22 results on '"Krych Ł"'
Search Results
2. Interactions between sleep and gut bacteria in healthy developing infants
- Author
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Schoch, S., Castro-Meija, J.L., Krych, L., Kot, W., Leng, B., Kohler, M., Huber, R., Rogler, G., Biedermann, L., Walser, J.-C., Nielsen, D., and Kurth, S.
- Published
- 2022
- Full Text
- View/download PDF
3. Gut microbial markers are associated with diabetes onset, regulatory imbalance, and IFN-γ level in NOD Mice
- Author
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Krych, Ł, primary, Nielsen, DS, additional, Hansen, AK, additional, and Hansen, CHF, additional
- Published
- 2015
- Full Text
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4. Restitution of gut microbiota in Ugandan children administered with probiotics ( Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB-12) during treatment for severe acute malnutrition.
- Author
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Castro-Mejía JL, O'Ferrall S, Krych Ł, O'Mahony E, Namusoke H, Lanyero B, Kot W, Nabukeera-Barungi N, Michaelsen KF, Mølgaard C, Friis H, Grenov B, and Nielsen DS
- Subjects
- Bacteria classification, Bacteria genetics, Bacteria growth & development, Bacteria isolation & purification, Bifidobacterium animalis, Child, Preschool, Diarrhea complications, Diarrhea diet therapy, Edema complications, Feces microbiology, Female, Humans, Infant, Lacticaseibacillus rhamnosus, Male, Severe Acute Malnutrition complications, Severe Acute Malnutrition microbiology, Uganda, Gastrointestinal Microbiome, Probiotics administration & dosage, Severe Acute Malnutrition therapy
- Abstract
Severe acute malnutrition (SAM) is a major challenge in low-income countries and gut microbiota (GM) dysbiosis may play a role in its etiology. Here, we determined the GM evolution during rehabilitation from SAM and the impact of probiotics ( Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB-12) supplementation. The GM (16S rRNA gene amplicon sequencing) of children admitted to hospital with SAM showed distinct composition over admission (e.g. Klebsiella spp., and Enterobacteriaceae spp.), discharge (e.g. Clostridiaceae spp., Veilonella dispar ) and follow-up (e.g. Lactobacillus ruminis, Blautia spp., Faecalibacterium prausnitzii ), reaching similar β- and α-diversity as healthy individuals. Children with diarrhea had reduced distribution of Bacteroidaceae, Lachnospiraceae, increased Enterobacteriaceae and Moraxellaceae, and lower α-diversity. Children suffering from edematous SAM had diminished proportion of Prevotellaceae, Lachnospiraceae, Ruminoccaceae and a higher α-diversity when compared to non-edematous SAM. Supplementation of probiotics did not influence β-diversity upon discharge or follow-up, but it increased ( p < .05) the number of observed species [SE: > 4.5]. Children where the probiotic species were detected had lower cumulative incidence ( p < .001) of diarrhea during the follow-up period compared to children receiving placebo and children receiving probiotics, but where the probiotics were not detected. The GM of children with non-edematous and edematous SAM differ in composition, which might have implications for future GM targeted treatments. Probiotics treatment reduced the cumulative incidence of diarrhea during the outpatient phase, with the strongest effect in children where the administered probiotics could be detected in the GM.
- Published
- 2020
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5. Physical fitness in community-dwelling older adults is linked to dietary intake, gut microbiota, and metabolomic signatures.
- Author
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Castro-Mejía JL, Khakimov B, Krych Ł, Bülow J, Bechshøft RL, Højfeldt G, Mertz KH, Garne ES, Schacht SR, Ahmad HF, Kot W, Hansen LH, Perez-Cueto FJA, Lind MV, Lassen AJ, Tetens I, Jensen T, Reitelseder S, Jespersen AP, Holm L, Engelsen SB, and Nielsen DS
- Subjects
- Aged, Aged, 80 and over, Aging physiology, Bacteria genetics, Body Composition, Cross-Sectional Studies, DNA, Bacterial genetics, DNA, Bacterial isolation & purification, Exercise physiology, Female, Humans, Life Style, Male, Metabolomics methods, Phenotype, Eating physiology, Energy Intake physiology, Gastrointestinal Microbiome genetics, Independent Living, Metabolome, Physical Fitness physiology
- Abstract
When humans age, changes in body composition arise along with lifestyle-associated disorders influencing fitness and physical decline. Here we provide a comprehensive view of dietary intake, physical activity, gut microbiota (GM), and host metabolome in relation to physical fitness of 207 community-dwelling subjects aged +65 years. Stratification on anthropometric/body composition/physical performance measurements (ABPm) variables identified two phenotypes (high/low-fitness) clearly linked to dietary intake, physical activity, GM, and host metabolome patterns. Strikingly, despite a higher energy intake high-fitness subjects were characterized by leaner bodies and lower fasting proinsulin-C-peptide/blood glucose levels in a mechanism likely driven by higher dietary fiber intake, physical activity and increased abundance of Bifidobacteriales and Clostridiales species in GM and associated metabolites (i.e., enterolactone). These factors explained 50.1% of the individual variation in physical fitness. We propose that targeting dietary strategies for modulation of GM and host metabolome interactions may allow establishing therapeutic approaches to delay and possibly revert comorbidities of aging., (© 2020 The Authors. Aging Cell published by the Anatomical Society and John Wiley & Sons Ltd.)
- Published
- 2020
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6. DNA enrichment and tagmentation method for species-level identification and strain-level differentiation using ON-rep-seq.
- Author
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Krych Ł, Castro-Mejía JL, Forero-Junco LM, Moesby DN, Mikkelsen MB, Rasmussen MA, Sykulski M, and Nielsen DS
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- Bacillus cereus genetics, High-Throughput Nucleotide Sequencing economics, Listeria monocytogenes genetics, Microbiological Techniques economics, Salmonella enterica genetics, Sequence Analysis, DNA economics, Species Specificity, Time Factors, DNA, Bacterial, High-Throughput Nucleotide Sequencing methods, Microbiological Techniques methods, Sequence Analysis, DNA methods
- Abstract
Despite the massive developments within culture-independent methods for detection of microorganisms during the last decade, culture-based methods remain a cornerstone in microbiology. Yet, the problem of rapid, accurate and inexpensive identification of bacterial isolates down to species/strain level remains unresolved. We have developed a new method for bacterial DNA enrichment and tagmentation allowing fast (<24 h) and cost-effective species level identification and strain level differentiation using the MinION portable sequencing platform (ON-rep-seq). DNA library preparation for 96 isolates takes less than 5 h and ensures highly reproducible distribution of reads that can be used to generate strain level specific read length counts profiles (LCp). We have developed a pipeline that by correcting reads error within peaks of LCp generates a set of high quality (>99%) consensus reads. Whereas, the information from high quality reads is used to retrieve species level taxonomy, comparison of LCp allows for strain level differentiation., Competing Interests: Competing interestsThe authors declare no competing interests., (© The Author(s) 2019.)
- Published
- 2019
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7. Targeting gut microbiota and barrier function with prebiotics to alleviate autoimmune manifestations in NOD mice.
- Author
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Hansen CHF, Larsen CS, Petersson HO, Zachariassen LF, Vegge A, Lauridsen C, Kot W, Krych Ł, Nielsen DS, and Hansen AK
- Subjects
- Animals, Autoimmunity physiology, Dietary Supplements, Female, Gastrointestinal Microbiome drug effects, Glucuronates therapeutic use, Mice, Mice, Inbred NOD, Oligosaccharides therapeutic use, Gastrointestinal Microbiome physiology, Prebiotics
- Abstract
Aims/hypothesis: Adopting a diet containing indigestible fibre compounds such as prebiotics to fuel advantageous bacteria has proven beneficial for alleviating inflammation. The effect of the microbial changes on autoimmunity, however, remains unknown. We studied the effects of prebiotic xylooligosaccharides (XOS) on pancreatic islet and salivary gland inflammation in NOD mice and tested whether these were mediated by the gut microbiota., Methods: Mother and offspring mice were fed an XOS-supplemented diet until diabetes onset or weaning and were compared with a control-fed group. Diabetes incidence was monitored, insulitis and sialadenitis were scored in histological sections from adult mice, and several metabolic and immune variables were analysed in mice before the development of diabetes. Gut barrier function was assessed using an in vivo FITC-dextran permeability test. The importance of XOS-mediated gut microbial changes were evaluated in antibiotic-treated mice fed either XOS or control diet or given a faecal microbiota transplant from test animals., Results: Diabetes onset was delayed in the XOS-fed mice, which also had fewer cellular infiltrations in their pancreatic islets and salivary glands. Interestingly, insulitis was most reduced in the XOS-fed groups when the mice were also treated with an antibiotic cocktail. There was no difference in sialadenitis between the dietary groups treated with antibiotics; the mice were protected by microbiota depletion regardless of diet. Faecal microbiota transplantation was not able to transfer protection. No major differences in glucose-insulin regulation, glucagon-like peptide-1, or short-chain fatty acid production were related to the XOS diet. The XOS diet did, however, reduce gut permeability markers in the small and large intestine. This was accompanied by a more anti-inflammatory environment locally and systemically, dominated by a shift from M1 to M2 macrophages, a higher abundance of activated regulatory T cells, and lower levels of induction of natural killer T cells and cytotoxic T cells., Conclusions/interpretation: Prebiotic XOS have microbiota-dependent effects on salivary gland inflammation and microbiota-independent effects on pancreatic islet pathology that are accompanied by an improved gut barrier that seems able to heighten control of intestinal diabetogenic antigens that have the potential to penetrate the mucosa to activate autoreactive immune responses.
- Published
- 2019
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8. Impact of Early Exposure to Cefuroxime on the Composition of the Gut Microbiota in Infants Following Cesarean Delivery.
- Author
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Kamal SS, Hyldig N, Krych Ł, Greisen G, Krogfelt KA, Zachariassen G, and Nielsen DS
- Subjects
- Bacteria drug effects, Female, Humans, Infant, Infant, Newborn, Microbial Sensitivity Tests, Pregnancy, Anti-Bacterial Agents pharmacology, Antibiotic Prophylaxis, Cefuroxime pharmacology, Cesarean Section, Feces microbiology, Gastrointestinal Microbiome drug effects
- Abstract
Objectives: To assess in mothers giving birth by cesarean delivery if prophylactic antibiotics administered either before skin incision or immediately after cutting the umbilical cord influences gut microbiota colonization and antibiotic susceptibility of the gut bacteria in the newborn., Study Design: Forty-two pregnant women scheduled for elective cesarean delivery were recruited at Odense University Hospital, Denmark, and randomly assigned to receive cefuroxime either before skin incision or immediately after the umbilical cord was cut. Fecal samples were collected from all infants at age 10 days and 9 months. Composition of the gut microbiota was determined by 16S ribosomal RNA gene amplicon high-throughput sequencing. Gram-positive cocci and Enterobacteriaceae were isolated and identified before antimicrobial susceptibility tests were performed by disk diffusion., Results: No clear difference in the composition of the gut microbiota was observed between infants whose mothers received cefuroxime before or after cesarean delivery at neither time point, though surprisingly at 9 months of age, but not at 10 days of age, the number of observed species was higher in infants where mothers received cefuroxime after cord clamping. No differences in antimicrobial susceptibility of Enterobacteriaceae, Enterococcus spp, and Staphylococcus spp were seen at 10 days., Conclusions: Timing of cefuroxime administration to mothers undergoing cesarean delivery does not have a major effect on the gut microbiota and bacterial antibiotic resistance traits in infants., Trial Registration: Clinicaltrials.gov: NCT02072798., (Copyright © 2019 Elsevier Inc. All rights reserved.)
- Published
- 2019
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9. Prebiotic Effect of Lycopene and Dark Chocolate on Gut Microbiome with Systemic Changes in Liver Metabolism, Skeletal Muscles and Skin in Moderately Obese Persons.
- Author
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Wiese M, Bashmakov Y, Chalyk N, Nielsen DS, Krych Ł, Kot W, Klochkov V, Pristensky D, Bandaletova T, Chernyshova M, Kyle N, and Petyaev I
- Subjects
- Adult, Aged, Bifidobacterium longum drug effects, Bifidobacterium longum metabolism, Female, Healthy Volunteers, Humans, Lactobacillus drug effects, Lactobacillus metabolism, Liver drug effects, Liver metabolism, Lycopene administration & dosage, Male, Middle Aged, Muscle, Skeletal drug effects, Muscle, Skeletal metabolism, Obesity metabolism, Obesity microbiology, Prebiotics administration & dosage, Skin drug effects, Skin metabolism, Chocolate, Gastrointestinal Microbiome drug effects, Obesity diet therapy
- Abstract
Lycopene rich food and dark chocolate are among the best-documented products with a broad health benefit. This study explored the systemic effect of lycopene and dark chocolate (DC) on gut microbiota, blood, liver metabolism, skeletal muscle tissue oxygenation and skin. 30 volunteers were recruited for this trial, 15 women and 15 men with a mean age of 55 ± 5.7 years and with moderate obesity, 30 < BMI < 35 kg/m
2 . They were randomized and divided into five equal interventional groups: three received different formulations of lycopene, one of them with a 7 mg daily dose and two with 30 mg; another group was given 10 g of DC with 7 mg lycopene embedded into its matrix, and the last group received 10 g DC. The trial was double-blinded for the three lycopene groups and separately for the 2 DC groups; the trial lasted for 1 month. By the end of the trial there were dose-dependent changes in the gut microbiota profile in all three lycopene groups with an increase of relative abundance of, e.g., Bifidobacterium adolescentis and Bifidobacterium longum . This was also accompanied by dose-dependent changes in the blood, liver metabolism, skeletal muscle and skin parameters. Consumption of DC resulted in increased relative abundance of, e.g., Lactobacillus and a reduction of corneocyte exfoliation. This is the first study which reports the prebiotic potential of lycopene and DC.- Published
- 2019
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10. Effect of the dietary polyacetylenes falcarinol and falcarindiol on the gut microbiota composition in a rat model of colorectal cancer.
- Author
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Kobaek-Larsen M, Nielsen DS, Kot W, Krych Ł, Christensen LP, and Baatrup G
- Subjects
- Animals, Colorectal Neoplasms drug therapy, Denmark, Diet, Male, Polyynes, RNA, Ribosomal, 16S, Rats, Rats, Inbred F344, Colorectal Neoplasms microbiology, Diynes pharmacology, Fatty Alcohols pharmacology, Gastrointestinal Microbiome drug effects
- Abstract
Objectives: (3R)-Falcarinol (FaOH) and (3R,8S)-falcarindiol (FaDOH) have previously been shown to reduce the number of neoplastic lesions and the growth rate of polyps in the colon of azoxymethane (AOM) treated rats. Based on previous investigations, it appears that different mechanisms of actions are involved in the antineoplastic effect of FaOH and FaDOH. One mechanism of action may be related to the antibacterial effect of FaOH and FaDOH and thus their effect on the gut microbiota. This study aimed to determine the effect of FaOH and FaDOH on gut microbiota composition of AOM treated rats., Results: Azoxymethane treated rats were fed either a standard rat diet or a rat diet supplemented with FaOH and FaDOH. The gut microbiota of AOM-induced rats was determined by 16S rRNA gene-amplicon sequencing. Analysis of fecal cecum samples demonstrated a significant gut microbiota change in rats receiving standard rat diet supplemented with FaOH and FaDOH compared with the control group that only received the rat diet. Comparison of the gut microbiota of rats who developed large neoplasms in the colon with rats without large neoplasms showed that the gut microbiota was significantly different in rats who developed large colon neoplasms compared to rats with no macroscopic colon neoplasms.
