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Biomedical subjects

M Kolar

Publications and source records attributed to M Kolar.

14 recordsLinked to original sources

Prevalence of extended-spectrum beta-lactamase-positive Klebsiella pneumoniae isolates in the Czech Republic.

This study focused on the prevalence and molecular biology of extended-spectrum beta-lactamase (ESBL)-positive Klebsiella pneumoniae isolates collected in the Czech Republic. Clinical material from patients hospitalised in 16 Czech hospitals in September 2004 was used to isolate K. pneumoniae strains. Strains were identified by standard identification procedures. Susceptibility of the strains to antibiotics was tested using a microdilution method. The double-disk synergy test and combination disk method were used to determine ESBL production. Molecular biology characteristics of ESBL-positive isolates were determined using genomic DNA isolation, XbaI restriction digestion and pulsed-field gel electrophoresis differentiation. The acquired restriction maps of individual isolates were compared using GelCompar II software and their relationships were determined. During the 3-week period, 483 K. pneumoniae strains causing clinically detectable diseases were isolated. Of these, 117 (24.2%) were determined to be ESBL-positive. The prevalence of ESBL-positive isolates was 38.9% in Intensive Care Units (ICUs) and 13.1% in standard wards. More than 50% of ESBL-positive isolates were treated effectively only with meropenem (98%), cefoperazone/sulbactam (61%) and amikacin (54%). Conversely, ESBL-negative isolates showed high susceptibility to all tested antibiotics (76-99%). Molecular biology analysis identified 18 clonal types containing two to six identical isolates. Seventeen clones usually contained isolates from only one hospital; isolates from two hospitals were identified only in one clone. Based on the abovementioned results, the prevalence of ESBL-positive K. pneumoniae isolates in the Czech Republic can be perceived as relatively high, especially in ICUs. Extensive spread of 'epidemic clones' within Czech hospitals and, to a limited extent, between them can be demonstrated.

Czech Republic↗

The influence of antibiotic use on the occurrence of vancomycin-resistant enterococci.

BACKGROUND: Several studies have documented the influence of antibiotic selective pressure, mainly from the use of glycopeptides, third-generation cephalosporins, quinolones and lincosamides, on the frequency of vancomycin-resistant enterococci (VRE) occurrence in hospitals. The aim of this study was to evaluate the relationship between VRE occurrence and antibiotic use in the Department of Hemato-Oncology of the Teaching Hospital in Olomouc (DHO), Czech Republic, over a 6-year period under standard and unchanged hygienic and epidemiological conditions. METHODS: During the period of 1998-2003, Enterococcus sp. strains and VRE were isolated by standard methods from clinical samples taken from DHO in-patients. The frequency of VRE occurrence was expressed as the number of isolated strains per 100 bed-days/year. DHO antibiotic consumption data were processed according to the anatomical therapeutic chemical (ATC)/defined daily dose (DDD) system valid in 2003 and expressed in defined daily dose per 100 bed-days (DDD/100 bed-days) for each year of the period. RESULTS: Since 1998, the occurrence of VRE decreased significantly (from 0.28 to 0.17 VRE/100 bed-days in 2001). Between 2001 and 2003, a significant (P < 0.05) increase from 0.17 to 0.38 was observed. The antibiotic use decreased from 205.2 in 1998 to 161.0 DDD/100 bed-days in 1999 and after an increase in 2001 (to 181.8 DDD/100 bed-days) it remained relatively stable. A significant decrease was observed in third-generation cephalosporins and quinolones (from 29.5 to 9.7 and from 42.2 to 30.2 DDD/100 bed-days respectively) between 1998 and 1999. In 2002-2003, the use of third-generation cephalosporins and glycopeptides increased substantially (from 10.1 to 13.9 and from 11.3 to 15.2 DDD/100 bed-days respectively). The Pearson correlation value was significantly positive (P < 0.05) for VRE occurrence and the use of glycopeptides and third-generation cephalosporins. CONCLUSIONS: While our study confirms the effect of use of glycopeptides and third-generation cephalosporins on occurrence of VRE, no influence of quinolones and lincosamides over the 6-year period was shown.

Anti-Bacterial Agents↗

Genotypic characterisation of vancomycin-resistant Enterococcus faecium isolates from haemato-oncological patients at Olomouc University Hospital, Czech Republic.

