PubMed Health⌕ Search

Biomedical subjects

H Neubauer

Publications and source records attributed to H Neubauer.

At least 37 records · Page 2Linked to original sources
PubMed2003

High grade and non-high grade ductal carcinoma in situ on dynamic MR mammography: characteristic findings for signal increase and morphological pattern of enhancement.

The objective of this review is to describe characteristic MR mammographic findings for signal increase and morphological patterns of enhancement in pure ductal carcinoma in situ (DCIS) and to differentiate between high grade and non-high grade lesions. The dynamic MR examination (1.5 T unit, contrast enhanced T(1) weighted two dimensional fast field echo, 96 ms repetition time, 5.0 ms echo time, 80 degrees flip angle) of 39 consecutive patients with pure DCIS was evaluated retrospectively. Categories were defined for signal increase (C1=normal, C2=slow, continuous, C3=strong initial and slow further increase, C4=strong initial increase followed by a plateau phenomenon, and C5=strong initial increase followed by a washout phenomenon) and morphological patterns (M0=no pattern observed, M1=linear or linear-branched, M2=segmental dotted or granular, M3=segmental homogeneous, and M4=focal spot-like). Time-intensity curves showing a C4 and C5 signal increase were considered suspicious for malignancy. All cases were correlated with histology. 62% of all tumours had a plateau or washout (C4, C5), 77% showed a strong initial signal increase (C3-C5). On evaluation of time-intensity curves alone MR mammography (MRM) findings were suspicious for malignancy in 62% of all DCIS cases. A segmental enhancement was found in 82% of all enhancing tumors and the M2 pattern in 73%. In a combined analysis of signal increase and morphology, 70% of non-high grade and 92% of high grade DCISs were correctly described as suspicious. The difference between non-high grade and high grade DCIS was not significant (p=0.148), while significant differences were found between G1 and G3 DCISs and between G1 and G2 DCISs (p<0.05). All G2 and G3 DCISs showed noticeable signal enhancement. The mean histological tumour size of non-high grade DCISs was smaller than that for high grade DCIS (p<0.05). The hallmark of DCIS on dynamic MRM was unilateral segmental enhancement, most commonly with a granular dotted morphology (M2). Hormone effects need to be considered as the main differential diagnosis. Signal enhancement kinetics similar to invasive carcinoma were seen in the majority of cases. A combined analysis of morphological pattern and signal enhancement considerably improved rate of detection. G2 and G3 DCISs were correctly diagnosed with a significantly higher rate of detection (92%) than G1 DCIS (53%) (p<0.05). Different average size of G1, G2 and G3 DCIS on pathology cannot be excluded as a reason for differences found. Normal MRM seems to exclude high grade DCIS.

Adult↗
PubMed2002

Characterisation of sucrose-negative Yersinia enterocolitica 4/O:3 isolates recovered from pig tonsils.

Sucrose-negative Yersinia enterocolitica isolates of bioserotype 4/O:3 have been recovered for the first time. They were found in 2% of the tonsils of clinically healthy fattening pigs. These sucrose-negative Y. enterocolitica isolates could not be differentiated from Y. kristensenii isolates using API 20E; thus, they were identified using PCR and sequencing. Using pulsed-field gel electrophoresis (PFGE). NotI profiles of sucrose-negative Y. enterocolitica 4/O:3 isolates showed a high similarity to sucrose-positive Y. enterocolitica 4/O:3 isolates. This study demonstrated that sucrose-negative Y. enterocolitica 4/O:3 isolates of porcine origin can harbour virulence genes; plasmid-encoded virulence markers were found in 8 out of 11 isolates and all isolates contained chromosomal-encoded virulence markers. Thus, the pathogenicity of sucrose-negative Yersinia isolates should always be assessed.

Animals↗
PubMed2002

Amplification of 'variola virus-specific' sequences in German cowpox virus isolates.

