A switch from originator-adalimumab to the biosimilar SB5 in patients with Crohn's disease: an analysis of two propensity score-matched cohorts.

BACKGROUND/AIMS: Originator-adalimumab, an established treatment for patients with Crohn's disease (CD), showed no difference in efficacy or adverse events versus adalimumab biosimilar SB5 (SB5-adalimumab) over 10 weeks (W) of treatment. To understand the long-term effectiveness of SB5-adalimumab in CD, patients switched from originator-adalimumab to SB5-adalimumab were compared with patients remaining on originator-adalimumab over 104 W. METHODS: Data on patients aged ≥18 years, diagnosed with CD and treated at ISCARE, were collected prospectively from July 2018 to January 2021. Primary outcome: clinical disease activity at W52, measured by Harvey-Bradshaw index (HBI). Secondary outcomes: C-reactive protein (CRP), faecal calprotectin (FC) and adalimumab concentrations at W10, 26, 52 and 104, and treatment persistence. To ensure comparable cohorts, patients were propensity score (PS)-matched for age, gender and disease activity. RESULTS: After matching, 54 patients remained per cohort. At W52, mean (SD) HBI score was 3.2 (2.5) for originator-adalimumab and 4.0 [3.6] for SB5-adalimumab (difference [95% CI] -0.78 [-2.8, 1.3]; n = 18/cohort); no clinically meaningful differences in CRP, FC or drug concentrations were noted. Kaplan-Meier's estimates (95% CI) of remaining on treatment were originator-adalimumab: 0.870 (0.785-0.965) versus SB5-adalimumab: 0.648 (0.533-0.789) at W52 and significantly lower for SB5-adalimumab versus originator-adalimumab (p < .001) over 104 W. Local skin reaction events/pain was the main reason for treatment discontinuation in the SB5-adalimumab cohort (n = 20/54 [37%]). CONCLUSIONS: These long-term results of CD patients receiving originator-adalimumab or following nonmedical switch to SB5-adalimumab show similar therapeutic effects on clinical disease activity, biological parameters and pharmacokinetic profile in both cohorts from 52 to 104 W. A separation in persistence was observed beyond W26, mainly due to differences in local reactions at the injection site.

Diagnosis and treatment of surgical skin and soft tissue infections - current status.

Surgical skin and soft tissue infections (SSTIs) result from microbial invasion of the skin and underlying soft tissues, often requiring surgical treatment. SSTIs encompass a variety of pathological conditions, ranging from frequent simple superficial skin infections with very good outcomes to rare, rapidly progressive necrotizing infections associated with long-lasting morbidity and high mortality. The document summarizes current knowledge of the diagnosis and therapy of these diseases and provides clinicians with current standards of care of these patients based on international guidelines. Additionally, regional specific aspects are also reflected, and thus in all cases, this paper on diagnostic-therapeutic management of individual clinical forms respects the actual clinical practice and epidemiology in the Czech Republic. The document has been prepared based on multidisciplinary consensus of experts from universities all over the Czech Republic.

Pre-booster and post-booster seroprevalence of measles antibodies among employees of the Olomouc University Hospital - a pilot study.

INTRODUCTION: The increase in measles cases in early 2019 led to the implementation of several preventive measures focused mainly on health care providers. The study aimed to evaluate the seroprevalence of measles antibodies among employees of a large hospital and, a year apart, the rate of seroconversion in a pilot sample of the revaccinated subjects. METHODS: In 3027 employees of the University Hospital Olomouc, specific immunoglobulin G levels were tested on a voluntary basis. Those with insufficient levels were offered a booster dose. About approximately one year after the booster dose, the same test was performed in a sample of 52 employees. RESULTS: Of the tested subjects with a mean age of 41.8 ± 9.2 years, 54.0% were seropositive. A higher proportion of seropositivity as well as higher absolute values of antibody titers were noted in those born before routine vaccination was introduced in 1969. A total of 80.9% of the seronegative subjects opted for a booster dose. Seroconversion occurred in 73.2% of retested subjects. The relative increase of post-booster antibody titers was moderately correlated with age (r = 0.47, p < 0.05). CONCLUSION: The proportion of seronegative employees of a large hospital reached 46.0%, being higher in younger individuals. Seroconversion occurred in 73.2% of booster dose recipients included in a pilot sample for reanalysis. A statistically significant correlation was noted between the relative increase of antibody titers and age.

Nosocomial pneumonias.

The paper provides clear definitions of the basic concepts of nosocomial pneumonias. Specifically, definitions and classifications of HAP and VAP, general treatment principles and specific recommended procedures for antibiotic therapy are given as applicable in the Czech Republic.

Highly variable vancomycin-resistant enterococci in the north-eastern part of the Czech Republic.

Vancomycin-resistant enterococci (VRE) are nosocomial pathogens of increasing medical importance. This study involved 121 VRE selectively obtained from a representative set of 1464 samples collected from various sources in the north-eastern part of the Czech Republic. In total, 119 VRE belonged to Enterococcus faecium and two to Enterococcus faecalis. All isolates of E. faecium were resistant to at least three antibiotic classes. The resistance genes vanA, erm(B), tet(M), tet(L), aac(3)-IIIa and aac(6')-aph(2'') were detected. We assigned the E. faecium to sequence types ST5, ST18, ST38, ST64, ST92, ST273, ST549 and ST640. In E. faecium isolates, we identified the presence of replicases rep20p LG 1 , rep2p RE 25 , rep17p RUM , rep21p VEF 1/2 and rep14p RI 1 , as well as relaxases relp EF 1 , relp LG 1 , relp CIZ 2 , relp RE 25 and relp RUM . The presence of the toxin-antitoxin system axe-txe was detected mainly among isolates of hospital origin. The A and D types of transposon Tn1546 were those occurring most frequently. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first extensive study of vancomycin-resistant enterococci of diverse origin in a single well-defined area of the Czech Republic. The isolates were investigated for their antibiotic resistance, epidemiological characteristics and plasmid characteristics. Based on the results obtained, we can make assumptions as to the ways that vancomycin resistance is disseminated throughout the environment including humans and animals.

Principles of antibiotic therapy.

The paper describes the basic sources and principles of antibiotic therapy in contemporary medicine in which the ability to treat bacterial infections may be lost. The main reason for that is the increasing resistance of bacterial pathogens to antibiotics. A possible solution is to implement a comprehensive program of antibiotic stewardship incorporating adequate consideration of indication and selection of antimicrobial agents including appropriate duration and way of administration. Another important component of the comprehensive approach to bacterial resistance and antibiotic therapy is adequately applied antibiotic prophylaxis in surgery.

Infliximab Biosimilar (Remsima™) in Therapy of Inflammatory Bowel Diseases Patients: Experience from One Tertiary Inflammatory Bowel Diseases Centre.

BACKGROUND: The evidence on the efficacy and safety of biosimilar infliximab (IFX) in patients with inflammatory bowel diseases (IBD) is sparse. METHODS: Consecutive IBD patients visiting our centre were included. One cohort composed of prospectively followed patients who were switched from original to biosimilar IFX between January and March 2015. The second cohort included retrospectively assessed anti-tumor necrosis factor α-naïve patients who started therapy between January 2015 and January 2016. Disease activity was assessed using standard clinical indices, endoscopic evaluation, and laboratory parameters (blood count, C-reactive protein (CRP) and fecal calprotectin (FC)). Trough levels and anti-drug antibodies (ATIs) were also measured. Patients were evaluated 56 weeks (W56) after switch and at week 14 (W14) and week 46 (W46) in the naïve cohort. RESULTS: Seventy-four IBD patients were switched to biosimilar IFX and 119 naïve patients newly initiated therapy with the preparation. Disease activity remained stable in a majority of switched patients (remission at week 0 (W0) vs. W56: 72.2 vs. 77.8%; median difference of both Harvey-Bradshaw index and Simple Clinical Colitis Activity Index between W0 and W56 was 0). When W0 and W56 were compared, no significant difference in CRP (4.3 ± 8.0 vs. 3.3 ± 3.8 mg/l; p = 0.89) and FC (135 ± 153 vs. 199 ± 225 µg/g; p = 0.17) was observed. In total, 92% of Crohn's disease (CD) and 83% of ulcerative colitis (UC) patients responded to induction therapy (W14) with biosimilar IFX. At W46, the response rate was 86% in CD and 64% in UC. Moreover, half of UC patients experienced mucosal healing at W14 and improvement of perianal disease occurred in 95% of CD at W46. In this cohort, clear steroid-sparing effect was observed. No increase in immunogenicity was found in switched patients (ATI positivity: 9.5 vs. 6.0%, p = 0.54) and the type and frequency of adverse events were comparable to the original preparation in both cohorts. CONCLUSION: Switching of IBD patients from original to biosimilar IFX is effective and safe.

