Biomedical subjects
H Schulze
Publications and source records attributed to H Schulze.
Thrombopoietin induces the generation of distinct Stat1, Stat3, Stat5a and Stat5b homo- and heterodimeric complexes with different kinetics in human platelets.
OBJECTIVE: Thrombopoietin (TPO) is the pivotal regulator of thrombocytopoiesis and megakaryocytopoiesis, and binding to its receptor c-Mpl leads to activation of at least two different signaling pathways: the Jak-Stat pathway and the Ras-MAPK pathway. Our aim was to elucidate which Stat-complexes are formed in TPO signal transduction in human blood platelets. MATERIALS AND METHODS: We used electrophoretic mobility shift assays (EMSA) in order to analyze the formation of distinct Stat complexes on two distinct oligonucleotide probes. Furthermore, we used immunoprecipitation and Western blotting of protein lysates from TPO-stimulated platelets. RESULTS: We found homodimers of Stat1alpha, Stat3, Stat5a, and Stat5b, as well as heterodimers of Stat1/Stat3 and Stat5a/Stat5b, but no Stat1/Stat5 or Stat3/Stat5 heterodimers are formed in platelets in response to TPO. Stat5 complexes bound to labeled DNA with a fast kinetic followed by Stat3 and Stat1. The adapter protein CrkL is present in DNA-bound Stat5 complexes and predominantly bound to Stat5b. The kinase ERK2 is also tyrosine phosphorylated after TPO-stimulation of platelets but this activation does not modulate the phosphorylation of the serine residues in the PXSP motif present in Stat1 and Stat3. CONCLUSION: Our findings thus emphasize the differential regulation of Stat1, Stat3, Stat5a, and Stat5b in platelets and may be an appropriate model of c-Mpl signaling in mega-karyopoiesis.
Objective evaluation of the morphology of human epididymal sperm heads.
Spermatozoa were obtained from nine epididymal regions of six epididymides taken from five men undergoing castration for prostatic carcinoma (53-76 years) and from one man with testicular cancer (38 years). Spermatozoa were obtained by mincing tissue in phosphate-buffered saline, making air dried smears and staining with Hemacolor. The percentage of sperm heads categorised subjectively as normal (of uniform shape) or otherwise was calculated for each region. This confirmed that grossly swollen sperm heads (previously shown to be artefacts) were only present in proximal regions of the duct. A computer-aided sperm morphology analyser (Sperm Class Analyzer(R)) was used to provide objective measurements of sperm head area, perimeter, length and width of the uniform sperm heads and revealed that there was a gradual and statistically significant decline in sperm head size upon maturation, as occurs in other species. There was no significant difference between the morphometric parameters of spermatozoa obtained from the distal cauda epididymis and those obtained from the ejaculates of young normozoospermic patients.
Induction of unscheduled DNA synthesis in primary human urothelial cells by the mycotoxin ochratoxin A.
Ochratoxin A (OTA) is a widespread contaminant in human staple food. Exposure of humans to this mycotoxin is a matter of concern because OTA is a known rodent carcinogen. As the urothelium is one target tissue of this mycotoxin, primary cultured human urothelial cells (HUC) from adults and children were used to analyze the induction of unscheduled DNA synthesis (UDS) by OTA. HUC were isolated from the ureters or renal pelves of two nephrectomized adults and of two children with ureteropelvic junction stenosis and cultured under serum-free conditions. After a confluency of 70-80% was reached, cell proliferation was suppressed by arginine-deficient medium (ADM), and UDS was assessed autoradiographically by 3H-thymidine incorporation upon exposure to OTA (10-2000 nM), ethyl methanesulfonate (EMS, 5 mM, positive control), or dimethyl sulfoxide (DMSO, 0.2%, solvent control). In control cultures the level of UDS was low. Exposure to EMS resulted in an induction of UDS (2-to 5-fold compared to control), thus allowing the sensitive detection of repair resulting from induction of DNA lesions in all four specimens, and demonstrating that repair of EMS-induced DNA lesions can take place under the chosen culture conditions. In two HUC cultures derived from adults, a significant induction of UDS was observed in the concentration range of 50-500 nM OTA. The highest fraction of cells in repair (CIR) was found at 50 nM OTA for the HUC from the older male (50% CIR). The maximum response in the other specimens from the adult female and the 7-year-old boy were seen at OTA concentrations of 500 and 250 nM, respectively. In contrast to all other specimens, no significant induction of UDS by OTA was found in the HUC cultures derived from an infant's urothelium. Signs of cytotoxicity were observed above 500 nM OTA in all cultures. The varying susceptibility toward OTA observed in vitro may hint at varying predispositions of individuals in vivo.
