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S Steiner

Publications and source records attributed to S Steiner.

At least 37 records · Page 2Linked to original sources
PubMed2000

Proteomics: applications in basic and applied biology.

The rapid evolution of proteomics has continued during the past year, with a series of innovations in the core technologies of two-dimensional electrophoresis and mass spectrometry, and a diversity of productive research programmes. Well-annotated proteomics databases are now emerging in a number of fields to provide a platform for systematic research, with particularly promising progress in clinical applications such as cardiology and oncology. Large-scale quantitative research, comparable in power and sensitivity to that achieved for gene expression, is thus becoming a reality at the protein level.

Databases, Factual↗
PubMed2000

Pharmaceutical proteomics.

Genomics and proteomics are today well established in drug discovery and, in combination with combinatorial chemistry and high-throughput screening, are helping to bring forward an unprecedented number of potential lead compounds. To avoid the generation of bottlenecks downstream in drug development, increasing pressure is arising to integrate these technologies into the development environment. Proteomics has demonstrated proof-of-concept in toxicology as shown by a number of successful applications in mechanistic toxicology and lead selection. The "technology wave" is now starting to impact the clinical phase of drug development. Expected benefits are optimized clinical trials based on the availability of biologically relevant markers of drug efficacy and safety.

Animals↗
PubMed1999

4-chloro-o-phenylenediamine induces a dose-related increase in G:C > T:A transversions and one major DNA adduct in the liver of Big Blue mice after 26 weeks in feed treatment.

The monocyclic aromatic amine 4-chloro-o-phenylenediamine (4-C-o-PDA), a known mutagen and mouse hepatocarcinogen, was tested for its in vivo mutagenic potential in the Big Blue transgenic mouse assay system. Genomic DNA was isolated from liver tissue of control and treated animals and lacI mutants were recovered. In an initial 2-week study 4-C-o-PDA was administered daily per os to groups of male and female C57BL/6 Big Blue mice at doses of 0 and 200 mg/kg for 2 weeks (on working days) followed by a treatment free expression time of 10 days. Only a weak increase in the mutant frequencies in females was observed. In a 26-week study, where 4-C-o-PDA was given to groups of male and female Big Blue mice in feed at dietary concentrations of 0, 5,000 and 10,000 ppm, 4-C-o-PDA was found to induce a pronounced dose-dependent increase in mutant frequencies in either sex. In the present work, we analyzed the mutation spectrum by automated DNA sequencing of lacI mutants from both studies. Following the 2-week administration of 4-C-oT:A transversions in both sexes. In addition, upon 26-week treatment with 4-C-o-PDA, one major DNA adduct was detected by 33P postlabelling and subsequent multidimensional thin layer chromatography. It is concluded that 4-C-oT:A transversions after 26 weeks in feed treatment. This result indicates that the sensitivity of the Big Blue transgenic assay system, in detecting a unique chemically induced mutation spectrum, is dependent on experimental parameters, such as treatment time. The data suggest that the formation of one major DNA adduct upon 4-C-o-PDA treatment may be critical for its mutagenicity.

Adenine↗
PubMed1999

[Development of evidence-based clinical practice guidelines; a model project integrating external evidence and clinical expertise].

BACKGROUND: Internationally, evidence-based clinical practice guidelines have been established for several years. In Germany, they are now gaining increasing importance. OWN STUDIES: The Institute of Health Economics and Clinical Epidemiology at the University of Cologne has developed a mostly standardized process for evidence-based guideline development. This process has been implemented in such a way that reproducibility, validity and practicability of guidelines can be assured: evidence from research (external evidence) has been searched and evaluated according to the criteria of the Agency of Health Care Policy and Research (AHCPR). Through cooperation with clinical experts, this evidence has been transferred into practicable recommendations. In addition, acceptance of guideline users and patients was improved by applying this method. CONCLUSION: Evidence-based guidelines are considered as an appropriate tool to assist medical decision-making of all stakeholders involved. The process will be demonstrated developing an evidence-based guideline for obesity treatment.

