustains circular economy principles by encouraging maximized recycling and minimized waste. Tomato processing by-products are abundant in carotenoids, which have several health-promoting properties, and their reintegration into functional food products represents a major interest for scientists and manufacturers. In the present study, carotenoids were recovered from tomato processing by-products based on the principles of green chemistry by using generally recognized as safe (GRAS) solvents, freeze-drying as pretreatment, and ultrasound in the recovery procedure. Spectrophotometric measurements and HPLC were used to identify and quantify total and individual carotenoids from the extracts. The highest values for lycopene (1324.89 µg/g dw) were obtained when ethyl lactate was applied as a solvent, followed by ethyl acetate with slightly smaller differences (1313.54 µg/g dw). The extracts obtained from freeze-dried samples presented significantly lower amounts of lycopene, indicating that carotenoids are highly susceptible to degradation during lyophilization. Flaxseed, grape seed, and hempseed oils were enriched with carotenoids and their rheological measurements showed favorable viscoelastic properties, especially hempseed and flaxseed oil, with viscosity under 50 mPa·s. Considering the results and the economic perspective of carotenoid recovery from tomato processing by-products, ethyl acetate is suitable, sustainable, and environmentally friendly for carotenoid extraction.

Natural Polyphenol Recovery from Apple-, Cereal-, and Tomato-Processing By-Products and Related Health-Promoting Properties.

Polyphenols of plant origin are a broad family of secondary metabolites that range from basic phenolic acids to more complex compounds such as stilbenes, flavonoids, and tannins, all of which have several phenol units in their structure. Considerable health benefits, such as having prebiotic potential and cardio-protective and weight control effects, have been linked to diets based on polyphenol-enriched foods and plant-based products, indicating the potential role of these substances in the prevention or treatment of numerous pathologies. The most representative phenolic compounds in apple pomace are phloridzin, chlorogenic acid, and epicatechin, with major health implications in diabetes, cancer, and cardiovascular and neurocognitive diseases. The cereal byproducts are rich in flavonoids (cyanidin 3-glucoside) and phenolic acids (ferulic acid), all with significant results in reducing the incidence of noncommunicable diseases. Quercetin, naringenin, and rutin are the predominant phenolic molecules in tomato by-products, having important antioxidant and antimicrobial activities. The present understanding of the functionality of polyphenols in health outcomes, specifically, noncommunicable illnesses, is summarized in this review, focusing on the applicability of this evidence in three extensive agrifood industries (apple, cereal, and tomato processing). Moreover, the reintegration of by-products into the food chain via functional food products and personalized nutrition (e.g., 3D food printing) is detailed, supporting a novel direction to be explored within the circular economy concept.

Antimicrobial Effects of Basil, Summer Savory and Tarragon Lyophilized Extracts in Cold Storage Sausages.

The problem of functional foods with bioactive components of natural origin is current for the food industry. Plant extracts rich in polyphenols with antioxidant and antimicrobial activity are a promising source for use in improving the quality and characteristics of fresh meat and meat products. In this context, the purpose of the present study was to evaluate the physico-chemical, microbiological, sensory properties of sausages prepared with the addition of lyophilized extract of basil, thyme or tarragon. For the beginning, the total amount of polyphenols, the antioxidant and antimicrobial activity of the extracts obtained from three spices were evaluated. In the sausages previously infected with Staphylococcus aureus and Escherichia coli it was observed that there is a much larger number of colonies of microorganisms in the control sample compared to the other samples within 24 and 48 h. Moreover, following the addition of sausage extracts, no changes were found regarding their sensory acceptability.

Recent advances in the biotechnological production of erythritol and mannitol.

Dietary habits that include an excess of added sugars have been strongly associated with an increased risk of obesity, heart disease, diabetes, and tooth decay. With this association in view, modern food systems aim to replace added sugars with low calorie sweeteners, such as polyols. Polyols are generally not carcinogenic and do not trigger a glycemic response. Furthermore, owing to the absence of the carbonyl group, they are more stable compared to monosaccharides and do not participate in Maillard reactions. As such, since polyols are stable at high temperatures, and they do not brown or caramelize when heated. Therefore, polyols are widely used in the diets of hypocaloric and diabetic patients, as well as other specific cases where controlled caloric intake is required. In recent years, erythritol and mannitol have gained increased importance, especially in the food and pharmaceutical industries. In these areas, research efforts have been made to improve the productivity and yield of the two polyols, relying on biotechnological manufacturing methods. The present review highlights the recent advances in the biotechnological production of erythritol and mannitol and summarizes the benefits of using the two polyols in the food and pharmaceutical industries.

An Overview of Gut Microbiota and Colon Diseases with a Focus on Adenomatous Colon Polyps.

It is known and accepted that the gut microbiota composition of an organism has an impact on its health. Many studies deal with this topic, the majority discussing gastrointestinal health. Adenomatous colon polyps have a high prevalence as colon cancer precursors, but in many cases, they are hard to diagnose in their early stages. Gut microbiota composition correlated with the presence of adenomatous colon polyps may be a noninvasive and efficient tool for diagnosis with a high impact on human wellbeing and favorable health care costs. This review is meant to analyze the gut microbiota correlated with the presence of adenomatous colon polyps as the first step for early diagnosis, prophylaxis, and treatment.

Walnut (Juglans regia L.) Septum: Assessment of Bioactive Molecules and In Vitro Biological Effects.

Walnut (Juglans regia L.) septum represents an interesting bioactive compound source by-product. In our study, a rich phenolic walnut septum extract, previously selected, was further examined. The tocopherol content determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed higher amounts of α-tocopherol compared to γ- and δ-tocopherols. Moreover, several biological activities were investigated. The in vitro inhibiting assessment against acetylcholinesterase, α-glucosidase, or lipase attested a real management potential in diabetes or obesity. The extract demonstrated very strong antimicrobial potential against Staphylococcus aureus, Pseudomonas aeruginosa and Salmonella enteritidis. It also revealed moderate (36.08%) and strong (43.27%) antimutagenic inhibitory effects against TA 98 and TA 100 strains. The cytotoxicity of the extract was assessed on cancerous (A549, T47D-KBluc, MCF-7) and normal (human gingival fibroblasts (HGF)) cell lines. Flow cytometry measurements confirmed the cytotoxicity of the extract in the cancerous cell lines. Additionally, the extract demonstrated antioxidant activity on all four cell types, as well as anti-inflammatory activity by lowering the inflammatory cytokines (interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-1 ß (IL-1ß)) evaluated in HGF cells. To the best of our knowledge, most of the cellular model analyses were performed for the first time in this matrix. The results prove that walnut septum may be a potential phytochemical source for pharmaceutical and food industry.

Spray drying and storage of probiotic-enriched almond milk: probiotic survival and physicochemical properties.

BACKGROUND: The combination of the nutritional profile of almond milk with the benefits of probiotic bacteria is an interesting development to meet the demand for sustainable and health-promoting food. Almond milk inoculated with probiotic Lactobacillus plantarum (ATCC8014) was spray dried and the almond, its milk, and powders were characterized physicochemically. Samples were characterized in terms of bacterial survival before and after atomization. Bacterial viability and total fatty acid changes were studied during 8 months' storage at 4 and 22 °C. RESULTS: Results showed adequate physicochemical properties and an optimal bacterial survival rate, maintaining almost the same values before and after the spray-drying operation. A decrease was observed in the cell viability for samples stored at 4 °C. However, the cell count was maintained above the minimum level suggested (107 living cells) to allow potential probiotic functionality for 8 months. On the other hand, the count cell of powders stored at 22 °C was below the minimum level required after 6 months. The fatty acids profile was not significantly (P > 0.05) affected by storage time and temperature. CONCLUSION: A new almond-based-product with probiotics was developed to meet consumer demands. Almond nutrients were recovered from almond milk powder and were found to be a good source of K and high in Mg and in monounsaturated fat. The viability of bacteria was assured during 8 months of storage at 4 °C and up to 6 months for samples stored at 22 °C. © 2020 Society of Chemical Industry.

Ultrasound-assisted Strecker synthesis of novel 2-(hetero)aryl-2-(arylamino)acetonitrile derivatives.

This work describes an efficient, simple, and ecofriendly sonochemical procedure for the preparation of new α-(arylamino)acetonitrile derivatives C-substituted with phenothiazine or ferrocene units. The synthetic protocol is based on the Strecker reaction of a (hetero)aryl aldimine substrate with trimethylsilyl cyanide (TMSCN) in poly(ethylene glycol) (PEG) solution. The advantages of the sonochemical versus the conventional α-(arylamino)acetonitrile synthesis are the significantly shorter reaction time (30 min instead of 72 hours), the higher purity and the easier separation of the product that precipitated from the reaction mixture in crystalline form as depicted by scanning electron microscopy (SEM) analysis. The single crystal X-ray diffraction analysis disclosed the arrangement of the α-(arylamino)acetonitrile molecules in the aggregated crystalline state as a racemic mixture. The mutagenic/antimutagenic potential for three representative derivatives containing phenothiazinyl, ferrocenyl, and phenyl units, respectively, was evaluated by the Ames Salmonella/microsome test using S. typhimurium TA98 and TA100 strains with and without metabolic activation. The preliminary screening results pointed out that the C-(hetero)aryl-α-(arylamino)acetonitrile derivatives can be considered genotoxically safe and possibly antimutagenic.

Gut Prevotella as a possible biomarker of diet and its eubiotic versus dysbiotic roles: a comprehensive literature review.

