The placenta plays a critical role in maternal-fetal nutrient transport and fetal protection against drugs. Creating physiological in vitro models to study these processes is crucial, but technically challenging. This study introduces an efficient cell model that mimics the human placental barrier using co-cultures of primary trophoblasts and primary human umbilical vein endothelial cells (HUVEC) on a Transwell®-based system. Monolayer formation was examined over 7 days by determining transepithelial electrical resistance (TEER), permeability of Lucifer yellow (LY) and inulin, localization of transport proteins at the trophoblast membrane (immunofluorescence), and syncytialization markers (RT-qPCR/ELISA). We analysed diffusion-based (caffeine/antipyrine) and transport-based (leucine/Rhodamine-123) processes to study the transfer of physiologically relevant compounds. The latter relies on the adequate localization and function of the amino-acid transporter LAT1 and the drug transporter P-glycoprotein (P-gp) which were studied by immunofluorescence microscopy and application of respective inhibitors (2-Amino-2-norbornanecarboxylic acid (BCH) for LAT1; cyclosporine-A for P-gp). The formation of functional monolayer(s) was confirmed by increasing TEER values, low LY transfer rates, minimal inulin leakage, and appropriate expression/release of syncytialization markers. These results were supported by microscopic monitoring of monolayer formation. LAT1 was identified on the apical and basal sides of the trophoblast monolayer, while P-gp was apically localized. Transport assays confirmed the inhibition of LAT1 by BCH, reducing both intracellular leucine levels and leucine transport to the basal compartment. Inhibiting P-gp with cyclosporine-A increased intracellular Rhodamine-123 concentrations. Our in vitro model mimics key aspects of the human placental barrier. It represents a powerful tool to study nutrient and drug transport mechanisms across the placenta, assisting in evaluating safer pregnancy therapies.
- Klíčová slova
- LAT1, P‐gp, co‐culture, endothelial cell, placental barrier, polarized monolayer, primary trophoblast, transport,
- MeSH
- biologické modely MeSH
- biologický transport MeSH
- endoteliální buňky pupečníkové žíly (lidské) * metabolismus MeSH
- inulin metabolismus MeSH
- isochinoliny MeSH
- kokultivační techniky MeSH
- leucin metabolismus MeSH
- lidé MeSH
- maternofetální výměna látek * MeSH
- P-glykoprotein metabolismus MeSH
- placenta * metabolismus MeSH
- rhodamin 123 metabolismus MeSH
- těhotenství MeSH
- trofoblasty * metabolismus MeSH
- Check Tag
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- inulin MeSH
- isochinoliny MeSH
- leucin MeSH
- lucifer yellow MeSH Prohlížeč
- P-glykoprotein MeSH
- rhodamin 123 MeSH
Inulin, an increasingly studied dietary fiber, alters intestinal microbiota. The aim of this study was to assess whether inulin decreases intestinal colonization by multidrug resistant E. coli and to investigate its potential mechanisms of action. Mice with amoxicillin-induced intestinal dysbiosis mice were inoculated with extended spectrum beta-lactamase producing E. coli (ESBL-E. coli). The combination of inulin and pantoprazole (IP) significantly reduced ESBL-E. coli fecal titers, whereas pantoprazole alone did not and inulin had a delayed and limited effect. Fecal microbiome was assessed using shotgun metagenomic sequencing and qPCR. The efficacy of IP was predicted by increased abundance of 74 taxa, including two species of Adlercreutzia. Preventive treatments with A. caecimuris or A. muris also reduced ESBL-E. coli fecal titers. Fecal microbiota of mice effectively treated by IP was enriched in genes involved in inulin catabolism, production of propionate and expression of beta-lactamases. They also had increased beta-lactamase activity and decreased amoxicillin concentration. These results suggest that IP act through production of propionate and degradation of amoxicillin by the microbiota. The combination of pantoprazole and inulin is a potential treatment of intestinal colonization by multidrug-resistant E. coli. The ability of prebiotics to promote propionate and/or beta-lactamase producing bacteria may be used as a screening tool to identify potential treatments of intestinal colonization by multidrug resistant Enterobacterales.
- Klíčová slova
- Escherichia coli, Inulin, intestinal colonization, microbiome, multi-drug resistance, prebiotics, resistome,
- MeSH
- amoxicilin * farmakologie MeSH
- antibakteriální látky farmakologie MeSH
- beta-laktamasy metabolismus genetika MeSH
- dysbióza mikrobiologie farmakoterapie MeSH
- Escherichia coli * účinky léků genetika MeSH
- feces * mikrobiologie MeSH
- infekce vyvolané Escherichia coli farmakoterapie mikrobiologie MeSH
- inulin * farmakologie metabolismus MeSH
- mnohočetná bakteriální léková rezistence * MeSH
- myši MeSH
- pantoprazol * farmakologie MeSH
- prebiotika aplikace a dávkování MeSH
- střevní mikroflóra * účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- amoxicilin * MeSH
- antibakteriální látky MeSH
- beta-laktamasy MeSH
- inulin * MeSH
- pantoprazol * MeSH
- prebiotika MeSH
AIM: The metabolic performance of the gut microbiota contributes to the onset of type 2 diabetes. However, targeted dietary interventions are limited by the highly variable inter-individual response. We hypothesized (1) that the composition of the complex gut microbiome and metabolome (MIME) differ across metabolic spectra (lean-obese-diabetes); (2) that specific MIME patterns could explain the differential responses to dietary inulin; and (3) that the response can be predicted based on baseline MIME signature and clinical characteristics. METHOD: Forty-nine patients with newly diagnosed pre/diabetes (DM), 66 metabolically healthy overweight/obese (OB), and 32 healthy lean (LH) volunteers were compared in a cross-sectional case-control study integrating clinical variables, dietary intake, gut microbiome, and fecal/serum metabolomes (16 S rRNA sequencing, metabolomics profiling). Subsequently, 27 DM were recruited for a predictive study: 3 months of dietary inulin (10 g/day) intervention. RESULTS: MIME composition was different between groups. While the DM and LH groups represented opposite poles of the abundance spectrum, OB was closer to DM. Inulin supplementation was associated with an overall improvement in glycemic indices, though the response was very variable, with a shift in microbiome composition toward a more favorable profile and increased serum butyric and propionic acid concentrations. The improved glycemic outcomes of inulin treatment were dependent on better baseline glycemic status and variables related to the gut microbiota, including the abundance of certain bacterial taxa (i.e., Blautia, Eubacterium halii group, Lachnoclostridium, Ruminiclostridium, Dialister, or Phascolarctobacterium), serum concentrations of branched-chain amino acid derivatives and asparagine, and fecal concentrations of indole and several other volatile organic compounds. CONCLUSION: We demonstrated that obesity is a stronger determinant of different MIME patterns than impaired glucose metabolism. The large inter-individual variability in the metabolic effects of dietary inulin was explained by differences in baseline glycemic status and MIME signatures. These could be further validated to personalize nutritional interventions in patients with newly diagnosed diabetes.
- MeSH
- diabetes mellitus 2. typu * MeSH
- inulin * metabolismus farmakologie MeSH
- lidé MeSH
- multiomika MeSH
- nadváha metabolismus MeSH
- obezita metabolismus MeSH
- průřezové studie MeSH
- studie případů a kontrol MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- pozorovací studie MeSH
- práce podpořená grantem MeSH
- Názvy látek
- inulin * MeSH
Fecal microbiota transfer may serve as a therapeutic tool for treating obesity and related disorders but currently, there is no consensus regarding the optimal donor characteristics. We studied how microbiota from vegan donors, who exhibit a low incidence of non-communicable diseases, impact on metabolic effects of an obesogenic diet and the potential role of dietary inulin in mediating these effects. Ex-germ-free animals were colonized with human vegan microbiota and fed a standard or Western-type diet (WD) with or without inulin supplementation. Despite the colonization with vegan microbiota, WD induced excessive weight gain, impaired glucose metabolism, insulin resistance, and liver steatosis. However, supplementation with inulin reversed steatosis and improved glucose homeostasis. In contrast, inulin did not affect WD-induced metabolic changes in non-humanized conventional mice. In vegan microbiota-colonized mice, inulin supplementation resulted in a significant change in gut microbiota composition and its metabolic performance, inducing the shift from proteolytic towards saccharolytic fermentation (decrease of sulfur-containing compounds, increase of SCFA). We found that (i) vegan microbiota alone does not protect against adverse effects of WD; and (ii) supplementation with inulin reversed steatosis and normalized glucose metabolism. This phenomenon is associated with the shift in microbiota composition and accentuation of saccharolytic fermentation at the expense of proteolytic fermentation.
