bacteria colon model
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Helminths and bacteria are major players in the mammalian gut ecosystem and each influences the host immune system and health. Declines in helminth prevalence and bacterial diversity appear to play a role in the dramatic rise of immune mediated inflammatory diseases (IMIDs) in western populations. Helminths are potent modulators of immune system and their reintroduction is a promising therapeutic avenue for IMIDs. However, the introduction of helminths represents a disturbance for the host and it is important to understand the impact of helminth reintroduction on the host, including the immune system and gut microbiome. We tested the impact of a benign tapeworm, Hymenolepis diminuta, in a rat model system. We find that H. diminuta infection results in increased interleukin 10 gene expression in the beginning of the prepatent period, consistent with induction of a type 2 immune response. We also find induction of humoral immunity during the patent period, shown here by increased IgA in feces. Further, we see an immuno-modulatory effect in the small intestine and spleen in patent period, as measured by reductions in tissue immune cells. We observed shifts in microbiota community composition during the patent period (beta-diversity) in response to H. diminuta infection. However, these compositional changes appear to be minor; they occur within families and genera common to both treatment groups. There was no change in alpha diversity. Hymenolepis diminuta is a promising model for helminth therapy because it establishes long-term, stable colonization in rats and modulates the immune system without causing bacterial dysbiosis. These results suggest that the goal of engineering a therapeutic helminth that can safely manipulate the mammalian immune system without disrupting the rest of the gut ecosystem is in reach.
- MeSH
- Bacteria klasifikace izolace a purifikace MeSH
- biodiverzita MeSH
- biologické modely * MeSH
- feces chemie MeSH
- fylogeneze MeSH
- Hymenolepis diminuta imunologie MeSH
- imunitní systém * MeSH
- imunoglobulin A analýza imunologie MeSH
- interakce hostitele a parazita imunologie MeSH
- interleukin-10 genetika metabolismus MeSH
- krysa rodu rattus MeSH
- potkani Wistar MeSH
- regulace genové exprese imunologie MeSH
- slezina imunologie MeSH
- střeva mikrobiologie parazitologie MeSH
- střevní mikroflóra * MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Bakteriálne infekcie kože a mäkkých tkanív zahŕňajú rozsiahlu problematiku a predstavujú najčastejšie vyskytujúce sa infekcie vo všetkých vekových skupinách. Obvykle ide o infekcie zmiešané vyvolané anaeróbnymi, mikroaerofilnými, fakultatívne anaeróbnymi a aeróbnymi baktériami. Pri patogenéze je možné v ložisku infekcie pozorovať prítomnosť aeróbnych baktérií, ktoré produktmi svojho metabolizmu a redukciou koncentrácie kyslíka umožnia pomnoženie striktne anaeróbnym patogénom. Úspešná terapia infekcií mäkkých tkanív je závislá od adekvátneho chirurgického prístupu, vhodnou antimikrobiálnou terapiou a adjuvantnej terapie infekcie mäkkých tkanív pomocou hyperbarickej oxygenoterapie (HBOT), t. j. dýchanie 100% kyslíka pod zvýšeným tlakom. V súčasnosti, aj napriek dostatočným empirickým znalostiam o benefitoch využitia HBOT pri liečbe infekcií mäkkých tkanív, neexistuje vhodný model pre detailnejšie štúdium interakcií organizmu a patogénov. Cieľom tohto projektu je vývoj in vitro modelu vhodného pre štúdium mechanizmov účinku antibiotík v kombinácii s HBOT, ako aj optimalizáciu používaných liečebných postupov.
Bacterial soft tissue infections represent the most common infections. Usually, they are caused by anaerobic, microaerophilic, facultative anaerobic and aerobic bacteria. In pathogenesis, a tissue is initially colonized by aerobic bacteria, which create a suitable environment for secondary colonization of anaerobic bacterial strains. Successful therapy of soft tissues infections consist of appropriate surgery treatment, targeted anti-microbial therapy and application of hyperbaric oxygen therapy (HBOT), i.e. breathing of pure oxygen under elevated ambient pressure. Nowadays, despite a lot of positive clinical evidence of HBOT in soft tissue infection treatment, no valid experimental model is established. The aim of our project was the development of an experimental model to study host-pathogen interaction during HBOT.