In 1995 a polymerase chain reaction (PCR) protocol describing the specific detection of variola virus, the causative agent of smallpox, was published by Knight and others. Virulent variola major strains could be differentiated from less virulent variola minor strains because of the distinct amplicon sizes. Here, we applied this PCR protocol to DNA from various orthopoxvirus isolates. There was no amplification with the orthopoxvirus species vaccinia, monkeypox, mousepox, or camelpox viruses. However, amplification was observed in six out of 15 cowpox virus strains investigated. The size of the amplicons corresponded exactly with the size described for variola minor strains and the nucleotide sequence identity accounted for 97%. Findings are discussed with respect to the evolution of orthopoxvirus species assuming that variola virus most probably stems from a rodent-transmitted cowpox virus-like progenitor.

Animals↗
PubMed2002

Strategies for PCR based detection of Burkholderia pseudomallei DNA in paraffin wax embedded tissues.

Recently, several cases of melioidosis imported to Europe have been reported. The diagnosis of the acute or chronic infection remains challenging. This report describes an optimised protocol for fast and reliable DNA preparation for use in two different polymerase chain reaction (PCR) assays, namely: (1) a seminested PCR assay targeting a genus specific sequence of the ribosomal protein subunit 21 (rpsU) gene and (2) a nested PCR assay targeting the gene encoding the filament forming flagellin (fliC). Various strains of Burkholderia spp, strains of closely related genera, and spleen tissue samples of experimentally infected mice were investigated. The combination of PCR and sequencing of the amplicons resulted in high sensitivity and specificity. These procedures may allow rapid, sensitive, and reliable detection of B pseudomallei DNA in routinely formalin fixed and paraffin wax embedded samples, thus providing a safe diagnostic tool and avoiding the cultivation of a risk group 3 agent. In addition, this method could be useful for retrospective histopathological investigations.

Animals↗
PubMed2001

Erythropoiesis in the absence of janus-kinase 2: BCR-ABL induces red cell formation in JAK2(-/-) hematopoietic progenitors.

The receptor-associated protein tyrosine kinase janus-kinase 2 (JAK2) is essential for normal red cell development and for erythropoietin receptor (EpoR) signaling. JAK2(-/-) embryos are severely deficient in erythropoiesis and die at an early stage of development from fetal anemia. The binding of erythropoietin (Epo) to the EpoR triggers the activation of JAK2, the phosphorylation of the EpoR, and the initiation of the EpoR signaling cascade. In addition to Epo binding to its receptor, signaling pathways downstream of the EpoR can also be stimulated by the BCR-ABL oncoprotein. This study explored whether JAK2 is required for BCR-ABL-mediated stimulation of erythropoiesis. Here, it is shown that JAK2 is constitutively tyrosine phosphorylated in cultured and primary erythroid cells expressing BCR-ABL. However, BCR-ABL effectively supports normal erythroid proliferation, differentiation, and maturation in JAK2-deficient fetal liver cells. Using mutants of BCR-ABL, this study shows that certain signaling pathways activated by BCR-ABL segments distinct from its tyrosine kinase domain are essential for rescue of erythropoiesis in JAK2(-/-) progenitors. The consequences of these multiple signaling pathways for normal erythroid development are discussed.

Amino Acid Substitution↗
PubMed2001

Temporal integration of sound pressure determines thresholds of auditory-nerve fibers.

Current propositions of the quantity of sound driving the central auditory system, specifically around threshold, are diverse and at variance with one another. They include sound pressure, sound power, or intensity, which are proportional to the square of pressure, and energy, i.e., the integral of sound power over time. Here we show that the relevant sound quantity and the nature of the threshold can be obtained from the timing of the first spike of auditory-nerve (AN) fibers after the onset of a stimulus. We reason that the first spike is triggered when the stimulus reaches threshold and occurs with fixed delay thereafter. By probing cat AN fibers with characteristic frequency tones of different sound pressure levels and rise times, we show that the differences in relative timing of the first spike (including latencies >100 msec of fibers with low spontaneous rates) can be well accounted for by essentially linear integration of pressure over time. The inclusion of a constant pressure loss or gain to the integrator improves the fit of the model and also accounts for most of the variation of spontaneous rates across fibers. In addition, there are tight correlations among delay, threshold, and spontaneous rate. First-spike timing cannot be explained by models based on a fixed pressure threshold, a fixed power or intensity threshold, or an energy threshold. This suggests that AN fiber thresholds are best measured in units of pressure by time. Possible mechanisms of pressure integration by the inner hair cell-AN fiber complex are discussed.