- Klíčová slova
- fecal microbiota transfer, inulin, liver steatosis, proteolytic fermentation, vegan microbiota,
- MeSH
- fekální transplantace MeSH
- glukosa farmakologie MeSH
- inulin farmakologie MeSH
- lidé MeSH
- myši MeSH
- potravní vláknina farmakologie MeSH
- střevní mikroflóra * MeSH
- vegani MeSH
- západní dieta MeSH
- ztučnělá játra * prevence a kontrola farmakoterapie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- glukosa MeSH
- inulin MeSH
- potravní vláknina MeSH
Fructooligosaccharides (FOS) are fructose-based oligosaccharides employed as additives to improve the food's nutritional and technological properties. The rhizosphere of plants that accumulate fructopolysaccharides as inulin has been revealed as a source of filamentous fungi. These fungi can produce FOS either by inulin hydrolysis or by biosynthesis from sucrose, including unusual FOS with enhanced prebiotic properties. Here, we investigated the ability of Fusarium solani and Neocosmospora vasinfecta to produce FOS from different carbon sources. Fusarium solani and N. vasinfecta grew preferentially in inulin instead of sucrose, resulting in the FOS production as the result of endo-inulinase activities. N. vasinfecta was also able to produce the FOS 1-kestose and 6-kestose from sucrose, indicating transfructosylating activity, absent in F. solani. Moreover, the results showed how these carbon sources affected fungal cell wall composition and the expression of genes encoding for β-1,3-glucan synthase and chitin synthase. Inulin and fructose promoted changes in fungal macroscopic characteristics partially explained by alterations in cell wall composition. However, these alterations were not directly correlated with the expression of genes related to cell wall synthesis. Altogether, the results pointed to the potential of both F. solani and N. vasinfecta to produce FOS at specific profiles.
- Klíčová slova
- Carbon source, Cell wall, Fructooligosaccharides, Fusarium, Neocosmospora,
- MeSH
- fruktosa metabolismus MeSH
- Fusarium * genetika metabolismus MeSH
- inulin * metabolismus MeSH
- oligosacharidy MeSH
- sacharosa metabolismus MeSH
- uhlík MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- fructooligosaccharide MeSH Prohlížeč
- fruktosa MeSH
- inulin * MeSH
- oligosacharidy MeSH
- sacharosa MeSH
- uhlík MeSH
Fructan-type inulin and fructo-oligosaccharides (FOS) are reserve polysaccharides that offer an interesting combination of nutritional and technological properties for food industry. Stevia rebaudiana is used commercially in the sweetener industry due to the high content of steviol glycosides in its leaves. With the proposal of using industrial waste, the objective of the present study was to isolate, characterize and evaluate the prebiotic activity of inulin and FOS from S. rebaudiana stems. The chemical characterization of the samples by GC-MS, NMR and off-line ESI-MS showed that it was possible to obtain inulin molecules from the S. rebaudiana stems with a degree of polymerization (DP) of 12, and FOS with a DP<6. The in vitro prebiotic assay of these molecules indicates a strain specificity in fermentation capacity of fructans as substrate. FOS molecules with a low DP are preferably fermented by beneficial microbiota than inulin molecules with higher DP.
