Biomedical subjects

D Hahn

Publications and source records attributed to D Hahn.

At least 109 records · Page 6Linked to original sources

PubMed1999

Analysis of bacterial communities in heavy metal-contaminated soils at different levels of resolution.

The impact of heavy metal contamination on soil bacterial communities was studied in soils amended for many years with sewage sludge contaminated with heavy metals to varying extents. At the broad level of resolution, DNA reassociation analysis indicated a dramatic decrease in bacterial diversity from 16{ omitted}000 bacterial genomes (g soil [wet wt])(-1) in the non-contaminated soil to 6400 bacterial genomes (g soil [wet wt])(-1) in soil with low metal amendments and only 2000 bacterial genomes (g soil [wet wt])(-1) in soil with high metal amendments. No differences between bacterial communities of these soils, however, were displayed in the %G+C profiles analysed by thermal denaturation. At a coarse level of characterisation, in situ hybridisation analysing larger phylogenetic groups of bacteria revealed a general decrease in the percentage of cells detected with probes ARCH915, BET42a, GAM42a, SRB385, CF319a, LGCb and HGC69a with increasing metal amendment. Only cells detected with probe ALF1b increased significantly from 3.1+/-0.8% of the cells detected by the domain-specific probe EUB338 in the non-contaminated soil to 6.5+/-1.3% in soil with high metal amendments. These shifts in populations of larger phylogenetic groups were largely confirmed by dot blot analysis of 16S and 23S rDNA clone libraries from bacteria in soil with low metal and high metal amendments, respectively. For a fine-level characterisation, 72 clones of 16S rDNA libraries were identified by comparative sequence analysis. A few sequences could not be assigned to the major taxa described. Most of the sequences were assigned to the Gram-positive bacteria with a high DNA G+C content (45%) and the alpha-subdivision of Proteobacteria (24%). However, only minor differences were seen between bacterial communities from the low and high metal soils. In the soil with high metal amendment, more sequences clustered to the alpha-subdivision of Proteobacteria, while in the low metal soil, more sequences clustered to the Gram-positive bacteria with a high DNA G+C content.

Journal Article↗

PubMed1999

Nonpolarized secretion of human meprin alpha in colorectal cancer generates an increased proteolytic potential in the stroma.

Epithelial cells of the normal human colonic mucosa secrete an astacin-type metalloprotease, meprin a (E. C. 3.4.24.18, N-benzoyl-L-tyrosyl-p-aminobenzoic acid hydrolase), into the intestinal lumen. We found that Caco-2 cells, a colon carcinoma cell line, expressed endogenous meprin alpha, which was secreted at both the basolateral and apical plasma membrane. The expression of meprin alpha in colorectal cancer was confirmed using Northern blot analysis. On tissue sections, a diversity of carcinoma cells with varying immunoreactivity for meprin alpha was observed. Western blots of a series of 11 paired samples of carcinomas and normal control colon tissue revealed that meprin alpha protein accumulated at significant levels in 6 carcinomas at Union International Contre le Cancer tumor stages I-IV. In contrast, the protease was never detected in normal control tissue samples. Meprin alpha zymogen was activated in the tumor tissue, as shown by a 3-fold increase in enzymatic activity. In conclusion, we describe a cancer-specific sorting of meprin alpha, leading to a redistribution with consecutively increased proteolytic activity in the tumor stroma. Because the protease is known to cleave extracellular matrix components in vitro, meprin a may contribute to tumor progression by facilitating migration, intravasation, and metastasis of carcinoma cells.

Blotting, Western↗

PubMed1999

Concentrations of human cardiac phosphorus metabolites determined by SLOOP 31P NMR spectroscopy.