Virulence and antibiotic resistance genes in Campylobacter spp. in the Czech Republic.

OBJECTIVE: Thermotolerant species of the genus Campy-lobacter are the important agents causing human foodborne infections throughout the world. The aims of this study were to evaluate the presence of nine putative virulence genes in Campylobacter spp. isolated from patients and from foods (poultry meat, pork liver), to determine the resistance of Campylobacter isolates to eight antibiotic agents and to detect four resistance genes.Matherial and methods: The presence of the virulence genes cdtA, cdtB, cdtC, virB11, ciaB, wlaN, iam, dnaJ and racR was detected by polymerase chain reaction (PCR) in 94 Campylobacter spp. isolates from humans and 123 campylobacters from foods. The phenotypic resistance to selected antimicrobial agents was tested with microdilution method in 82 human isolates and 91 food isolates. The isolates with antibiograms were tested for the presence of blaOXA-61, tet(O), aph-3-1 and cmeB genes by PCR with specific primers. RESULTS: In both human and food C. jejuni isolates the preva-lence of the studied virulence genes, especially dnaJ, racR, ciaB genes and the toxigenic genes cdtA, cdtB, cdtC, was considerably higher than in C. coli isolates. The only exception was the iam gene identified in only C. coli. The tested isolates of both C. jejuni and C. coli were highly resistant to quinolone antibiotics. Additionally, C. coli was also more resistant to erythromycin, streptomycin and, in case of isolates from pork liver, to tetracycline. High prevalence rates of genes encoding antibiotic resistance was noted for the blaOXA-61 and tet(O) genes in both Campylobacter species. CONCLUSIONS: The presented study is the first to assess the presence of genes for virulence and resistance to antibiotics in thermotolerant Campylobacter spp. isolated from humans and foods in the Czech Republic. The resistance of Campylobacter isolates to eight antibiotic agents was also assessed. The prevalence of genes responsible for virulence and resistance is rather varied in thermotolerant Campylobacter spp.

Detection of the etiological agents of hospital-acquired pneumonia - validity and comparison of different types of biological sample collection: a prospective, observational study in intensive care patients.

BACKGROUND: There is still a lack of evidence as to which method of biological sample collection is optimal for identifying bacterial pathogens causing hospital-acquired pneumonia (HAP). Much effort has been made to find an easy and valid approach to be used in clinical practice. METHODS: The primary endpoint of this prospective, observational study was to determine the predictive value of oropharyngeal swab (OS) and gastric aspiration (GA) as simple and non-invasive methods for diagnosing HAP. Their efficacy was compared to endotracheal aspiration (ETA) and protected specimen brushing (PSB), the standard methods approved for HAP diagnosis. RESULTS: Initially, 56 patients were enrolled. Significant amounts of bacterial pathogens were detected in 48 patients (79 isolates) in Round A and in 39 patients (45 isolates) in Round B (after 72 hours). The sensitivity rates were: ETA 98%, PSB 31%, OS 64% and GA 67% in Round A and ETA 87%, PSB 32%, OS 74% and GA 42% in Round B. Strains of 12 bacterial species were identified in the samples. The three most common etiological agents (both rounds together) were Klebsiella pneumoniae (23.7%), Burkholderia multivorans (21.1%) and Pseudomonas aeruginosa (15.8%). CONCLUSIONS: Blind ETA is an optimum method for obtaining biological samples for identification of etiological agents causing HAP in intubated patients. Microbial etiological agents were more frequently detected in ETA samples than in those collected by PSB. If ETA/PSB results are negative, samples may be collected by OS and/or GA as these techniques followed ETA in terms of the frequency of pathogen detection.

Clonality of Bacterial Pathogens Causing Hospital-Acquired Pneumonia.

Hospital-acquired pneumonia (HAP) is one of the most serious complications in patients staying in intensive care units. This multicenter study of Czech patients with HAP aimed at assessing the clonality of bacterial pathogens causing the condition. Bacterial isolates were compared using pulsed-field gel electrophoresis. Included in this study were 330 patients hospitalized between May 1, 2013 and December 31, 2014 at departments of anesthesiology and intensive care medicine of four big hospitals in the Czech Republic. A total of 531 bacterial isolates were obtained, of which 267 were classified as etiological agents causing HAP. Similarity or identity was assessed in 231 bacterial isolates most frequently obtained from HAP patients. Over the study period, no significant clonal spread was noted. Most isolates were unique strains, and the included HAP cases may therefore be characterized as mostly endogenous. Yet there were differences in species and potential identical isolates between the participating centers. In three hospitals, Gram-negative bacteria (Enterobacteriaceae and Pseudomonas aeruginosa) prevailed as etiological agents, and Staphylococcus aureus was most prevalent in the fourth center.

[Detection of cell death markers as a tool for bacterial antimicrobial susceptibility testing]. / Detekcia markerov bunkovej smrti ako nástroj na testovanie citlivosti baktérií na antibiotiká.

Antimicrobial resistance among nosocomial pathogens has emerged as one of the most important health care problems in the new millennium. In this review, we present new methods for bacterial antimicrobial susceptibility testing, based on the detection of antibiotic-mediated cell death markers that could provide valuable alternatives to existing phenotypic approaches in the very near future.

[Effect of lipophosphonoxins on inhibition of bacterial colonization of bone cements]. / Vliv lipofosfonoxinu na inhibici bakteriální kolonizace kostních cementu.

OBJECTIVE: The study aimed at determining the ability of lipophosphonoxin DR5026 to inhibit the formation of bacterial biofilm on the bone cement surface and assessing potential development of bacterial resistance. MATERIAL AND METHODS: Bone cement (Hi-Fatigue Bone Cement 2x40, aap Biomaterials GmbH, Germany) was polymerized with lipophosphonoxin DR5026. Cement samples were cultured using bacterial suspension containing Staphylococcus epidermidis CCM7221 at an inoculum density of 106 CFU/mL. After three, 24, and 48 hours of incubation at 35 °C, the number of bacteria adhered to the sample was measured and their growth curve was plotted. In 14 cycles, strains of Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, and Streptococcus agalactiae were exposed to subinhibitory concentrations of DR5026 and the minimum inhibitory concentrations (MICs) were determined. RESULTS: After three hours of culture in the bacterial inoculum with an initial concentration of 106 CFU/mL, the number of colonies isolated from the cement sample treated with DR5026 was smaller by two orders of magnitude when compared to a control cement sample. After 24 and 48 hours of incubation, the number of CFU remained at 50 in the treated cement, whereas 109 CFU were cultured from control cement samples. The plotted growth curves for bacteria adhered to cements clearly showed the inhibitory effect of lipophosphonoxin on their growth and multiplication, particularly after 48 hours. Following 14 cycles of repeated exposure to subinhibitory concentrations of DR5026, no increase in MICs was noted in the tested strains. CONCLUSION: Lipophosphonoxin DR5026 used to treat bone cement was found to have antibacterial effects and to inhibit the formation of bacterial biofilm. Repeated exposure of the tested bacteria to subinhibitory concentrations of the above lipophosphonoxin did not induce their resistance or increase their MICs.Key words: bone cement - joint replacement infections - lipophosphonoxins - antibacterial effect - biofilm.

Degradability of hexachlorocyclohexanes in water using ferrate (VI).

Regarding environmental pollution, the greatest public and scientific concern is aimed at the pollutants listed under the Stockholm Convention. These pollutants are not only persistent but also highly toxic with a high bioaccumulation potential. One of these pollutants, γ-hexachlorocyclohexane (γ-HCH), has been widely used in agriculture, which has resulted in wide dispersion in the environment. Remediation of this persistent and hazardous pollutant is difficult and remains unresolved. Of the many different approaches tested, to date, none has used ferrates. This is unexpected as ferrates are generally believed to be an ideal chemical reagent for water treatment due to their strong oxidation potential and the absence of harmful by-products. In this paper, the degradation/transformation of HCHs by ferrates under laboratory conditions was studied. HCH was degraded during this reaction, producing trichlorobenzenes and pentachlorocyclohexenes as by-products. A detailed investigation of pH conditions during Fe(VI) application identified pH as the main factor affecting degradation. We conclude that ferrate itself is unreactive with HCH and that high pH values, produced by K2O impurity and the reaction of ferrate with water, are responsible for HCH transformation. Finally, a comparison of Fe(VI) with Fe(0) is provided in order to suggest their environmental applicability for HCH degradation.

[The prevalence of nasal carriage of Staphylococcus aureus aureus and methicillin-resistant S. aureus (MRSA) among general medicine students of the Palacky University Olomouc]. / Prevalence výskytu nosního nosicství kmenu Staphylococcus aureus a meticilin rezistentních kmenu S. aureus (MRSA) u studentu vseobecného lékarství LF UP v Olomouci.

INTRODUCTION: Staphylococcus aureus is one of the major agents of both healthcare- and community-associated infections. Nasal carriage of S. aureus has been reported in approximately 37% of healthy persons. OBJECTIVES: 1. To investigate the prevalence of nasal carriage of S. aureus and MRSA in healthy persons of the age group 18-26 years (general medicine students of the Palacký University Olomouc). This type of information is not available in the Czech Republic. 2. To find whether the prevalence of nasal carriage strains of S. aureus and MRSA varies over the years of studies. 3. To compare the Year 1 (baseline) and Year 5 (after a long, close contact with the indoor environment of health-care facilities) general medicine students for nasal carriage of S. aureus and MRSA strains. MATERIALS AND METHODS: The prevalence study was conducted for over 12 months in 2012 and 2013. Nasal swabs were taken from the general medicine students participating in the study. Each subject was sampled once. RESULTS: In total, 307 samples were collected: 206 samples from the Year 1 students and 101 from the Year 5 students. Sixty-one (about 32%) Year 1 students and 32 (30%) Year 5 students were positive for S. aureus. Of them, 94 (30.6%) were males and 213 (69.4%) were females. CONCLUSIONS: The observed prevalence of nasal S. aureus carriage is consistent with the literature data. No statistically significant difference in the prevalence of nasal carriage of S. aureus was found between the Year 1 and Year 5 general medicine students.

Prevalence study on carbapenemase-producing Escherichia coli and Klebsiella pneumoniae isolates in Czech hospitals--results from Czech Part of European Survey on Carbapenemase--Producing Enterobacteriaceae (EuSCAPE).

OBJECTIVE: One of the most important threats of current medicine is the spread of multiresistant Gram-negative bacteria. We report here data from a six-month prevalence study on carbapenemase-producing K. pneumoniae and E. coli performed in Czech hospitals participating on European Survey on Carbapenemase-Producing Enterobacteriaceae (EuSCAPE). METHODS: Ten hospitals covering all regions of the Czech Republic were selected. During the study period (1st November 2013 to 30th April 2014), first ten carbapenem non-susceptible isolates of K. pneumoniae or E. coli isolated from non-surveillance specimens (i.e., blood, lower respiratory tract secretions, urine, puncture fluids, and wound secretions) of single successive patients were collected. Successive carbapenem-susceptible isolates of the same species were also preserved as controls. Susceptibility to 15 antibiotics was determined using EUCAST recommendations. Carbapenemase activity was detected by MALDI-TOF MS meropenem hydrolysis assay. Positive isolates were subjected for molecular typing (multi-locus sequence typing, identification of carbapenemase gene). RESULTS: During the study period, thirty non-susceptible isolates (K. pneumoniae n=28, E. coli n=2) were identified in 5 hospitals. Only two of them were confirmed to be carbapenemase producers. A NDM-1-producing K. pneumoniae ST11 was recovered from a patient, transferred from Ukraine, being injured during a Maidan revolution. The second isolate, an OXA-48-producing K. pneumoniae, belonging to ST101, was recovered from a patient admitted to a hospital for an ischemic stroke. CONCLUSIONS: This study again confirmed that the Czech Republic still belongs to the countries with low prevalence of carbapenemase-producing Enterobacteriaceae (CPE). Cases of CPE are usually restricted to an import from high-prevalence countries or countries with unknown epidemiological situation.

