Effects of fluvastatin and coenzyme Q10 on skeletal muscle in normo- and hypercholesterolaemic rats.

Myalgia and muscle weakness may appreciably contribute to the poor adherence to statin therapy. Although the pathomechanism of statin-induced myopathy is not completely understood, changes in calcium homeostasis and reduced coenzyme Q10 levels are hypothesized to play important roles. In our experiments, fluvastatin and/or coenzyme Q10 was administered chronically to normocholesterolaemic or hypercholaestherolaemic rats, and the modifications of the calcium homeostasis and the strength of their muscles were investigated. While hypercholesterolaemia did not change the frequency of sparks, fluvastatin increased it on muscles both from normocholesterolaemic and from hypercholesterolaemic rats. This effect, however, was not mediated by a chronic modification of the ryanodine receptor as shown by the unchanged ryanodine binding in the latter group. While coenzyme Q10 supplementation significantly reduced the frequency of the spontaneous calcium release events, it did not affect their amplitude and spatial spread in muscles from fluvastatin-treated rats. This indicates that coenzyme Q10 supplementation prevented the spark frequency increasing effect of fluvastatin without having a major effect on the amount of calcium released during individual sparks. In conclusion, we have found that fluvastatin, independently of the cholesterol level in the blood, consistently and specifically increased the frequency of calcium sparks in skeletal muscle cells, an effect which could be prevented by the addition of coenzyme Q10 to the diet. These results support theories favouring the role of calcium handling in the pathophysiology of statin-induced myopathy and provide a possible pathway for the protective effect of coenzyme Q10 in statin treated patients symptomatic of this condition.

Fitness cost associated with resistance to fluoroquinolones is diverse across clones of Klebsiella pneumoniae and may select for CTX-M-15 type extended-spectrum ß-lactamase.

Lowered fitness cost associated with resistance to fluoroquinolones was recently demonstrated to influence the clonal dynamics of methicillin-resistant Staphylococcus aureus (MRSA) in the health care setting. We investigated whether or not a similar mechanism impacts Klebsiella pneumoniae. The fitness of K. pneumoniae isolates from major international hospital clones (ST11, ST15, ST147) already showing high-level resistance to fluoroquinolones and of strains from three minor clones (ST25, ST274, ST1028) in which fluoroquinolone resistance was induced in vitro was tested in a propagation assay. Strains from major clones showed significantly less fitness cost than three of four fluoroquinolone-resistant derivatives of minor clone isolates. In addition, plasmids with CTX-M-15 type extended-spectrum ß-lactamase (ESBL) genes were all retained in both major and minor clone isolates, irrespective of the strains' level of fluoroquinolone resistance, while each plasmid harboring SHV-type ESBLs had been lost during the induction of resistance. Major clone K. pneumoniae strains harbored more amino acid substitutions in the quinolone resistance determining regions (QRDRs) of the gyrA and parC genes than minor clone isolates. The presence of an active efflux system could be demonstrated in all fluoroquinolone-resistant derivatives of originally SHV-producing minor clone isolates but not in any CTX-M-15-producing strain. Further investigations are needed to expand and confirm our findings on a larger sample. In addition, a long-term observation of our ciprofloxacin-resistant minor clone isolates is required in order to elucidate whether or not they are capable of restoring their fitness while concomitantly retaining high minimum inhibitory concentration (MIC) values.

Varying fitness cost associated with resistance to fluoroquinolones governs clonal dynamic of methicillin-resistant Staphylococcus aureus.

The purpose of this study was to investigate the impact of fluoroquinolone resistance on the existence and dynamic of MRSA clones. Resistance to ciprofloxacin was induced in strains of community-acquired (CA) MRSA from various sequence types and the fitness cost suffered by mutant derivatives measured in a propagation assay. In addition, the fitness of fluoroquinolone resistant health care-associated (HA) MRSA isolates from major clones prevalent in Hungary were compared with each other and with those of the CA-MRSA derivatives. The genetic background of fluoroquinolone resistance and fitness cost in CA-MRSA was investigated. The fitness cost observed in the CA-MRSA derivatives proved diverse; the derivatives of the ST30-MRSA-IV strain suffered significantly greater fitness cost than those of the ST8-MRSA-IV and ST80-MRSA-IV isolates. Strains from the New York-Japan (ST5-MRSA-II), South German (ST228-MRSA-I) and EMRSA-15 (ST22-MRSA-IV) HA-MRSA clones proved more viable than CA-MRSA derivatives with similar MIC values to ciprofloxacin and HA-MRSA strains from the Hungarian/Brazilian clone (ST239-MRSA-III). Our strains from the New York-Japan, South-German and EMRSA-15 clones seem to have a competitive edge over the tested CA-MRSA isolates in the health care setting. The greater fitness observed in our New York-Japan and South-German strains could account for the replacement by them of the Hungarian/Brazilian clone in Hungary about ten years ago. Alterations in relevant genes were detected. The Ser80 → Phe mutation in the grlA gene may have seriously compromised viability. Surprisingly silent nucleotide substitutions in the grlB gene seemed to impact fitness in derivatives of the ST30-MRSA-IV isolate.