This study describes the first molecular characterisation of clinical isolates of vancomycin-resistant enterococci (VRE) in the Czech Republic. Of 2647 patient isolates of Enterococcus spp. from 1997-2002, 121 (4.6%) were identified as VRE. The most common isolates were VanA+ Enterococcus faecium (78%) and VanB+ Enterococcus faecalis (10%). In addition, five VanA+ E. faecium isolates were obtained from environmental and staff sampling. Macrorestriction analysis of SmaI restriction fragment length polymorphism was performed for 54 VanA+ E. faecium clinical isolates and the five VanA+ E. faecium environmental isolates. Thirty-two unique restriction endonuclease patterns were identified, including two predominant clonal types represented by five or more isolates. Two environmental VanA+ E. faecium isolates were closely related to two patient isolates, which had an identical SmaI macrorestriction pattern. The results indicated potential survival of strains in the hospital environment and possible subsequent transmission to hospitalised patients.

Cancer Care Facilities↗

Can bioimpedance determine the volume of distribution of antibiotics in sepsis?

The relationship between the volume of distribution, assessed according to the two-compartmental pharmacokinetic model, and extracellular water estimated by bioimpedance was studied in mechanically ventilated patients with sepsis and capillary leak. A prospective observational study was performed in a twenty-bed general intensive care unit in the university hospital. Patients received either vancomycin (n = 16) or netilmicin (n = 12) for more than 48 hours. Those with ascites, pleural effusion, on renal replacement therapy or with haemodynamic instability were excluded. Serum concentrations of drugs were taken for pharmacokinetic analysis before, 1 hour and 4 hours after the 30 minute infusion. Bioimpedance measurement was performed at the time of the third sampling. The protocol was repeated after 24 hours. Fluid balance during the 24 hour interval was recorded. Extracellular water was increased and represented 45.6 to 46.6% of total body water Fluid balance correlated with the change of extracellular water (r = 0.82, P < 0.0001) and total body water (r = 0.74, P < 0.0001). Volumes of distribution of vancomycin (0.677 +/- 0.339 l/kg) and netilmicin (0.505 +/- 0.172 l/kg) were increased compared to normal values. A correlation was demonstrated between volume of distribution (Vd(area)) of vancomycin and extra cellular water/total body ratio (r = 0.70, P < 0.0001). The central compartment distribution volume (V1) of netilmicin correlated with extracellular water/total body water ratio (r = 0.60, P < 0.003). Serum concentrations above the recommended therapeutic range were detected in 81.2% of patients on vancomycin and in 50% of patients on netilmicin. Increased volumes of distribution can be estimated by the bioimpedance measurements but are not associated with requirements for higher dosage of the glycopeptide or aminoglycoside antibiotics.

Anti-Bacterial Agents↗

Urinary tract infection: the urological perspective.

Urinary tract infection in babies often presents with non specific symptoms and signs. It must be considered in the differential diagnosis of a febrile sick baby or any baby with failure to thrive. In a significant proportion of babies with recurrent urinary tract infection, urological abnormality is demonstrable. The diagnosis of urinary tract infection is confirmed by the presence of pus cells and growth of microorganism in a fresh urine specimen. The voidoing history and detailed ultrasound examination in the baby including a post void evaluation of the upper and lower urinary tract can lead to the possible diagnosis in the majority. Antibiotic therapy for urinary tract infection should be followed by consideration about the need for urinary drainage at the appropriate level, particularly in cases where resolution is delayed despite antibiotics. There is a role for prophylactic antibiotics after the resolution of acute infection, at least until detailed evaluation several weeks after the acute episode, has excluded any abnormality. Micturating cytourethrogram is usually done under antibiotic cover and better done in centres with facility for fluoroscopic examination. Isotope studies have to be evaluated in the light of inherent limitations. Transient urodynamic abnormality of the urinary bladder in infancy is being increasingly recognized and should be the subject of evaluation in specific circumstances. A multidisciplinary team improves the quality of investigations, subsequent interpretation and long term care of these children. A large majority of urological abnormality in infancy and early childhood can be effectively managed by endocopic procedures. Temporary urinary diversion may be required in a small proportion. Urological reconstruction should be approached with caution, after a detailed analysis of the evolving urological tract in a baby. An aggressive approach to early diagnosis and appropriate treatment of urinary tract infection and any underlying abnormality, has been shown to significantly reduce the incidence of chronic renal failure in long term longitudinal studies in children.

Child↗

Chemically prepared silver electrode for determination of N-acetyl-L-cysteine by flow-injection potentiometry.

This paper describes the use of the silver electrode by means of chemical pretreatment of the electrode surface with mercuric(II) chloride solution and potassium iodide solution in flow injection analysis (FIA). The electrode is used as a potentiometric sensor for the indirect determination of NAC in a carrier stream containing iodine. A one-channel flow system that consists of a peristaltic pump, injection valve, a silver wire electrode and a saturated calomel reference electrode (SCE) was used. Some typical FIA parameters such as flow rate, tube length and composition of the carrier stream were varied. The electrode is further characterised by a constant linear response within the concentration range for NAC between 4.0 x 10(-6) and 1.0 x 10(-3) M at the slope of 60.6 +/- 1.0 mV/p(NAC). Some pharmaceutical products containing NAC were also tested. These results can be compared to the results obtained by the direct potentiometric titrations with silver nitrate and are also in good agreement with values declared by pharmaceutical manufacturers.

Acetylcysteine↗

[Developing a network for psychosomatic medicine and psychotherapy in Austria].

In Austria additional in-patient treatment facilities for psychosomatic patients are currently established. In this context an advisory board of the planned psychosomatic clinics suggests to develop a network for psychosomatics and psychotherapy. This suggestion focuses on four goals: further integration of specialised psychosomatic care into general medical care; further promotion of general psychosomatic care in primary health care; increasing acceptance of psychotherapy among psychosomatic patients in need of psychotherapeutic interventions; decreasing the number of chronic psychosomatic disorders by early and adequate treatment. The proposed network should involve three levels of cooperation: the primary care level, the specialised psychosomatic-psychotherapeutic level and the institutional level of planning future structural developments according to the actual needs and demands. Such a network should also facilitate a common approach regarding the admission of patients, in-patient treatment and post-treatment care. A need inventory and an implementation analysis involving all levels of cooperation as well as the patients perspective are regarded as a prerequisite for the implementation of the network.

Aftercare↗

The occurrence of vancomycin-resistant enterococci in hematological patients in relation to antibiotic use.

Very important bacterial pathogens found in hematological patients at present are vancomycin-resistant enterococci (VRE). The main goal of this retrospective study was to assess their occurrence in relation to antibiotic use. We isolated 1918 Enterococcus strains, in toto, 138 (7.2%) of which proved to be VRE. The VRE most frequently identified were Enterococcus faecium VanA (77%) and Enterococcusfaecalis VanB (12%), mostly isolated from stools (57%). Comparing the development of the selection pressure of antibiotics and percentage of VRE in each period of observation, an effect of the administration of each antibiotic group on the occurrence of VRE can be presumed. A reduction in the administration of third generation cephalosporins, glycopeptides and fluoroquinolones and its replacement by penicillin antibiotics combined with inhibitors of bacterial beta-lactamases, contributed to the cessation of VRE incidence and succeeding reduced occurrence from 15.1% in the second half of 1998 to 6.1% in the first half of 2000.

Anti-Bacterial Agents↗

Potentiometric flow-injection determination of vitamin C and glutathione with a chemically prepared tubular silver electrode.

This paper describes the preparation and use of a tubular electrode by means of chemical pretreatment of a silver tube with mercuric (II) chloride solution and iodide solution in flow injection analysis (FIA). The electrode was used as a potentiometric sensor for the indirect determination of vitamin C and glutathione in a carrier stream containing iodine. A simple FIA system that consists of a peristaltic pump, an injection T valve, a tubular silver electrode and a saturated calomel reference electrode (SCE) was used. Some typical FIA parameters such as flow rate, tube length, sample volume and composition of the carrier stream were varied. After optimisation of these parameters, the electrode was further characterised by a constant linear response within the concentration range for the vitamin C between 5 x 10(-5) and 5 x 10(-3) M at the slope of 60.5 +/- 1.0 mV/p (vitamin C). Glutathione has a linear concentration range between 5 x 10(-5) and 10(-2) M at a slope of 55.2 +/- 1.0 mV/p (glutathione). The experimental slope was in good agreement with the theoretical values. Some pharmaceutical products containing vitamin C were also tested. These results can be compared to the results obtained by the standard volumetric method for the determination of vitamin C and are also in good agreement with values declared by pharmaceutical manufacturers.

Ascorbic Acid↗

Effects of resistance vs. aerobic training combined with an 800 calorie liquid diet on lean body mass and resting metabolic rate.

OBJECTIVE: Utilization of very-low-calorie diets (VLCD) for weight loss results in loss of lean body weight (LBW) and a decrease in resting metabolic rate (RMR). The addition of aerobic exercise does not prevent this. The purpose of this study was to examine the effect of intensive, high volume resistance training combined with a VLCD on these parameters. METHODS: Twenty subjects (17 women, three men), mean age 38 years, were randomly assigned to either standard treatment control plus diet (C+D), n = 10, or resistance exercise plus diet (R+D), n = 10. Both groups consumed 800 kcal/day liquid formula diets for 12 weeks. The C+D group exercised 1 hour four times/week by walking, biking or stair climbing. The R+D group performed resistance training 3 days/week at 10 stations increasing from two sets of 8 to 15 repetitions to four sets of 8 to 15 repetitions by 12 weeks. Groups were similar at baseline with respect to weight, body composition, aerobic capacity, and resting metabolic rate. RESULTS: Maximum oxygen consumption (Max VO2) increased significantly (p<0.05) but equally in both groups. Body weight decreased significantly more (p<0.01) in C+D than R+D. The C+D group lost a significant (p<0.05) amount of LBW (51 to 47 kg). No decrease in LBW was observed in R+D. In addition, R+D had an increase (p<0.05) in RMR O2 ml/kg/min (2.6 to 3.1). The 24 hour RMR decreased (p<0.05) in the C+D group. CONCLUSION: The addition of an intensive, high volume resistance training program resulted in preservation of LBW and RMR during weight loss with a VLCD.

Adipose Tissue↗

Development of a six station knee wear simulator and preliminary wear results.

In order to assess the wear performance of different designs of total knee replacements (TKR), a six station multi-axis knee simulator has been designed, built and commissioned. The most important features of a knee simulator are representative angles of flexion-extension synchronized with a dynamically applied load, and a combination of rolling and sliding motion. The simulator typically applies flexion-extension of 0-65, anterior-posterior translation of up to 15 mm, a dynamic load of up to 5.0 kN, and operates at 1.0 Hz. The loads and motions are applied using computer controlled servohydraulic actuators and hence their profiles are easily modified. A preliminary wear test has been conducted using a Kinemax (Howmedica, United Kingdom) TKR. The test was conducted in 30 per cent bovine serum which was changed every 150,000 cycles, at which time the bearing surfaces were examined and the UHMWPE tibial component was weighed. Over eight million cycles, a tibial wear rate of 2.62 mg/10(6) cycles was measured. The mild wear observed was characterized by burnishing and slight scratching in the anterior posterior direction. These observations are broadly in line with both in vitro and ex vivo studies reported in the literature for this type of prosthesis. Delamination wear sometimes observed in vivo was not seen.

Animals↗

Molecular characterization and functional analysis in Aspergillus nidulans of the 5'-region of the Penicillium chrysogenum isopenicillin N synthetase gene.

The isopenicillin N synthetase gene (pcbC) was isolated from a genomic library of Penicillium chrysogenum BC39813, a penicillin production strain. The nucleotide sequence, including 555 bp upstream of the translation start site was determined. Various deletions within the pcbC 5'-region were constructed and linked to the Escherichia coli lacZ gene. An Aspergillus nidulans argB strain was transformed with DNA of these constructions. The region essential for promoter function could be localized between positions -307 and -89 by analyzing beta-galactosidase expression of transformants containing a single copy of the corresponding plasmid integrated at the homologous argB locus. A region responsible for regulatory effects concerning nitrogen metabolism was identified by determining beta-galactosidase activities in cell-lysates of transformants cultivated under varying growth conditions. Two major transcription start sites at positions -131 and -132, as well as a further upstream located site at position -397 +/- 1 could be located by primer extension studies employing RNA isolated from P. chrysogenum BC39813.

Amino Acid Sequence↗

Transformation of Penicillium chrysogenum using dominant selection markers and expression of an Escherichia coli lacZ fusion gene.

An industrial Penicillium chrysogenum strain was transformed using two dominant selection markers, namely the bacterial gene for phleomycin resistance (ble) fused to a fungal promoter, and the acetamidase (amdS) gene from Aspergillus nidulans. Transformation frequencies of up to 20 transformants per microgram of DNA were obtained with the ble system. With the amdS marker the frequency was up to 120 transformants. Cotransformation was very efficient when using amdS as a selection marker. The introduction of pAN5-41B, a plasmid carrying the Escherichia coli lacZ gene fused to the strong glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter from A. nidulans, resulted in the formation of blue colonies on XGal plates indicating expression of the lacZ fusion gene in P. chrysogenum. A more detailed analysis of expression levels in several transformants showed that up to 6% of the total amount of soluble protein consists of the beta-galactosidase fusion protein.

Amidohydrolases↗