Risk factors for relapse in stage I non-seminomatous germ-cell tumors: preliminary results of the German Multicenter Trial. German Testicular Cancer Study Group.
Risk factor analysis to identify low-risk patients for occult metastatic disease (vascular invasion, percentage embryonal carcinoma, MIB-I proliferation rate) yields reliable results if performed by experts. A correct prediction is possible at the 90% level. Similar accuracy, however, may be achieved if the computed tomography (CT) staging is optimized and the evaluation performed by an experienced investigator. The combination of both methods (biological risk factor analysis and CT staging) may virtually exclude the risk of relapse in a limited number of patients. However, so far, no risk factor that is able to reliably predict occult metastatic disease or relapse in clinical state I patients has been identified in prospective trials. The preliminary results of the current German Multicenter Trial suggest an inferior value of prediction for low-risk patients if risk factor analysis and/or CT staging is performed in non-specialized centers.
Thrombopoietin acts synergistically on Ca(2+) mobilization in platelets caused by ADP or thrombin receptor agonist peptide.
Thrombopoietin (TPO) is the main regulator of megakaryopoiesis and influences also the function of mature platelets. TPO has been shown to synergize in multiple platelet activation processes induced by various agonists. Our aim was to elucidate whether TPO affects calcium signaling during platelet activation processes. TPO demonstrated a synergistic effect on the exocytosis induced by suboptimal doses of adenosine diphosphate (ADP) and the thrombin receptor agonist peptide (TRAP). We detected synergistic effects of TPO on the ADP or TRAP induced Ca(2+) mobilization in a small range of very low agonist concentrations. The TPO synergism on Ca(2+) mobilization and CD62P expression was measurable in different, nonoverlapping ranges of ADP or TRAP concentrations. Sustaining the agonist-induced calcium signal with thapsigargin led to a detectable TPO synergism in CD62P expression even in agonist concentrations in which the synergism only occurs in Ca(2+) signaling without thapsigargin.
Discrimination learning of amplitude modulated tones in Mongolian gerbils.
Discrimination learning of 100% sinusoidal amplitude modulated tones (AM) was investigated in adult Mongolian gerbils using a footshock motivated shuttle box avoidance go/no go paradigm. AM stimuli to be discriminated had identical carrier frequency (2 kHz) but differed in modulation frequency (fm) by one octave. Six groups of gerbils were trained to discriminate AM-pairs with fm ranging from 10 to 640 Hz. Learning proceeded faster and discrimination performance was slightly better for low fm, up to 80 Hz, than for high fm, above 80 Hz. These results may be related to cortical AM encoding (Schulze, H. and Langner, G., J. Comp. Physiol. A, 181 (1997) 651-663), which is temporal for low fm (synchrony code) and spatial for high fm (rate-place code). This may implicate different neuronal learning strategies or distinct behavioral meanings influencing the discrimination training.
Enhancement of cation diffusion rates across the 410-kilometer discontinuity in Earth's mantle
Rates of cation diffusion (magnesium, iron, and nickel) have been determined in olivine and its high-pressure polymorph, wadsleyite, at 9 to 15 gigapascals and 1100 degrees to 1400 degreesC for compositions that are relevant to Earth's mantle. Diffusion in olivine becomes strongly dependent on composition at high pressure. In wadsleyite, diffusion is one to two orders of magnitude faster than in olivine, depending on temperature. Homogenization of mantle heterogeneities (chemical mixing) and mineral transformations involving a magnesium-iron exchange will therefore occur considerably faster in the transition zone than at depths of less than 410 kilometers.
[Occupational risk factors for bladder carcinoma. A case control study].
Aim of this case-control study, performed on 412 male bladder cancer cases and 414 controls with benign prostatic hyperplasia in a former area of coal, iron and steel industries in Germany, was to identify occupations with an increased bladder cancer risk. In bladder cancer cases, smokers were overrepresented (58.3%) compared to controls (35.2%). The percentage of patients who had stopped smoking for at least 10 years did not differ in cases (10.2%) and controls (9.7%). Significantly elevated smoking-adjusted bladder cancer odds ratios (MH) were observed in painters and lacquers (MH 2.24, 95% CI 1.07-5.13), chemistry-related occupations (MH 2.44, 95% CI 1.05-5.67), coke plant workers (MH 2.89, 95% CI 1.16-7.16) and hard coal miners (MH 2.33, 95% CI 1.52-3.58). Significantly decreased smoking-adjusted bladder cancer odds ratios (MH) were observed in businessmen (MH 0.64, 95% CI 0.45-0.92) and office personnel (MH 0.58, 95% CI 0.41-0.81). In these two groups a relevant exposure to occupational bladder carcinogens is not likely.
Serum levels of thrombopoietin, IL-11, and IL-6 in pediatric thrombocytopenias.
We measured serum levels of thrombopoietin (TPO), interleukin (IL)-11, and IL-6 in 90 different samples from 67 pediatric patients with thrombocytopenia (TP). The cytokine levels were determined by enzyme-linked immunosorbent assays (ELISA), and the biological activity of TPO was measured using a cell line transfected with human c-mpl. In patients with impaired megakaryocytopoiesis, as found in diseases such as aplastic anemia, amegakaryocytic TP, or TP with absent radii, we found TPO levels which were highly elevated compared with normal values (mean=261 AU/ml, n=52, vs. 22 AU/ml in healthy controls). In contrast, patients suffering from idiopathic thrombocytopenic purpura (mean=16 AU/ml, n=31) or platelet function defects (mean=23 AU/ml, n=7) demonstrated normal TPO levels. The biological activity tested in the bioassay correlated well with the ELISA data. However, sera of some patients with amegakaryocytic TP demonstrated a remarkably higher biological activity of TPO than expected from the ELISA data. Within the different groups there was no correlation between platelet counts and TPO levels. Only 27% of all samples had elevated levels of IL-11 (mean=450 pg/ml, n=20). Elevated IL-6 serum levels were detected in only 13% of all samples analyzed (mean=42 pg/ml, n=12). We conclude that megakaryocytopoiesis is regulated mainly by TPO, that it is dependent on the platelet and the megakaryocytic mass, and that IL-11 plays an additional role in supporting the platelet production. IL-6 does not appear to be up-regulated in children with thrombocytopenia.
Auditory cortical responses to amplitude modulations with spectra above frequency receptive fields: evidence for wide spectral integration.
Auditory neurons typically respond to a restricted range of frequencies and amplitudes of pure tone stimuli. These findings have led to the concept of the classical frequency receptive field. Over the last few years evidence has accumulated that stimuli outside the frequency and amplitude boundaries of a neuron's receptive field can influence responses to stimuli inside the classical receptive field. We could recently show that sinusoidally amplitude-modulated pure tones could excite cortical neurons although all of their spectral components were above the spectral range of pure tones effective to excite the neuron. This result demonstrated that neurons in the auditory cortex integrate over spectral ranges that are much wider than is evident from responses to pure tones. Here, using sinusoidally amplitude-modulated pure tone stimuli we determine electrophysiologically the high-frequency boundaries of the spectral integration capabilities of auditory cortical neurons in anaesthetized Mongolian gerbils under normal conditions and under the influence of the microiontophoretically applied GABAA-receptor antagonist bicuculline. Our results demonstrate that some auditory cortical neurons integrate over the gerbil's entire audible spectrum. Therefore, the classical excitatory frequency receptive field of an auditory cortical neuron, as determined with pure tone stimuli, cannot provide a satisfactory description of its spectral integrative properties.
