Biomedical subjects

Heinrich Neubauer

Publications and source records attributed to Heinrich Neubauer.

At least 19 recordsLinked to original sources

PubMed2006

Spatial organization of EEG coherence during listening to consonant and dissonant chords.

Theories of harmony state that the contribution of both sensory and cognitive components is important for musical consonance perception. The aims of the present study were to analyze (a) functional intra- and inter-hemispheric connectivity associated with listening to consonant and dissonant chords using EEG coherence method; (b) relationships between affective responsiveness, sensory aspects of perceived consonance and associated brain connectivity. We identified two lines of inter-hemispheric connectivity in the theta band; one localized anterior being sensitive to consonance and one localized posterior sensitive to dissonance. Stronger right intra-hemispheric connectivity for consonance than dissonance in the theta band was associated with higher pleasantness ratings. The relationship between sensory aspects of perceived consonance and left intra-hemispheric connectivity found in theta-2 was interpreted as processing of vertical harmony without emotional involvement. The stronger connectivity along the axis "left anterior-right posterior" for dissonance than consonance in the alpha-1 band is discussed as a correlate of novelty processing. By introducing a "auditory object dissociation" hypothesis we suggest to extend the present concept of harmony perception. We believe that "auditory object dissociation" is a component of "sensory dissonance."

Acoustic Stimulation↗

PubMed2006

Correcting for false alarms in a simple reaction time task.

Simple reaction times (RTs) constitute an important source of information and tool in human and animal psychophysics, in cognitive neuroscience, and in the clinic. We measure simple RTs to auditory stimuli, in a high signal rate vigilance design, to examine the possibility that simple RT may be used as a tool to study mechanisms of temporal summation at absolute threshold. By means of catch trials, we monitor the subjects' tendencies to produce false alarms, that is reactions not controlled by the reaction stimulus. Here we examine the possibility that a model of a race between a stimulus-controlled reaction process and a false alarm process can account for the numbers as well as for the timing of early and late RTs on stimulus trials in our experiments. We show that the responses in both tails of our empirical RT distributions on stimulus trials are indeed correctly predicted by the race model and can be considered false alarms. This suggests that the race model might also provide a reasonable description of the way in which false alarms and stimulus-controlled reactions combine when they overlap in time. We examine the magnitudes of the estimated effects of false alarms on several parameters of the RT distributions by application of the race model. The analyses reveal that the effects not only vary with false alarm rate but also with stimulus parameters. Our data suggest that the race model may provide a theoretically reasonable and easy means of correcting for false alarms in simple RT paradigms and thus may constitute a useful alternative to the common practice of truncation.

Adult↗

PubMed2006

Comparison of absolute thresholds derived from an adaptive forced-choice procedure and from reaction probabilities and reaction times in a simple reaction time paradigm.

An understanding of the auditory system's operation requires knowledge of the mechanisms underlying thresholds. In this work we compare detection thresholds obtained with a three-interval-three-alternative forced-choice paradigm with reaction thresholds extracted from both reaction probabilities (RP) and reaction times (RT) in a simple RT paradigm from the same listeners under otherwise nearly identical experimental conditions. Detection thresholds, RP, and RT to auditory stimuli exhibited substantial variation from session to session. Most of the intersession variation in RP and RT could be accounted for by intersession variation in a listener's absolute sensitivity. The reaction thresholds extracted from RP were very similar, if not identical, to those extracted from RT. On the other hand, reaction thresholds were always higher than detection thresholds. The difference between the two thresholds can be considered as the additional amount of evidence required by each listener to react to a stimulus in an unforced design on top of that necessary for detection in the forced-choice design. This difference is inversely related to the listener's probability of producing false alarms. We found that RT, once corrected for some irreducible minimum RT, reflects the time at which a given stimulus reaches the listener's reaction threshold. This suggests that the relationships between simple RT and loudness (reported in the literature) are probably caused by a tight relationship between temporal summation at threshold and temporal summation of loudness.

Adult↗

PubMed2006

Real-time PCR using hybridization probes for the rapid and specific identification of Francisella tularensis subspecies tularensis.