- MeSH
- antibakteriální látky * terapeutické užití MeSH
- Bacteria klasifikace účinky léků MeSH
- bakteriální infekce farmakoterapie terapie MeSH
- hyperbarická oxygenace * MeSH
- infekce měkkých tkání * klasifikace mikrobiologie terapie MeSH
- kombinovaná terapie MeSH
- lidé MeSH
- techniky in vitro MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- přehledy MeSH
The aim of this study was to develop a multiple-unit dosage system that released model drug into the colon, and also to evaluate the efficiency of the dosage form in human volunteers. The developed system combines pH-, time- and biodegradable polymer-based mechanisms for drug targeting to the colon. Pellet cores containing caffeine as model drug and chitosan and microcrystalline cellulose as excipients were prepared by the extrusion/spheronization method. The prepared pellets were film coated with a pH-dependent polymer, Eudragit FS 30 D. The coating total weight gain was 28.83% (w/w). Thanks to the application of an outer enteric film and the multiple unit design of the dosage form, the variability in gastric emptying was overcome, and a colon-specific targeting relied on the reproducibility of a small intestinal transit time, which was reported to be 3 ± 1 hours. A biodegradable polymer in the pellet core, chitosan, ensured the site-specific release of the model drug due to its solubility at the lower pH of the colonic region and by its biodegradability from the bacteria present. The efficiency of the system was confirmed by the in vivo testing of human saliva. The time of the first appearance of caffeine into the saliva, T(lag), was used as a parameter to estimate the disintegration time of the pellets into the gastrointestinal tract. The caffeine appeared in the saliva within 6.95 ± 1.12 hours (T(lag)) in 9 volunteers. A comparison of the reported colon arrival times indicates that the developed system is applicable to colonic drug delivery.
- MeSH
- aplikace orální MeSH
- celulosa aplikace a dávkování chemie MeSH
- chitosan aplikace a dávkování chemie MeSH
- dospělí MeSH
- farmaceutická chemie metody MeSH
- implantované léky aplikace a dávkování chemie farmakokinetika MeSH
- kofein aplikace a dávkování chemie MeSH
- kolon metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- lékové formy MeSH
- lékové transportní systémy metody MeSH
- lidé MeSH
- pomocné látky aplikace a dávkování chemie MeSH
- reprodukovatelnost výsledků MeSH
- rozpustnost MeSH
- sliny metabolismus MeSH
- vyprazdňování žaludku účinky léků MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Fluorescent tagging of nodule bacteria forming symbioses with legume host plants represents a tool for vital tracking of bacteria inside the symbiotic root nodules and monitoring changes in gene activity. The constitutive expression of heterologous fluorescent proteins, such as green fluorescent protein (GFP), also allows screening for nodule occupancy by a particular strain. Imaging of the fluorescence signal on a macro-scale is associated with technical problems due to the robustness of nodule tissues and a high level of autofluorescence. SCOPE: These limitations can be reduced by the use of a model species with a fine root system, such as Vicia tetrasperma. Further increases in the sensitivity and specificity of the detection and in image resolution can be attained by the use of a fluorescence scanner. Compared with the standard CCD-type cameras, the availability of a laser source of a specified excitation wavelength decreases non-specific autofluorescence while the photomultiplier tubes in emission detection significantly increase sensitivity. The large scanning area combined with a high resolution allow us to visualize individual nodules during the scan of whole root systems. Using a fluorescence scanner with excitation wavelength of 488 nm, a band-pass specific emission channel of 532 nm and a long-pass background channel of 555 nm, it was possible to distinguish nodules occupied by a rhizobial strain marked with one copy of cycle3 GFP from nodules colonized by the wild-type strain. CONCLUSIONS: The main limitation of the current plant model and GFP with the wild-type emission peak at 409 nm is a sharp increase in root autofluorescence below 550 nm. The selectivity of the technique can be enhanced by the use of red-shifted fluorophores and the contrasting labelling of the variants, provided that the excitation (482 nm) and emission (737 nm) maxima corresponding to root chlorophyll are respected.
BACKGROUND: Although irritable bowel syndrome (IBS) is a symptom-based diagnosis, clinicians' management of and communication about the disorder is often hampered by an unclear conceptual understanding of the nature of the problem. We aimed to elucidate an integrated explanatory model (EM) for IBS from the existing literature for pragmatic use in the clinical setting. METHODS: Systematic and exploratory literature searches were performed in PubMed to identify publications on IBS and EMs. KEY RESULTS: The searches did not identify a single, integrated EM for IBS. However, three main hypotheses were elucidated that could provide components with which to develop an IBS EM: (i) altered peripheral regulation of gut function (including sensory and secretory mechanisms); (ii) altered brain-gut signaling (including visceral hypersensitivity); and (iii) psychological distress. Genetic polymorphisms and epigenetic changes may, to some degree, underlie the etiology and pathophysiology of IBS and could increase the susceptibility to developing the disorder. The three model components also fit into one integrated explanation for abdominal symptoms and changes in stool habit. Additionally, IBS may share a common pathophysiological mechanism with other associated functional syndromes. CONCLUSIONS & INFERENCES: It was possible to elucidate an integrated, three-component EM as a basis for clinicians to conceptualize the nature of IBS, with the potential to contribute to better diagnosis and management, and dialog with sufferers.
- MeSH
- dysbióza komplikace patofyziologie MeSH
- gastrointestinální trakt inervace patofyziologie sekrece MeSH
- genetická predispozice k nemoci MeSH
- lidé MeSH
- nervový přenos MeSH
- polymorfismus genetický MeSH
- psychický stres komplikace psychologie MeSH
- signální transdukce MeSH
- somatosenzorické poruchy komplikace patofyziologie MeSH
- střevní mikroflóra MeSH
- syndrom dráždivého tračníku etiologie patofyziologie psychologie MeSH
- vzdělávání pacientů jako téma * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Cíl práce: Cílem práce bylo upozornit na závažnost výskytu anaerobních bakterií skupiny Bacteroides fragilis (BAFR) group u lidí, zvláště v klinických materiálech z oblasti břicha, a na zjištění souvislostí s výskytem rakoviny tlustého střeva a konečníku. Již dříve byla vyslovena domněnka o korelaci mezi výskytem bakterií z rodu Bacteroides u pacientů preferujících masitou stravu a rakovinným onemocněním tenkého a především tlustého střeva a konečníku. Vzhledem k tomu, že bakteroidy ze skupiny BAFR group jsou významnými producenty endotoxinů, bylo k porovnání bakterií, izolovaných z různých klinických vzorků pacientů s karcinomem tlustého střeva, rekta a dalšími afekcemi v dutině břišní, použito měření a statistické vyhodnocení tvorby endotoxinů u jednotlivých kmenů izolovaných druhů bakteroidů. Materiál a metody: Produkce endotoxinů byla stanovena u bakterií ze skupiny BAFR group (B. fragilis, B. thetaiotaomicron, B. distasonis a B. vulgatus), izolovaných z klinického materiálu pacientů s karcinomem rekta, tlustého a tenkého střeva, a porovnána se vzorky pacientů se zánětlivými afekcemi (anální absces, apendicitis, absces kůže) po jejich kultivaci v anaerobním i mikroaerofilním prostředí (s 5 % kyslíku). Tvorba endotoxinů byla prokazována kvantitativně pomocí gelové metody Pyrosate LAL testu (Limulus Amoebocyte Lysate Test, Biogenix, ČR) u 4 druhů bakteroidů ze skupiny BAFR group, kultivovaných v anaerobním a mikroaerofilním prostředí. Ve všech odebraných vzorcích bylo vyšetřeno vždy 5 kmenů každého izolovaného druhu bakteroidů (celkem 140 kmenů BAFR). Množství endotoxinu bylo detekováno v EU/ml – Endotoxin Units. Výsledky: Produkce endotoxinů u bakterií kultivovaných v mikroaerofilním prostředí byla několikrát vyšší než v případě produkce u bakterií kultivovaných v přísně anaerobním prostředí. Tyto rozdíly v produkci endotoxinů byly statisticky významné ( F1,269 = 160, p < 0,0001). Pokud se týká vlivu kyslíku na produkci endotoxinů, tak produkce endotoxinu byla 2,5 krát vyšší po kultivaci bakteroidů v mikroaerofilních (v průměru 889,1 EU/ml) než v anaerobních (358,2 EU/ml) podmínkách, a to nezávisle na druhu bakteroidu a diagnóze. Tyto výsledky svědčí o tom, že množství volného objemu kyslíku v prostředí má vliv na množství vytvářeného endotoxinu. Závěr: Výsledky produkce endotoxinů produkovaných těmito bakteriemi svědčí o tom, že produkce endotoxinů u bakterií kultivovaných v mikroaerofilním prostředí byla několikrát vyšší než v případě produkce u těchže bakterií kultivovaných v přísně anaerobním prostředí.
Objective: The aim of the study was to draw attention to the risk posed by anaerobic bacteria of the Bacteroides fragilis (BAFR) group, isolated particularly from abdominal lesions, and to assess the possible role of these species in colorectal cancer. A correlation has previously been suggested between the detection of the bacteria of the genus Bacteroides in patients on a meat-based diet and intestinal and, in particular, colorectal cancer. Given that the species of the BAFR group are major producers of endotoxins, measurements and statistical analysis of endotoxin production were used to compare the Bacteroides strains isolated from clinical specimens of patients with colon cancer, rectal cancer, and other abdominal lesions. Materials and Methods: Endotoxin production was detected in bacterial strains of the BAFR group (B. fragilis, B. thetaiotaomicron, B. distasonis, and B. vulgatus) isolated from clinical specimens of patients with rectal cancer, colon cancer, and intestinal cancer and was compared with that in strains from samples of patients with inflammatory conditions (anal abscess, appendicitis, skin abscess, etc.) under anaerobic and microaerophilic (with 5% of oxygen) culture conditions. The production of endotoxins was detected quantitatively using the Pyrosate LAL assay kit (Limulus Amoebocyte Lysate Test, BIOGENIX, CR) in four species of the BAFR group after anaerobic and microaerophilic culture. Five strains of each isolated Bacteroides species from each type of specimens were tested (in total 140 BAFR strains). The amount of endotoxin was given in endotoxin units per ml (EU/ml). Results: Endotoxin production by bacteria under microaerophilic culture conditions was several times higher in comparison with strictly anaerobic culture. The difference was statistically significant (F1.269 = 160, p <0.0001). As regards the effect of oxygen on endotoxin production, the amount of endotoxins produced under microaerophilic culture conditions (average 889.1 EU/ml) was 2.5 times as high as that observed under anaerobic culture conditions (358.2 EU/ml), regardless of the bacteroides species and diagnosis. These results suggest that the amount of free oxygen in the environment affects the amount of endotoxin generated by the Bacteroides strains. Conclusion: The results show that endotoxin production by the Bacteroides strains under microaerophilic culture conditions is several times as high as that under strictly anaerobic culture conditions.
- MeSH
- Bacteroides fragilis * izolace a purifikace MeSH
- endotoxiny * analýza MeSH
- kolorektální nádory MeSH
- kultivační média chemie MeSH
- kultivační techniky metody MeSH
- kultivované buňky chemie metabolismus MeSH
- kyslík * metabolismus MeSH
- lidé MeSH
- lineární modely MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- srovnávací studie MeSH
BACKGROUND: The aim was to analyze the influence of intestinal microbiota on the development of intestinal inflammation. We used the model of chronic inflammation that develops spontaneously in the colon of conventional severe combined immunodeficiency (SCID) mice restored with the CD45 RB(high) subset of CD4+T cells isolated from the spleen of normal BALB/c mice. METHODS: A CD4+CD45RB(high) subpopulation of T cells was purified from the spleen of conventional BALB/c mice by magnetic separation (MACS) and transferred into immunodeficient SCID mice. Germ-free (GF) SCID mice or SCID mice monoassociated with Enterococcus faecalis, SFB (segmented filamentous bacteria), Fusobacterium mortiferum, Bacteroides distasonis, and in combination Fusobacterium mortiferum + SFB or Bacteroides distasonis + SFB were used as recipients. SCID mice were colonized by a defined cocktail of specific pathogen-free (SPF) bacteria. Mice were evaluated 8-12 weeks after the cell transfer for clinical and morphological signs of inflammatory bowel disease (IBD). RESULTS: After the transfer of the CD4+CD45RB(high) T-cell subpopulation to SCID mice severe colitis was present in conventional animals and in mice colonized with a cocktail of SPF microflora plus SFB. Altered intestinal barrier in the terminal ileum of mice with severe colitis was documented by immunohistology using antibodies to ZO-1 (zona occludens). CONCLUSIONS: Only SFB bacteria together with a defined SPF mixture were effective in triggering intestinal inflammation in the model of IBD in reconstituted SCID mice, while no colitis was detected in GF mice or in mice colonized either with SPF microflora or monoassociated only with SFB or colonized by Bacteroides distasonis + SFB or Fusobacterium mortiferum + SFB.