Human cardiac 31P nuclear magnetic resonance (NMR) spectra are usually quantified in relative terms, i.e., the ratio of metabolite signals is calculated. If 31P NMR spectroscopy of the heart is to emerge as a clinically relevant diagnostic modality, reliable quantification of absolute concentrations of 31P metabolites is required. We applied spectral localization with optimal point spread function (SLOOP) 31P NMR spectroscopy to measure absolute concentrations of phosphocreatine (PCr) and adenosine triphosphate (ATP) in human myocardium. The accuracy of the quantification was first validated in a phantom study. Seven healthy volunteers (aged 19-29 years) were then examined at 1.5 T using a nominal spatial resolution of 25 mL. SLOOP allowed us to obtain localized spectra from compartments anatomically matched to the left ventricular wall. The a priori knowledge of the anatomical structure was obtained from 1H images. The spatially varying effects of saturation, off-resonance, and sensitivity were considered during the reconstruction process. Metabolites were quantified with reference to an external 31P standard. Concentrations of 9.0 +/- 1.2 and 5.3 +/- 1.2 mmol/kg wet wt (mean +/- SD, n = 9) were determined for PCr and ATP in normal heart, respectively. The influence of nuclear Overhauser enhancement on metabolite quantification is discussed.

Adenosine Triphosphate↗

PubMed1999

Fast T1 mapping on a whole-body scanner.

A method for the fast acquisition of quantitative T1 maps is presented. It is based on the acquisition of a series of snapshot fast low-angle shot (FLASH) images after inversion of the magnetization. A drawback of this method is the necessity for a sufficiently long relaxation delay before each inversion pulse, leading to long experimental times if averaging or segmentation is required. Thus implementation of this method on a whole-body scanner is problematic, as the longer gradient rise times provide prolonged repetition times, which makes segmentation indispensable to provide a sufficient temporal resolution. We present a modification of the method that allows for reduction in the intermediate relaxation delays and thus for considerably reduced experimental durations.

Artifacts↗

PubMed1999

Signal intensities in FLASH-EPI-hybrid sequences.

Theoretical considerations on the signal-to-noise ratio (SNR) in FLASH-EPI-Hybrid imaging were published previously. The purpose of this work was to investigate in vivo the signal intensities in Hybrid images as a function of sequence specific parameters. In detail, the SNR as a function of the number of echoes m per RF excitation, the excitation flip angle alpha, and the dependence on the tissue relaxation times T1 and T2* were studied. In eight healthy subjects brain and abdominal Hybrid images were acquired where m and alpha were changed independently. Signal intensities in human brain, liver, and kidney were evaluated for each Hybrid experiment. Additionally, T1 and T2* values of these tissue types were quantified to allow for a comparison with the theory. An excellent agreement between calculated and measured signal behavior was found. The theory was therefore validated in vivo and can thus be used to optimize the signal-to-noise in Hybrid experiments.

Brain↗

PubMed1999

Differential enumeration and in situ localization of microorganisms in the hindgut of the lower termite mastotermes darwiniensis by hybridization with rRNA-targeted probes

We examined the abundance and spatial distribution of major phylogenetic groups of the domain Bacteria in hindguts of the Australian lower termite Mastotermes darwiniensis by using in situ hybridization with group-specific, fluorescently labeled, rRNA-targeted oligonucleotide probes. Between 32.0 +/- 7.2% and 52.3 +/- 8.2% of the DAPI-stained cells in different hindgut fractions were detected with probe EUB338, specific for members of the domain Bacteria. About 85% of the prokaryotic cells were associated with the flagellates of the thin-walled anterior region (P3a) and the thick wall of the posterior region (P3b/P4) of the hindgut, as shown by DAPI staining. At most, half of the EUB338-detected cells hybridized with one of the other probes that targeted a smaller assemblage within the bacterial domain. In most fractions, cells were found in varying numbers with probe ALF1b, which targeted members of the alpha-Proteobacteria, whereas substantial amounts of sulfate-reducing bacteria, gram-positive bacteria with a high DNA G+C content and members of the Cytophaga-Flavobacterium cluster of the Cytophaga-Flavobacterium-Bacteroides (CFB) phylum could be detected only in the wall fraction of P3b/P4. This clearly indicates that the hindgut microhabitats differ in the composition of their microbial community. In situ hybridization of cryosections through the hindgut showed only low numbers of bacteria attached to the P3a wall. In contrast, the wall of P3b was densely colonized by rod- and coccus-shaped bacteria, which could be assigned to the Cytophaga-Flavobacterium cluster of the CFB phylum and to the group of gram-positive bacteria with a high DNA G+C content, respectively. Oxygen concentration profiles determined with microelectrodes revealed steep oxygen gradients both in P3a and P3b. Oxygen was consumed within 100 &mgr;m below the gut surface, and anoxic conditions prevailed in the central portions of both gut regions, indicating that oxygen consumption in the hindgut does not depend on the presence of a biofilm on the hindgut wall.

Journal Article↗

PubMed1999

Identification of DNA sequences controlling light- and chloroplast-dependent expression of the lhcb1 gene from Chlamydomonas reinhardtii.

In Chlamydomonas reinhardtii, expression of the lhcb1 gene encoding a chlorophyll-a/b-binding protein of photosystem II is highly regulated by light, inhibitors of chlorophyll synthesis, as well as by circadian rhythms. In light/dark synchronized cultures, the rapid increase of lhcb1 mRNA levels during the light phase is regulated primarily at the transcriptional level. We have used the arylsulphatase (ars) reporter gene to analyze the lhcb1 5' upstream sequences for the presence of light-responsive elements. In transformants carrying chimeric reporter genes, accumulation of lhcb1/ars mRNA is markedly stimulated by light, with a time course similar to that of transcripts from the endogenous lhcb1 gene. Promoter deletion studies revealed that a 255-bp fragment of the lhcb1 5' upstream region is sufficient to confer proper light regulation on the promoterless ars gene. Moreover, the region between positions -255 and -122 with respect to the start site of translation were found to contain one or more light-responsive elements. Strikingly, these sequences also seem to be involved in chloroplast-dependent lhcb1 gene expression as indicated by Northern analyses of transformants with photo-oxidatively damaged chloroplasts. This suggests that both light- and chloroplast-dependent expression of the lhcb1 gene are mediated by the same cis-acting elements.

Animals↗

PubMed1999

Spiral CT angiography of renal arteries: comparison with angiography.

A prospective study was carried out to determine the accuracy of spiral CT angiography (CTA) in the detection of renal artery stenosis (RAS). Eighty-two patients with arterial hypertension underwent CTA and digital subtraction angiography (DSA) to exclude RAS. For CTA a contrast medium bolus of 100-150 ml (flow rate 3 ml/s) was injected. A 24 or 40 s CTA was started at the origin of the superior mesenteric artery after a delay time determined by test bolus injection (collimation = 2 mm, pitch = 1/1.5). For stenosis detection transverse images supported by maximum intensity projections (MIP) or multiplanar reconstruction projections were used. Of 197 renal arteries examined (including 33 accessory arteries), 34 RAS were visualized using DSA. With CTA, one hemodynamic RAS was missed and one additional hemodynamic RAS was found. Sensitivity/specificity was calculated to be 94 %/98 %. For hemodynamically relevant RAS (> 50 %) the sensitivity/specificity was 96 %/99 %. CTA additionally depicted five adrenal masses. The high accuracy rate of RAS detection thus allows the use of CTA as a screening method in patients with arterial hypertension to exclude a renovascular cause.

Adult↗

PubMed1999

Intralobar lung sequestration with systemic coronary arterial supply.

Pulmonary sequestration is a rare anomaly. An accurate pre-operative evaluation of its vascular supply is essential for the surgeon's operative approach. We describe here an intrapulmonary sequestration with vascular arterial supply via the left circumflex and the right coronary artery. This case demonstrates that if aortography is unrevealing, then a coronary source should be considered in the preoperative search for the arterial supply to a pulmonary sequestration. Moreover, pulmonary sequestration should be listed in the differential diagnosis of aberrant coronary arteries.

Bronchopulmonary Sequestration↗

PubMed1999

Multivariate analysis of the adjustment of the colour duplex unit for the differential diagnosis of lymph node alterations.

