BACKGROUND: Exposure of critically ill patients to antibiotics lead to intestinal dysbiosis, which often manifests as antibiotic-associated diarrhoea. Faecal microbiota transplantation restores gut microbiota and may lead to faster resolution of diarrhoea. METHODS: Into this prospective, multi-centre, randomized controlled trial we will enrol 36 critically ill patients with antibiotic-associated diarrhoea. We will exclude patients with ongoing sepsis, need of systemic antibiotics, or those after recent bowel surgery or any other reason that prevents the FMT. Randomisation will be in 1:1 ratio. Patients in the control group will receive standard treatment based on oral diosmectite. In the intervention group, patients will receive, in addition to the standard of care, faecal microbiota transplantation via rectal tube, in the form of a preparation mixed from 7 thawed aliquots (50 mL) made from fresh stool of 7 healthy unrelated donors and quarantined deep frozen for 3 to 12 months. Primary outcome is treatment failure defined as intervention not delivered or diarrhoea persisting at day 7 after randomisation. Secondary outcomes include safety measures such as systemic inflammatory response, adverse events, and also diarrhoea recurrence within 28 days. Exploratory outcomes focus on gut barrier function and composition of intestinal microbiota. DISCUSSION: Faecal microbiota transplantation has been effective for dysbiosis in non-critically ill patients with recurrent C. difficile infections and it is plausible to hypothesize that it will be equally effective for symptoms of dysbiosis in the critically ill patients. In addition, animal experiments and observational data suggest other benefits such as reduced colonization with multi-drug resistant bacteria and improved gut barrier and immune function. The frozen faeces from unrelated donors are immediately available when needed, unlike those from the relatives, who require lengthy investigation. Using multiple donors maximises graft microbiota diversity. Nonetheless, in vulnerable critically ill patients, Faecal microbiota transplantation might lead to bacterial translocation and unforeseen complications. From growing number of case series it is clear that its off label use in the critically ill patients is increasing and that there is a burning need to objectively assess its efficacy and safety, which this trial aims. TRIAL REGISTRATION: www.clinicaltrials.gov (NCT05430269).
- MeSH
- antibakteriální látky * škodlivé účinky terapeutické užití MeSH
- dysbióza terapie mikrobiologie MeSH
- feces mikrobiologie MeSH
- fekální transplantace * metody škodlivé účinky MeSH
- klinické zkoušky, fáze II jako téma MeSH
- kritický stav * MeSH
- lidé MeSH
- multicentrické studie jako téma MeSH
- prospektivní studie MeSH
- průjem * terapie mikrobiologie MeSH
- randomizované kontrolované studie jako téma MeSH
- střevní mikroflóra * účinky léků MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- protokol klinické studie MeSH
BACKGROUND: Faecal microbiota transplantation (FMT) is a developing therapy for disorders related to gut dysbiosis. Despite its growing application, standardised protocols for FMT filtrate preparation and quality assessment remain undeveloped. The viability of bacteria in the filtrate is crucial for FMT's efficacy and for validating protocol execution. We compared two methods-in vitro cultivation and membrane integrity assessment-for their accuracy, reproducibility and clinical applicability in measuring bacterial viability in frozen FMT stool filtrate. METHODS: Bacterial viability in stool filtrate was evaluated using (i) membrane integrity through fluorescent DNA staining with SYTO9 and propidium iodide, followed by flow cytometry and (ii) culturable bacteria counts (colony-forming units, CFU) under aerobic or anaerobic conditions. RESULTS: Using different types of samples (pure bacterial culture, stool of germ-free and conventionally bred mice, native and heat-treated human stool), we refined the bacterial DNA staining protocol integrated with flow cytometry for assessment of bacterial viability in frozen human stool samples. Both the membrane integrity-based and cultivation-based methods exhibited significant variability in bacterial viability across different FMT filtrates, without correlation. The cultivation-based method showed a mean coefficient of variance of 30.3%, ranging from 7.4% to 60.1%. Conversely, the membrane integrity approach yielded more reproducible results, with a mean coefficient of variance for viable cells of 6.4% ranging from 0.2% to 18.2%. CONCLUSION: Bacterial viability assessment in stool filtrate using the membrane integrity method offers robust and precise data, making it a suitable option for faecal material evaluation in FMT. In contrast, the cultivation-dependent methods produce inconsistent outcomes.
- MeSH
- Bacteria izolace a purifikace MeSH
- feces * mikrobiologie MeSH
- fekální transplantace * metody MeSH
- lidé MeSH
- mikrobiální viabilita * MeSH
- myši MeSH
- průtoková cytometrie * metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
BACKGROUND: Faecal microbiota transplantation (FMT) is an established treatment for Clostridioides difficile infection and is under investigation for other conditions. The availability of suitable donors and the logistics of fresh stool preparation present challenges, making frozen, biobanked stools an attractive alternative. AIMS: This study aimed to evaluate the long-term viability of bacterial populations in faecal samples stored at -80°C for up to 12 months, supporting the feasibility of using frozen grafts for FMT. METHODS: Fifteen faecal samples from nine healthy donors were processed, mixed with cryoprotectants and stored at -80°C. Samples were assessed at baseline and after 3, 6 and 12 months using quantitative culturing methods to determine the concentration of live bacteria. RESULTS: Quantitative analysis showed no significant decrease in bacterial viability over the 12-month period for both aerobic and anaerobic cultures (p = 0.09). At all timepoints, the coefficients of variability in colony-forming unit (CFU) counts were greater between samples (102 ± 21% and 100 ± 13% for aerobic and anaerobic cultures, respectively) than the variability between measurements of the same sample (30 ± 22% and 30 ± 19%). CONCLUSIONS: The study confirmed that faecal microbiota can be preserved with high viability in deep-freeze storage for up to a year, making allogenic FMT from biobanked samples a viable and safer option for patients. However, a multidonor approach may be beneficial to mitigate the risk of viability loss in any single donor sample.
- MeSH
- feces * mikrobiologie MeSH
- fekální transplantace * metody MeSH
- kryoprezervace metody MeSH
- lidé MeSH
- mikrobiální viabilita * MeSH
- zmrazování MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
The collet (root-hypocotyl junction) region is an important plant transition zone between soil and atmospheric environments. Despite its crucial importance for plant development, little is known about how this transition zone is specified. Here we document the involvement of the exocyst complex in this process. The exocyst, an octameric tethering complex, participates in secretion and membrane recycling and is central to numerous cellular and developmental processes, such as growth of root hairs, cell expansion, recycling of PIN auxin efflux carriers and many others. We show that dark-grown Arabidopsis mutants deficient in exocyst subunits can form a hair-bearing ectopic collet-like structure above the true collet, morphologically resembling the true collet but also retaining some characteristics of the hypocotyl. The penetrance of this phenotypic defect is significantly influenced by cultivation temperature and carbon source, and is related to a defect in auxin regulation. These observations provide new insights into the regulation of collet region formation and developmental plasticity of the hypocotyl.
Cíl práce: Kandidémie patří mezi poměrně časté infekční komplikace u kriticky nemocných pacientů. V posledním desetiletí se díky novým terapeutickým přístupům (zejména u některých skupin pacientů) mění spektrum kvasinek, které infekci způsobují. Předkládaná multicentrická studie si dala za úkol získat aktuální informace o epidemiologické situaci týkající se invazivních kandidóz (IC) v České republice. Materiál a metody: Do souboru byla zařazena data pacientů s kultivačně pozitivním nálezem kvasinky v krvi z 11 mikrobiologických pracovišť v České republice v rozmezí let 2012–2015. Byla hodnocena incidence, zastoupení jednotlivých kvasinkových druhů podle sledovaných let, podle věku a podle odborností. Dále byl posouzen vliv kolonizace a nakonec zda IC předcházela přítomnost toxigenního kmene Clostridium difficile ve stolici. U části izolovaných kmenů byly standardní metodikou stanoveny minimální inhibiční koncentrace (MIC) systémových antimykotik. Výsledky: V průběhu sledovaného období bylo v zúčastněných laboratořích zachyceno 921 kvasinkových kmenů od 886 pacientů. Celková incidence IC na 1 000 přijatých pacientů byla 0,40 (rozmezí 0,21–1,22 podle pracovišť). Takřka polovina izolátů spadala do druhu Candida albicans (49,7 %), následovaná Candida glabrata (15,3 %) a Candida parapsilosis (11,2 %). Na onkologických odděleních výrazně převažovaly non-albicans druhy (71,6 %), na rozdíl od chirurgických (40,4 %) nebo interních (52,0 %) oborů. Více než 70,0 % pacientů bylo v době pozitivní kultivace hospitalizováno na jednotkách intenzivní péče (JIP) a 65,0 % nemocných bylo kolonizováno stejným druhem kvasinky, který byl později nalezen v hemokultuře. Pouze u 5,1 % pacientů z celkového počtu byla potvrzena předchozí přítomnost toxigenního kmene C. difficile ve stolici. Celkem bylo otestováno 56 kmenů kvasinek (z 921) a byla zaznamenána zvýšená MIC u azolů pro C. glabrata. Závěr: Incidence kandidémií v České republice se ve sledovaném období výrazně neměnila a po C. albicans je druhou nejčastěji izolovanou kvasinkou v krvi C. glabrata.
Background: Candidemia is a severe and often life-threatening infection frequently occurring in critically ill patients. During the last decade, new therapeutic and prophylactic strategies influenced (at least in some patient subgroups) the epidemiological situation and the spectrum of causative Candida strains. The present multicentre study aimed to assess the current epidemiological situation of Candida strains causing invasive candidiasis (IC) in patients of tertiary care hospitals in the Czech Republic. Material and methods: Clinical and microbiological data on patients with bloodstream yeast isolates collected in 11 tertiary care hospitals in the Czech Republic between 2012 and 2015 were analysed. The incidence of cases and species distribution were assessed by study year, age, and specialty. Moreover, an association with the Candida colonization and presence of toxigenic strains of Clostridium difficile in stool prior to blood culture positivity was analysed. For some of the strains, minimum inhibitory concentrations (MICs) of systemic antifungals were determined using standard methods. Results: A total of 886 episodes of candidemia (921 yeast strains) were identified during the study period. The overall incidence per 1000 admissions was 0.40 (range 0.21–1.22 depending on the hospital). Almost half of the isolates belonged to the species Candida albicans (49.7 %), followed by Candida glabrata (15.3 %) and Candida parapsilosis (11.2 %). Non-albicans species of Candida significantly predominated in oncology wards (71.6 %) as compared to surgery (40.4 %) or internal medicine (52.0 %) departments. More than 70.0 % of patients stayed in intensive care units at the time of positive culture; in 65.0 % of patients, colonization with the same yeast species preceded blood culture positivity. In only 5.1 % of all patients, the previous presence of toxigenic strains of Clostridium difficile in stool was found. Fifty-six of the 921 yeast strains were tested for antifungal susceptibility, with an increase in MICs to azoles being observed for C. glabrata. Conclusion: The incidence of candidemia in the Czech Republic did not vary significantly between 2012 and 2015, and C. glabrata was the second most common yeast species after C. albicans isolated from blood.
- MeSH
- antifungální látky MeSH
- Candida klasifikace patogenita MeSH
- incidence MeSH
- infekce spojené se zdravotní péčí * MeSH
- kandidemie * diagnóza epidemiologie krev MeSH
- lidé MeSH
- mikrobiální testy citlivosti metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- multicentrická studie MeSH
- Geografické názvy
- Česká republika MeSH
RAB GTPases are important directional regulators of intracellular vesicle transport. Membrane localization of RAB GTPases is mediated by C-terminal double geranylgeranylation. This post-translational modification is catalyzed by the alpha-beta-heterodimer catalytic core of RAB geranylgeranyl transferase (RAB-GGT), which cooperates with the RAB escort protein (REP) that presents a nascent RAB. Here, we show that RAB-geranylgeranylation activity is significantly reduced in two homozygous mutants of the major Arabidopsis beta-subunit of RAB-GGT (AtRGTB1), resulting in unprenylated RAB GTPases accumulation in the cytoplasm. Both endocytosis and exocytosis are downregulated in rgtb1 homozygotes defective in shoot growth and morphogenesis. Root gravitropism is normal in rgtb1 roots, but is significantly compromised in shoots. Mutants are defective in etiolation and show constitutive photomorphogenic phenotypes that cannot be rescued by brassinosteroid treatment, similarly to the det3 mutant that is also defective in the secretory pathway. Transcriptomic analysis revealed an upregulation of specific RAB GTPases in etiolated wild-type plants. Taken together, these data suggest that the downregulation of the secretory pathway is interpreted as a photomorphogenic signal in Arabidopsis.
- MeSH
- Arabidopsis genetika růst a vývoj metabolismus MeSH
- gravitropismus MeSH
- prenylace MeSH
- proteiny huseníčku genetika metabolismus MeSH
- rekombinantní fúzní proteiny genetika metabolismus MeSH
- transferasy genetika metabolismus MeSH
- výhonky rostlin genetika růst a vývoj MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Vyd. 1. 134 s. : il. ; 24 cm
- MeSH
- dětská psychologie MeSH
- domovy s denní péčí MeSH
- pěstounství MeSH
- rodinné vztahy MeSH
- sociální péče o dítě MeSH
- výchova dítěte MeSH
- vývoj dítěte MeSH
- vztahy mezi rodiči a dětmi MeSH
- Publikační typ
- beletrie MeSH
- Geografické názvy
- Česká republika MeSH
- Konspekt
- Druhy sociální pomoci a služeb
- NLK Obory
- sociologie
- psychologie, klinická psychologie
- NLK Publikační typ
- vzpomínky
60 s. : il. ; 20 cm
- MeSH
- dětská psychologie MeSH
- pěstounství MeSH
- sociální péče o dítě MeSH
- Publikační typ
- beletrie MeSH
- Konspekt
- Druhy sociální pomoci a služeb
- NLK Obory
- sociologie
- psychologie, klinická psychologie
- NLK Publikační typ
- vzpomínky