[Prevalence and characteristics of thermotolerant Campylobacter spp. in the human food chain]. / Výskyt a charakteristika termotolerantních kampylobakteru v potravinovém retezci cloveka.

OBJECTIVES: To monitor the prevalence of Campylobacter spp. in poultry in slaughterhouses, poultry and pork liver at retail, and cows milk in Moravia. To determine the resistance of animal isolates to selected antibiotics; and to compare it with an antibiogram of human strains. MATERIAL AND METHODS: Throughout the year 2013, the following samples were collected in the South Moravian and Olomouc Regions: mixed samples of broiler cecal contents in slaughterhouses, fresh and frozen chickens and pork liver at retail, and raw cows milk from vending machines. The samples were both qualitatively and quantitatively analyzed for the presence of Campylobacter spp. The isolates recovered were tested for resistance to antibiotics. For comparison, antimicrobial resistance was also studied in human isolates from the same regions. RESULTS: A total of 41.8% of the tested food samples were found to contain Campylobacter spp.. The most contaminated (73.2%) were fresh chickens. Campylobacter spp. were not detected in raw cows milk samples. The isolates showed high levels of resistance to quinolone antibiotics and, in the case of C. coli, also to tetracycline and streptomycin. CONCLUSION: The studied commodities were frequently contaminated with Campylobacter spp. The levels of contamination (in CFU/g) varied between commodities and so, evidently, did the real risk for human infections. When antibiotic therapy is needed, quinolone antibiotics cannot be used. Adherence to high standards of consumer safe food handling is crucial for the prevention of diseases.

[Basic principles of diagnosis and treatment of secondary peritonitis - recommendations of experts with the support of SIS]. / Základní principy diagnostiky a lécby sekundární peritonitidy - doporucení odborníku s podporou SIS.

Secondary peritonitis is the most common cause of inflammatory acute abdomen treated at general surgery departments. Only early and correct diagnosis may improve the prognosis of these patients. The authors compiled an interdisciplinary review of the basic principles of diagnosis and treatment of secondary peritonitis, which reflects current findings supported by evidence-based medicine. The work is based on published international literature but also shares opinions and experiences of the selected specialists. The presented work in its extent is not meant to substitute an in-depth study of the issue, but to allow a basic and quick review of the topic.

Quantifying acyl-chain diversity in isobaric compound lipids containing monomethyl branched-chain fatty acids.

Compound lipids comprise a diverse group of metabolites present in living systems, and metabolic- and environmentally-driven structural distinctions across this family is increasingly linked to biological function. However, methods for deconvoluting these often isobaric lipid species are lacking or require specialized instrumentation. Notably, acyl-chain diversity within cells may be influenced by nutritional states, metabolic dysregulation, or genetic alterations. Therefore, a reliable, validated method of quantifying structurally similar even-, odd-, and branched-chain acyl groups within intact compound lipids will be invaluable for gaining molecular insights into their biological functions. Here we demonstrate the chromatographic resolution of isobaric lipids containing distinct combinations of straight-chain and branched-chain acyl groups via ultra-high-pressure liquid chromatography (UHPLC)-mass spectrometry (MS) using a C30 liquid chromatography column. Using metabolically-engineered adipocytes lacking branched-keto acid dehydrogenase A (Bckdha), we validate this approach through a combination of fatty acid supplementation and metabolic tracing using monomethyl branched-chain fatty acids and valine. We observe resolution of numerous isobaric triacylglycerols and other compound lipids, demonstrating the resolving utility of this method. This approach strengthens our ability to quantify and characterize the inherent diversity of acyl chains across the lipidome.

[Microbial and pharmacological characteristics of VancogenX]. / Mikrobiologické a farmakologické vlastnosti kostního cementu VancogenX.

PURPOSE OF THE STUDY: Prosthetic joint infection is a feared complication in total hip arthroplasty. The use of antibiotic-impregnated bone cement is the important part of preventive and therapeutic strategies. At present a number of commercial bone cements are available and support of their use by the results of experimental trials and clinical studies has varied. In relation to this issue we studied essential microbiological and pharmacological characteristics of VancogenX in comparison with gentamicin-loaded bone cement used conventionally. MATERIAL AND METHODS: Based on a previously described protocol, we tested pellets of four commercial bone cements, namely, Hi-Fatigue G, Palacos R+G, VancogenX, and Palacos R as a control. Bone cement was prepared in a vacuum-mixing system. The strains used for bacterial load included Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Pseudomonas aeruginosa and Escherichia coli. Each cement was tested for its antimicrobial and antibiofilm activities and the results were evaluated by standard methods. In addition, we investigated time-related release of gentamicin and vancomycin from the bone cements tested. RESULTS: All antibiotic-loaded cement pellets were able to prevent growth of the bacterial strains tested. The bactericidal effect lasted for several days in relation to the bacterial species and cement used, with the exception of S. epidermidis whose growth was inhibited by gentamicin-loaded cement only for one day. The antibiotic-loaded pellets also prevented the formation of a biofilm for 24 hours at least. The major amount of vancomycin (32.915 mg/l) was released from VancogenX to MH medium within 24 hours and the last measureable amount (4.327 mg/l) was recorded at 48 hours after the start of the experiment. In physiological saline the highest level of vancomycin was 139.852 mg/ml measured at 24 hours, and the antibiotic was detectable at a level of 2.334 mg/ml as late as 8 days after the experiment started. Release of gentamicin from VancogenX was as follows: the 24-hour level was higher in MH medium than physiological saline (178 versus 131.4 mg/ml); however, gentamicin was still detectable in physiological saline at 192 hours after the start of the experiment while no gentamicin was found in MH medium after 72 hours. DISCUSSION: The antimicrobial effect of VancogenX bone cement was not an unexpected finding since both gentamicin and vancomycin have been used with bone cement for a long time and their combination is optimal in terms of preventing prosthetic joint infection. However, there is a disputable issue of poor release of vancomycin from bone cement. In MH medium we were able to detect the vancomycin released from VancogenX only for two days after the initial rapid elution while its release into physiological saline seemed to be slower but much longer. It is possible that more vancomycin is released from bone cement, but this amount is immediately bound to proteins in the cement vicinity and this process is not detectable by any analytical method. CONCLUSIONS: The bone cement VancogenX showed excellent antimicrobial and antibiofilm properties and can be recommended for use in orthopaedic practice. Therapy of prosthetic joint infection is the main indication. Extension of the indication to the prevention of prosthetic joint infection in high-risk patients should be preceded by biomechanical studies demonstrating that the cement is appropriate for long-term implant fixation.

[Rapid identification of ESBL--positive clinical samples using real-time PCR method]. / Rychlá identifikace ESBL-pozitivních klinických vzorku metodou real-time PCR.

OBJECTIVES: A new method has been developed for detecting genes determining the extended-spectrum beta-lactamase (ESBL) phenotype directly from patients' clinical material. The method enables detection of the bla(CTX-M) gene encoding CTX-M beta-lactamases and the bla(SHV) gene variants with real-time PCR technology using locked nucleic acid oligonucleotides. MATERIAL AND METHODS: In this pilot study, tracheal aspirates obtained from patients with mechanical ventilation hospitalized at Department of Anaesthesiology and Resuscitation of the University Hospital in Olomouc between 1st March and 30th December 2010 period were tested. Each sample was identified with standard microbiological procedures including phenotypic determination of ESBL-positive enterobacteria. At the same time, each sample was analyzed for the presence of nucleic acids (DNA) which encode CTX-M and SHV ESBL using real-time PCR. RESULTS: 150 samples of tracheal aspirates from 71 patients were included into testing. In the set, 13 (8.7%) ESBL-positive samples were identified by culture methods while 27 (18 %) positive samples were identified by the real-time PCR method. Of the 27 PCR-positive samples, 24 were positive for the bla(CTX) gene; in 2 samples, the ESBL bla(SHV) gene was detected, and both genes were present in 1 sample. All culture-positive samples were also PCR-positive for the presence of bla(CTX) and/or bla(SHV) sequences. CONCLUSIONS: The new real-time PCR assay is likely to shorten the time for detection of enterobacteria producing SHV and CTX-M beta-lactamases from 48 to 6 hours. It enables ESBL-positive enterobacteria determination in tracheal aspirates of patients suffered from life-threatening nosocomial pneumonia where the early introduction of adequate antimicrobial treatment plays the important role.