infekce a stres v raném věku během zranitelného období vývoje cns souvisejí se zvýšeným rizikem vzniku neuropsychiatrických poruch, jako je schizofrenie. na základě teorie dvojího zásahu u schizofrenie je hypoteticky ke spuštění poruchy nutný ještě druhý inzult rùzné modality. neonatální aplikace lipopolysacharidu (lPs, známého také jako endotoxin) u potkanù je uznávaným modelem vedoucím k behaviorálnímu deficitu a neurokognitivním poruchám na podkladě imunitní stimulace v raném věku a používá se jako neurovývojový model psychózy. V této práci jsme na modelu potkana studovali, jak neonatální podání lPs spolu s dysbiózou vyvolanou chronickou léčbou antibiotiky jako druhým zásahem na základě teorie dvojího zásahu ovlivní metabolické dráhy pomocí necílené metabolomické analýzy. hodnotili jsme rozdíly mezi metabolomy kontrolní a testovací skupiny (vystavené lPs a/nebo léčbě antibiotiky), abychom zjistili rozdíly mezi jejich metabolitovými profily. Vzorky skupiny léčené lPs vykazovaly pozoruhodné metabolomické změny ve srovnání s kontrolními subjekty. Jako nejvíce ovlivněné divergentní dráhy byly určeny dráhy pyrimidinového metabolismu a dráhy biosyntézy pantothenátu a koenzymu a (coa); p-hodnota upravená podle gama je 0,04275 a 0,04278. Léčba ATB nevedla k žádným změnám metabolomu, a naopak maskovala účinky vyvolané aplikací lPs. S odkazem na tyto údaje jsme schopni naznačit, že neonatální výzva LPS hraje významnou roli v rozkladu pyrimidinového metabolismu a drah biosyntézy pantothenátu a coa, které by mohly být identifikovány jako metabolické dráhy zapojené do patogeneze schizofrenie. Vzhledem k pøedběžné povaze naší studie je nutné naše zjištění ověøit v budoucím výzkumu schizofrenie.
infections and stress in early life, during a vulnerable development period of the cns, are related to an increased risk of the development of neuropsychiatric disorders such as schizophrenia. Based on the dual-intervention theory of schizophrenia, a second insult modality is hypothesized to be required to trigger the disorder. neonatal lipopolysaccharide (lPs, also known as endotoxin) treatment in rats is an acknowledged model for inducing the behavioral deficits and neurocognitive impairments caused by an early-life immune challenge and is used as a neurodevelopmental model of psychosis. in the present work, we studied in the rat model how the neonatal lPs administration along with dysbiosis induced by chronic antibiotic treatment as a second hit based on the dual hit theory will affect metabolic pathways by an untargeted metabolomic analysis. We have evaluated differences between the metabolome of control and test groups (exposed to lPs and/or antibiotic treatment) to determine differences between their metabolite profiles. the lPs-treated group samples have exhibited notable metabolomic changes compared to the control subjects. the most affected divergent pathways were determined as Pyrimidine metabolism and Pantothenate and coenzyme a (coa) biosynthesis pathways (the gamma-adjusted p-values are 0.04275 and 0.04278, respectively). the atB treatment did not result in any changes in metabolome and, on the contrary, masked the effects induced by lPs treatment. referring to these data, we are able to suggest that a neonatal lPs challenge plays a significant role in the discomposing Pyrimidine metabolism and Pantothenate and coa biosynthesis pathways, which could be identified as metabolism pathways involved in schizophrenia pathogenesis. Due to the preliminary nature of our study, it is necessary to confirm our findings in future schizophrenia research
Ulcerative colitis (UC) is an inflammatory bowel disease, and intestinal bacteria are implicated in the pathogenesis of this disorder. The administration of aminosalicylates (5-ASA) is a conventional treatment that targets the mucosa, while fecal microbial transplantation (FMT) is a novel treatment that directly targets the gut microbiota. The aim of this study was to identify changes in fecal bacterial composition after both types of treatments and evaluate clinical responses. Sixteen patients with active left-sided UC underwent enema treatment using 5-ASA (n = 8) or FMT (n = 8) with a stool from a single donor. Fecal microbiota were analyzed by 16S rDNA high-throughput sequencing, and clinical indices were used to assess the efficacy of treatments. 5-ASA therapy resulted in clinical remission in 50% (4/8) of patients, but no correlation with changes in fecal bacteria was observed. In FMT, remission was achieved in 37.5% (3/8) of patients and was associated with a significantly increased relative abundance of the families Lachnospiraceae, Ruminococcaceae, and Clostridiaceae of the phylum Firmicutes, and Bifidobacteriaceae and Coriobacteriaceae of the phylum Actinobacteria. At the genus level, Faecalibacterium, Blautia, Coriobacteria, Collinsela, Slackia, and Bifidobacterium were significantly more frequent in patients who reached clinical remission. However, the increased abundance of beneficial taxa was not a sufficient factor to achieve clinical improvement in all UC patients. Nevertheless, our preliminary results indicate that FMT as non-drug-using method is thought to be a promising treatment for UC patients.
- MeSH
- analýza hlavních komponent MeSH
- aplikace lokální MeSH
- Bacteria klasifikace MeSH
- biodiverzita MeSH
- dárci tkání MeSH
- diskriminační analýza MeSH
- dospělí MeSH
- feces mikrobiologie MeSH
- fekální transplantace * MeSH
- lidé středního věku MeSH
- lidé MeSH
- mesalamin aplikace a dávkování farmakologie terapeutické užití MeSH
- senioři MeSH
- střevní mikroflóra * účinky léků MeSH
- ulcerózní kolitida farmakoterapie mikrobiologie terapie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- randomizované kontrolované studie MeSH
A feeding study was performed to monitor the effect of chitosan intake on the fecal microbiota of ten healthy human subjects. Diversity of microflora was monitored during 8 weeks including 4 weeks of chitosan supplementations. Using denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene amplicons and quantitative PCR method we revealed possible changes originating in the overall bacterial composition and also in the subpopulation of Bifidobacterium group. DGGE profiles displayed high complexity and individuality for each subject. Considerable variations in the composition of band patterns were observed among different persons. A raised level of fecal Bacteroides in response to chitosan intake was found in all samples. Bifidobacterium levels following chitosan intake increased or remain unchanged. Non-significant increase was, surprisingly, found in the numbers of butyrate-producing bacteria.
- MeSH
- Bacteroides genetika izolace a purifikace MeSH
- Bifidobacterium genetika izolace a purifikace MeSH
- biodiverzita MeSH
- chitosan metabolismus MeSH
- denaturace nukleových kyselin MeSH
- DNA bakterií genetika MeSH
- dospělí MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- experimenty na lidech MeSH
- feces mikrobiologie MeSH
- financování organizované MeSH
- lidé středního věku MeSH
- lidé MeSH
- polymerázová řetězová reakce MeSH
- ribozomální DNA genetika MeSH
- RNA ribozomální 16S genetika MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
The anaerobic fungus Anaeromyces mucronatus KF8 grown in batch culture on M10 medium with rumen fluid and microcrystalline cellulose as carbon source produced a broad range of enzymes requisite for degradation of plant structural and storage saccharides including cellulase, endoglucanase, xylanase, alpha-xylosidase, beta-xylosidase, alpha-glucosidase, beta-glucosidase, beta-galactosidase, mannosidase, cellobiohydrolase, amylase, laminarinase, pectinase and pectate lyase. These enzymes were detected in both the intra- and extracellular fractions, but production into the medium was prevalent with the exception of intracellular beta-xylosidase, chitinases, N-acetylglucosaminidase, and lipase. Xylanase activity was predominant among the polysaccharide hydrolases. Extracellular production of xylanase was stimulated by the presence of cellobiose and oat spelt xylan. Zymogram of xylanases of strain KF8 grown on different carbon sources revealed several isoforms of xylanases with approximate molar masses ranging from 26 to 130 kDa.
- MeSH
- anaerobióza MeSH
- celobiosa metabolismus MeSH
- celulosa metabolismus MeSH
- financování organizované MeSH
- fungální proteiny biosyntéza MeSH
- glykosidhydrolasy biosyntéza chemie klasifikace MeSH
- kultivační média chemie MeSH
- molekulová hmotnost MeSH
- Neocallimastigales enzymologie fyziologie MeSH
- xylany metabolismus MeSH
The diversity of anaerobic fungi was evaluated in cow semiliquid manure obtained from input homogenizing tank of biogas plant. Among three sets of tested primers, the combination of fungal universal ITS1F and Neocallimastigales specific Neo QPCR Rev primers was selected and used for the construction of clone library. Eighty-four new complete internal transcribed spacers (ITS1) and partial 5.8S rDNA sequences generated within this study were analyzed by Bayesian inference and assigned to an existing order of the Neocallimastigales. Sixty-seven% of sequences were affiliated with Cyllamyces, 24% with Piromyces, 7% with Anaeromyces, only 2% with Neocallimastix, and no sequences with Orpinomyces. According to Bayesian analysis the genus Caecomyces was polyphyletic and disappeared from the presented ITSbased phylogram. This study gave a first insight into the diversity of anaerobic fungi in cow manure, where the prevalence of fungi with bulbous morphology was indicated.
- MeSH
- anaerobióza MeSH
- biodiverzita MeSH
- DNA fungální genetika chemie MeSH
- DNA primery genetika MeSH
- financování organizované MeSH
- fungální RNA genetika MeSH
- fylogeneze MeSH
- geny rRNA MeSH
- hnůj mikrobiologie MeSH
- houby MeSH
- mezerníky ribozomální DNA genetika chemie MeSH
- molekulární sekvence - údaje MeSH
- ribozomální DNA genetika chemie MeSH
- RNA ribozomální 5.8S genetika MeSH
- sekvenční analýza DNA MeSH
- shluková analýza MeSH
- skot MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
The influence of geographic location, season, age, and part of the digestive tract on bacterial diversity was evaluated on intestinal microflora of honeybees, wasps, and cockroaches using DGGE analysis. PCR-DGGE analyses with universal bacterial primers targeting 200-bp region of the 16S rDNA gene afforded the profile of complex bacterial DNA; specific primers were used to determine the profile of bifidobacteria whose concentration in digestive tract was determined by real-time PCR. Selected PCR products were identified by sequencing. The microflora of the bees exhibited little variations among the hives from distant locations. Their bifidobacterial population formed 2.8-8.4 % of total bacteria and was very homogeneous. The total gut microflora of wasps was also homogeneous, only two samples being affected by the season or the location; on the other hand, wasp bifidobacterial population was very heterogeneous. Cockroaches showed the highest variations in microflora composition, the age and diet being the ultimate factors; bifidobacteria counts also varied among tested individuals (0.1-34.1 % of total bacteria). Our results suggest that nutrition habits are the strongest factor affecting the insect microflora, giving higher variations to omnivorous species.
- MeSH
- Bacteria genetika izolace a purifikace klasifikace MeSH
- Bifidobacterium genetika izolace a purifikace MeSH
- DNA bakterií analýza izolace a purifikace MeSH
- genetická variace MeSH
- hmyz mikrobiologie MeSH
- polymerázová řetězová reakce metody MeSH
- ribozomální DNA analýza MeSH
- RNA ribozomální 16S genetika MeSH
- sršňovití mikrobiologie MeSH
- střeva mikrobiologie MeSH
- švábi mikrobiologie MeSH
- včely mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
Chitinolytic systems of anaerobic polycentric rumen fungi of genera Orpinomyces and Anaeromyces were investigated in three crude enzyme fractions - extracellular, cytosolic and cell-wall. Endochitinase was found as a dominant enzyme with highest activity in the cytosolic fraction. Endochitinases of both genera were stable at pH 4.5-7.0 with optimum at 6.5. The Orpinomyces endochitinase was stable up to 50 degrees C with an optimum for enzyme activity at 50 degrees C; similarly, Anaeromyces endochitinase was stable up to 40 degrees C with optimum at 40 degrees C. The most suitable substrate for both endochitinases was fungal cell-wall chitin. Enzyme activities were inhibited by Hg(2+) and Mn(2+), and activated by Mg(2+) and Fe(3+). Both endochitinases were inhibited by 10 mmol/L SDS and activated by iodoacetamide.
- MeSH
- anaerobióza MeSH
- bachor metabolismus mikrobiologie MeSH
- chitin metabolismus MeSH
- chitinasy chemie metabolismus MeSH
- Chytridiomycota enzymologie klasifikace růst a vývoj MeSH
- fungální proteiny genetika MeSH
- koncentrace vodíkových iontů MeSH
- Neocallimastigales enzymologie klasifikace růst a vývoj MeSH
- stabilita enzymů MeSH
- teplota MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
The fecal short-chain fatty acids concentration was higher (154 +/- 46.9 mmol/L) in childhood patients than in healthy children (96.6 +/- 19.2 mmol/L). On the other hand, pH values were nonsignificantly lower in patients stool (6.78 +/- 0.75 vs. children 7.42 +/- 0.74). Using denaturing gradient gel electrophoresis specific for total bacteria, lactobacilli and bifidobacteria the microbial population was characterized in fecal samples and in duodenal biopsies. Bacteria adhering to duodenal biopsies were not dominating in stool samples. More than 50 % of detected bacterial species belonged to as yet uncultured strains.
- MeSH
- Bacteria genetika izolace a purifikace klasifikace MeSH
- biopsie MeSH
- celiakie mikrobiologie MeSH
- dítě MeSH
- DNA bakterií analýza izolace a purifikace MeSH
- duodenum mikrobiologie MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- feces chemie mikrobiologie MeSH
- kojenec MeSH
- koncentrace vodíkových iontů MeSH
- kyseliny mastné těkavé analýza MeSH
- lidé MeSH
- mladiství MeSH
- polymerázová řetězová reakce metody MeSH
- předškolní dítě MeSH
- střeva mikrobiologie MeSH
- Check Tag
- dítě MeSH
- kojenec MeSH
- lidé MeSH
- mladiství MeSH
- předškolní dítě MeSH
The common feature of all chytridiomycetous fungi, aerobic as well as anaerobic, is an abundance of chitin in their cell wall. The genes coding for chitinases have therefore been widely used as phylogenetic markers in ascomycetes. As their utility for Chytridiomycetes has not been determined we chose the gene encoding an enzyme involved in chitin degradation and energy metabolism, the beta-(1,4)-N-acetylglucosaminidase (nag1). Primer pair Nag-forward and Nag-reverse was used to create PCR product from 5 strains of anaerobic and 7 strains of aerobic chytrids. However, Blast search of sequenced amplicons showed that these primers are specific only for fungus Emericella nidulans. Amino acid alignment of Nag1 proteins of fungal, protozoal and bacterial origin available in GenBank database was therefore performed. Five amino acid regions were found to be conserved enough to serve as a suitable domain for the design of a set of primers for the universal amplification of the nag1 gene in the Neocallimastigales fungi.
- MeSH
- acetylglukosaminidasa genetika chemie MeSH
- aerobióza MeSH
- anaerobióza MeSH
- Chytridiomycota enzymologie genetika MeSH
- DNA fungální analýza izolace a purifikace MeSH
- DNA primery genetika MeSH
- fungální proteiny genetika MeSH
- houby enzymologie genetika klasifikace růst a vývoj MeSH
- polymerázová řetězová reakce MeSH
- techniky amplifikace nukleových kyselin metody MeSH
The fatty acid (FA) composition of fresh mycelia of anaerobic rumen fungi was determined. The fatty acids methyl esters (FAME) of six strains belonging to four genera (Neocallimastix, Caecomyces, Orpinomyces, Anaeromyces) and one unknown strain were analyzed by gas chromatography. All studied fungi possess the same FAs but differences were found in their relative concentrations. The FA profile of anaerobic fungi comprises carbon chains of length ranging from 12 to 24; the most common fatty acids were stearic (C(18:0)), arachidic (C(20:0)), heneicosanoic (C(21:0)), behenic (C(22:0)), tricosanoic (C(23:0)) and lignoceric (C(24:0)) with relative amount representing >4% of total FA. Significant differences were determined for heptadecanoic, oleic, behenic and tricosanoic acids. Rumen anaerobic fungi can contain very long chain fatty acids; we found unsaturated fatty acids including cis-11-eicosenoic (C(20:1)), cis-11,14-eicosadienoic (C(20:2)), erucic (C(22:1n9)), cis-13,16-docosadienoic (C(22:2)) and nervonic (C(24:1)) acids in very small amounts but their presence seems to be unique for anaerobic fungi.