- Klíčová slova
- Chlorotrimethylsilane (PubChem CID: 6397), Ethanol (PubChem CID: 702), Fructo-oligosaccharides, Fructose (PubChem CID: 5984), Glucose (PubChem CID: 5793), Hexamethyldisilazane (PubChem CID: 13838), Industrial waste, Inulin, Inulin from chicory (PubChem CID: 16219508), Myo-inositol (PubChem CID: 892), Prebiotic activity, Pyridine (PubChem CID: 1049)., Stevia rebaudiana, Trifluoroacetic acid (PubChem CID: 6422),
- MeSH
- fermentace * MeSH
- inulin chemie metabolismus MeSH
- konformace sacharidů MeSH
- mikrobiota * MeSH
- průmyslový odpad analýza MeSH
- Stevia chemie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- inulin MeSH
- průmyslový odpad MeSH
BACKGROUND/AIM: Chemopreventive activity of a new probiotic strain Lactobacillus plantarum LS/07 (PRO) and prebiotic oligofructose-enriched inulin (PRE) in rat mammary carcinogenesis induced by procarcinogen 7,12-dimethylbenz[a]anthracene has been reported before. This study evaluated the anticancer and immunomodulatory efficacy of PRO, PRE, PRO+PRE (PRO/PRE) and combination with melatonin (PRO+PRE+MEL) in a rat model, when breast cancer was induced by a direct-acting carcinogen N-nitroso-N-methylurea (NMU). MATERIALS AND METHODS: Daily administration of PRO (at a dose of 8.4×10(8) colony-forming units (c.f.u.)/rat), PRE (in the diet, 20 g/kg) and MEL (in tap water, 20 mg/l) started 14 days before the first NMU dose and lasted for 16 weeks. RESULTS: Although tumor growth was not altered, a marked decrease in the ratio of high-/low-grade carcinomas and in tumoral Ki-67 expression was found after PRO+PRE treatment; melatonin augmented these effects. PRO+PRE+MEL combination enhanced CD4(+) and CD8(+) T-cell tumor infiltration induced by PRO/PRE and increased CD25(+)FoxP3(+) regulatory T-cells in tumors. CONCLUSION: In mammary carcinogenesis, Lactobacillus plantarum LS/07 and inulin exert prodifferentiating, antiproliferative and immunomodulatory activities, which are significantly amplified by melatonin co-administration.
- Klíčová slova
- Breast cancer, Lactobacillus plantarum, N-nitroso-N-methylurea, melatonin, oligofructose-enriched inulin, prebiotic, probiotic,
- MeSH
- experimentální nádory mléčných žláz farmakoterapie MeSH
- imunologické faktory farmakologie MeSH
- interleukin-6 fyziologie MeSH
- inulin farmakologie MeSH
- krysa rodu Rattus MeSH
- Lactobacillus plantarum * MeSH
- melatonin farmakologie MeSH
- methylnitrosomočovina MeSH
- potkani Sprague-Dawley MeSH
- probiotika farmakologie MeSH
- protinádorové látky farmakologie MeSH
- transformující růstový faktor beta1 fyziologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- imunologické faktory MeSH
- interleukin-6 MeSH
- inulin MeSH
- melatonin MeSH
- methylnitrosomočovina MeSH
- protinádorové látky MeSH
- transformující růstový faktor beta1 MeSH
Pfaffia glomerata (Amaranthaceae) is popularly known as "Brazilian ginseng." Previous studies have shown that fructose is the major carbohydrate component present in its roots. Inulin-type fructans, polymers of fructose, are the most widespread and researched prebiotics. Here, we isolated and chemically characterized inulin extracted from P. glomerata roots and investigated its potential prebiotic effect. Fructans were isolated and their structures were determined using colorimetric, chromatography, polarimetry, and spectroscopic analysis. The degree of polymerization (DP) was determined, and an in vitro prebiotic test was performed. The structure of inulin was confirmed by chromatography and spectroscopic analysis and through comparison with existing data. Representatives from the genera Lactobacillus and Bifidobacterium utilized inulin from P. glomerata, because growth was significantly stimulated, while this ability is strain specific. The results indicated that inulin extracted from P. glomerata roots represents a promising new source of inulin-type prebiotics.
