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Keywords = in vitro two-stage simulated gastrointestinal digestion

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21 pages, 506 KB  
Article
Fermented Milk Supplemented with Sodium Butyrate and Inulin: Physicochemical Characterization and Probiotic Viability Under In Vitro Simulated Gastrointestinal Digestion
by Katarzyna Szajnar, Małgorzata Pawlos, Magdalena Kowalczyk, Julita Drobniak and Agata Znamirowska-Piotrowska
Nutrients 2025, 17(13), 2249; https://doi.org/10.3390/nu17132249 - 7 Jul 2025
Cited by 1 | Viewed by 3228
Abstract
Background/Objectives: Probiotics are increasingly recognized for their role in managing gastrointestinal disorders through modulation of gut microbiota. Restoring microbial balance remains a therapeutic challenge. Recent strategies combine probiotics, inulin, and sodium butyrate as synergistic agents for gut health. This study aimed to evaluate [...] Read more.
Background/Objectives: Probiotics are increasingly recognized for their role in managing gastrointestinal disorders through modulation of gut microbiota. Restoring microbial balance remains a therapeutic challenge. Recent strategies combine probiotics, inulin, and sodium butyrate as synergistic agents for gut health. This study aimed to evaluate the effects of milk supplementation with inulin and sodium butyrate on physicochemical properties, sensory characteristics, and the survival of selected probiotic strains during in vitro simulated gastrointestinal digestion. Methods: Fermented milk samples were analyzed for color, pH, titratable acidity, and syneresis. A trained sensory panel evaluated aroma, texture, and acceptability. Samples underwent a standardized in vitro digestion simulating oral, gastric, and intestinal phases. Viable probiotic cells were counted before digestion and at each stage, and survival rates were calculated. Results: Physicochemical and sensory attributes varied depending on probiotic strain and supplementation. Inulin and the inulin–sodium butyrate combination influenced syneresis and acidity. Lacticaseibacillus casei 431 and Lactobacillus johnsonii LJ samples showed the highest viable counts before digestion. Two-way ANOVA confirmed that probiotic strain, supplementation type, and their interactions significantly affected bacterial survival during digestion (p < 0.05). Conclusions: The addition of inulin and sodium butyrate did not impair probiotic viability under simulated gastrointestinal conditions. The effects on product characteristics were strain-dependent (Bifidobacterium animalis subsp. lactis BB-12, L. casei 431, L. paracasei L26, L. acidophilus LA-5, L. johnsonii LJ). These findings support the use of inulin–butyrate fortification in dairy matrices to enhance the functional potential of probiotic foods targeting gut health. Full article
(This article belongs to the Special Issue Probiotics, Postbiotics, Gut Microbiota and Gastrointestinal Health)
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12 pages, 576 KB  
Article
Exploring the Bioaccessibility of Roasted Japanese Green Tea: Impact of Simulated Gastrointestinal Digestion
by Wei Qin and Sunantha Ketnawa
Foods 2025, 14(2), 311; https://doi.org/10.3390/foods14020311 - 17 Jan 2025
Cited by 3 | Viewed by 3169
Abstract
In this study, the effects were explored of digestive enzymes and pH on the bioaccessibility of polyphenols, flavonoids, and antioxidant activities in Hojicha (roasted green tea, RT) infusions during simulated in vitro digestion. Roasting modifies its polyphenolic profile and reduces bitterness, making it [...] Read more.
In this study, the effects were explored of digestive enzymes and pH on the bioaccessibility of polyphenols, flavonoids, and antioxidant activities in Hojicha (roasted green tea, RT) infusions during simulated in vitro digestion. Roasting modifies its polyphenolic profile and reduces bitterness, making it a popular variation of green tea. In this study, RT was used for assessing how the roasting-induced changes influenced the tea’s bioaccessibility and stability under digestive conditions. A two-step gastrointestinal digestion model was applied to mimic real digestion. Total polyphenol content (TPC), total flavonoid content (TFC), and antioxidant activity (DPPH, ABTS, FRAP, and MIC) were measured at different digestion stages. Gastric conditions led to a 2.07-fold reduction in TPC and a 4.27-fold reduction in TFC. Digestive enzymes enhanced bioactive compound stability, with TPC and TFC bioaccessibility reaching 56% and 25% in the simulated digestion with digestive enzymes (MD) group, compared to 52% and 20% in the without digestive enzymes (WOE) group. Antioxidant activities were also better preserved, with antioxidant activity retention at 31% in the MD samples versus 19% in the WOE. These findings emphasize the key role of digestive enzymes in maintaining the antioxidant potential of roasted green tea during digestion, providing insight into future research on roasting methods and tea functionality for product development. Full article
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11 pages, 1760 KB  
Article
A Novel In Vivo Method Using Caenorhabditis elegans to Evaluate α-Glucosidase Inhibition by Natural Products for Type 2 Diabetes Treatment
by María Pilar de Torre, José Luis Vizmanos, Rita Yolanda Cavero and María Isabel Calvo
Pharmaceuticals 2024, 17(12), 1685; https://doi.org/10.3390/ph17121685 - 13 Dec 2024
Viewed by 1698
Abstract
Background: Non-insulin-dependent diabetes mellitus, or type 2 diabetes, is one of the diseases of greatest concern worldwide, and research into natural compounds that are capable of regulating glycemia and insulin resistance is therefore gaining importance. In the preclinical stages, Caenorhabditis elegans is considered [...] Read more.