Tularemia is a plague-like infection caused by Francisella (F.) tularensis classified as a biological warfare agent. F. tularensis subsp. tularensis is the most virulent subspecies demanding rapid diagnosis. Typing systems for this fastidious bacterium to the subspecies level are laborious and time consuming. Therefore, the aim of this study was to develop a real-time PCR for the rapid and specific identification of F. tularensis subsp. tularensis. The specificity of the assay was determined using a comprehensive panel of Francisella strains, clinically relevant bacteria, and DNA preparations of potential hosts. F. tularensis subsp. tularensis was specifically detected but no other organisms. The range of linearity was determined to be 100 fg to 10 ng, the lower limit of detection was 25 fg of DNA (13 genome equivalents). An internal amplification control PCR system targeting lambda phage DNA was included. Neither the internal amplification control nor host DNA influenced the cycle threshold values obtained for F. tularensis subsp. tularensis. In conclusion, we have developed a highly sensitive and specific assay that can be integrated into real-time PCR-based identification procedures for biological agents. This is a major diagnostic improvement, as all other methods for the specific identification of F. tularensis subsp. tularensis are more time consuming.

Animals↗

PubMed2006

Molecular characterization of Coxiella burnetii isolates by infrequent restriction site-PCR and MLVA typing.

BACKGROUND: Coxiella burnetii, the causative agent of Q fever, has a wide host range. Few epidemiological tools are available, and they are often expensive or not easily standardized across laboratories. In this work, C. burnetii isolates from livestock and ticks were typed using infrequent restriction site-PCR (IRS-PCR) and multiple loci variable number of tandem repeats (VNTR) analysis (MLVA). RESULTS: By applying IRS-PCR, 14 C. burnetii isolates could be divided into six groups containing up to five different isolates. Clustering as deduced from MLVA typing with 17 markers provided an increased resolution with an excellent agreement to IRS-PCR, and with the plasmid type of each strain. MLVA was then applied to 28 additional C. burnetii isolates of different origin and 36 different genotypes were identified among the 42 isolates investigated. The clustering obtained is in agreement with published Multiple Locus Sequence Typing (MLST) data. Two panels of markers are proposed, panel 1 which can be confidently typed on agarose gel at a lower cost and in any laboratory setting (10 minisatellite markers with a repeat unit larger than 9 bp), and panel 2 which comprises 7 microsatellites and provides a higher discriminatory power. CONCLUSION: Our analyses demonstrate that MLVA is a powerful and promising molecular typing tool with a high resolution and of low costs. The consistency of the results with independent methods suggests that MLVA can be applied for epidemiological studies. The resulting data can be queried on a dedicated MLVA genotyping Web service.

Animals↗

PubMed2006

Evaluation and selection of tandem repeat loci for a Brucella MLVA typing assay.

BACKGROUND: The classification of Brucella into species and biovars relies on phenotypic characteristics and sometimes raises difficulties in the interpretation of the results due to an absence of standardization of the typing reagents. In addition, the resolution of this biotyping is moderate and requires the manipulation of the living agent. More efficient DNA-based methods are needed, and this work explores the suitability of multiple locus variable number tandem repeats analysis (MLVA) for both typing and species identification. RESULTS: Eighty tandem repeat loci predicted to be polymorphic by genome sequence analysis of three available Brucella genome sequences were tested for polymorphism by genotyping 21 Brucella strains (18 reference strains representing the six 'classical' species and all biovars as well as 3 marine mammal strains currently recognized as members of two new species). The MLVA data efficiently cluster the strains as expected according to their species and biovar. For practical use, a subset of 15 loci preserving this clustering was selected and applied to the typing of 236 isolates. Using this MLVA-15 assay, the clusters generated correspond to the classical biotyping scheme of Brucella spp. The 15 markers have been divided into two groups, one comprising 8 user-friendly minisatellite markers with a good species identification capability (panel 1) and another complementary group of 7 microsatellite markers with higher discriminatory power (panel 2). CONCLUSION: The MLVA-15 assay can be applied to large collections of Brucella strains with automated or manual procedures, and can be proposed as a complement, or even a substitute, of classical biotyping methods. This is facilitated by the fact that MLVA is based on non-infectious material (DNA) whereas the biotyping procedure itself requires the manipulation of the living agent. The data produced can be queried on a dedicated MLVA web service site.

Animals↗

PubMed2006

Detection of the reemerging agent Burkholderia mallei in a recent outbreak of glanders in the United Arab Emirates by a newly developed fliP-based polymerase chain reaction assay.