Dissemination of ST274 Klebsiella pneumoniae epidemic clone in newborn and adult hospital settings harbouring SHV-2A or CTX-M-15 type extended spectrum ß-lactamases-producing known plasmids.

Molecular epidemiology and genetic features of an extended-spectrum ß-lactamase (ESBL) producing Klebsiella pneumoniae epidemic clone (KP-EC) with elevated ciprofloxacin MIC (minimum inhibitory concentration) values from multiple nosocomial outbreaks and sporadic cases between 2006 and 2008 in Hungary were investigated. As a result of continuous monitoring of ESBL-producing KP-ECs, 27 isolates collected from five healthcare facilities were selected for macrorestriction profile analysis by PFGE (pulsed field gel electrophoresis). Of these, 12 strains were isolated from adult inpatients, while 15 strains were from newborns. The MIC values for several antibiotics were determined by agar dilution technique. Molecular typing was further performed by PCR (polymerase chain reaction) and sequencing of several antibiotic resistance genes, plasmid profile analysis, transfer of resistance determinants and multilocus sequence typing (MLST). All isolates showed moderate resistance to ciprofloxacin (MICs ranged from 0.5 to 8 mg L(-1)). PFGE revealed the existence of only one genetic cluster defined as EC IV. PstI digestion of plasmid DNA revealed two highly diverse restriction patterns in "adult" and "newborn" isolates corresponding to plasmids from the Hungarian Epidemic Clone and plasmids isolated from a neonatal nosocomial outbreak in 1998, respectively. Sequence analysis of ß-lactamase genes from plasmids of 14 selected isolates detected bla SHV-2a in strains isolated exclusively from newborns and bla CTX-M-15 in strains isolated exclusively from adult inpatients. MLST established that strains of the PFGE cluster belonged to a novel sequence type ST274. ESBL-producing K. pneumoniae isolates belonging to the novel sequence type ST274 appeared in the newborn and adult hospital settings in Hungary and acquired SHV-2a or CTX-M-15 type enzymes, respectively. Thus, a new antimicrobial resistance strategy for successful conformation to distinct hospital settings was found.

Investigation of extended-spectrum beta-lactamase-producing Klebsiella pneumoniae outbreaks in Hungary between 2005 and 2008.

Fourteen outbreaks in Hungary between 2005 and 2008 caused by extended-spectrum beta-lactamase-producing Klebsiella pneumoniae (ESBL-KP) were epidemiologically investigated and the isolated pathogens were characterized by molecular techniques. Ten of the fourteen outbreaks occurred in adult wards and four in neonatal units affecting a total number of 73 patients. The 54% [40] of the patients developed bloodstream infections and 21.9%-21.9% [16] pneumonia and surgical site infections, respectively. The overall rate of mortality proved high: 36.9% [27]. Outbreaks in adults affected more patients, had higher attack rates, were more prolonged in duration and had a 6.9-fold higher mortality rate than outbreaks observed in neonates. The outbreaks in neonates were caused by SHV-type ESBL-producing klebsiellae, while in the "adult outbreaks" exclusively CTX-M-type ESBL-KP strains were involved. While the outbreak strains isolated from neonatal units could be assigned to a variety of pulsotypes, the previously described K. pneumoniae epidemic clones, ST15 and ST147, could be identified among the pathogens causing outbreaks in adult units.

Risk factors and outcomes for bloodstream infections with extended-spectrum beta -lactamase-producing Klebsiella pneumoniae ; Findings of the nosocomial surveillance system in Hungary.