The gut microbiota has a profound impact on human health. Emerging data show that dietary patterns are associated with different communities of bacterial species within the gut. Prevotella species have been correlated with plant-rich diets, abundant in carbohydrates and fibres. Dysbiosis within the gut ecosystem has been associated with the development of non-communicable diseases such as obesity, the metabolic syndrome, inflammatory bowel disease, irritable bowel syndrome, colorectal cancer, type 1 diabetes, allergies and other diseases. The purpose of this comprehensive literature review was to evaluate the available data on the impact of diet on the Prevotella genus, as a dietary fibre fermenter in the gut as well as its implications as a potential biomarker for homeostasis or disease state through its metabolite signature. Studies were identified by conducting PubMed, Web of Science Core Collection and Google Scholar electronic searches. We found eighty-five publications reporting the impact of dietary patterns on gut microbial communities, including Prevotella or Prevotella/Bacteroides ratio in particular. Moreover, the role of Prevotella species on health status was also evaluated. Prevotella possess a high genetic diversity, representing one of the important groups found in the oral cavity and large intestine of man. The gut commensal Prevotella bacteria contribute to polysaccharide breakdown, being dominant colonisers of agrarian societies. However, studies also suggested a potential role of Prevotella species as intestinal pathobionts. Further metagenomic studies are needed in order to reveal health- or disease-modulating properties of Prevotella species in the gut.

Solidago graminifolia L. Salisb. (Asteraceae) as a Valuable Source of Bioactive Polyphenols: HPLC Profile, In Vitro Antioxidant and Antimicrobial Potential.

Solidago species are often used in traditional medicine as anti-inflammatory, diuretic, wound-healing and antimicrobial agents. Still, the bioactive compounds and biological activities of some species have not been studied. The present work aimed to investigate the polyphenolic profile and the biological properties of Solidago graminifolia L. Salisb., a poorly explored medicinal plant. The hydroalcoholic extracts from aerial parts were evaluated for total phenolic content (TPC), total flavonoid content (TFC) and the polyphenolic compounds were investigated by HPLC-MS. The antioxidant potential in vitro was determined using DPPH and FRAP assays. Antibacterial and antifungal effects were evaluated by dilution assays and MIC, MBC and MFC were calculated. The results showed that Solidago graminifolia aerial parts contain an important amount of total phenolics (192.69 mg GAE/g) and flavonoids (151.41 mg RE/g), with chlorogenic acid and quercitrin as major constituents. The hydroalcoholic extracts showed promising antioxidant and antimicrobial potential, with potent antibacterial activity against Staphylococcus aureus and important antifungal effect against Candida albicans and C. parapsilosis. The obtained results indicated that the aerial parts of Solidago graminifolia could be used as novel resource of phytochemicals in herbal preparations with antioxidant and antimicrobial activities.

Comparative Phytochemical Profile, Antioxidant, Antimicrobial and In Vivo Anti-Inflammatory Activity of Different Extracts of Traditionally Used Romanian Ajuga genevensis L. and A. reptans L. (Lamiaceae).

Several Ajuga species are used in Romanian folk medicine for their antioxidant, antimicrobial and anti-inflammatory properties, to treat pain, fever or arthritis. Still, the active compounds responsible for these effects and their mechanism of action are scarcely known. This research was designed to investigate the phytochemical profile (e.g. iridoids, polyphenolic compounds, phytosterols), as well as the biological potential (antioxidant, antibacterial, antifungal, anti-inflammatory properties) of two selected Ajuga species collected from different regions of Romanian spontaneous flora. The main compounds identified in A. reptans aerial parts extracts were 8-O-acetylharpagide, isoquercitrin and ß-sitosterol, whilst in A. genevensis were 8-O-acetylharpagide, luteolin and campesterol. The extracts were screened for their antioxidant potential using different methods (DPPH, TEAC, EPR) and the results showed a good activity, in accordance with the polyphenol content (18-26 mg GAE/g dw). The antifungal activity on the tested strains was good. The determination of few parameters linked with the inflammatory mechanism allowed the assessment of in vivo anti-inflammatory potential. Ajuga reptans and A. genevensis ethanol extracts had anti-inflammatory activity through lowering the oxidative stress, phagocytosis, PMN and total leukocytes. The best anti-oxidative and anti-inflammatory activity was observed for the Ajuga reptans 100 mg dw/mL extract when compared with diclofenac, thus the dose could be correlated with the pharmacological effect. These findings provide substantial evidence that both selected Ajuga species have the potential to be valued as sources of phytochemicals in effective anti-inflammatory herbal preparations.

Warfarin-Capped Gold Nanoparticles: Synthesis, Cytotoxicity, and Cellular Uptake.