Evidence-Based Medicine↗
PubMed1999

Congenital asplenia detected in a 60 year old patient with septicemia.

A 60 year-old female who had never been seriously ill, was brought to the emergency ward after she had been found somnolent at home. She developed renal failure, meningitis, respiratory distress and disseminated intravascular coagulation. Overwhelming septicemia was evident, and streptococcus pneumoniae was isolated from blood and cerebrospinal fluid. Surprisingly, peripheral blood smears showed numerous Howell-Jolly-bodies, indicating severe impairment of splenic function. On abdominal ultrasound, CT-scan, and scintigraphy no spleen could be detected. There was no history of abdominal surgery. Apparently, congenital asplenia, which was not noticed until the age of 60, was responsible for the patient's life-threatening septicemia. We suggest that, in cases of severe septicemia, the examination of a blood smear is useful to detect functional (or congenital) asplenia.

Bacteremia↗
PubMed1999

Compact organization of rRNA genes in the filamentous fungus Ashbya gossypii.

The rDNA cluster in the phytopathogenic fungus Ashbya gossypii consists of approximately 50 tandem repeat units of 8197 bp. Each unit carries a gene for the 35S pre-rRNA, processed into 18S, 5.8S and 25S rRNA, and a divergently transcribed gene for 5S rRNA. The well-characterized rDNA of the yeast Saccharomyces cerevisiae is the only other example of a completely sequenced rDNA unit (9137 bp) carrying both a 35S pre-rRNA and a 5S rRNA gene. The coding regions for the 5S, 5.8S, 18S and 25S rRNAs are 95-100% identical whereas transcribed and non-transcribed spacers show 43-66% sequence identity. Functionally characterized rDNA and rRNA elements of S. cerevisiae can be unambiguously recognized in the A. gossypii sequence, including the RNA polymerase-I transcription start site, two Reb1p enhancer binding sites and numerous recognition sequences for rRNA modification and processing. In addition to these functionally characterized sequences eight highly conserved elements from 10 to 71 bp were detected in the over 600-bp transcribed region upstream of the 18S rRNA gene which most likely play as yet uncharacterized functions at the DNA or RNA level. In addition to this work we started to identify A. gossypii homologs of S. cerevisiae nucleolar proteins involved in rDNA maturation.

Amino Acid Sequence↗
PubMed1999

[Influence of pulmonary hemodynamics on right ventricular ejection fraction in chronic obstructive pulmonary disease].

Right ventricular dysfunction is common in patients with chronic obstructive pulmonary disease. Right ventricular function might be influenced by the afterload, which depends on pulmonary artery pressure (PAP) and pulmonary vascular resistance (PVR). To evaluate the influence of the right ventricular afterload on right ventricular performance, we investigated 30 patients with chronic obstructive pulmonary disease without clinical signs or history of left heart failure or coronary heart disease. The study includes lung function tests, analysis of blood gases and right heart catheterisation. RV function was assessed by a thermodilution technique using a pulmonary artery catheter equipped with a rapid response thermistor (produced by Baxter, USA). There are 9 patients with normal, 12 with latent and 9 with fixed pulmonary hypertension. Median RVEF was measured to be 33.3% (19-44%). There was a significant correlation between RVEF and PAP (r = -0.66; p < 0.0001) and RVEF and PVR (r = -0.54; p < 0.0018). RVEF was not directly influenced by lungfunction or pulmonary capillary wedge pressure (PCWP). Under treadmill exercise RVEF and cardiac index increased without a change of PCWP. A low RVEF at rest seems to be a predictive value for a reduced exercise capacity. A reduced RVEF has a predictive value of pulmonary hypertension with a sensitivity of 66% in patients with unstable and 89% in patients with lasting pulmonary hypertension. In a subgroup of 6 cases treadmill exercise led to a RVEF decrease. These patients showed no difference in afterload, blood gases or lung function-tests compared with the total group. In conclusion, right ventricular ejection fraction seems to be influenced by PVR and PAP which determinate the right ventricular afterload. The validity of the method depends on the severity of pulmonary hypertension, and hence measurement of RVEF might not provide a reliable estimation of pulmonary arterial pressure in patients suffering from mild to moderate pulmonary hypertension.