Intranodal angioarchitecture was used as criterion for the differential diagnosis of lymph node alterations by colour duplex sonography. The influence of the adjustment of the duplex unit on the diagnostic accuracy and on the rate of lymph nodes with detectable intranodal flow signals was tested. A total of 243 superficial lymph nodes in 127 patients were included in a retrospective study. Diagnoses were approved by histopathology (n = 173) or clinical follow-ups (n = 70 reactive lymph nodes). The intranodal angioarchitecture and sonomorphology were assessed. We used nonparametric discriminant analysis in order to define appropriate settings for transducer frequency, flow mode, power and threshold which result in the best diagnostic quality. Of the lymph nodes, 87 % displayed flow signals using a threshold of >/= 13 and the flow modes low or medium. Vascularisation was detected only in 47 % of the nodes assessed with different adjustments. Diagnostic accuracy dropped from 94 to 68-84 % if threshold was < 13 or power was less than -11 dB. An insufficient adjustment of the colour duplex unit decreases the chance of detecting intranodal flow signals and impairs the diagnostic quality of colour duplex sonography. Flow mode, power and threshold are the most important parameters which have to be adjusted properly.

Adolescent↗

PubMed1999

Current value of double-contrast pharyngography and of computed tomography for the detection and for staging of hypopharyngeal, oropharyngeal and supraglottic tumors.

In light of recent endoscopic techniques the current value of double-contrast pharyngography (DCP) and of CT for detection and staging of hypo-, oropharyngeal, and supraglottic tumors is evaluated. The DCP of 151 patients and CT obtained from 99 of these patients were retrospectively analyzed in a double-blinded manner. We used a standard protocol which comprised all relevant anatomical subregions. Results were compared with direct microlaryngoscopy (DL), indirect laryngoscopy (IL), and post-operative histopathological findings. Sensitivity and specificity of DCP was 75.0 % and 86.7 %, respectively. The DCP and IL techniques together yielded a higher sensitivity (96.7 %) than each method separately. Sensitivity and specificity of CT was 87.5 and 100 %, respectively. In 74.7 % CT provided correct staging. Subregional analysis revealed that the results of DCP and CT depend highly on the localization of the tumor. Our results indicate that DCP represents an important screening method for diagnosing hypo-, oropharyngeal, and supraglottic tumors to complete IL and DL. We show that CT is a reliable method for preoperative staging, although small superficial tumors may occasionally be missed by this method.

Administration, Oral↗

PubMed1999

Enrichment, phylogenetic analysis and detection of a bacterium that performs enhanced biological phosphate removal in activated sludge.

Activated sludge communities which performed enhanced biological phosphate removal (EBPR) were phylogenetically analyzed by 16S rRNA-targeted molecular methods. Two anaerobic-aerobic sequencing batch reactors were operated with two different carbon sources (acetate vs. a complex mixture) for three years and showed anaerobic-aerobic cycles of polyhydroxybutyrate- (PHB) and phosphate-accumulation characteristic for EBPR-systems. In situ hybridization showed that the reactor fed with the acetate medium was dominated by bacteria phylogenetically related to the Rhodocyclus-group within the beta-Proteobacteria (81% of DAPI-stained cells). The reactor with the complex medium was also predominated by this phylogenetic group albeit at a lesser extent (23% of DAPI-stained cells). More detailed taxonomic information on the dominant bacteria in the acetate-reactor was obtained by constructing clone libraries of 16S rDNA fragments. Two different types of Rhodocyclus-like clones (R1 and R6) were retrieved. Type-specific in situ hybridization and direct rRNA-sequencing revealed that R6 was the type of the dominant bacteria. Staining of intracellular polyphosphate- and PHB-granules confirmed that the R6-type bacterium accumulates PHB and polyphosphate just as predicted by the metabolic models for EBPR. High similarities to 16S rDNA fragments from other EBPR-sludges suggest that R6-type organisms were present and may play an important role in EBPR in general. Although the R6-type bacterium is closely related to the genus Rhodocyclus, it did not grow phototrophically. Therefore, we propose a provisional new genus and species Candidatus Accumulibacter phosphatis.