Acoustic Stimulation↗
PubMed2001

Comparison of closely related orthopoxvirus isolates by random amplified polymorphic DNA and restriction fragment length polymorphism analysis.

The reliability and reproducibility of random amplified polymorphic DNA analysis (RAPD) was compared with restriction fragment length polymorphism (RFLP) by analysing three virus strains isolated from zoo animals in Berlin and three isolates which were cultivated from pets from Northern Germany. The RAPD technique was evaluated as a reliable tool with good reproducibility of the patterns for each virus strain investigated. Problems of interpretation due to inconsistent intensity of bands in different polymerase chain reaction runs may arise for less experienced personnel. The RAPD analysis can be performed within one working day and needs less DNA compared with RFLP so costs will be reduced. The obvious advantage of RFLP is that the pattern can be traced to the recognition site of the restriction enzyme whereas the RAPD primer sequence is not present in the orthopoxvirus genome at all. To the authors knowledge, the RAPD technique has never been applied in DNA viruses before and they conclude that this technique is a useful tool for the discrimination of closely related cowpoxviruses.

Animals↗
PubMed2001

A highly specific one-step PCR - assay for the rapid discrimination of enteropathogenic Yersinia enterocolitica from pathogenic Yersinia pseudotuberculosis and Yersinia pestis.

Based on differences within the yopT-coding region of Yersinia. enterocolitica, Y pseudotuberculosis and Y pestis, a rapid and sensitive one-step polymerase chain reaction assay with high specificity for pathogenic Y enterocolitica was developed. By this method pathogenic isolates of Y enterocolitica can be easily identified and discriminated from other members of this genus. The entire coding sequence of the yopT effector gene of Y. pseudotuberculosis Y36 was determined.

Animals↗
PubMed2001

Comparison of different methods for the isolation of Burkholderia cepacia DNA from pure cultures and waste water.

DNA from Burkholderia cepacia was prepared from suspensions of pure cultures and artificially contaminated waste water. The efficacy of four standard methods (lysis buffer containing proteinase K, phenol/chloroform/isoamylalcohol extraction, microwave treatment, heat treatment) and six commercially available kits (Puregene, High Pure PCR Template Preparation Kit, InstaGene, QIAamp Tissue Kit, DNAzol, Elu-Quik) was compared in terms of sensitivity in a subsequent PCR. The results showed that a simple and inexpensive procedure using a lysis buffer containing proteinase K was superior to all other methods tested.

Burkholderia cepacia↗
PubMed2001

Study on the pathophysiology of experimental Burkholderia pseudomallei infection in mice.