Female↗
PubMed1998

Cyclosporine A-induced decrease in calbindin-D 28 kDa in rat kidney but not in cerebral cortex and cerebellum.

Recently, we reported that in rat, cyclosporine A (CsA) markedly decreases the levels of calbindin-D (CABP-D) 28 kDa in kidney. CABP-D 28 kDa is a calcium-binding protein which is highly expressed in calcium-transporting tissues such as kidney or brain. In this study, we investigated whether, in addition to the kidney, CsA also has an effect on CABP-D 28 kDa in rat brain. Three groups of male Wistar rats received 15 mg/kg/day or 50 mg/kg/day of CsA orally for 12 days, whereas controls received vehicle solution for the same period. CABP-D 28-kDa protein and CsA were quantified in homogenates of kidney, cerebral cortex and cerebellum, and the localization of CABP-D 28 kDa was assessed in the different tissue sections by immunohistochemistry. In kidney, CABP-D 28 kDa was strongly and dose dependently decreased, and was located in tubular epithelial cells. In brain, CABP-D 28 kDa was not changed and was mainly located in pyramidal cells of the cortex and in cerebellum exclusively in Purkinje cells. High CsA concentrations were measured in kidney, more than 17-fold greater than those found in cortex. In cerebellum, CsA was below the limit of detection. These data suggest that at clinically relevant doses, CsA may not affect CABP-D 28-kDa levels in brain.

Animals↗
PubMed1998

The cyclosporine A-induced decrease in rat renal calbindin-D28kDa protein as a consequence of a decrease in its mRNA.

Cyclosporine A (CsA) is a potent immunosuppressant with the drawback of renal side-effects. We recently reported that relatively high doses of CsA markedly decreased the calcium-binding protein calbindin-D28kDa in kidneys of male Wistar rats, and showed that this decrease could be associated with some of the drug-induced adverse renal effects. To investigate the events leading to this decrease, the calbindin-D28kDa mRNA level in kidneys of rats treated with 15 or 50 mg/kg/day CsA for 12 days was analysed by reverse transcription followed by polymerase chain reaction. At both doses, a marked dose-dependent decrease in the calbindin-D28kDa mRNA level was found, one very similar to the decrease measured in the calbindin-D28kDa protein abundance. Thus, the CsA-mediated down-regulation of the renal calbindin-D28kDa protein is most likely the result of a decrease in the calbindin-D28kDa mRNA level.

Animals↗
PubMed1998

New insights into cyclosporine A nephrotoxicity by proteome analysis.

Using two-dimensional gel electrophoresis (2-DE), we recently discovered an association between decreased calcium-binding protein, calbindin-D 28 kDa, urinary calcium wasting and intratubular corticomedullary calcifications in rat kidney. This observation prompted us to investigate kidney tissues of other species, including man. In this paper we show that in dogs and monkeys, which are generally devoid of cyclosporine A (CsA)-mediated nephrotoxicity, renal calbindin levels were not affected by the CsA treatment whereas in CsA-treated human kidney-transplant recipients with renal vascular or tubular toxicity, a marked decrease in renal calbindin-D 28 kDa protein level was found in most of the kidney biopsy sections. The present results strongly suggest that calbindin is a marker for CsA-nephrotoxicity. The discovery of calbindin-D 28 kDa being involved in CsA toxicity has evolved from the application of 2-DE and has not been reported previously, proving that proteomics can provide essential information in mechanistic toxicology. Considering the current improvements in proteome methods it is expected that high throughput proteomics will become an indispensable tool in preclinical safety testing.