- Klíčová slova
- Amaranthaceae, Inulin, Pfaffia glomerata, Prebiotic,
- MeSH
- Amaranthaceae chemie MeSH
- chemická precipitace MeSH
- inulin chemie izolace a purifikace farmakologie MeSH
- kořeny rostlin chemie MeSH
- Lactobacillus účinky léků růst a vývoj MeSH
- polymerizace MeSH
- prebiotika MeSH
- rostlinné extrakty chemie izolace a purifikace farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- inulin MeSH
- prebiotika MeSH
- rostlinné extrakty MeSH
The polysaccharide inulin has great importance in the food and pharmaceutical industries. The degree of polymerization (DP) of inulin influences important properties, such as, solubility, thermal stability, sweetness power and prebiotic activity. Molecules with a high degree of polymerization are obtained through physical techniques for enrichment of the inulin chains because they are not commonly obtained from plants extract. Gas chromatography/Mass Spectrometry and (1)H Nuclear Magnetic Resonance analysis showed that inulin from Stevia rebaudiana roots has a degree of polymerization (DPn 28) higher than the value of DPn 12-15 for inulins from other plant species. Furthermore, the methodology of freeze/thaw to enrich the chains allowed us to increase the DP, similarly to other methodologies used for the enrichment of inulin chains. The prebiotic assays confirm that inulin from S. rebaudiana has a high DP. The combined use of these molecules with low degree of polymerization fructans seems to be advantageous to prolong the prebiotic effect in the colon. Our results suggest that S. rebaudiana roots are a promising source of high degree polymerization inulins.
- Klíčová slova
- Fructooligosaccharides, Inulin, Polymerization degree, Prebiotic, Stevia rebaudiana,
- MeSH
- Bifidobacterium růst a vývoj metabolismus MeSH
- fermentace MeSH
- inulin chemie izolace a purifikace MeSH
- kultivační média MeSH
- Lactobacillus růst a vývoj metabolismus MeSH
- polymerizace MeSH
- prebiotika MeSH
- Stevia chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- inulin MeSH
- kultivační média MeSH
- prebiotika MeSH
This review emphasises the fact that studies of acetone-butanol-ethanol (ABE) fermentation by solventogenic clostridia cannot be limited to research on the strain Clostridium acetobutylicum ATCC 824. Various 1-butanol producing species of the genus Clostridium, which differ in their patterns of product formation and abilities to ferment particular carbohydrates or glycerol, are described. Special attention is devoted to species and strains that do not produce acetone naturally and to the utilisation of lactose, inulin, glycerol and mixtures of pentose and hexose carbohydrates. Furthermore, process-mapping tools based on different principles, including flow cytometry, DNA microarray analysis, mass spectrometry, Raman microscopy, FT-IR spectroscopy and anisotropy of electrical polarisability, which might facilitate fermentation control and a deeper understanding of ABE fermentation, are introduced. At present, the methods with the greatest potential are flow cytometry and transcriptome analysis. Flow cytometry can be used to visualise and capture cells within clostridial populations as they progress through the normal cell cycle, in which symmetric and asymmetric cell division phases alternate. Cell viability of a population of Clostridium pasteurianum NRRL B-598 was determined by flow cytometry. Transcriptome analysis has been used in various studies including the detection of genes expressed in solventogenic phase, at sporulation, in the stress response, to compare expression patterns of different strains or parent and mutant strains, for studies of catabolite repression, and for the detection of genes involved in the transport and metabolism of 11 different carbohydrates. Interestingly, the results of transcriptome analysis also challenge our earlier understanding of the role of the Spo0A regulator in initiation of solventogenesis in C. acetobutylicum ATCC 824. Lastly, the review describes other significant recent discoveries, including the deleterious effects of intracellular formic acid accumulation in C. acetobutylicum DSM 1731 cells on the metabolic switch from acidogenesis to solventogenesis and the development of a high-cell density continuous system using Clostridium saccharoperbutylacetonicum N1-4, in which 1-butanol productivity of 7.99 g/L/h was reached.
- MeSH
- aceton metabolismus MeSH
- butanoly metabolismus MeSH
- Clostridium cytologie genetika metabolismus MeSH
- ethanol metabolismus MeSH
- fermentace * MeSH
- glycerol metabolismus MeSH
- hexosy metabolismus MeSH
- inulin metabolismus MeSH
- laktosa metabolismus MeSH
- pentosy metabolismus MeSH
- průtoková cytometrie MeSH
- Ramanova spektroskopie MeSH
- sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
- spektroskopie infračervená s Fourierovou transformací MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- aceton MeSH
- butanoly MeSH
- ethanol MeSH
- glycerol MeSH
- hexosy MeSH
- inulin MeSH
- laktosa MeSH
- pentosy MeSH