Burkholderia pseudomallei is the etiological agent of melioidosis, a potentially fatal disease occurring in man and animals. The aim of this study was to investigate the pathophysiological course of experimental melioidosis, and to identify the target organs, in an animal model. For this purpose SWISS mice were infected intraperitoneally with the virulent strain B. pseudomallei 6068. The bacterial load of various organs was quantified daily by bacteriological analysis and by an enzyme-linked immunosorbent assay (ELISA) based on a monoclonal antibody specific to B. pseudomallei exopolysaccharide (EPS). Electron microscopic investigation of the spleen was performed to locate the bacteria at the cellular level. In this model of acute melioidosis, B. pseudomallei had a marked organ tropism for liver and spleen, and showed evidence of in vivo growth with a bacterial burden of 1.6x10(9) colony forming units (CFU) per gram of spleen 5 days after infection with 200 CFU. The highest bacterial loads were detected in the spleen at all time points, in a range from 2x10(6) to 2x10(9) CFU g(-1). They were still 50-80 times greater than the load of the liver at the time of peak burden. Other investigated organs such as lungs, kidneys, and bone marrow were 10(2)-10(4)-fold less infected than the spleen, with loads ranging from 3x10(2) to 3x10(6) CFU g(-1). The heart and the brain were sites of a delayed infection, with counts in a range from 10(3) to 10(7) times lower than bacterial counts in the spleen. The EPS-specific ELISA proved to be highly sensitive, particularly at the level of those tissues in which colony counting on agar revealed low contamination. In the blood, EPS was detected at concentrations corresponding to bacterial loads ranging from 8x10(3) to 6x10(4) CFU ml(-1). Electron microscopic examination of the spleen revealed figures of phagocytosis, and the presence of large numbers of intact bacteria, which occurred either as single cells or densely packed into vacuoles. Sparse figures suggesting bacterial replication were also observed. In addition, some bacteria could be seen in vacuoles that seemed to have lost their membrane. These observations provide a basis for further investigations on the pathogenesis of the disease.

Animals↗
PubMed2000

Evaluation of a Yersinia adhesion gene (yadA) specific PCR for the identification of enteropathogenic Yersinia enterocolitica.

A total of 101 Yersinia enterocolitica strains was investigated with a PCR assay [Blais and Phillipe, Food Control, 6 (1995) 211-214] targeting the Yersinia adhesin gene (yadA) responsible for autoagglutination. Compared to the autoagglutination test the PCR assay has a specificity of 100% but a sensitivity of only 70%. This failure might be caused by the sequence heterogeneity of yadA.

Adhesins, Bacterial↗
PubMed2000

Identification of Yersinia enterocolitica within the genus Yersinia.

In this report we describe a PCR strategy for the unambigous identification of biochemically presumptive typed Yersinia (Y.) enterocolitica. A total of 269 isolates belonging to ten species of the genus Yersinia were investigated. In a first PCR only isolates classified as Y. enterocolitica (n = 113) gave rise to a specific amplification resulting in a sensitivity and a specificity of 100%. By sequencing the 269 amplicons of a second pan-Yersinia PCR spanning a distinct 16S rRNA gene region, 20 different sequence clusters could be identified within the genus. By this, Y. enterocolitica isolates of American and European origin could be distinguished safely and already described sequence clusters of the species Y. frederiksenii were confirmed. New 16S rRNA gene sequence clusters were detected for the species Y. frederiksenii, Y. intermedia, Y. mollaretii, Y. aldovae, Y. kristensenii, and Y. rohdei.

Animals↗
PubMed2000

Partial impairment of cytokine responses in Tyk2-deficient mice.

To assess the role of the Janus kinase (Jak) family member Tyk2, we have generated Tyk2-/- mice. In contrast to other Jaks, where inactivation leads to a complete loss of the respective cytokine receptor signal, Tyk2-/- mice display reduced responses to IFNalpha/beta and IL-12 and a selective deficiency in Stat3 activation in these pathways. Unexpectedly, IFNgamma signaling is also impaired in Tyk2-/- mice. Tyk2-/- macrophages fail to produce nitric oxide upon lipopolysaccharide induction. Tyk2-/- mice are unable to clear vaccinia virus and show a reduced T cell response after LCMV challenge. These data imply a selective contribution of Tyk2 to the signals triggered by various biological stimuli and cytokine receptors.

Animals↗
PubMed2000

Yersinia enterocolitica 16S rRNA gene types belong to the same genospecies but form three homology groups.

The species Yersinia (Y.) enterocolitica consists of biochemically and serologically heterogeneous strains. A vernacular nomenclature divides these strains in 'European' and 'American' bioserotypes. We investigated six strains of each group by DNA-DNA hybridization, determination of G + C mol% content and sequence alignment studies. Based on different DNA-DNA hybridization values and the 16S rRNA gene sequences a division into two Yersinia enterocolitica subspecies is justified. We propose the names Yersinia enterocolitica subsp. enterocolitica for strains belonging to the 16S rRNA gene type represented by the Type strain ATCC 9610 and Yersinia enterocolitica subsp. palearctica for strains belonging to the 16S rRNA gene type of strain Y11 (DSMZ13030).