A polymerase chain reaction (PCR) assay targeting the flagellin P (fliP)-I S407A genomic region of Burkholderia mallei was developed for the specific detection of this organism in pure cultures and clinical samples from a recent outbreak of equine glanders. Primers deduced from the known fliP-IS407A sequence of B. mallei American Type Culture Collection (ATCC) 23344(T) allowed the specific amplification of a 989-bp fragment from each of the 20 B. mallei strains investigated, whereas other closely related organisms tested negative. The detection limit of the assay was 10 fg for purified DNA of B. mallei ATCC 23344(T). B. mallei DNA was also amplified from various tissues of horses with a generalized B. mallei infection. The developed PCR assay can be used as a simple and rapid tool for the specific and sensitive detection of B. mallei in clinical samples.

Animals↗

PubMed2006

Brucella endocarditis in prosthetic valves.

Human brucellosis is a multiple organ disease that presents with fever and is most often transmitted via contaminated, unpasteurized goat milk and cheese. In chronic cases, focal complications (eg, spondylitis, neurobrucellosis and endocarditis) are frequently seen. Although the disease may be severely debilitating, the mortality rate is low. Fatal cases are often due to endocarditis. Because Brucella endocarditis is a rare complication (2% to 5%), therapeutic considerations are based on single-case experiences only. Therapy includes long-term antibiotic treatment using combinations of various antimicrobial drugs and surgical valve replacement when required. A case of Brucella endocarditis complicated by the infection of two valvular prostheses implanted after involvement of the mitral and aortic valve due to rheumatic fever is described. The patient was successfully treated by a medical and surgical approach. Therapeutic strategies in Brucella endocarditis are discussed in light of the current literature.

Anti-Bacterial Agents↗

PubMed2006

Genotyping of Ochrobactrum anthropi by recA-based comparative sequence, PCR-RFLP, and 16S rRNA gene analysis.

A recA-PCR restriction fragment length polymorphism assay was developed to study intraspecies variation among Ochrobactrum anthropi. Primers deduced from the known recA gene sequence of the genetically closely related genus Brucella allowed the specific amplification of a 1065 bp recA fragment from each of the 38 O. anthropi and the eight Brucella strains investigated. RecA was also amplified from the type strains of O. intermedium, O. tritici, and O. lupini but could not be generated from O. grignonense and O. gallinifaecis. Subsequent comparative recA sequence- and HaeIII-recA restriction fragment length polymorphism analysis identified nine different genospecies among the tested 38 O. anthropi isolates, whereas the recA sequences of the Brucella spp. were indistinguishable. Furthermore, Brucella spp., O. anthropi, O. intermedium, and O. tritici were clearly separated from each other by means of their recA sequences and HaeIII restriction patterns. Five strains of uncertain species status listed in the Culture Collection University of Göteborg bacterial culture collection as O. anthropi were characterized by recA analysis, and their phylogenetic position within the Brucella-Ochrobactrum group was determined. In summary, recA-sequence analysis provides a new reliable molecular subtyping tool to study the phylogeny of the Ochrobactrum taxon at both the inter- and intraspecies level.

Animals↗

PubMed2006

Development of a 5'-nuclease real-time PCR assay targeting fliP for the rapid identification of Burkholderia mallei in clinical samples.

BACKGROUND: Burkholderia mallei is a potential biological agent that causes glanders or farcy in solipeds, a disease notifiable to the Office International des Epizooties (OIE). The number of reported outbreaks has increased steadily during the last decade, but diagnosis is hampered by the low bacterial load in infected tissues and excretions. METHODS: We developed a B. mallei-specific 5'-nuclease real-time PCR assay that targets the fliP gene of B. mallei and includes an internal amplification control. Specificity was assessed with 19 B. mallei strains, 27 Burkholderia pseudomallei strains, other Burkholderia strains of 29 species, and clinically relevant non-Burkholderia organisms. RESULTS: Amplification products were observed in all B. mallei strains but in no other bacteria. The linear range of the B. mallei real-time PCR covered concentrations from 240 pg to 70 fg of bacterial DNA/reaction. The detection limit was 60 fg of B. mallei DNA. The clinical applicability of the assay was demonstrated by use of organ samples from diseased horses of a recent outbreak that was reported to the OIE by the United Arab Emirates in 2004. CONCLUSIONS: Compared with conventional PCR, our rapid 5'-nuclease real-time PCR assay for the specific identification of B. mallei has a lower risk of carryover contamination and eliminates the need for post-PCR manipulations. This real-time PCR assay also shortens the turnaround time for results and has the potential for automation.

Animals↗

PubMed2005

Immunoproteomic characterization of Brucella abortus 1119-3 preparations used for the serodiagnosis of Brucella infections.

The diagnosis of brucellosis is mainly based on the detection of anti-LPS antibodies. Due to substantial similarity of the O-polysaccharide of Brucella LPS to that of various other Gram-negative bacteria, serological tests of samples containing high amounts of LPS lack specificity. Hence, the development of assays based on more specific protein antigens is an essential subject in brucellosis research. The aim of this study was proteomic characterization of various antigen preparations of the diagnostic reference strain Brucella abortus 1119-3 and the identification of immunogenic proteins suitable for serological assays. Seventeen out of 383 protein spots of B. abortus 1119-3 were identified to be immunogenic by 2-D immunoblotting. These immunogenic spots were assigned to 6 proteins by MALDI-MS and nLC-ESI-MS/MS: Cu-Zn SOD, BCSP31, L7/L12, GroEL, GroES, and DnaK. All immunogenic proteins were present in three different antigen preparations investigated, i.e. native antigen, standard agglutination and commercially available agglutination antigen. 2-D immunoblotting of bacteria cross-reacting with Brucellae in agglutination tests proved that cross-reactivity of proteins is negligible. Surface enhanced laser desorption/ionization mass spectrometry (SELDI-MS) spectra also differentiated B. abortus clearly from cross-reacting bacteria. The combination of SELDI-MS analysis with the specificity of antibody binding will improve the identification of Brucella specific immunogenic proteins.

Animals↗

PubMed2005

Failure of a short-term antibiotic therapy for human brucellosis using ciprofloxacin. A study on in vitro susceptibility of Brucella strains.

BACKGROUND: Human brucellosis is characterized byfocal complications, chronic courses, and therapeutic failures. METHODS: In a relapsed case of brucellosis after short-term antibiotic therapy using doxycycline and ciprofloxacin two Brucella strains were isolated, before and after treatment. In vitro susceptibilities of both isolates were determined by E tests including a great variety of antibiotics. In a killing rate experiment the bactericidal activities of doxycycline, streptomycin, rifampin and ciprofloxacin as single agents and in combinations were determined. RESULTS: Lowest MIC values were measured for doxycycline and ciprofloxacin. MICs did not change under therapy. Streptomycin alone exhibited the most effective killing within 6 h, whereas the other single agents did not show bactericidal activity. Doxycycline plus ciprofloxacin was the most active combination in vitro. CONCLUSION: Routine susceptibility testing of Brucellae is not obligatory as most of the 'traditional' anti-Brucella antibiotics are active in vitro and bactericidal efficacy may differ in vivo.

Anti-Infective Agents↗

PubMed2005

Detection of Chromobacterium violaceum by multiplex PCR targeting the prgI, spaO, invG, and sipB genes.

Based on the recently completed genomic sequence of Chromobacterium violaceum American Type Culture Collection (ATCC) 12472 a multiplex PCR assay targeting the prgI, spaO, invG, and sipB genes of the Salmonella SPI-1 homologue type-III secretion system was developed. PCR products of 255bp (prgI), 749bp (spaO), 1685bp (invG), and 1752bp (sipB) were successfully amplified simultaneously in a single reaction with all Chr. violaceum strains investigated whereas other bacteria tested negative. The detection limit for pure cultures in multiplex PCR analysis was 100CFU. The developed assay significantly improves rapid identification of Chr. violaceum and allows its differentiation from closely related organisms.

Animals↗

PubMed2005

Growth characteristics of Bacillus anthracis compared to other Bacillus spp. on the selective nutrient media Anthrax Blood Agar and Cereus Ident Agar.

Anthrax Blood Agar (ABA) and Cereus Ident Agar (CEI) were evaluated as selective growth media for the isolation of Bacillus anthracis using 92 B. anthracis and 132 other Bacillus strains from 30 species. The positive predictive values for the identification of B. anthracis on ABA, CEI, and the combination of both were 72%, 71%, and 90%, respectively. Thus, less than 10% of all species were misidentified using both nutrient media. Species which might be misidentified as B. anthracis were B. cereus, B. mycoides, and B. thuringiensis. Particularly, 30% of B. weihenstephanensis strains were misidentified as B. anthracis.