Aerobiosis↗

PubMed1999

Detection of myocardial viability by low-dose dobutamine Cine MR imaging.

The purpose of this work was to test the diagnostic value of dobutamine stress magnetic resonance imaging (MRI) for predicting recovery of regional myocardial contractility after revascularization. Cardiac wall motion abnormalities are due to either non-viable and/or scarred, or viable, but hibernating, myocardial tissue. Dobutamine stress leads to increased systolic wall thickening only in viable myocardium. Twenty-five patients with akinetic or dyskinetic myocardial regions were examined with a Cine FLASH-2D sequence at rest and during dobutamine stress (10 microg/kg/min). Patients were re-examined at rest 3, and in case of persisting wall motion defects, 6 months after revascularization. Criterion of viability was increasing end-systolic wall thickening during stress and/or at follow-up. Akinetic regions related either to the LAD (n = 19) or to the RCA (n = 6) were judged viable if > or = 50% of the affected segments improved. MR studies were completed in all subjects without arrhythmia or need for early terminations due to symptoms. Sensitivity, specificity, and positive predictive value for the prediction of myocardial viability were 61%, 90%, and 87% for the segment-related analysis, and 76%, 100%, and 100% for the patient-related analysis based on coronary artery distribution, respectively. Dobutamine stress MRI allows to predict global functional recovery of akinetic myocardial regions after revascularization with a high positive predictive value and high specificity.

Adult↗

PubMed1999

Cine MR imaging after myocardial infarction--assessment and follow-up of regional and global left ventricular function.

Myocardial infarction often leads to regional wall motion defects and in case of large defects to remodeling of the left ventricle. With this study, changes in regional and global myocardial function of 12 patients 3 weeks after myocardial infarction and after revascularization therapy were determined using MRI. Cine MRI was performed at study entry at rest and during low-dose dobutamine stimulation. All patients were re-examined at rest 3 and 6 months after the revascularization, including analysis of wall thickening and of left ventricular end-diastolic volume index (LVEDVI), end-systolic volume index (LVESVI), ejection fraction (LVEF), and mass index. After revascularization. 6 patients with stress-induced improvement of regional wall thickening recovered, 4 patients without improvement did not, but 2 patients without stress-induced improvement of wall thickening also recovered. Concerning global cardiac function, patients with mainly improved regional wall motion also showed a lower LVESVI and a higher LVEF than patients without improvement of regional contractility 6 months after revascularization in comparison to study entry. In conclusion, improvement of global myocardial function after revascularization is higher in patients with improved contractility in the infarcted region. The extent of the response of regions with wall motion defects to dobutamine stress correlates with the actual improvement after revascularization, and, therefore, dobutamine stress MRI may be helpful in selecting patients that will have a higher benefit from a revascularization therapy.

Adult↗

PubMed1999

Secretion of human meprin from intestinal epithelial cells depends on differential expression of the alpha and beta subunits.

Human meprin (N-benzoyl-l-tyrosyl-p-aminobenzoic acid hydrolase, EC 3.4.24.18), an astacin-type metalloprotease, is expressed by intestinal epithelial cells as a dimeric protein complex of alpha and beta subunits. In transfected cells, intracellular proteolytic removal of the membrane anchor from the alpha subunit results in its secretion, while the beta subunit and alpha/beta heterodimers are retained at the cell membrane. We investigated the consequence of differential intracellular processing of alpha and beta subunits in the human small and large intestine using subunit-specific immunohistochemistry, in situ hybridization and biosynthetic studies in organ culture. In the ileum, both subunits localize to the brush-border membrane of villus enterocytes. In contrast, the beta subunit is not expressed in the colon, which leads to the secretion of the alpha subunit. We conclude that differential expression of meprin alpha and beta subunits is a unique means of targeting the proteolytic activity of the alpha subunit either to the brush-border membrane in the ileum or to the lumen in the colon, suggesting dual functions of cell-associated and luminal meprin. Meprin alpha and beta subunits are also coexpressed in distinct lamina propria leukocytes, suggesting an additional role for this protease in leukocyte function in the intestinal mucosa.