Animals↗
PubMed1998

Efficient identification of point mutations by automated DNA sequencing of artificial heterozygote samples.

DNA sequencing templates of individual point mutants of the lacI target gene were amplified by polymerase chain reaction (PCR). By mixing the PCR fragments from two individual mutants in a defined ratio, samples of artificial heterozygous composition were prepared. These samples were then submitted to automated DNA sequencing. The simultaneous, visual comparison of the mixed mutant traces using a graphics program efficiently revealed all heterozygous positions. Based on the individual intensities of the heterozygous base signals the identified point mutations could be assigned to the corresponding mutants. This efficient approach doubles the sample throughput for both the sequencing reactions and the gel electrophoresis using an automated DNA sequencing system.

Animals↗
PubMed1998

Changes in the liver protein pattern of female Wistar rats treated with the hypoglycemic agent SDZ PGU 693.

SDZ PGU 693 acts as a hypoglycemic agent by stimulating glucose utilisation in insulin-sensitive peripheral tissues, such as skeletal muscle and fat. In a 28 day toxicity study the compound was found to induce hepatocellular hypertrophy in Wistar rats treated with 300 mg/kg/day. To gain insights into the pathomechanism of these alterations, aliquots of liver samples from control and treated female Wistar rats were separated by two-dimensional protein gel electrophoresis and the digitized images of the protein patterns were searched for protein abundance changes. Significant treatment-related quantitative changes (P < 0.001) were found in 29 liver proteins. Major increases were observed in several microsomal proteins, including NADPH cytochrome P-450 reductase, cytochrome b5 and serine protease inhibitor. The changes in the cytochrome related enzymes, both known co-factors of the P-450 enzyme system, strongly suggest that SDZ PGU 693 induces microsomal proliferation and induction of the P-450 enzyme system. Decreases were observed in a series of mitochondrial proteins, such as F1ATPase-delta subunit and ornithine aminotransferase precursor as well as in several cytosolic proteins such as the liver fatty acid binding protein, arylsulfotransferase and the senescence marker protein-30. The changes in F1ATPase-delta subunit and liver fatty acid binding protein together suggest a down-regulation of the mitochondrial liver fatty acid metabolism, likely reflecting the pharmacological action of the compound. These results show that SDZ PGU 693 produces a complex pattern of gene expression changes which give insights into the molecular mechanisms of both its pharmacological action and a toxic response.

Animals↗
PubMed1998

[Results and complications of fiber bronchoscopy in HIV positive patients].

Fibreoptic bronchoscopy is an established diagnostic procedure for HIV-associated pulmonary infections. We retrospectively evaluated the diagnostic effectivity and safety of fibreoptic bronchoscopy with bronchoalveolar lavage (BAL) and transbronchial biopsy (TBB) in 153 patients with late-stage HIV infection and clinical signs of pulmonary infection or abnormal chest radiograph. Bronchoscopy leads to diagnosis in 82.4% and changed therapy in 54%. 45 patients (30%) were found to have pneumocystis carinii pneumonia (PCP), the most common bronchoscopic finding, followed by bacterial lung disease (29.3%). BAL had a sensitivity of 78% for PCP. Diagnostic yield of BAL for PCP was higher in patients without previous treatment (positive results in 82%) with regard to PCP independend of the prior treatment. Serious complication occurred in 22 cases (pneumothorax: 6 (3.9%), bleeding: 12 (7.8%), hypoxaemia: 4 (2.6%)). High serum levels of lactate dehydrogenase (LDH) correlated with pulmonary complications like pneumothorax. Age, sex and kind of pulmonary infection did not influence complication rates. 6 (3.9%) episodes of spontaneous pneumothorax occurred in the further course, 3 of them concurrently with PCP or prior history of PCP. We conclude that fibreoptic bronchoscopy is of great value for diagnosing pulmonary infection in HIV-seropositive patients. TBB provides incremental diagnostic information not available from BAL, especially in patients pretreated with cotrimoxazol or pentamidin. For that reason we believe that TBB should be performed in these patients.