Currently, research studies on nanoparticle cytotoxicity, uptake or internalization into the body's cells are of great interest for the improvement of diagnostic and therapeutic applications. We report here the synthesis and characterization of very stable novel warfarin-capped gold nanoparticles with an average diameter of 54 ± 10 nm which were prepared using sodium warfarin as a reducing agent. The nanoparticles were tested in terms of cytotoxicity and cellular internalization in vitro on two cell lines: normal lung fibroblast HFL-1 and human retinal pigment epithelial D407 cells. Our results showed that the normal lung fibroblast HFL-1 cells were more sensitive to the nanoparticle treatment compared to the human retinal pigment epithelial D407 cells. Moreover, any signs of potential cytotoxicity occurred during the first 24 h of treatment, the cellular viability remaining largely unchanged for longer exposure times. Transmission electron microscopy and dark field hyperspectral imaging revealed that the nanoparticles were effectively delivered and released to the HFL-1 and D407 cells' cytoplasm. Our results provide valuable information to further investigate sodium warfarin-capped gold nanoparticles for possible biological applications.

Valorification of crude glycerol for pure fractions of docosahexaenoic acid and ß-carotene production by using Schizochytrium limacinum and Blakeslea trispora.

The goal of this research is the investigation of a way to maximize the production of docosahexaenoic acid (DHA) and ß-carotene by optimizing the culture conditions of their sources, microalgae Schizochytrium limacinum and fungus Blakeslea trispora respectively, in a fermentation medium. The influencing factors in the fermentation process for producing DHA and ß-carotene have proven to be: the concentration of carbon source (different glycerol crude and pure concentrations) for both of them, and in particular temperature for DHA and pH for ß-carotene. Testing the effect of these parameters was determined: biomass, DHA and ß-carotene concentration. The highest production by S. limacinum was obtained at 25 °C, while using a quantity of 90 g/L of glycerol (crude or pure) as a carbon source. Temperature was the main factor that influenced the biosynthesis of DHA. The quantification of DHA was made by GC-MS chromatography, followed by a purification process, with the end result of DHA in pure phase. The maximum quantities for ß-carotene production were obtained with pH 7 and 60 g/L of crude glycerol. The results highlight the possibility of using crude glycerol as a low-cost substrates for growth of microalgae S. limacinum and of fungus B. trispora in order to obtain the crucial molecules: docosahexaenoic acid and ß-carotene.

In Vitro Transcriptome Response to a Mixture of Lactobacilli Strains in Intestinal Porcine Epithelial Cell Line.

BACKGROUND: Food and feed supplements containing microorganisms with probiotic potential are of increasing interest due to their healthy promoting effect on human and animals. Their mechanism of action is still unknown. Using a microarray approach, the aim of this study was to investigate the differences in genome-wide gene expression induced by a mixture of three Lactobacillus strains (L. rhamnosus, L. plantarum, and L. paracasei) in intestinal porcine epithelial cells (IPEC-1) and to identify the genes and pathways involved in intestinal barrier functions. METHODS: Undifferentiated IPEC-1 cells seeded at a density of 2.0 × 105/mL in 24-wells culture plates were cultivated at 37 °C and 5% CO2 until they reached confluence (2⁻3 days). Confluent cells monolayer were then cultivated with 1 mL of fresh lactobacilli (LB) mixture suspension prepared for a concentration of approximately 3.3 × 107 CFU/mL for each strain (1 × 108 CFU/mL in total) for 3 h and analyzed by microarray using Gene Spring GX v.11.5. RESULTS: The functional analysis showed that 1811 of the genes modulated by LB treatment are involved in signaling (95% up-regulation, 121 genes with a fold change higher than 10). The most enhanced expression was registered for AXIN2 (axis inhibition protein 2-AXIN2) gene (13.93 Fc, p = 0.043), a negative regulator of ß-catenin with a key role in human cancer. LB affected the cellular proliferation by increasing 10 times (Fc) the NF1 gene encoding for the neurofibromin protein, a tumor suppressor that prevent cells from uncontrolled proliferation. The induction of genes like serpin peptidase inhibitor, clade A member 3 (SERPINA 3), interleukin-20 (IL-20), oncostatin M(OSM), granulocyte-macrophage colony-stimulating factor (GM-CSF), and the suppression of chemokine (C-X-C motif) ligand 2/macrophage inflammatory protein 2-alpha (CXCL-2/MIP-2), regulator of G-protein signaling 2 (RGS2), and of pro-inflammatory interleukin-18 (IL-18) genes highlights the protective role of lactobacilli in epithelial barrier function against inflammation and in the activation of immune response. CONCLUSION: Gene overexpression was the predominant effect produced by lactobacilli treatment in IPEC-1 cells, genes related to signaling pathways being the most affected. The protective role of lactobacilli in epithelial barrier function against inflammation and in the activation of immune response was also noticed.