Suramin treatment in hormone- and chemotherapy-refractory prostate cancer.
OBJECTIVES: Suramin, a polysulfonated naphtylurea with anti-growth factor activity, was used in the treatment of metastatic, hormone- and chemotherapy-refractory prostate cancer. Recent studies have proved the effect of suramin on prostate cancer. METHODS: Between March 1990 and January 1994, 27 patients with metastatic prostate cancer were enrolled in this study. Treatment regimen consisted of a loading phase, allowing patients to reach suramin serum levels between 180 and 250 microg/mL using a suramin dose of 1.4 g/m2 at 3-day intervals. Constant suramin serum levels were maintained by a 0.5 to 1-g/m2 dose every 7 to 10 days. Because previous studies showed suramin to have serious toxicity, compromised organ status was excluded by repeated examinations. RESULTS: Six patients did not complete the suramin loading phase because of side effects and were removed from the study. With an average cumulative suramin dose of 14.2 g, 33% of the assessable patients (7 of 21) experienced a more than 50% reduction of prostate-specific antigen (PSA) and/or alkaline phosphatase (AP) serum levels. Mean survival in these suramin-responsive patients was 495 days. Two of these patients experienced a remarkable reduction of metastases in bone scan examinations. Another 48% of the patients (10 of 21) had essentially unchanged AP and PSA serum levels during suramin treatment, indicating stable disease. Mean survival of these patients was 341 days. In 4 patients undergoing suramin treatment, continuous clinical progression of the disease was observed (mean survival 79 days). Toxicity was less or comparable to prior reported studies; the most common side effects were polyneuropathy, allergic skin rash, and vortex keratopathy. CONCLUSIONS: Suramin has limited, but significant, efficacy even in chemotherapy- and hormone-refractory prostate cancer, without serious toxicity.
[High-flow priapism following blunt perineal trauma. Interventional therapy].
A 10-year old boy developed an arteriocavernosal fistula with a permanent penile erection (high-flow priapism) after a blunt perineal trauma. The diagnosis was established by Doppler ultrasound, blood gas analysis and angiography. A conservative treatment attempt with penile cooling and puncture of the cavernosal corpera with blood aspiration and etilefrine instillation failed to resolve the priapism. An embolization of both bulbocavernosal arteries was followed by an immediate detumescence. Normal functional erection was preserved. No recurrence was observed during a follow-up of 6 months.
Guanidinated protein test meals with higher concentration of soybean trypsin inhibitors increase ileal recoveries of endogenous amino acids in pigs.
The amino acid concentrations of cornstarch-based guanidinated unprocessed (UGM) and autoclaved (AGM) Nutrisoy (defatted soy flour) protein test meals were compared with the respective unguanidinated Nutrisoy diets. Endogenous ileal recoveries and true digestibilities of amino acids were determined in six growing pigs, fitted with a simple T-cannula at the distal ileum, fed the guanidinated protein test meals. The UGM and AGM contained 13.4 (high) and 3.0 (low) g/kg dry matter of soybean trypsin inhibitors (SBTI), respectively. The experiment was a two-period cross-over design with each period lasting 15 d. On d 14 of each period, the pigs were fed the guanidinated test meals followed by 24 h continuous collection of digesta. Concentrations of crude protein and most of the amino acids in the test meals were higher than in the respective diets. Apparent ileal amino acid digestibilities of the test meals did not differ (P > 0.05) from reported values for the respective diets and were higher (P < 0.05) by 22.7 (cysteine) to 61.3 (tyrosine) percentage units for AGM compared with UGM. The ileal recoveries of endogenous amino acids in AGM-fed pigs were lower (P < 0.05) than UGM-fed pigs. Values ranged from -0.10 (arginine) to 0.64 (aspartate + asparagine) and from 0.84 (histidine) to 2.61 (tyrosine) g/kg dry matter intake for AGM- and UGM-fed pigs, respectively. True ileal amino acid digestibilities for AGM were higher (P < 0.05) than UGM with differences ranging from 12.7 (tyrosine) to 38.3 (leucine) percentage units. In conclusion, ileal recoveries of endogenous amino acids were increased in pigs fed guanidinated protein test meals with the higher concentration of SBTI.