AIDS-Related Opportunistic Infections↗
PubMed1997

Decrease in kidney calbindin-D 28kDa as a possible mechanism mediating cyclosporine A- and FK-506-induced calciuria and tubular mineralization.

The use of the immunosuppressant cyclosporine A (CsA) is limited by its adverse renal effects. Most recently, we reported that the drug markedly decreases the levels of the calcium-binding protein calbindin-D 28kDa in kidneys of male Wistar rats. In the present study, the potential relationship between drug-induced nephrotoxicity and the decrease in kidney calbindin-D 28kDa was investigated. Four groups of male Wistar rats were treated for 10 or 31 days with either the immunosuppressant CsA (50 mg/kg/day), FK-506 (5 mg/kg/day), rapamycin (5 mg/kg/day) or with the nonimmunosuppressive cyclosporine derivative 3'keto-[Bmt1]-[Val2]-CsA (SDZ PSC-833) (50 mg/kg/day), and the effects on calcium homeostasis, kidney histology and renal calbindin-D 28kDa were examined. Similar effects were found with CsA and FK-506; both drugs strongly reduced kidney calbindin-D 28kDa protein levels, increased urine calcium excretion, caused intratubular calcification, and induced basophilic tubules. In contrast, rapamycin and SDZ PSC-833 caused no decrease in renal calbindin-D 28kDa levels, no noticeable alterations in calcium metabolism, and no renal calcification. The results provide evidence for a link between decreased renal calbindin, increased calcium urine excretion, and intratubular kidney calcification. The present data show no correlation between the decrease in renal calbindin and the induction of basophilic tubules; however, it needs to be investigated if these apparently independent kidney effects may have a common origin upstream of calbindin expression.

Animals↗
PubMed1997

[Prognostically relevant parameters in patients with coronary heart disease, arterial hypertension and sleep apnea disorders].

Patients with untreated sleep apnea syndrome have a higher cardiovascular mortality. It is not known which mechanisms lead to this increase in mortality and whether it is independent from the often associated coronary heart disease and systemic hypertension. In 48 consecutive patients with coronary heart disease confirmed by angiography, exercise-ECG, Holter-ECG, echocardiography, spirometric tests, analysis of ventricular late potentials, heart rate variability and a test for sleep-disordered breathing with a screening device were performed. Seventeen patients showed disordered breathing during sleep (obstructive sleep apnea) with a desaturation index of > or = 10 (mean desaturation index 17.3 +/- 9.3 vs. 2.6 +/- 3.1 in the patients without sleep-disordered breathing). There are no significant differences in age (58.9 +/- 6.1 vs. 59.7 +/- 7.6 years), body-mass-index (28.6 +/- 3.7 vs. 27.7 +/- 3.3 kg/m2), left ventricular ejection fraction (57.2 +/- 13.6 vs. 64.0 +/- 14.6%), forced expiratory volume in 1 second/vital capacity 95.4 +/- 13.9 vs. 92.9 +/- 11.2% predicted, heart rate variability (standard deviation of the RR-intervals 39.4 +/- 29.4 vs. 37.2 +/- 17.0 ms), the frequency of premature ventricular beats over 24 h and at night, the frequency of multivessel disease (71 vs. 68%), additional hypertension 53 vs. 48%), status postmyocardial infarction (47 vs. 48%) and positive late potential analysis (24 vs. 13%). There were no ST segment depressions during the night. Patients with coronary heart disease and mild sleep-disordered breathing show no significant differences in the investigated parameters compared with patients without obstructive sleep apnea or sleep-disordered breathing.

Adult↗