Background: Non-insulin-dependent diabetes mellitus, or type 2 diabetes, is one of the diseases of greatest concern worldwide, and research into natural compounds that are capable of regulating glycemia and insulin resistance is therefore gaining importance. In the preclinical stages, Caenorhabditis elegans is considered a promising in vivo model for research into this disease. Most studies have been carried out using daf-2 mutant strains and observing changes in their phenotype rather than directly measuring the effects within the worms. Methods: We evaluated the in vitro α-glucosidase inhibition of two oral formulations of Origanum vulgare before and after a simulated gastrointestinal digestion process. After confirming this activity, we developed a method to measure α-glucosidase inhibition in vivo in the C. elegans wild-type strain. Results: The crude extract showed a similar IC50 value to that of acarbose (positive control), before and after gastrointestinal digestion. Formulation 1 also showed no differences with the positive control after digestion (111.86 ± 1.26 vs. 110.10 ± 1.00 µL/mL; p = 0.282). However, formulation 2 showed a higher hypoglycemic activity (59.55 ± 0.85 µL/mL; p < 0.05). The IC50 values obtained in the in vivo assays showed results that correlated well with the in vitro results, so the proposed new method of direct quantification of the in vivo activity seems suitable for directly quantifying the effects of this inhibition without the need to measure changes in the phenotype. Conclusion: A novel, simple and reliable method has been developed to directly determine pharmacological activities in an in vivo model of wild-type C. elegans. This allows the hypoglycemic activity to be directly attributed to in vivo treatment without quantifying phenotypic changes in mutant strains that may arise from other effects, opening the door to a simple analysis of in vivo pharmacological activities. Full article
(This article belongs to the Section Natural Products)
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12 pages, 700 KB  
Article
Probiotic Milk and Oat Beverages with Increased Protein Content: Survival of Probiotic Bacteria Under Simulated In Vitro Digestion Conditions
by Małgorzata Pawlos, Katarzyna Szajnar and Agata Znamirowska-Piotrowska
Nutrients 2024, 16(21), 3673; https://doi.org/10.3390/nu16213673 - 29 Oct 2024
Cited by 3 | Viewed by 4512
Abstract
Background: The increasing prevalence of plant-based dietary preferences, driven by lactose intolerance, allergies, and adherence to vegan diets, has necessitated the exploration of alternative food matrices for probiotic delivery. Objectives: This study aimed to evaluate the effects of whey protein isolate, pea protein [...] Read more.
Background: The increasing prevalence of plant-based dietary preferences, driven by lactose intolerance, allergies, and adherence to vegan diets, has necessitated the exploration of alternative food matrices for probiotic delivery. Objectives: This study aimed to evaluate the effects of whey protein isolate, pea protein isolate, and soy protein isolate on the viability of L. casei and L. johnsonii during simulated in vitro gastrointestinal digestion. Furthermore, the study investigated the impact of two distinct matrices—cow’s milk and an oat-based beverage—on the survival of these probiotic strains. Fermented products were prepared using cow’s milk and an oat-based beverage as matrices, with simulated digestion performed following a seven-day storage period at 5 °C. The in vitro digestion model encompassed oral, gastric, and small intestinal phases, with probiotic viability assessed using the plate-deep method at each stage. Methods: Before digestion, L. casei exhibited higher populations than L. johnsonii in both matrices. Including 3% soy and pea protein, isolates promoted the growth of L. casei in both fermented milk and oat beverages. However, a marked reduction in probiotic viability was observed during the gastric phase, with L. casei counts decreasing by 6.4–7.8 log cfu g−1 in fermented milk and 3.1–4 log cfu g−1 in oat beverages, while L. johnsonii demonstrated similar reductions. Conclusion: These findings underscore the protective role of dairy components on probiotic viability, while the oat-based matrix exhibited a reduced capacity for sustaining probiotic populations throughout digestion. Future research should focus on optimizing plant-based matrices to enhance probiotic stability during gastrointestinal transit. Full article
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12 pages, 300 KB  
Article
Exploring the Chemical Composition of Female Zucchini Flowers for Their Possible Use as Nutraceutical Ingredient
by Luigi Castaldo, Sonia Lombardi, Luana Izzo and Alberto Ritieni
Antioxidants 2023, 12(12), 2108; https://doi.org/10.3390/antiox12122108 - 13 Dec 2023
Cited by 2 | Viewed by 2203
Abstract
The zucchini (Cucurbita pepo L.) plant is well known for its fruits; however, its edible flowers appear to contain several active molecules, including polyphenols, which display poor bioaccessibility after gastrointestinal digestion (GiD). This study explores the bioaccessibility of polyphenols and antioxidant capacity [...] Read more.