Animals↗
PubMed2000

Serodiagnosis of human plague by an anti-F1 capsular antigen specific IgG/IgM ELISA and immunoblot.

Plague is a re-emerging disease endemic in at least 24 countries. Non-endemic countries should be able to confirm plague to prevent outbreaks due to imported cases. We established a combination of a IgG/IgM screening ELISA and a confirmation immunoblot employing F1 capsular antigen (CA) for the serodiagnosis of plague in countries where yersiniosis is present. The ELISA and the immunoblot assay showed a specificity of 96.1% and 100% among sera from healthy German blood donors. This group had a seroprevalence of 39% of anti-yersinia outer protein (YOP) antibodies obviously caused by previous Y. enterocolitica infection. The ELISA detected anti-F1 CA antibodies in 22 and the immunoblot in 20 out of 26 sera of plague vaccinees. Five control sera from bacteriologically confirmed plague cases from Madagascar reacted positively. It can be concluded that anti-YOP antibodies do not affect assays based on purified F1 CA.

Antibodies, Bacterial↗
PubMed2000

A combination of different polymerase chain reaction (PCR) assays for the presumptive identification of Yersinia pestis.

A combination of four polymerase chain reaction (PCR) assays targeting the Yersinia pestis-specific plasmoidal genes of the fraction 1 capsular antigen and plasminogen activator/coagulase, the gene of the V antigen of the Yersinia virulence plasmid, and the chromosomal 16S rRNA gene was evaluated for the identification of Y. pestis isolates. All four assays were subjected to the same sample preparation technique, reagents and cycling conditions. Eighteen Y. pestis, 66 Y. pseudotuberculosis, 40 Y. enterocolitica strains, the type strains of the other Yersinia species, and 20 other pathogenic bacterial strains were investigated. By using the proposed combination of PCR assays all Y. pestis strains were identified correctly. The applicability of this combination of PCR assays was demonstrated by the detection of Y. pestis DNA in spiked tissues from Rattus norwegicus and fleas (Xenopsylla cheopis and Ctenocephalides spp.). As little as 60 genome equivalents were detected. This system is applicable for monitoring Y. pestis and its vectors in enzootic natural foci and in the diagnosis of plague in humans and animals.

Amino Acid Sequence↗
PubMed2000

A review on melioidosis with special respect on molecular and immunological diagnostic techniques.

Melioidosis is an 'emerging' tropical disease which causes diagnostic problems in endemic and especially in nonendemic areas e.g. Germany when imported. In the last decade, many efforts have been made to develop new molecular and immunological techniques for diagnostic use. However, an actual comprehensive review does not exist. Apart from classical microbiological procedures (microscopy, culture and biochemical identification) efforts have been made to identify Burkholderia pseudomallei using specific antibodies and PCR. The direct antigen detection can be done within a few hours leading to diagnosis days before cultural proof. ELISA and immunoblot techniques were examined for their ability to replace indirect hemagglutination and immunofluorescence test in serology. The diagnostic value of serological procedures for early detection of melioidosis is limited, however. The rapid and reliable diagnosis of melioidosis is required for an adequate onset of therapy. But no evaluated test kit based on the detection of specific antibodies, specific antigens, or on the amplification of species-specific DNA sequences is commercially available up to now. Even PCR testing--primers can easily be ordered by many gene technology companies--can not be recommended as B. pseudomallei DNA for positive controls is not available. Therefore, only microscopy and biochemical identification systems like the API 20NE can be used in the routine laboratory up to now. At this point, it has to be stressed that B. pseudomallei is a level 3 agent in many countries e.g. Germany and that only laboratories with high containment are allowed to handle it. In all cases the help of an experienced reference laboratory is adviced.

Burkholderia pseudomallei↗