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P Müller

Publications and source records attributed to P Müller.

At least 199 records · Page 11Linked to original sources

Transverse movement of spin-labeled phospholipids in the plasma membrane of a hepatocytic cell line (HepG2): implications for biliary lipid secretion.

The redistribution of spin-labeled phospholipid analogs across the plasma membrane of HepG2 cells, either in suspension or grown as monolayers, was investigated. After incorporation into the outer membrane leaflet spin-labeled aminophospholipids phosphatidylserine (PS) and phosphatidylethanolamine (PE) moved rapidly to the inner monolayer, whereas the analog of phosphatidylcholine (PC) disappeared more slowly from the outer leaflet. The fast, inward movement of the aminophospholipids was abolished after adenosine triphosphate (ATP)-depletion of cells, suggesting the presence of an aminophospholipid translocase in the plasma membrane of these cells. Compared with human red blood cells, the activity of the aminophospholipid translocase is two orders of magnitude higher in HepG2 cells. From these data, a transverse phospholipid asymmetry can be inferred with the aminophospholipids mainly concentrated on the inner monolayer and the choline-containing phospholipids on the outer leaflet. The relevance of the enrichment of PC in the outer membrane leaflet for the formation and composition of the bile is discussed.

Adenosine Triphosphate↗

[Early clinical experiences with MR-guided laser-induced thermotherapy (LITT) of liver metastases in preoperative care].

PURPOSE: To evaluate the LITT-induced changes with the aid of MRT and correlate these with histopathological findings. MATERIAL AND METHODS: Five patients with solitary colorectal liver metastases were treated by means of MR-guided LITT before liver resection. Application time and energy of the Nd:YAG laser (1064 nm) was 10-20 minutes and 4.5-8.8 W. MRT monitoring during the LITT was carried out with temperature-sensitive T1 weighted sequences (FLASH-2-D, turbo FLASH). The extent of the induced necrosis as seen on MR was compared with the unfixed specimen and with the histopathological findings. RESULTS: The extent of necrosis visible by MRT correlated with the histopathological findings with an accuracy of 95.3% +/- 4.2%. Following single treatments (three cases) the metastases suffered a reduction of 24%-55% of their original volume. In two patients a second application produced laser-induced necrosis of 78% and 98% of volume. In these two patients a temperature sound was used for measuring regional heating and showed an exact correlation with MR thermometry. CONCLUSION: The results of pre-operative MR-guided LITT indicates the potential of this form of treatment for obtaining reproducible tumor necrosis of liver metastases.

Aged↗

Liver ischemia and reperfusion induces a systemic inflammatory response through Kupffer cell activation.

To study the role of Kupffer cells (KC) as a cellular source of proinflammatory cytokines in hepatic ischemia/reperfusion, Sprague-Dawley rats were subjected to 20 min global hepatic ischemia. Sham-operated animals served as controls. Blood levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1 alpha (IL-1 alpha), and interleukin 6 (IL-6) were determined after 10, 30, 60, 120, and 240 min of reperfusion and compared with spontaneous cytokine release by KC isolated after 60 min of reperfusion. Hepatic ischemia/reperfusion resulted in an enhanced (p < .01) spontaneous release of TNF-alpha (+482%), IL-1 alpha (+33%), and IL-6 (+175%) by KC. Kinetic analysis of cytokinemia revealed an early increase (p < .01) of TNF-alpha and IL-1 alpha within minutes upon reperfusion, while an elevation of IL-6 serum levels was observed with a delay of 2 h. Early cytokinemia was associated with dysfunction/injury of the liver, lung, and kidney after 4 and 24 h of reperfusion, respectively. These data indicate that hepatic ischemia/reperfusion results in Kupffer cell activation and increased cytokine levels, which may produce systemic inflammation and may be responsible for tissue injury locally and on remote sites.

Animals↗

[Treatment of pelvic endometriosis with goserelin, an LHRH agonist].