Crystal structure of cytochrome P-450cam complexed with the (1S)-camphor enantiomer.
The crystal structure of cytochrome P-450cam complexed with the enantiomer (1S)-camphor has been solved to 1.8 angstroms resolution and compared with the structure of the (1R)-camphor P-450cam complex. The overall protein structure is the same for both enantiomer complexes. However, the orientation of the substrates in the heme pocket differs. In contrast to (1R)-camphor, the (1S)-enantiomer binds in at least two orientations. The major binding mode of (1S)-camphor resembles the one of the (1R)-enantiomer in that there is a hydrogen bond between Tyr-96 and the quinone group of camphor, and the 10-methyl group points towards the I-helix. The binding differs in that C-5 is not at a position suitable for hydroxylation. In the other orientation (1S)-camphor is not hydrogen bonded, but C-5 is located suitably for hydroxylation.
Thrombopoietin in patients with congenital thrombocytopenia and absent radii: elevated serum levels, normal receptor expression, but defective reactivity to thrombopoietin.
The pathophysiology of thrombocytopenia in the syndrome of thrombocytopenia with absent radii (TAR) is not yet understood. We examined thrombopoietin (TPO) serum levels and the in vitro reactivity of platelets to TPO in five patients affected with TAR syndrome. We found elevated TPO serum levels in all patients tested, excluding a TPO production defect as cause for thrombocytopenia in TAR syndrome. In addition, we found similar expression of the TPO receptor c-Mpl on the surface of platelets from TAR patients (5 of 5) and a similar molecular weight of the receptor as compared with healthy controls (4 of 4). Platelet response to adenosine diphosphate or thrombin receptor agonist peptide SFLLRN (TRAP) was normal in TAR patients. However, in contrast to results with healthy controls we could show absence of in vitro reactivity of platelets from TAR patients to recombinant TPO as measured by testing TPO synergism to adenine diphosphate and TRAP in platelet activation. TPO induced tyrosine phosphorylation of platelet proteins was completely absent (3 of 4) or markedly decreased (1 of 4). Our results indicate that defective megakaryocytopoiesis/thrombocytopoiesis in TAR syndrome is not caused by a defect in TPO production but a lack of response to TPO in the signal transduction pathway of c-Mpl.
Mobility of norbornane-type substrates and water accessibility in cytochrome P-450cam.
The behaviour of norbornane-type substrates bound to oxidised cytochrome P-450cam (CYP 101) in 60% (w/w) glycerol-containing phosphate buffer was investigated using electronic absorption spectroscopy. The high-pressure dependence study revealed that the value of the spin-state reaction-volume change decreased from -70 to -22.8 cm3/mol with decreasing high-spin state content from 99 to 63%. Simultaneously, the values for the enthalpy and entropy determined from the low-temperature dependence of the spin-state transition decreased from 73.7 to 24.3 kJ/mol and from 310.4 to 88.9 J/mol K, respectively. Under our experimental conditions the pH-value of the buffer remained at low temperatures and high pressures in the range of pH 7-8, in which no pH-value-induced spin-state conversion occurred. Therefore, the secondary effect of the temperature and pressure-induced pH change can be disregarded as being responsible for the observed spin-state transition effects. Substrate dissociation constants were determined. From the temperature-jump experiments (297 K to 180 K) we found a higher mobility in the active site for the substrates in the sequence (1R)-camphor, (1S)-camphor, camphane, (1R)- and (1S)-camphorquinone, norcamphor, and norbornane. Our findings can be explained by the incomplete fit of the methyl groups of the norbornane-type substrate to the protein, in particular to the I-helix, predominantly determining the substrate mobility and water accessibility to the protein.
[Benign prostatic hyperplasia].
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