Colon↗

PubMed1999

N-Benzoyl-L-tyrosyl-p-aminobenzoic acid hydrolase beta (human meprinbeta). A 13-amino-acid sequence is required for proteolyticprocessing and subsequent secretion.

N-Benzoyl-L-tyrosyl-p-aminobenzoic acid hydrolase or human meprin (PPH) is a brush-border membrane enzyme of small intestinal epithelial cells. It is a type I integral membrane protein composed of two disulphide-bridged subunits (alpha and beta). PPH and its homologous counterparts in rodents belong to the astacin family of zinc-metalloendopeptidases. Although the amino-acid sequence of the beta subunits is 80-90% identical in these three species, processing is different. Expression of PPHbeta in simian virus 40-transformed African green monkey kidney cells (COS-1) and Madin Darby canine kidney (MDCK) cells results in its cell surface localization and secretion, whereas mouse meprinbeta is only found at the plasma membrane. To investigate proteolytic processing of PPHbeta and to identify the cleavage site, different C-terminal domains of wild-type PPHbeta were exchanged with the homologous domains of mouse meprinbeta. We identified a 13-amino-acid sequence (QIQLTPAPSVQDL) necessary for cleavage and subsequent secretion of PPHbeta. Using brefeldin A, the site of processing was identified as being after passage through the Golgi compartment. Proteolytic processing of PPHbeta thus provides a means for secretion of alphabeta heterodimers.

Amino Acid Sequence↗

PubMed1999

[CINE-MRT for the study of the effects of regional left ventricular wall motion disorders on global heart function after a myocardial infarct and revascularization].

PURPOSE: To determine changes in global cardiac function and mass caused by infarct-associated regional wall motion abnormalities and to compare the changes after revascularization in patients with and without improvement of regional contractility. MATERIALS AND METHODS: 21 patients with regional left ventricular wall motion abnormalities and associated coronary artery stenoses requiring revascularization were examined with a Cine FLASH-2D sequence 26 +/- 12 days after their first myocardial infarction and re-examined three months after revascularization. Regional contractility and volumes and masses of both ventricles were determined. RESULTS: After revascularization, regional wall motion improvement led to decreased left ventricular volumes and improved ejection fractions, whereas patients with persisting wall motion defects showed unchanged left ventricular functional parameters. Comparing both groups of patients, the patients with improvement of regional contractility revealed lower end-systolic volumes and higher ejection fractions at follow-up. Cardiac masses and right ventricular parameters were not different, patients with a depressed right ventricular ejection fraction showed improvement at follow-up. DISCUSSION: After myocardial infarction, revascularization of the infarct-related coronary artery leads to an improvement of left ventricular function only if there is also an improvement of regional contractility. An effect on right ventricular function was not observed three months after the first small- or middle-sized myocardial infarction.

Adult↗

PubMed1999

[The quantification of energy-rich phosphates in healthy and damaged heart muscle by SLOOP 31P-MR spectroscopy. Spatial localization with optimal pointspread function].

PURPOSE: A quantitative 31P-MR-spectroscopic technique was used to assess the energy metabolism in healthy and diseased myocardium. METHODS: 31P spectra were acquired on a 1.5 T scanner using a 3D-chemical shift imaging technique. Based on the anatomical information provided by 1H images, SLOOP (Spatial Localization with Optimal Pointspread Function) allows to obtain spectra from defined compartments. With SLOOP a free voxel shape with adaption to anatomic structures, e.g. the myocardium, is possible. Absolute values for phosphocreatine (PCr) and adenosine triphosphate (ATP) were determined using an external standard. RESULTS: 31P-spectra showed only minimal contamination by surrounding tissue. The standard deviation for the determined values of healthy volunteers was low. Compared to healthy volunteers, reduced PCr and ATP concentrations were seen for dilative cardiomyopathies and coronary artery disease and unchanged concentrations were observed for hypertensive heart disease. CONCLUSION: 31P-MR spectroscopy with SLOOP allows a non-invasive, quantitative analysis of cardiac energy metabolism.

Adenosine Triphosphate↗