Agar↗

PubMed2005

Genotyping of Chromobacterium violaceum isolates by recA PCR-RFLP analysis.

Intraspecies variation of Chromobacterium violaceum was examined by comparative sequence - and by restriction fragment length polymorphism analysis of the recombinase A gene (recA-PCR-RFLP). Primers deduced from the known recA gene sequence of the type strain C. violaceum ATCC 12472(T) allowed the specific amplification of a 1040bp recA fragment from each of the 13 C. violaceum strains investigated, whereas other closely related organisms tested negative. HindII-PstI-recA RFLP analysis generated from 13 representative C. violaceum strains enabled us to identify at least three different genospecies. In conclusion, analysis of the recA gene provides a rapid and robust nucleotide sequence-based approach to specifically identify and classify C. violaceum on genospecies level.

Bacterial Typing Techniques↗

PubMed2005

Antimicrobial susceptibilities of Austrian Francisella tularensis holarctica biovar II strains.

The antibiotic susceptibilities of 50 Francisella tularensis subsp. holarctica biovar II strains isolated from hares and human patients from the eastern part of Austria were examined. Minimum inhibitory concentrations (MICs) of 24 antimicrobial agents were determined using Eteststrade mark on cysteine heart agar plates supplemented with 10% sheep blood. All isolates were sensitive to tetracyclines, aminoglycosides, quinolones, chloramphenicol and rifampicin. Resistance was observed in all isolates against macrolides, penicillins and aztreonam. Bacteria were resistant to cephalosporins and carbapenems, except for 8% of strains investigated that were susceptible or intermediately susceptible. Our in vitro susceptibility data can be applied for the detection and comparison of resistance development and to provide in vitro data for the guidance of therapy.

Aminoglycosides↗

PubMed2005

Rapid presumptive identification of Burkholderia pseudomallei with real-time PCR assays using fluorescent hybridization probes.

Burkholderia pseudomallei (the etiologic agent of melioidosis) can cause pyogenic or granulomatous lesions in almost any organ. Septicemia has a case fatality rate of >40%. Early diagnosis and appropriate antibiotic therapy are crucial for survival, but cultivation, biochemical identification, and conventional PCR of B. pseudomallei are time consuming. We established real-time PCR assays using fluorescent hybridization probes targeting the 16S rDNA, the flagellin C (fliC) and the ribosomal protein subunit S21 (rpsU) genes. The test sensitivity and specificity were assessed with a representative panel of 39 B. pseudomallei, 9 B. mallei, 126 other Burkholderia strains of 29 species, and 45 clinically relevant non-Burkholderia organisms. The detection limit for the 16S rDNA, fliC, and rpsU assay was 40, 40, and 400 genome equivalents per reaction, however, in spiked blood samples it was 300, 300, and 3000, respectively. Specificity, positive and negative predictive value of the assays was 100%. In conclusion, we recommend the use of the 16S rDNA and/or fliC real-time PCR assays for the rapid identification of B. mallei and B. pseudomallei in positive blood cultures or from suspicious bacterial colonies.

Burkholderia pseudomallei↗

PubMed2005

Attempted passive prophylaxis with a monoclonal anti-Burkholderia pseudomallei exopolysaccharide antibody in a murine model of melioidosis.

Melioidosis is a severe gram-negative infection caused by the facultative intracellular bacterium Burkholderia pseudomallei, which is responsible for a broad spectrum of symptoms in both humans and animals. No licensed vaccine currently exists. This study evaluated the protective effect of a monoclonal antibody (Mab Ps6F6) specific to B. pseudomallei exopolysaccharide in an outbred murine model of sub-acute melioidosis. When administered before the infectious challenge, Ps6F6 significantly increased resistance to infection and restrained bacterial burden in the spleen over a 30-days period. Patterns of IFN-gamma production were similar in the treated and non treated groups of mice. However, Ps6F6 lowered IFN-gamma levels over the duration of the assay period, except on day 1, suggesting a transient and rapid production of IFN-gamma under Ps6F6 control. Minor but persisting increases occurred in IL-12 levels while TNF-alpha was detected only in the controls at the later stages of infection. No IL-10 secretion was detected in both groups of mice. These data suggest that passive prophylaxis with Mab Ps6F6 provide a moderate and transient induction of inflammatory responses in infected mice but failed to trigger a sterilizing protective immunity.

Animals↗