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- 2018
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11. Gut microbiota recovery and immune response in ampicillin-treated mice.
- Author
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Castro-Mejía JL, Jakesevic M, Fabricius NF, Krych Ł, Nielsen DS, Kot W, Bendtsen KM, Vogensen FK, Hansen CHF, and Hansen AK
- Subjects
- Animals, Cytokines, Mice, Microbiota, Ampicillin pharmacology, Anti-Bacterial Agents pharmacology, Gastrointestinal Microbiome immunology
- Abstract
Ampicillin is applied in rodents to induce a temporarily depleted microbiota. To elucidate whether bacteria are just temporarily suppressed or fully eliminated, and how this affects the re-colonisation process, we compared the microbiota and immune system in conventionally housed untreated mice with newly weaned ampicillin treated mice subsequently housed in either a microbe containing environment or in an isolator with only host associated suppressed bacteria to recolonize the gut. Two weeks ampicillin treatment induced a seemingly germ-free state with no bacterial DNA to reveal. Four weeks after treatment caeca were still significantly enlarged in both treated groups, but bacteria re-appeared even in isolator housed mice. While some suppressed bacteria were able to recover and even dominate the community, the abundances and composition were far from the untreated mice and differed between isolator and conventional housing. The treatment reduced the innate cytokine expressions at least for three weeks after treatment, and had a non-lasting reducing impact on the regulatory T cells, and a more lasting impact on the natural killer T cells. We conclude that temporary ampicillin treatment suppresses the majority but does not eliminate all the gut microbiota members. The re-colonisation process is as such influenced by both suppressed host associated bacteria and by environmental bacteria. Treated mice do not re-obtain a complex gut microbiota comparable to untreated mice, and the immune response and gut morphology reflect this. This is a concern when comparing host parameters sensitive to microbial regulation after an antibiotic-induced temporarily "germ-free" state., (Copyright © 2018 Elsevier Ltd. All rights reserved.)
- Published
- 2018
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12. Long-term Western diet fed apolipoprotein E-deficient rats exhibit only modest early atherosclerotic characteristics.
- Author
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Rune I, Rolin B, Lykkesfeldt J, Nielsen DS, Krych Ł, Kanter JE, Bornfeldt KE, Kihl P, Buschard K, Josefsen K, Fels JJ, Mortensen A, Christoffersen B, Kirk RK, and Hansen AK
- Subjects
- Animals, Aorta drug effects, Aorta pathology, Atherosclerosis microbiology, Body Weight drug effects, Diet, High-Fat adverse effects, Female, Gastrointestinal Microbiome, Insulin Resistance, Liver pathology, Oxidative Stress drug effects, Rats, Time Factors, Apolipoproteins E deficiency, Atherosclerosis metabolism, Atherosclerosis pathology, Diet, Western adverse effects
- Abstract
In the apolipoprotein E-deficient mouse, the gut microbiota has an impact on the development of atherosclerosis, but whether such correlations are also present in rats requires investigation. Therefore, we studied female SD-Apoe
tm1sage (Apoe-/- ) rats fed either a Western diet or a low-fat control diet with or without gluten, which is known to promote gut microbiota changes, until 20 weeks of age. We hypothesized that the manifestation of atherosclerosis would be more severe in Apoe-/- rats fed the Western high-fat diet, as compared with rats fed the low-fat diet, and that atherosclerosis would be accelerated by gluten. Both Western diet-feeding and gluten resulted in significant changes in gut microbiota, but the microbiota impact of gluten was transient. Compared with Apoe-/- rats fed a low-fat diet, Western diet-fed Apoe-/- rats were heavier and became glucose intolerant with increased levels of oxidative stress. They developed early fatty streak lesions in their aortic sinus, while there was no evidence of atherosclerosis in the thoracic aorta. No conclusions could be made on the impact of gluten on atherosclerosis. Although Western diet-fed Apoe-/- rats exhibited a more human-like LDL dominated blood lipid profile, signs of obesity, type 2 diabetes and cardiovascular disease were modest.- Published
- 2018
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13. Have you tried spermine? A rapid and cost-effective method to eliminate dextran sodium sulfate inhibition of PCR and RT-PCR.