The zucchini (Cucurbita pepo L.) plant is well known for its fruits; however, its edible flowers appear to contain several active molecules, including polyphenols, which display poor bioaccessibility after gastrointestinal digestion (GiD). This study explores the bioaccessibility of polyphenols and antioxidant capacity within zucchini flower extracts during simulated GiD. Two nutraceutical formulations, non-acid-resistant (NAcR) and acid-resistant (AcR) capsules containing an aqueous extract of zucchini flowers, were employed in this investigation. Additionally, high-resolution mass spectrometry (Q-Orbitrap HRMS) was utilized for a comprehensive analysis of their polyphenolic constituents. Predominantly, rutin and isorhamnetin-3-rutinoside were the most prevalent compounds detected in the samples (514.62 and 318.59 mg/kg, respectively). Following in vitro GiD, the extract encapsulated in AcR capsules exhibited enhanced bioaccessibility during both the duodenal (189.2 and 162.5 mg GAE/100 g, respectively) and colonic stages (477.4 and 344.7 mg GAE/100 g, respectively) when compared with the extract encapsulated in NAcR capsules. This suggests that gastric acidity adversely impacted the release of polyphenols from NAcR capsules. In conclusion, the aqueous zucchini flower extract emerges as a promising and readily accessible source of dietary polyphenols. Moreover, the utilization of AcR capsules presents a potential nutraceutical formulation strategy to improve polyphenol bioaccessibility, enhancing its applicability in promoting health and well-being. Full article
(This article belongs to the Special Issue Plant Materials and Their Antioxidant Potential)
17 pages, 1364 KB  
Article
Effect of the Production Parameters and In Vitro Digestion on the Content of Polyphenolic Compounds, Phenolic Acids, and Antiradical Properties of Innovative Snacks Enriched with Wild Garlic (Allium ursinum L.) Leaves
by Kamila Kasprzak-Drozd, Tomasz Oniszczuk, Iwona Kowalska, Jarosław Mołdoch, Maciej Combrzyński, Marek Gancarz, Bohdan Dobrzański, Adrianna Kondracka and Anna Oniszczuk
Int. J. Mol. Sci. 2022, 23(22), 14458; https://doi.org/10.3390/ijms232214458 - 21 Nov 2022
Cited by 16 | Viewed by 2937
Abstract
A new type of corn snack has been created containing additions of wild garlic (Allium ursinum L.). This medicinal and dietary plant has a long tradition of use in folk medicine. However, studies on wild garlic composition and activity are fairly recent [...] Read more.
A new type of corn snack has been created containing additions of wild garlic (Allium ursinum L.). This medicinal and dietary plant has a long tradition of use in folk medicine. However, studies on wild garlic composition and activity are fairly recent and scarce. This research aimed to investigate the influence of the screw speed and A. ursinum amounts on the antiradical properties as well as the content of polyphenolic compounds and individual phenolic acids of innovative snacks enriched with wild garlic leaves. The highest radical scavenging activity and content of polyphenols and phenolic acids were found in the snacks enriched with 4% wild garlic produced using screw speed 120 rpm. The obtained findings demonstrated that snacks enriched with wild garlic are a rich source of polyphenolic compounds. Since the concentration of such compounds is affected by many factors, e.g., plant material, presence of other compounds, and digestion, the second aim of this study was to determine radical scavenging activity, the content of polyphenols, and individual phenolic acids of snacks after in vitro simulated gastrointestinal digestion. Using an in vitro two-stage model, authors noted a significant difference between the concentration of polyphenolic compounds and the polyphenol content of the plant material before digestion. Full article
(This article belongs to the Collection Feature Papers in Bioactives and Nutraceuticals)
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11 pages, 1320 KB  
Article
Persistence of Anti SARS-CoV-2 Antibodies in Breast Milk from Infected and Vaccinated Women after In Vitro-Simulated Gastrointestinal Digestion
by Joaquim Calvo-Lerma, Pierre Bueno-Llamoga, Christine Bäuerl, Erika Cortés-Macias, Marta Selma-Royo, Francisco Pérez-Cano, Carles Lerin, Cecilia Martínez-Costa and Maria Carmen Collado
Nutrients 2022, 14(10), 2117; https://doi.org/10.3390/nu14102117 - 19 May 2022
Cited by 11 | Viewed by 3641
Abstract
Breastfeeding is key for infant development and growth. Breast milk contains different bioactive compounds including antibodies. Recent studies have demonstrated the presence of breast milk SARS-CoV-2 antibodies after maternal infection and vaccination. However, the potential impact on the infant has not been explored [...] Read more.