OBJECTIVE: To determine the safety and effectiveness of ZOLADEX depot (goserelin) in the therapy of endometriosis (study No. 9393CZ/0001). MATERIAL AND METHODS: A multicentric prospective open study of 60 women with endometriosis. R-AFS score of pelvic endomemtriosis was established in patients suffering from pelvalgia, dyspareunia and dysmenorrhoea during pre-treatment laparoscopy. The effect of Zoladex depot applied subcutaneously were evaluated by gynaecological and laboratory examinations, including assessment of subjective complaints every four weeks in the course of six months. Laparoscopic check-up was carried out immediately after terminating the therapy. RESULTS: (1) The average total score of subjective complaints decreased by 90% after 24 weeks of treatment. (2) Treatment was successful in 67% patients, i.e. laparoscopic examination showed a decrease of the R-AFS score by at least 50%. In 28% treated women visible endometrial foci disappeared completely. (3) The most frequent side-effects of therapy are associated with hypoestrogenic effects of GnRH agonists. Vasomotor instability, flushes and night hyperidrosis occurred in 93% and 90% treated women, respectively. CONCLUSION: Goserelin acetate is a synthetic GnRH agonist suitable for safe and effective therapy of pelvic endometriosis. The administration of 3.6 mg goserelin every day for 24 weeks reduced markedly the size of endomerial foci and painful symptoms.

Adult↗

The influence of external surface potential and transmembrane potential on the passive transbilayer movement of phospholipids in the red blood cell membrane.

The passive transbilayer movement of spin-labelled analogues of phosphatidyl-choline (PC), phosphatidyl-ethanolamine (PE), and phosphatidyl-serine (PS) in red blood cell membranes was investigated at physiological and low ionic strength of the extracellular solution. Passive transbilayer movement of aminophospholipids PS and PE was measured in ATP-depleted cells. To discriminate between a possible surface potential and a transmembrane potential effect, NaCl in physiological ionic strength solution was replaced either by sucrose or by Na-tartrate (constant osmolarity). Neither in sucrose (low ionic strength) nor in Na-tartrate media a significant change of the translocation rate of the phospholipids was observed. From these results in can be concluded that changes of the external surface potential as well as of the transmembrane potential do not affect the passive transbilayer movement of phospholipids in human red blood cells.

Adenosine Triphosphate↗

Evidence for endothelin-1-mediated vasoconstriction in severe chronic heart failure.

Heart failure is commonly associated with high plasma concentrations of endothelin-1, a powerful vasoconstrictor produced by endothelium. The role of endogenously released endothelin-1 in the maintenance of vascular tone in chronic heart failure was assessed by acute administration of an endothelin receptor antagonist, bosentan. 24 patients with chronic heart failure received randomly and double blind two intravenous infusions of either placebo or bosentan (100 mg followed after 60 min by 200 mg). Systemic haemodynamics and plasma endothelin-1 and big-endothelin-1 concentrations were determined before and repeatedly during the 120 min observation period. Baseline endothelin-1 and big-endothelin-1 concentrations, which were above the normal range in all patients, correlated directly with the extent of pulmonary hypertension, with left and right heart filling pressures, and with pulmonary vascular resistance and inversely with cardiac index. Compared with placebo, bosentan reduced mean arterial pressure by 7.7% (95% CI 7.1-9.7), pulmonary artery pressure by 13.7% (10.5-16.9), right atrial pressure by 18.2% (12.0-24.4), and pulmonary artery wedged pressure by 8.6% (5.3-12.0); it increased cardiac index by 13.6% (9.1-18.2), decreased systemic vascular resistance by 16.5% (13.2-19.8), and decreased pulmonary vascular resistance by 33.2% (22.4-44.0). Heart rate did not change. Plasma endothelin-1 concentrations rose more than twofold from baseline in bosentan recipients while big-endothelin-1 concentrations were unchanged. These findings indicate that, in patients with chronic heart failure who have high circulatory endothelin-1 concentrations, this peptide contributes to maintenance of vascular tone. The acute haemodynamic effects of bosentan suggest that chronic endothelin antagonism could be beneficial in such patients.

Adult↗

An improved assay for measuring the transverse redistribution of fluorescent phospholipids in plasma membranes.

The internalization of fluorescent 7-nitro-2,1,3-benzoxadiazol-4-yl (NBD)-phospholipids from the plasma membrane can be assessed by the irreversible quenching of analogues in the outer leaflet by dithionite. Here we have utilized this assay to follow the redistribution of short-chain C6-NBD-sphingomyelin and C6-NBD-phosphatidylserine from the cell membrane of human gingiva fibroblasts. The significant uptake of dithionite across the plasma membrane and the subsequent reduction of NBD-analogues exposed to the cytoplasmic lumen does not allow an accurate measurement of the amount of internalized lipid probes even at low temperature. We could show that a precise determination can be achieved by extraction of analogues remaining in the exoplasmic half by a short pretreatment with bovine serum albumin prior to addition of dithionite. The fluorescence of those analogues localized to the cytoplasmic lumen was slowly destroyed by permeating dithionite. The fluorescence of those NBD-probes which are localized in the inner layer of intracellular vesicles remained almost unaffected in the time course of the assay. Thus, this approach allows to distinguish between different routes of internalization of NBD-phospholipids from the plasma membrane.

4-Chloro-7-nitrobenzofurazan↗

[Status of ranitidine in eradication of Helicobacter pylori].

Permanent removal of Helicobacter pylori leads to a dramatic drop in the recurrence rate of peptic ulcers. The NIH Consensus Conference recommends that Hp-positive patients with duodenal or gastric ulcer should undergo eradication treatment. To date, however, there is no consensus on what constitutes optimal eradication treatment. Presently available therapy may be triple (bismuth plus two antibiotics), dual (acid inhibition and one antibiotic) or modified triple therapy (acid inhibition and two antibiotics). Among numerous treatment proposals, results gained with the H2 blocker ranitidine stand out, on account of the double-blind nature of the studies and the adequate number of cases involved. With modified triple therapy (300 mg ranitidine nocte and 3 times 750 mg amoxicillin daily) eradication rates ranging from 85 to 90%, and with dual therapy (600 mg ranitidine and 2 g amoxicillin daily) 60 to 65%, are obtained.

Amoxicillin↗

HPV DNA and p53 alterations in oropharyngeal carcinomas.

We have examined a series of 37 oropharyngeal squamous cell carcinomas for the presence of HPV 6/11, 16, and 18 DNA by polymerase chain reaction (PCR)/Southern blotting and for p53 alterations by immunohistochemistry and mutation screening with temperature gradient gel electrophoresis (TGGE). HPV sequences were found in a total of 26 of 37 cancers (70.3%), most frequently HPV 16 (20/37) followed by HPV 18 (11/37). Double infections with HPV 16 and 18 were present in 5 tumours. p53 accumulation was detectable immunohistochemically in 21 of 37 carcinomas (56.8%). There were remarkable differences in the distribution of immunoreactive tumour cells in relation to the tumour grade. A mutation screening for p53 by TGGE, directed to the amplified exons 5-8, revealed p53 mutations in 14 of 37 carcinomas (37.8%). Mutations in two different exons were present in 3 tumours, 11 tumours being hit once. Exon 7 was mutated in 6 carcinomas, exons 5 and 8 in 4 cases, and exon 6 in 3 cases. When grouping the tumours with p53 mutation according to their HPV state, HPV-positive cases showed slightly more mutations (11/26) than HPV-negative cases (3/11). Only 5 of 37 carcinomas (13.5%) contained neither HPV DNA nor p53 alterations. Our results indicate that high-risk HPV and p53 mutations frequently coexist in oropharyngeal carcinomas, in contrast to genital tumours, notably carcinomas of the cervix uteri. This may reflect different pathways in carcinogenesis in squamous cell epithelium from different sites.

Adult↗

[MR-guided laser-induced thermotherapy in tumors of the head and neck region: initial clinical results].

PURPOSE: To analyse the value of MR-guided laser-induced thermotherapy (LITT) for palliative treatment of recurrent tumours of the head and neck region. MATERIAL AND METHODS: 8 patients with recurrent tumours of the head and neck region (squamous cell carcinomas n = 6, pleomorphic adenomas n = 2) underwent MR-controlled LITT. A 7 French laser applicator was inserted under local anaesthesia into the centre of the recurrent tumour. A Nd:YAG laser with a wavelength of 1064 nm was used. Therapy was monitored on-line using special MR thermosequences. RESULTS: Preinterventional contrast-enhanced MRI revealed a recurrent tumour of the head and neck region for all eight patients. All patients tolerated the procedures well under local anaesthesia, with no clinically relevant side effects. The MR thermosequences depicted up to 15 mm diameter areas of less signal near the laser tip. Postinterventional contrast-enhanced MRI revealed hypovascularized areas due to the resulting coagulative necrosis. Coagulative necrosis of 4 cc to 28 cc occurred in all patients, and a reduction of clinical symptoms was achieved in five. CONCLUSION: The results obtained indicate that minimal invasive LITT can be a therapeutic alternative for palliative treatment of head and neck tumours.

Adenoma, Pleomorphic↗

Combined FISH painting and harlequin staining for cell cycle-controlled chromosome analysis in human lymphocytes.

We present a simple method that allows scoring of FISH-painted chromosomes exclusively in the first division human lymphocytes. It consists of a combination of FISH with chromosome-specific libraries and a differential sister chromatid staining after BrdU treatment of lymphocyte cultures. The method allows a precise quantification of induced chromosome damage for human biodosimetry.

Cell Cycle↗

Chromosome painting in highly irradiated Chernobyl victims: a follow-up study to evaluate the stability of symmetrical translocations and the influence of clonal aberrations for retrospective dose estimation.

Follow-up fluorescence in situ hybridization (FISH) measurements of symmetrical translocations were performed in peripheral blood lymphocytes from 12 highly irradiated victims of the Chernobyl nuclear power plant accident biannually, between September 1991 and July 1994, to investigate the persistence of these aberration type with time post-exposure. Translocations were determined using biotin-labelled painting DNA probes for human chromosomes 1, 4 and 12 and a digoxigenin-labelled alpha-satellite pancentromeric DNA probe. In 11 of 12 cases the translocation frequencies remained fairly constant during the observation period, which allows to generate comparable dose estimates on the various sampling times. In one case (no. 9) the existence of a cell clone containing the consistent chromosome rearrangement t(1;13) (q25;q14) was identified using FISH in rehybridized slides with a digoxigenin-labelled painting DNA probe for chromosome 13 and a separate G-banding analysis. To obtain reliable dose estimates, total translocation frequency has to be corrected for the high contribution (16.5-23.5%) of this clonal translocation.

Accidents↗

A TnphoA insertion within the Bradyrhizobium japonicum sipS gene, homologous to prokaryotic signal peptidases, results in extensive changes in the expression of PBM-specific nodulins of infected soybean (Glycine max) cells.

Bradyrhizobium japonicum mutant 132 was obtained by random TnphoA mutagenesis of strain 110spc4. A 6.5 kb BamHI kanamycin-resistance-coding DNA fragment of mutant 132 was used as a hybridization probe to clone the corresponding wild-type fragment. DNA sequence analysis of a 3213 bp BamHI-ClaI fragment revealed that three open reading frames (ORFs) were encoded in the same orientation. Based on sequence similarities to other proteins in the database, the second ORF was called sipS (signal peptidase). The TnphoA insertion in mutant 132 was found to be in frame near the 3' end of sipS. The resulting SipS-PhoA hybrid polypeptide was shown to be expressed in free-living B. japonicum and in Escherichia coli cultures. An immunoblot analysis with a polyclonal antibody directed against the alkaline phosphatase revealed the appearance of a weak signal of a 70 kDa polypeptide both in B. japonicum and in E. coli, in agreement with the calculated size of the hybrid polypeptide. A much stronger 52 kDa band was also detected. Mutant 132 was specifically disturbed in the interaction with soybean (Glycine max) when the bacteria were released from the infection threads. The bacteroids were not stably maintained within the symbiosome. Numerous vesicles were found in the plant cytosol, which finally resulted in the formation of large vacuoles within the infected nodule cells. Immunohistochemical analyses with antibodies directed against nodulins of the peribacteroid membrane indicated a lower expression of these proteins. The mutant phenotype was genetically complemented by a 4.4 kb BamHI fragment including sipS. Subfragments thereof also complemented a temperature-sensitive E. coli lepB mutant, demonstrating that the B. japonicum fragment was functionally replacing Lepts in E. coli. Genetic studies suggested that the three genes are organized in one common operon which is expressed from a promoter upstream of the sequenced region. Inactivation of the gene downstream of sipS did not result in a detectable phenotype.

Alkaline Phosphatase↗

Recurrent nasopharyngeal tumors: preliminary clinical results with interventional MR imaging--controlled laser-induced thermotherapy.

PURPOSE: To evaluate magnetic resonance (MR) imaging-controlled laser-induced thermotherapy (LITT) in the treatment of recurrent head and neck tumors. MATERIALS AND METHODS: Six patients with recurrent nasopharyngeal tumors (squamous cell carcinoma [n = 4], pleomorphic adenoma [n = 2]) underwent LITT with local anesthesia. A 7-F introducing sheath was inserted into the center of the tumor followed by a specially designed laser emitter. Therapy was monitored on-line with MR thermometry, and the amount of necrosis was estimated with dynamic and static contrast material-enhanced sequences. RESULTS: All procedures were well tolerated with use of local anesthesia, with no clinically relevant side effects. MR thermometry depicted up to 15-mm-diameter areas of less signal intensity near the laser tip. Coagulative necrosis was achieved in all patients (volume range, 4-28 cm3), and clinical symptoms were reduced in four. CONCLUSION: MR imaging-controlled LITT may be a safe, minimally invasive alternative in the treatment of recurrent head and neck tumors.

Adenoma, Pleomorphic↗