- Author
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Krych Ł, Kot W, Bendtsen KMB, Hansen AK, Vogensen FK, and Nielsen DS
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- Animals, DNA analysis, DNA isolation & purification, Dextran Sulfate chemistry, Disease Models, Animal, Humans, Mice, Polymerase Chain Reaction standards, Polynucleotide 5'-Hydroxyl-Kinase drug effects, RNA analysis, RNA isolation & purification, RNA, Ribosomal, 16S genetics, Reverse Transcriptase Polymerase Chain Reaction standards, Time Factors, Dextran Sulfate antagonists & inhibitors, Polymerase Chain Reaction methods, Reverse Transcriptase Polymerase Chain Reaction methods, Spermine chemistry
- Abstract
The Dextran Sulfate Sodium (DSS) induced colitis mouse model is commonly used to investigate human inflammatory bowel disease (IBD). Nucleic acid extracts originating from these animals are often contaminated with DSS, which is a strong inhibitor of many enzymatic based molecular biology reactions including PCR and reverse-transcription (RT). Methods for removing DSS from nucleic acids extracts exist for RNA, but no effective protocol for DNA or cDNA is currently available. However, spermine has previously been shown to be an effective agent for counteracting DSS inhibition of polynucleotide kinase, which led to the hypothesis, that spermine could be used to counteract DSS inhibition of PCR and RT. We investigated the means of adding spermine in an adequate concentration to PCR based protocols (including qPCR, two-step RT-qPCR, and amplicon sequencing library preparation) to remove DSS inhibition. Within the range up to 0.01g/L, spermine can be added to PCR/qPCR or RT prophylactically without a significant reduction of reaction efficiency. Addition of spermine at the concentration of 0.08g/L can be used to recover qualitative PCR signal inhibited by DSS in concentrations up to 0.32g/L. For optimal quantitative analysis, the concentration of spermine requires fine adjustment. Hence, we present here a simple fluorometric based method for adjusting the concentration of spermine ensuring an optimal efficiency of the reaction exposed to an unknown concentration of DSS. In conclusion, we demonstrate a cost effective and easy method to counteract DSS inhibition in PCR and two-step RT-qPCR. Fixed or fine-tuned concentrations of spermine can be administered depending on the qualitative or quantitative character of the analysis., (Copyright © 2017 Elsevier B.V. All rights reserved.)
- Published
- 2018
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14. Immunological effects of reduced mucosal integrity in the early life of BALB/c mice.
- Author
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Bendtsen KM, Hansen CHF, Krych Ł, Skovgaard K, Kot W, Vogensen FK, and Hansen AK
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- Ampicillin adverse effects, Ampicillin pharmacology, Animals, Anti-Bacterial Agents pharmacology, Antigens, Bacterial immunology, Antigens, Bacterial metabolism, Colon drug effects, Colon immunology, Dextran Sulfate, Diet, Female, Intestinal Mucosa drug effects, Intestinal Mucosa growth & development, Killer Cells, Natural drug effects, Killer Cells, Natural immunology, Lipopolysaccharides blood, Lipopolysaccharides immunology, Lymph Nodes drug effects, Lymph Nodes immunology, Mice, Inbred BALB C, Models, Animal, Natural Killer T-Cells drug effects, Natural Killer T-Cells immunology, Permeability, Random Allocation, Spleen drug effects, Spleen immunology, T-Lymphocytes, Regulatory drug effects, T-Lymphocytes, Regulatory immunology, Toll-Like Receptor 4 metabolism, Gastrointestinal Microbiome drug effects, Gastrointestinal Microbiome physiology, Immune Tolerance, Intestinal Mucosa immunology, Intestinal Mucosa microbiology
- Abstract
Certain stimuli at the gut barrier may be necessary in early life to establish a proper balance of immune tolerance. We evaluated a compromised barrier in juvenile mice in relation to microbiota and local and systemic immunity. BALB/c mice were treated with a low dose of dextran sulfate sodium (DSS) with or without ampicillin and lipopolysaccharide (LPS) to clarify the importance of microbial antigens and interaction between microbial-associated patterns and toll-like receptors. The barrier breach resulted in increased plasma LPS, which was highest in mice treated simultaneously with ampicillin. Adding LPS in the food reduced its levels in plasma. Regulatory T cells were acutely increased in mesenteric lymph nodes (MLN) and spleen during DSS treatment regardless of simultaneous ampicillin treatment. In contrast, NK T and NK cells decreased in MLN and in spleen. This acute DSS effect was reflected in fold changes of haptoglobin and Il1a in colon, and this was also more pronounced in mice simultaneously treated with ampicillin. On day 1 post-treatment, major upregulations of Ifng, Foxp3, Il1b, Il2, and Il6 genes in colon were only observed in the mice simultaneously treated with ampicillin. A two-fold upregulation of colonic Foxp3 and Il1a was evident 25 days post-treatment. DSS skewed the microbiota in favor of Gram negative phyla. Therefore, increased permeability induced tolerogenic immunity independent of microbiota, and this was enhanced by LPS stimulation.
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- 2017
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15. Minimal short-term effect of dietary 2'-fucosyllactose on bacterial colonisation, intestinal function and necrotising enterocolitis in preterm pigs.
- Author
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Cilieborg MS, Bering SB, Østergaard MV, Jensen ML, Krych Ł, Newburg DS, and Sangild PT
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- Animals, Enterocolitis, Necrotizing diet therapy, Enterocolitis, Necrotizing microbiology, Microvilli enzymology, Swine, Trisaccharides administration & dosage, Enterocolitis, Necrotizing veterinary, Intestines drug effects, Premature Birth, Swine Diseases microbiology, Trisaccharides pharmacology
- Abstract
Human milk decreases the risk of necrotising enterocolitis (NEC), a severe gastrointestinal disease that occurs in 5-10 % of preterm infants. The prebiotic and immune-modulatory effects of milk oligosaccharides may contribute to this protection. Preterm pigs were used to test whether infant formula enriched with α1,2-fucosyllactose (2'-FL, the most abundant oligosaccharide in human milk) would benefit gut microbial colonisation and NEC resistance after preterm birth. Caesarean-delivered preterm pigs were fed formula (Controls, n 17) or formula with 5 g/l 2'-FL (2'-FL, n 16) for 5 d; eight 2'-FL pigs (50 %) and twelve Controls (71 %) developed NEC, with no difference in lesion scores (P=0·35); 2'-FL pigs tended to have less anaerobic bacteria in caecal contents (P=0·22), but no difference in gut microbiota between groups were observed by fluorescence in situ hybridisation and 454 pyrosequencing. Abundant α1,2-fucose was detected in the intestine with no difference between groups, and intestinal structure (villus height, permeability) and digestive function (hexose absorption, brush border enzyme activities) were not affected by 2'-FL. Formula enrichment with 2'-FL does not affect gut microbiology, digestive function or NEC sensitivity in pigs within the first few days after preterm birth. Milk 2'-FL may not be critical in the immediate postnatal period of preterm neonates when gut colonisation and intestinal immunity are still immature.
- Published
- 2016
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16. Provision of Amniotic Fluid During Parenteral Nutrition Increases Weight Gain With Limited Effects on Gut Structure, Function, Immunity, and Microbiology in Newborn Preterm Pigs.
- Author
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Østergaard MV, Shen RL, Støy AC, Skovgaard K, Krych Ł, Leth SS, Nielsen DS, Hartmann B, Bering SB, Schmidt M, and Sangild PT
- Subjects
- Animals, Cesarean Section veterinary, Enterocolitis, Necrotizing, Female, Gastric Inhibitory Polypeptide metabolism, Gastrointestinal Motility, Gastrointestinal Tract growth & development, Gastrointestinal Tract microbiology, Gestational Age, Glucagon-Like Peptide 2 metabolism, Immunity, Pregnancy, Weight Gain, Amniotic Fluid, Animals, Newborn growth & development, Gastrointestinal Tract physiology, Parenteral Nutrition veterinary, Premature Birth veterinary, Sus scrofa
- Abstract
Background: Small enteral boluses with human milk may reduce the risk of subsequent feeding intolerance and necrotizing enterocolitis in preterm infants receiving parenteral nutrition (PN). We hypothesized that feeding amniotic fluid, the natural enteral diet of the mammalian fetus, will have similar effects and improve growth and gastrointestinal (GI) maturation in preterm neonates receiving PN, prior to the transition to milk feeding., Materials and Methods: Twenty-seven pigs, delivered by cesarean section at ~90% of gestation, were provided with PN and also fed boluses with amniotic fluid (AF; n = 13, 24-72 mL/kg/d) or no oral supplements (nil per os [NPO]; n = 14) until day 5 when blood, tissue, and fecal samples were collected for analyses., Results: Body weight gain was 2.7-fold higher in AF vs NPO pigs. AF pigs showed slower gastric emptying, reduced meal-induced release of gastric inhibitory peptide and glucagon-like peptide 2, changed gut microbiota, and reduced intestinal permeability. There were no effects on GI weight, percentage mucosa, villus height, plasma citrulline, hexose absorptive capacity, and digestive enzymes. Intestinal interleukin (IL)-1β levels and expression of IL1B and IL8 were increased in AF pigs, while blood biochemistry and amino acid levels were minimally affected., Conclusion: Enteral boluses of AF were well tolerated in the first 5 days of life in preterm pigs receiving PN. Enteral provision of AF before the initiation of milk feeding may stimulate body growth and improve hydration in preterm infants receiving PN. Furthermore, it may improve GI motility and integrity, although most markers of GI maturation remain unchanged., (© 2015 American Society for Parenteral and Enteral Nutrition.)
- Published
- 2016
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17. Treatment with a Monoclonal Anti-IL-12p40 Antibody Induces Substantial Gut Microbiota Changes in an Experimental Colitis Model.
- Author
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Castro-Mejía J, Jakesevic M, Krych Ł, Nielsen DS, Hansen LH, Sondergaard BC, Kvist PH, Hansen AK, and Holm TL
- Abstract
Background and Aim. Crohn's disease is associated with gut microbiota (GM) dysbiosis. Treatment with the anti-IL-12p40 monoclonal antibody (12p40-mAb) has therapeutic effect in Crohn's disease patients. This study addresses whether a 12p40-mAb treatment influences gut microbiota (GM) composition in mice with adoptive transfer colitis (AdTr-colitis). Methods. AdTr-colitis mice were treated with 12p40-mAb or rat-IgG2a or NaCl from days 21 to 47. Disease was monitored by changes in body weight, stool, endoscopic and histopathology scores, immunohistochemistry, and colonic cytokine/chemokine profiles. GM was characterized through DGGE and 16S rRNA gene-amplicon high-throughput sequencing. Results. Following 12p40-mAb treatment, most clinical and pathological parameters associated with colitis were either reduced or absent. GM was shifted towards a higher Firmicutes-to-Bacteroidetes ratio compared to rat-IgG2a treated mice. Significant correlations between 17 bacterial genera and biological markers were found. The relative abundances of the RF32 order (Alphaproteobacteria) and Akkermansia muciniphila were positively correlated with damaged histopathology and colonic inflammation. Conclusions. Shifts in GM distribution were observed with clinical response to 12p40-mAb treatment, whereas specific GM members correlated with colitis symptoms. Our study implicates that specific changes in GM may be connected with positive clinical outcomes and suggests preventing or correcting GM dysbiosis as a treatment goal in inflammatory bowel disease.
- Published
- 2016
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18. Colonic Lesions, Cytokine Profiles, and Gut Microbiota in Plasminogen-Deficient Mice.
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Vestergaard B, Krych Ł, Lund LR, Jørgensen BP, Hansen L, Jensen HE, Nielsen DS, and Hansen AK
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- Animals, Colitis genetics, Colitis microbiology, Colitis pathology, Colon microbiology, Colon pathology, Feces microbiology, Genetic Predisposition to Disease, Male, Mice, 129 Strain, Mice, Knockout, Necrosis, Phenotype, Plasminogen genetics, Rectal Prolapse genetics, Rectal Prolapse microbiology, Rectal Prolapse pathology, Time Factors, Wound Healing, Colitis metabolism, Colon metabolism, Cytokines metabolism, Gastrointestinal Microbiome, Inflammation Mediators metabolism, Plasminogen deficiency, Rectal Prolapse metabolism
- Abstract
Plasminogen-deficient (FVB/NPan-plg(tm1Jld), plg(tm1Jld)) mice, which are widely used as a wound-healing model, are prone to spontaneous rectal prolapses. The aims of this study were 1) to evaluate the fecal microbiome of plg(tm1Jld) mice for features that might contribute to the development of rectal prolapses and colonic inflammation and 2) to assess the relevance of the inflammatory phenotype to the variability in wound healing in this model. The (plgtm1Jld) mice exhibited delayed wound healing, and they could be divided into 3 distinct groups that differed according to the time until wound closure. Colonic lesions in plg(tm1Jld) mice, which were characterized by necrotizing ulcerations and cystically dilated glands, were restricted to the intermediate and distal parts of the colon. The cytokine profile was indicative of chronic tissue damage, but the genetic modification did not change the composition of the gut microbiota, and none of the clinical or biochemical parameters correlated with the gut microbiota composition.
- Published
- 2015
19. Early gradual feeding with bovine colostrum improves gut function and NEC resistance relative to infant formula in preterm pigs.
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Shen RL, Thymann T, Østergaard MV, Støy AC, Krych Ł, Nielsen DS, Lauridsen C, Hartmann B, Holst JJ, Burrin DG, and Sangild PT
- Subjects
- Amino Acids blood, Animals, Cattle, Cytokines metabolism, Female, Gastric Inhibitory Polypeptide metabolism, Glucagon-Like Peptide 2 metabolism, Intestines pathology, Pregnancy, Swine, Bottle Feeding, Colostrum metabolism, Enterocolitis, Necrotizing veterinary, Intestinal Mucosa metabolism
- Abstract
It is unclear when and how to start enteral feeding for preterm infants when mother's milk is not available. We hypothesized that early and slow advancement with either formula or bovine colostrum stimulates gut maturation and prevents necrotizing enterocolitis (NEC) in preterm pigs, used as models for preterm infants. Pigs were given either total parenteral nutrition (TPN, n = 14) or slowly advancing volumes (16-64 ml·kg(-1)·day(-1)) of preterm infant formula (IF, n = 15) or bovine colostrum (BC, n = 13), both given as adjunct to parenteral nutrition. On day 5, both enteral diets increased intestinal mass (27 ± 1 vs. 22 ± 1 g/kg) and glucagon-like peptide 2 release, relative to TPN (P < 0.05). The incidence of mild NEC lesions was higher in IF than BC and TPN pigs (60 vs. 0 and 15%, respectively, P < 0.05). Only the IF pigs showed reduced gastric emptying and gastric inhibitory polypeptide release, and increased tissue proinflammatory cytokine levels (IL-1β and IL-8, P < 0.05) and expression of immune-related genes (AOAH, LBP, CXCL10, TLR2), relative to TPN. The IF pigs also showed reduced intestinal villus-to-crypt ratio, lactose digestion, and some plasma amino acids (Arg, Cit, Gln, Tyr, Val), and higher intestinal permeability, compared with BC pigs (all P < 0.05). Colonic microbiota analyses showed limited differences among groups. Early feeding with formula induces intestinal dysfunction whereas bovine colostrum supports gut maturation when mother's milk is absent during the first week after preterm birth. A diet-dependent feeding guideline may be required for newborn preterm infants., (Copyright © 2015 the American Physiological Society.)