Breastfeeding is key for infant development and growth. Breast milk contains different bioactive compounds including antibodies. Recent studies have demonstrated the presence of breast milk SARS-CoV-2 antibodies after maternal infection and vaccination. However, the potential impact on the infant has not been explored yet. As a first step, we aimed at assessing the potential persistence of SARS-CoV-2 IgA and IgG antibodies from infected and vaccinated women in the gastrointestinal tract of the infants by means of an in vitro-simulated gastrointestinal digestion approach. Breast milk samples from 10 lactating women receiving mRNA vaccination against SARS-CoV-2 (n = 5 with BNT162b2 mRNA and n = 5 with mRNA-1273) and also, COVID-19 infected (n = 5) were included. A control group with women with no exposure to the virus (n = 10 pre-pandemic) were also studied. The presence of IgA and IgG SARS-CoV-2 antibody levels was determined by ELISA after the gastric and intestinal stages. The impact of digested antibodies on infant gut microbiota was tested by simulating colonic fermentation with two different fecal inoculums: infants from vaccinated and non-vaccinated mothers. Specific gut microbial groups were tested by targeted qPCR. In vitro infant gastrointestinal digestion significantly decreased the levels of both anti-SARS-CoV-2 IgA and IgG. However, both remained resistant in all the study groups except in that evaluating breast milk samples from infected women, in which IgG was degraded below the cut-off values in the intestinal phase. No effect of the antibodies on microbiota were identified after digestion. In conclusion, antibody levels against SARS-CoV-2 are reduced after in vitro-simulated gastrointestinal tract but remain present, so a positive biological effect could be expected from this infant immunization pathway. Full article
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12 pages, 295 KB  
Article
Chemical Composition, In Vitro Bioaccessibility and Antioxidant Activity of Polyphenolic Compounds from Nutraceutical Fennel Waste Extract
by Luigi Castaldo, Luana Izzo, Stefania De Pascale, Alfonso Narváez, Yelko Rodriguez-Carrasco and Alberto Ritieni
Molecules 2021, 26(7), 1968; https://doi.org/10.3390/molecules26071968 - 31 Mar 2021
Cited by 43 | Viewed by 4343
Abstract
Fennel (Foeniculum vulgare Mill.) waste contains a broad range of bioactive molecules, including polyphenols, which have poor bioaccessibility during gastrointestinal digestion. This work aimed to investigate the bioaccessibility of total phenolic compounds and the antioxidant capacity during simulated gastrointestinal digestion using two [...] Read more.
Fennel (Foeniculum vulgare Mill.) waste contains a broad range of bioactive molecules, including polyphenols, which have poor bioaccessibility during gastrointestinal digestion. This work aimed to investigate the bioaccessibility of total phenolic compounds and the antioxidant capacity during simulated gastrointestinal digestion using two nutraceutical formulations based on non-acid-resistant (NAR) and acid-resistant (AR) capsules containing aqueous-based extracts from fennel waste. Moreover, to obtain a comprehensive investigation of the polyphenolic constituents of the fennel waste extract, a high-resolution mass spectrometry (Q-Orbitrap) analysis was performed. Notably, chlorogenic acids, such as 4-caffeoylquinic acid and 3,4-dicaffeoylquinic acid, were the most detected compounds found in assayed samples (1.949 and 0.490 mg/g, respectively). After in vitro gastrointestinal digestion, the extract contained in AR capsules displayed higher bioaccessibility in both the duodenal and colonic stages (1.96 and 5.19 mg GAE/g, respectively) than NAR capsules (1.72 and 3.50 mg GAE/g, respectively), suggesting that the acidic gastric conditions negatively affected the polyphenol compounds released from the NAR capsules. Therefore, the aqueous extract of fennel waste could be proposed as an innovative and easily available source of dietary polyphenols. Furthermore, the use of an AR capsule could improve the polyphenol bioaccessibility and can be proposed as a nutraceutical formulation. Full article
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