Risk factors for and outcomes of bloodstream infections (BSIs) caused by ESBL-producing and by ESBL-non-producing Klebsiella pneumoniae were compared in a four-year multicenter study in Hungary. One hundred ESBL-positive and one hundred ESBL-negative patients were included as cases and controls. Investigated risk factors were related to demographics, comorbid conditions, treatments, invasive procedures, surgery prior bacteremia, presence of additional nosocomial infections and preceding hospital admission within a year. Measured outcomes were crude mortality, mortality related to infection and delay in introducing appropriate therapy (DAT). Though some risk factors for infection (admission to intensive care units, having central venous and/or urinary catheter, mechanical ventilation) were shared by both groups, in other respects cases and controls were found to differ substantially. The 36 percent of patients with BSIs with ESBL-producing Klebsiella died versus 23 percent of controls (odds ratio [OR]: 2.5; 95% confidence interval [CI]: 1.0-5.4; p = 0.02). The 18 percent of deaths in cases versus 9% in controls could be attributed to infection (OR: 5.0; 95% CI: 1.5-16.2; p = 0.006). Cases more often received previous antibiotic therapy than controls (OR: 2.7; 95% CI: 1.1-6.7; p = 0.02) and delay in the introduction of appropriate antibiotic treatment was observed in 44% of cases versus 19% of controls (OR: 3.4; 95% CI: 1.6-7.3; p = 0.001). The results demonstrate that BSIs caused by ESBL-producing K. pneumoniae are related to previous antibiotic therapy and are associated with a high rate of mortality that is often linked to delay in the introduction of appropriate antibiotic therapy. This confirms that besides infection control measures the early identification and antibiotic resistance profiling of the infecting pathogen is salient in the control of BSIs caused by ESBL-producing K. pneumoniae .

Broadening spectrum of bovine spongiform encephalopathies.

Until recently the etiology of bovine spongiform encephalopathy (BSE) was considered uniform. The infectious agent was thought to be a single strain of prion (posttranslationally altered form of normal prion protein: PrPSc) retaining its biochemical and biological characteristics during interspecies transmission. However, alternate PrPSc signatures through large-scale screening have recently been detected. In addition, genetic alterations governing susceptibility to prion infection and a mutation (E211K) capable of eliciting spontaneous BSE have been demonstrated. Thus, the spectrum of BSEs have broadened and three PrPSc variants (BSE-C, BSE-H and BSE-L) are now defined. Moreover, a new condition resembling BSE, idiopathic brainstem neuronal chromatolysis (IBNC), has been described that may also turn out to be a prion disease. Since one of the new BSE variants, L-type BSE, proved highly pathogenic detection and further characterization of the new conditions are essential.

Characterisation of the first VIM metallo-beta-lactamase-producing Pseudomonas aeruginosa clinical isolate in Serbia.

From the Central-East European region the first VIM metallo-beta-lactamase (MBL) producing Pseudomonas aeruginosa strains were published from Croatia, Poland and Hungary. The aim of this study was to assess the contribution of MBL-production to carbapenem-resistance among P. aeruginosa clinical isolates in the Military Medical Academy (MMA) in Belgrade, Serbia between August 2004 and September 2007. Only one P. aeruginosa isolate with strain number 722 proved MBL-positive that harboured a novel class 1 integron with a bla(VIM-2)-like cassette in the first position, followed by orfD, a putative gene with unknown function. Our data indicate that MBL-producing strains occur at a prevalence of less than 1% among imipenem-nonsusceptible P. aeruginosa clinical isolates in this Belgrade hospital. The newly identified VIM MBL-producing P. aeruginosa strain 722 could be assigned to serotype O11, and it was panresistant to all antimicrobials tested. The isolate displayed sequence type ST235 by multilocus sequence typing which is the founder sequence type of the previously identified international clonal complex CC11 that already contains bla(VIM)-positive isolates from Italy, Greece, Sweden, Hungary and Poland. In conclusion, this is the first report of VIM MBL-producing P. aeruginosa from Serbia and also of the occurrence of such isolates belonging to the international clonal complex CC11 in this country.

Replacement of methicillin-resistant Staphylococcus aureus clones in Hungary over time: a 10-year surveillance study.

The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in Hungary has been increasing and is now close to 20% among invasive isolates of S. aureus. In order to understand the evolution of MRSA in Hungary, two collections of isolates were studied: 22 representatives of a collection of 238 MRSA isolates recovered between 1994 and 1998, and a collection of 299 MRSA isolates recovered between 2001 and 2004. The isolates were first characterised by pulsed-field gel electrophoresis (PFGE) and were distributed into 19 different PFGE patterns. Representatives of each pattern were further characterised by spa typing, multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing. The Hungarian clone that was predominant in 1994-1998 (PFGE E, ST239-III) had almost disappeared in 2003-2004, being replaced by the Southern German clone (PFGE B, ST228-I) and the New York/Japan epidemic clone (PFGE A, ST5-II), which represented c. 85% of the 2001-2004 isolates. Thus, this study describes, for the first time, the co-dominance and extensive spread of the New York/Japan clone in a European country.

Molecular characterisation of invasive Streptococcus pyogenes isolates from Hungary obtained in 2004 and 2005.

Our aim was to characterise by molecular techniques group A streptococci isolated from invasive infections in Hungary in 2004-2005. Twenty-six nonduplicate invasive GAS isolates were selected and examined. The mortality rate proved high (52.3%) for those cases (n = 21) where data were available. Predominant emm types were emm1 (n = 13, 50%) and emm80 (n = 5, 19.2%), but other M types (emm4, emm28, emm66, emm81.1, emm82, emm84) were also identified. Eight different PFGE types were distinguished, and each emm type showed an individual PFGE pattern. Our results show that--similarly to results obtained in several other countries--emm type 1 strains predominate among invasive GAS isolates, and that emm 1 type strains recovered from severe streptococcal infections were associated with the presence of the speA gene. The rate for macrolide resistance proved low: only two isolates showed elevated MICs for erythromycin.

Occurrence and regional distribution of SHV-type extended-spectrum beta-lactamases in Hungary.

In order to determine the presence and geographical distribution of SHV-type extended-spectrum beta-lactamase genes among Enterobacteriaceae strains in Hungary, isolates from 25 microbiology laboratories throughout the country were collected between January 2002 and August 2003 and examined. Sequencing of the genes showed that SHV-5 and SHV-2a are the dominant SHV-types in extended-spectrum beta-lactamase-producing Enterobacteriaceae strains in this country. The SHV-2 gene, which is prevalent in many European countries, was not detected, but one isolate carried the SHV-12 gene. The results show that these genes are circulating among Enterobacteriaceae strains in Hungary and indicate that strict infection control measures are warranted in order to prevent their spread.

Characterization of extended-spectrum beta-lactamase (TEM-52)-producing strains of Salmonella enterica serovar typhimurium with diverse resistance phenotypes.

Two Salmonella enterica serovar Typhimurium strains from different clonal origins, both producing an extended-spectrum beta-lactamase (TEM-52), were isolated from a patient. This enzyme was encoded on a single plasmid and was found at very low levels in one strain, while being encoded on multiple plasmids and in multiple different EcoRI fragments in the other strain.

Is the pathogen of prion disease a microbial protein?

Though considerable circumstantial evidence suggests that the pathogen of prion disease is proteinaceous, it has not yet been conclusively identified. Epidemiological observations indicate that a microbial vector is responsible for the transmission of natural prion disease in sheep and goats and that the real causative agent may correspond to a structural protein of that microorganism. The microbial protein should resemble prion protein (PrP) and may replicate itself in the host by using mammalian DNA. A similar phenomenon was already described with a protein antigen of the ameba Naegleria gruberi. The various serotypes of the microbial protein may account for the existence of scrapie strains. It is proposed that many microbial proteins may be capable of replicating themselves in mammalian cells eliciting and sustaining thereby degenerative and/or autoimmune reactions subsequent to infections with microorganisms.

[Severe genital mycoplasma infection following cesarean section]. / Genitális mycoplasmák által okozott súlyos fertözés sectio caesareát követöen.

Authors report a serious case of post-caesarean delivery endometritis caused, probably exclusively, by genital mycoplasmas: Ureaplasma urealyticum and Mycoplasma hominis. The initial treatment of the patient with various penicillins, ceftriaxone, gentamicin, metronidazole and nystatine proved ineffective. Subsequently, as microbiological tests turned out positive for genital mycoplasmas, a therapy of doxycyclin was introduced and a full recovery could be attained. Authors' experience is consistent with the observation of American scientists that U. urealyticum is an important pathogen in post-caesarean delivery endometritis. Since the carriage of U. urealyticum in women is frequent in Hungary, it is suggested that microbiological investigations related to sectio caesarea always include tests for genital mycoplasmas.

Immunologic and molecular biologic studies of prion proteins in bovine spongiform encephalopathy.

Bovine spongiform encephalopathy (BSE) is a transmissible neurodegenerative disease. Six brain regions from 11 cattle were examined for the presence of the abnormal isoform of the prion protein (PrPBSE). The highest concentrations of PrPBSE were found in the brain stem, where the greatest degree of spongiform change was observed. Molecular cloning of the bovine PrP gene showed that it encodes a protein of 256 or 264 amino acids with five or six Gly:Pro-rich octarepeats, respectively, in contrast to all other mammalian PrP genes, which encode only five octarepeats. The bovine PrP gene is single copy, and the entire open-reading frame lies within a single exon. Since the transmission of prions across species seems to be restricted by differences in PrP sequence, the high degree of homology between sheep and bovine PrP (98%) correlates with the proposed cause of BSE.