- Published
- 2015
- Full Text
- View/download PDF
20. Phytase-producing capacity of yeasts isolated from traditional African fermented food products and PHYPk gene expression of Pichia kudriavzevii strains.
- Author
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Greppi A, Krych Ł, Costantini A, Rantsiou K, Hounhouigan DJ, Arneborg N, Cocolin L, and Jespersen L
- Subjects
- Gene Expression Regulation, Fungal, Phosphates metabolism, Phytic Acid metabolism, Real-Time Polymerase Chain Reaction, Yeasts isolation & purification, 6-Phytase genetics, 6-Phytase metabolism, Food Microbiology, Pichia enzymology, Pichia genetics, Yeasts enzymology, Yeasts genetics
- Abstract
Phytate is known as a strong chelate of minerals causing their reduced uptake by the human intestine. Ninety-three yeast isolates from traditional African fermented food products, belonging to nine species (Pichia kudriavzevii, Saccharomyces cerevisiae, Clavispora lusitaniae, Kluyveromyces marxianus, Millerozyma farinosa, Candida glabrata, Wickerhamomyces anomalus, Hanseniaspora guilliermondii and Debaryomyces nepalensis) were screened for phytase production on solid and liquid media. 95% were able to grow in the presence of phytate as sole phosphate source, P. kudriavzevii being the best growing species. A phytase coding gene of P. kudriavzevii (PHYPk) was identified and its expression was studied during growth by RT-qPCR. The expression level of PHYPk was significantly higher in phytate-medium, compared to phosphate-medium. In phytate-medium expression was seen in the lag phase. Significant differences in gene expression were detected among the strains as well as between the media. A correlation was found between the PHYPk expression and phytase extracellular activity., (Copyright © 2015 Elsevier B.V. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
21. A Review of Applied Aspects of Dealing with Gut Microbiota Impact on Rodent Models.
- Author
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Hansen AK, Krych Ł, Nielsen DS, and Hansen CH
- Subjects
- Animals, Rats, Gastrointestinal Microbiome physiology, Models, Animal, Rodentia microbiology
- Abstract
The gut microbiota (GM) affects numerous human diseases, as well as rodent models for these. We will review this impact and summarize ways to handle this challenge in animal research. The GM is complex, with the largest fractions being the gram-positive phylum Firmicutes and the gram-negative phylum Bacteroidetes. Other important phyla are the gram-negative phyla Proteobacteria and Verrucomicrobia, and the gram-positive phylum Actinobacteria. GM members influence models for diseases, such as inflammatory bowel diseases, allergies, autoimmunity, cancer, and neuropsychiatric diseases. GM characterization of all individual animals and incorporation of their GM composition in data evaluation may therefore be considered in future protocols. Germfree isolator-housed rodents or rodents made virtually germ free by antibiotic cocktails can be used to study diverse microbial influences on disease expression. Through subsequent inoculation with selected strains or cocktails of microbes, new "defined flora" models can yield valuable knowledge on the impact of the GM, and of specific GM members and their interactions, on important disease phenotypes and mechanisms. Rodent husbandry and microbial quality assurance practices will be important to ensure and confirm appropriate and research relevant GM., (© The Author 2015. Published by Oxford University Press on behalf of the Institute for Laboratory Animal Research. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
- Published
- 2015
- Full Text
- View/download PDF
22. Beyond genetics. Influence of dietary factors and gut microbiota on type 1 diabetes.
- Author
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Nielsen DS, Krych Ł, Buschard K, Hansen CH, and Hansen AK
- Subjects
- Animals, Diabetes Mellitus, Type 1 genetics, Diabetes Mellitus, Type 1 virology, Gastrointestinal Tract parasitology, Gastrointestinal Tract virology, Humans, Diabetes Mellitus, Type 1 etiology, Diabetes Mellitus, Type 1 microbiology, Diet adverse effects, Gastrointestinal Tract microbiology, Microbiota
- Abstract
Type 1 diabetes (T1D) is an autoimmune disease ultimately leading to destruction of insulin secreting β-cells in the pancreas. Genetic susceptibility plays an important role in T1D etiology, but even mono-zygotic twins only have a concordance rate of around 50%, underlining that other factors than purely genetic are involved in disease development. Here we review the influence of dietary and environmental factors on T1D development in humans as well as animal models. Even though data are still inconclusive, there are strong indications that gut microbiota dysbiosis plays an important role in T1D development and evidence from animal models suggests that gut microbiota manipulation might prove valuable in future prevention of T1D in genetically susceptible individuals., (Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
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