- MeSH
- antigeny CD45 aplikace a dávkování MeSH
- CD4-pozitivní T-lymfocyty imunologie MeSH
- financování organizované MeSH
- hybridizace in situ fluorescenční MeSH
- hyperplazie MeSH
- hypertrofie patologie MeSH
- kolitida imunologie mikrobiologie prevence a kontrola MeSH
- mikroskopie elektronová rastrovací MeSH
- modely nemocí na zvířatech MeSH
- myši inbrední BALB C MeSH
- myši SCID MeSH
- myši MeSH
- převzatá imunita MeSH
- průtoková cytometrie MeSH
- slezina imunologie MeSH
- střevní sliznice mikrobiologie ultrastruktura MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
SCOPE: This multi-omic study investigates the bidirectional interactions between gut microbiota and silymarin metabolism, highlighting the differential effects across various age groups. Silymarin, the extract from Silybum marianum (milk thistle), is commonly used for its hepatoprotective effects. METHODS AND RESULTS: An in vitro fermentation colon model was used with microbiota from 20 stool samples obtained from healthy donors divided into two age groups. A combination of three analytical advanced techniques, namely proton nuclear magnetic resonance (1H NMR), next-generation sequencing (NGS), and liquid chromatography-mass spectrometry (LC-MS) was used to determine silymarin microbial metabolites over 24 h, overall metabolome, and microbiota composition. Silymarin at a low diet-relevant dose of 50 μg mL-1 significantly altered gut microbiota metabolism, reducing short-chain fatty acid (acetate, butyrate, propionate) production, glucose utilization, and increasing alpha-diversity. Notably, the study reveals age-related differences in silymarin catabolism. Healthy elderly donors (70-80 years) exhibited a significant increase in a specific catabolite associated with Oscillibacter sp., whereas healthy young donors (12-45 years) showed a faster breakdown of silymarin components, particularly isosilybin B, which is associated with higher abundance of Faecalibacterium and Erysipelotrichaceae UCG-003. CONCLUSION: This study provides insights into microbiome functionality in metabolizing dietary flavonolignans, highlighting implications for age-specific nutritional strategies, and advancing our understanding of dietary (poly)phenol metabolism.
- MeSH
- dítě MeSH
- dospělí MeSH
- feces mikrobiologie MeSH
- fermentace MeSH
- kolon * mikrobiologie metabolismus účinky léků MeSH
- kyseliny mastné těkavé metabolismus MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- silymarin * farmakologie MeSH
- střevní mikroflóra * účinky léků fyziologie MeSH
- zdraví dobrovolníci pro lékařské studie MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Metagenomic approaches are currently being used to decipher the genome of the microbiota (microbiome), and, in parallel, functional studies are being performed to analyze the effects of the microbiota on the host. Gnotobiological methods are an indispensable tool for studying the consequences of bacterial colonization. Animals used as models of human diseases can be maintained in sterile conditions (isolators used for germ-free rearing) and specifically colonized with defined microbes (including non-cultivable commensal bacteria). The effects of the germ-free state or the effects of colonization on disease initiation and maintenance can be observed in these models. Using this approach we demonstrated direct involvement of components of the microbiota in chronic intestinal inflammation and development of colonic neoplasia (i.e., using models of human inflammatory bowel disease and colorectal carcinoma). In contrast, a protective effect of microbiota colonization was demonstrated for the development of autoimmune diabetes in non-obese diabetic (NOD) mice. Interestingly, the development of atherosclerosis in germ-free apolipoprotein E (ApoE)-deficient mice fed by a standard low-cholesterol diet is accelerated compared with conventionally reared animals. Mucosal induction of tolerance to allergen Bet v1 was not influenced by the presence or absence of microbiota. Identification of components of the microbiota and elucidation of the molecular mechanisms of their action in inducing pathological changes or exerting beneficial, disease-protective activities could aid in our ability to influence the composition of the microbiota and to find bacterial strains and components (e.g., probiotics and prebiotics) whose administration may aid in disease prevention and treatment.
- MeSH
- autoimunitní nemoci etiologie mikrobiologie MeSH
- gastrointestinální trakt mikrobiologie MeSH
- gnotobiologické modely MeSH
- imunita MeSH
- lidé MeSH
- metagenom imunologie MeSH
- modely nemocí na zvířatech MeSH
- nádory etiologie mikrobiologie MeSH
- sliznice imunologie MeSH
- zánět etiologie mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH