• MeSH
• Bacteria růst a vývoj   MeSH
• buněčná diferenciace MeSH
• enzymová indukce MeSH
• enzymová represe MeSH
• enzymy biosyntéza   MeSH
• genetika MeSH
• molekulární biologie MeSH

Intact cell MALDI-TOF mass spectrometry is a rapid tool for the identification and classification of microorganisms, now widely used even in clinical laboratories. However, its distinctive power is not sufficient for some closely-related species. The genus Cronobacter, formerly known as Enterobacter sakazakii, contains such species. In this work, a new method for the differentiation of five Cronobacter species is presented involving the tryptic digestion of cytoplasmatic proteins followed by MALDI mass spectrometry analysis. A database was developed for use in Bruker Biotyper software including 52 reference spectra and tested on a set of 45 samples with an overall accuracy of about 80%. The possibility of measurement automation and the short time and low cost requirements of this method compared to those of biochemical tests or PCR methods make it a supplementary option to intact cell MALDI, providing additional information about the differentiation of problematic species.

• MeSH
• bakteriologické techniky metody   MeSH
• Cronobacter sakazakii izolace a purifikace   MeSH
• Cronobacter izolace a purifikace   MeSH
• polymerázová řetězová reakce metody   MeSH
• software MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice metody   MeSH
• techniky typizace bakterií metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Routinely used biomarkers of bacterial etiology of infection, such as C-reactive protein and procalcitonin, have limited usefulness for evaluation of infections since their expression is enhanced by a number of different conditions. Therefore, several inflammatory cytokines and chemokines were analyzed with sera from patients hospitalized for moderate bacterial and viral infectious diseases. In total, 57 subjects were enrolled: 21 patients with community-acquired bacterial infections, 26 patients with viral infections, and 10 healthy subjects (control cohorts). The laboratory analyses were performed using Luminex technology, and the following molecules were examined: IL-1Ra, IL-2, IL-4, IL-6, IL-8, TNF- α , INF- γ , MIP-1 β , and MCP-1. Bacterial etiology of infection was associated with significantly (P < 0.001) elevated serum concentrations of IL-1Ra, IL-2, IL-6, and TNF- α in comparison to levels observed in the sera of patients with viral infections. In the patients with bacterial infections, IL-1Ra and IL-8 demonstrated positive correlation with C-reactive protein, whereas, IL-1Ra, TNF- α , and MCP-1 correlated with procalcitonin. Furthermore, elevated levels of IL-1Ra, IL-6, and TNF- α decreased within 3 days of antibiotic therapy to levels observed in control subjects. The results show IL-1Ra as a potential useful biomarker of community-acquired bacterial infection.

• MeSH
• antagonista receptoru pro interleukin 1 krev   MeSH
• bakteriální infekce krev   MeSH
• biologické markery krev   MeSH
• C-reaktivní protein metabolismus   MeSH
• chemokiny krev   MeSH
• cytokiny krev   MeSH
• dospělí MeSH
• infekce získané v komunitě krev   MeSH
• interleukin-2 krev   MeSH
• interleukin-4 krev   MeSH
• interleukin-6 krev   MeSH
• interleukin-8 krev   MeSH
• kalcitonin krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• proteinové prekurzory krev   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• TNF-alfa krev   MeSH
• Check Tag
• 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
• práce podpořená grantem MeSH

Toxin-antitoxin systems (TAS) emerged more than 25 years ago and developed as an important field in molecular microbiology. TAS are autoregulated operons coding a stable toxin and an unstable antitoxin found in plasmids and chromosomes of Bacteria and Archaea. The conditional activation of their toxins interferes with cell growth/viability and, depending on the context, can influence plasmid maintenance, stress management, bacterial persistence, cell differentiation and, likely, bacterial virulence. This review summarizes recent results on the parD system of plasmid R1 and on the chromosomal relBE systems found in Escherichia coli and in Streptococcus pneumoniae with a focus on the RNase activity of their toxins, their regulation and their biomedical applications and implications.

• MeSH
• antitoxiny genetika   imunologie   metabolismus   MeSH
• bakteriální toxiny genetika   imunologie   metabolismus   MeSH
• biotechnologie metody   trendy   MeSH
• buňky - růstové procesy genetika   imunologie   MeSH
• financování organizované MeSH
• inhibitory syntézy proteinů imunologie   metabolismus   MeSH
• lidé MeSH
• mikrobiologie trendy   MeSH
• molekulární biologie metody   trendy   MeSH
• plazmidy genetika   MeSH
• ribonukleasy genetika   imunologie   toxicita   MeSH
• ribozomy genetika   imunologie   MeSH
• RNA genetika   imunologie   toxicita   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

Timely knowledge of the bacterial etiology and localization of infection are important for empirical antibiotic therapy. Thus, the goal of this study was to evaluate routinely used biomarkers together with novel laboratory parameters in the diagnosis of infection. METHODS: In this prospective study, 54 adult patients with bacterial infections admitted to the Department of Infectious Diseases were included. For comparison, 27 patients with viral infections were enrolled. In these patients, white blood cell (WBC) counts, differential blood counts, serum levels of procalcitonin (PCT), IL-1β, IL-6, IL-8, IL-10, IL-12, TNF-α, IFN-γ, soluble CD14 (sCD14), heparin-binding protein (HBP), cortisol (Cort), and monocyte surface expression of TLR2, TLR4, HLA-DR, and CD14 were analyzed. Also, these biomarkers were evaluated in 21 patients with acute community-acquired bacterial pneumonia (CABP), as well as in 21 patients with pyelonephritis and urosepsis. RESULTS: The highest sensitivity and specificity (expressed as the area under the curve [AUC]) for bacterial infection were observed in serum concentration of PCT (0.952), neutrophil and lymphocyte counts (0.852 and 0.841, respectively), and serum levels of HBP (0.837), IL-6 (0.830), and Cort (0.817). In addition, the serum levels of IFN-γ and Cort were significantly higher and IL-8 levels were lower in CABP when compared to pyelonephritis or urosepsis. CONCLUSIONS: From the novel potential biomarkers, only PCT demonstrated superiority over the routine parameters in the differentiation of bacterial from viral infections. However, some of the novel parameters should be further evaluated in larger and better characterized cohorts of patients in order to find their clinical applications.

• MeSH
• bakteriální infekce krev   diagnóza   MeSH
• biologické markery MeSH
• dospělí MeSH
• infekce získané v komunitě krev   diagnóza   MeSH
• kalcitonin krev   diagnostické užití   MeSH
• lidé středního věku MeSH
• lidé MeSH
• močové ústrojí mikrobiologie   MeSH
• prediktivní hodnota testů MeSH
• prospektivní studie MeSH
• proteinové prekurzory krev   diagnostické užití   MeSH
• pyelonefritida krev   diagnóza   MeSH
• senzitivita a specificita MeSH
• sepse krev   diagnóza   MeSH
• stupeň závažnosti nemoci MeSH
• virové nemoci krev   diagnóza   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Geografické názvy
• Česká republika MeSH

The aim of this study was to evaluate the serum levels of calprotectin and calgranulin C and routine biomarkers in patients with bacterial sepsis (BS). The initial serum concentrations of calprotectin and calgranulin C were significantly higher in patients with BS (n = 66) than in those with viral infections (n = 24) and the healthy controls (n = 26); the level of calprotectin was found to be the best predictor of BS, followed by the neutrophil-lymphocyte count ratio (NLCR) and the level of procalcitonin (PCT). The white blood cell (WBC) count and the NLCR rapidly returned to normal levels, whereas PCT levels normalized later and the increased levels of calprotectin, calgranulin C, and C-reactive protein persisted until the end of follow-up. Our results suggest that the serum levels of calprotectin are a reliable biomarker of BS and that the WBC count and the NLCR are rapid predictors of the efficacy of antimicrobial therapy.

• MeSH
• bakteriální infekce krev   diagnóza   MeSH
• biologické markery krev   MeSH
• C-reaktivní protein analýza   MeSH
• dospělí MeSH
• kinetika MeSH
• leukocytární L1-antigenní komplex krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• neutrofily cytologie   MeSH
• počet leukocytů MeSH
• počet lymfocytů MeSH
• protein S100A12 krev   MeSH
• senioři MeSH
• senzitivita a specificita MeSH
• sepse krev   diagnóza   MeSH
• virové nemoci krev   diagnóza   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

OBJECTIVE: Pulmonary endarterectomy (PEA) is an effective and potentially curative treatment for chronic thrombo-embolic pulmonary hypertension (CTEPH). The postoperative course after PEA is accompanied by a number of complications, which contribute to the high rate of early postoperative mortality. Markers allowing the early detection of infectious complication during the postoperative period may be of major clinical importance. The aim of the prospective study was to analyse a predictive value of five inflammatory markers to recognise inflammatory complications accompanying PEA before the first clinical signs of infection. METHODS: Eighty-two patients with CTEPH, who underwent PEA using cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA), were included into the study. Procalcitonin (PCT), tumour necrosis factor-α, interleukin (IL)-6, IL-8 and C-reactive protein arterial concentrations were measured before sternotomy and repeatedly up to 72h after the end of surgery. Haemodynamic parameters, infectious and non-infectious complications were recorded. RESULTS: Postoperative course was uncomplicated in 59/82 patients (group 1). Fourteen out of 82 patients (group 2) developed an infection in the first 3 days after surgery (bronchopneumonia, n = 9; bacterial sepsis, n = 5). Nine out of 82 patients (group 3) developed non-infectious complications in the same period. PCT and IL-6 were the only significant independent predictors of infection in days 1-3 after PEA. The area under receiver operating characteristic (ROC) curve calculated for PCT to predict postoperative infection was 0.83 (95% confidence interval (CI): 0.74-0.92) compared with 0.74 (95% CI: 0.68-0.81) for IL-6. With the cut-off 2.3 ng ml(-1), the test characteristics of PCT were as follows: sensitivity, 86%; specificity, 83%; negative predictive value, 92%; and positive predictive value, 84%. CONCLUSIONS: The increase in PCT and IL-6 may allow patients at increased risk of infection after PEA to be identified, allowing earlier institution of antibiotic treatment. These changes that occur before infection can be detected clinically. This finding may make the daily monitoring of PCT post-PEA useful.

• MeSH
• arteria pulmonalis chirurgie   MeSH
• bakteriální infekce krev   diagnóza   etiologie   MeSH
• biologické markery krev   MeSH
• C-reaktivní protein metabolismus   MeSH
• časná diagnóza MeSH
• endarterektomie metody   MeSH
• epidemiologické metody MeSH
• interleukin-6 krev   MeSH
• kalcitonin krev   MeSH
• kardiopulmonální bypass MeSH
• lidé středního věku MeSH
• lidé MeSH
• mediátory zánětu krev   MeSH
• plicní embolie komplikace   chirurgie   MeSH
• plicní hypertenze etiologie   chirurgie   MeSH
• pooperační komplikace krev   diagnóza   MeSH
• pooperační péče metody   MeSH
• proteinové prekurzory krev   MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: The aim of this prospective study was to evaluate the role of serum IL-6 as a potential predictive biomarker of postoperative complications (POC) in elective colorectal surgery. METHOD: A total of 115 patients underwent colorectal surgery for malignancy. IL-6 was measured on the first and third postoperative days (POD1, POD3), and C-reactive protein (CRP) was measured on the POD3. POC was analysed in subgroups according to Clavien‒Dindo (CD), antibiotic (ATB) treatment, intensive care unit (ICU) and hospital length of stay. The predictive power of variables for evaluated endpoints was analysed using receiver-operating characteristic (ROC) analysis and described by area under the curve (AUC). ROC analysis was adopted for the identification of optimal cut-offs. Histological analysis was performed to verify IL-6 production by the tumour. RESULTS: Out of 115 patients who were analysed, 42% had POC. Patients with POC had significantly higher serum levels of IL-6 on POD1 (p < 0.001) and POD3 (p < 0.001). IL-6 early on POD1 as a predictor of antibiotic treatment, ICU stay and hospital stay (AUC 0.818; 0.811; 0.771) did not significantly differ from the AUC of CRP late on POD3 (0.879; 0.838, 0.752). A cut-off IL-6 value of 113 pg/ml on POD1 and 180.5 pg/ml on POD3 in severe complications (CD > 3a) resulted in 75% and 72% sensitivity, 78.6% and 99% specificity, negative predictive value 96.4% and 97% and positive predictive value 29% and 88.9%. CONCLUSION: The serum level of interleukin-6 can predict severe (CD > 3a) POC early on POD1. On POD3, IL-6 is superior to CRP in terms of high positive predictive power of severe POC. Interestingly, the advantage of IL-6 on POD1 is early prediction of the need for antibiotic treatment, ICU stay and hospital stay, which is comparable to the CRP serum level late on the third POD.

• MeSH
• antibakteriální látky MeSH
• biologické markery MeSH
• C-reaktivní protein analýza   MeSH
• interleukin-6 * MeSH
• kolorektální chirurgie * MeSH
• lidé MeSH
• pooperační komplikace etiologie   MeSH
• prospektivní studie MeSH
• ROC křivka MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

An investigation was made of the adhesion, growth and differentiation of osteoblast-like MG-63 and Saos-2 cells on titanium (Ti) and niobium (Nb) supports and on TiNb alloy with surfaces oxidized at 165°C under hydrothermal conditions and at 600°C in a stream of air. The oxidation mode and the chemical composition of the samples tuned the morphology, topography and distribution of the charge on their surfaces, which enabled us to evaluate the importance of these material characteristics in the interaction of the cells with the sample surface. Numbers of adhered MG-63 and Saos-2 cells correlated with the number of positively-charged (related with the Nb2O5 phase) and negatively-charged sites (related with the TiO2 phase) on the alloy surface. Proliferation of these cells is correlated with the presence of positively-charged (i.e. basic) sites of the Nb2O5 alloy phase, while cell differentiation is correlated with negatively-charged (acidic) sites of the TiO2 alloy phase. The number of charged sites and adhered cells was substantially higher on the alloy sample oxidized at 600°C than on the hydrothermally treated sample at 165°C. The expression values of osteoblast differentiation markers (collagen type I and osteocalcin) were higher for cells grown on the Ti samples than for those grown on the TiNb samples. This was more particularly apparent in the samples treated at 165°C. No considerable immune activation of murine macrophage-like RAW 264.7 cells on the tested samples was found. The secretion of TNF-α by these cells into the cell culture media was much lower than for either cells grown in the presence of bacterial lipopolysaccharide, or untreated control samples. Thus, oxidized Ti and TiNb are both promising materials for bone implantation; TiNb for applications where bone cell proliferation is desirable, and Ti for induction of osteogenic cell differentiation.

• MeSH
• biologické markery metabolismus   MeSH
• buněčná adheze účinky léků   MeSH
• buněčná diferenciace účinky léků   MeSH
• buněčné linie MeSH
• kolagen typu I metabolismus   MeSH
• lidé MeSH
• lipopolysacharidy farmakologie   MeSH
• makrofágy cytologie   účinky léků   metabolismus   MeSH
• myši MeSH
• osteoblasty cytologie   účinky léků   metabolismus   MeSH
• osteokalcin metabolismus   MeSH
• oxidace-redukce MeSH
• povrchové vlastnosti MeSH
• proliferace buněk účinky léků   MeSH
• slitiny chemie   farmakologie   MeSH
• statická elektřina MeSH
• tkáňové podpůrné struktury * MeSH
• TNF-alfa farmakologie   MeSH
• vysoká teplota MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: CD157 has been recently reported as a useful glycosylphosphatidylinositol (GPI)-linked marker for the detection of paroxysmal nocturnal hemoglobinuria (PNH) clones in patients with suspected paroxysmal nocturnal hemoglobinuria by flow cytometry as it targets both neutrophils and monocytes. The aim of this study is to test the feasibility of a non-fluorescent aerolysin (FLAER) approach and propose an alternative for laboratories, where FLAER is not available. METHODS: We validated a non-FLAER-based single-tube, 6-color assay targeting the GPI-linked structures CD157, CD24, and CD14. We determined its performance characteristics on 20 PNH patient samples containing a variety of clone sizes and compared results with a previously validated FLAER-based approach. RESULTS: Coefficient of variation (CV) for intra-/interassay precision analyses ranged from 0.1%/0.2% to 3.02%/7.58% for neutrophils and from 0.10%/0.3% to 5.39%/6.36% for monocytes. Coefficient of determination (r2 ) for linear regression analysis of PNH clones from 20 patients ranging from 0.06% to 99.7% was 0.99 in all cases, Wilcoxon ranks test showed no statistically significant differences (P > 0.05), Bland-Altman analysis demonstrated performance agreement with mean bias ranging from 0.06 to 0.2. CONCLUSION: Our results confirm very good performance characteristics for both intra- and interassay precision analyses, favorable correlation, and agreement between the FLAER and non-FLAER-based approaches, using the CD157 GPI marker. Our experience suggests that a rapid and cost-effective simultaneous evaluation of PNH neutrophils and monocytes by flow cytometry without using FLAER is possible in areas where FLAER may not be widely available. © 2016 International Clinical Cytometry Society.

• MeSH
• ADP-ribosylcyklasa imunologie   metabolismus   MeSH
• antigen CD24 imunologie   metabolismus   MeSH
• antigeny CD14 imunologie   metabolismus   MeSH
• bakteriální toxiny MeSH
• biologické markery metabolismus   MeSH
• CD antigeny imunologie   metabolismus   MeSH
• cytotoxické proteiny tvořící póry MeSH
• GPI-vázané proteiny imunologie   metabolismus   MeSH
• lidé MeSH
• monocyty imunologie   metabolismus   MeSH
• neutrofily imunologie   metabolismus   MeSH
• paroxysmální hemoglobinurie imunologie   metabolismus   MeSH
• průtoková cytometrie metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH