Use of inhaled iloprost in a case of pulmonary hypertension during pediatric congenital heart surgery.
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Administration, Inhalation↗
Biomedical subjects
Matthias Müller
Publications and source records attributed to Matthias Müller.
Changes in biochemical and biophysical surfactant properties with cardiopulmonary bypass in children.
OBJECTIVE: The aim of the present study was to characterize pulmonary surfactant properties in children undergoing cardiovascular surgery with cardiopulmonary bypass. DESIGN: Prospective clinical trial. SETTING: University hospital pediatric intensive care unit. PATIENTS: Fifty pediatric patients with congenital cardiac defects undergoing cardiovascular surgery with (n = 35) and without (n = 15) cardiopulmonary bypass procedure. INTERVENTIONS: Tracheal aspirates were collected by saline lavage during routine suctioning before (baseline) and after cardiopulmonary bypass, as well as 4, 8, and 24 hrs after admission to the pediatric intensive care unit. MEASUREMENTS AND MAIN RESULTS: Total protein and phospholipid concentrations were assessed in native tracheal aspirates, in large surfactant aggregates, and in small surfactant aggregates. Phospholipid profiles and phosphatidylcholine fatty acids; surfactant apoproteins SP-A, SP-B, and SP-C (enzyme-linked immunosorbent assay); and surface activity (Pulsating Bubble Surfactometer) were all analyzed in large surfactant aggregates. With cardiopulmonary bypass, an initial increase in total protein content was followed by an increase in phospholipid concentration in tracheal aspirates. Large surfactant aggregates decreased 4 hrs after cardiopulmonary bypass (4 hrs, 22.6 +/- 5.6%; mean +/- SEM; p<.01 compared with baseline, 55.4 +/- 9.2%) but recovered within 24 hrs. The phospholipid-protein ratio of large surfactant aggregates 24 hrs after cardiopulmonary bypass (1.2 +/- 0.2; p<.01) was significantly decreased compared with baseline (2.9 +/- 0.6). The relative amount of phosphatidylglycerol content in the large surfactant aggregates-fraction dropped linearly over time but other phospholipids remained mainly unchanged. Phosphatidylcholine fatty acid profiles remained unaffected by cardiopulmonary bypass. The relative content of SP-B and SP-C in large surfactant aggregates increased approximately three-fold compared with baseline. Altogether, our findings with recovered large surfactant aggregate/small surfactant aggregate ratios and increased phospholipid in tracheal aspirates after 24 hrs represent an approximately ten-fold increase in large surfactant aggregate-associated SP-B and SP-C compared with baseline. Only minor changes were detected in biophysical properties of large surfactant aggregates throughout the observation period. CONCLUSIONS: Cardiopulmonary bypass procedure in children induces profound changes in the surfactant system involving both phospholipid and protein components; biophysical function may have been maintained by compensatory increase in SP-B and SP-C.
Cardiopulmonary Bypass↗
Description of new mutations in the rpoB gene in rifampicin-resistant Neisseria meningitidis selected in vitro in a stepwise manner.
Fourteen meningococcal strains were selected towards rifampicin resistance in a stepwise manner in vitro; final MICs were between 8 and >256 microg ml(-1). Sequence analysis of a 295 bp subgenic fragment of the RNA polymerase beta-subunit (rpoB) gene from the original and the fully resistant strains revealed that, with one exception, the strain pairs differed by just one position in the deduced amino acid sequence. Transformation of a PCR-amplified subgenic rpoB fragment harbouring the mutated site into a susceptible strain demonstrated the resistance-conferring mechanism.
Antibiotics, Antitubercular↗
Oligomeric properties and signal peptide binding by Escherichia coli Tat protein transport complexes.
The Escherichia coli Tat apparatus is a protein translocation system that serves to export folded proteins across the inner membrane. The integral membrane proteins TatA, TatB and TatC are essential components of this pathway. Substrate proteins are directed to the Tat apparatus by specialized N-terminal signal peptides bearing a consensus twin-arginine sequence motif. Here we have systematically examined the Tat complexes that can be purified from overproducing strains. Our data suggest that the TatA, TatB and TatC proteins are found in at least two major types of high molecular mass complex in detergent solution, one consisting predominantly of TatA but with a small quantity of TatB, and the other based on a TatBC unit but also containing some TatA protein. The latter complex is shown to be capable of binding a Tat signal peptide. Using an alternative purification strategy we show that it is possible to isolate a TatABC complex containing a high molar excess of the TatA component.
Arginine↗
Overlap between emotional blunting, depression, and extrapyramidal symptoms in schizophrenia.
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Basal Ganglia Diseases↗
Application of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for monitoring the digestion of phosphatidylcholine by pancreatic phospholipase A(2).
Different methods were established for monitoring the phospholipase A(2)(PLA(2)) activity but all of them are rather cumbersome and time consuming. In this paper we have investigated the suitability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the determination of the PLA(2) activity. Phosphatidylcholine (PC) was digested with pancreatic PLA(2) under different conditions, i.e., various Ca(2+), PC, and PLA(2) concentrations. The digestion products were analyzed by MALDI-TOF MS and the concentration of lysophosphatidylcholine (LPC)-generated upon PLA(2) digestion-was determined by the application of an internal standard (known concentration) and by a comparison of their signal-to-noise ratios. The results clearly demonstrate that the LPC concentration determined from the MALDI-TOF mass spectra correlates directly with the activity of the applied enzyme. Additionally, LPC concentration increased with an increase in Ca(2+), as well as in the PC concentration. A single MALDI-TOF mass spectrum provides immediate information on the digestion products as well as on the residual substrate without requirements for any previous derivatization. MALDI-TOF MS can be easily and simply applied for monitoring the PLA(2) activity and we assume that this method might also be useful for other types of phospholipases.
Animals↗
Separate analysis of twin-arginine translocation (Tat)-specific membrane binding and translocation in Escherichia coli.
The twin-arginine translocation (Tat) pathway exports those precursor proteins to the periplasmic space of bacteria that harbor a twin-arginine (RR) consensus motif in their signal sequences. We have reproduced translocation of several Tat substrates into inside-out plasma membrane vesicles from Escherichia coli. Translocation proceeding at an efficiency of up to 20% occurs specifically via the Tat pathway as indicated by (i) its requirement for elevated levels of the TatABC proteins in the membrane vesicles, (ii) competition by an intact twin-arginine signal peptide, and (iii) susceptibility toward dissipation of the transmembrane H(+) gradient. The latter treatment, while blocking translocation, still allows for functional membrane association of Tat precursors. This is shown by the finding that translocation of isolated membrane-bound Tat precursor is restored upon re-energization of the vesicles.
Amino Acid Sequence↗
Epidemiological dynamics of hepatitis C virus among 747 German individuals: new subtypes on the advance.
This study demonstrates the dynamics in the epidemiology of hepatitis C virus subtypes. Subtypes 3a and 4a have become increasingly prevalent in patients where an infection within recent years can be assumed. Evidence is presented that the subtypes observed among younger patients can spread rapidly and lead to significant changes in the subtype distribution.
Adolescent↗
Syntheses of the Uranium Complexes [U{N(SiMe(3))(2)}(2){N(SiMe(3))(SiMe(2)CH(2)B(C(6)F(5))(3))}] and [U{C(Ph)(NSiMe(3))(2)}(2){&mgr;(3)-BH(4)}(2)]. Determination of Hydrogen Positions by Single-Crystal X-ray and Neutron Diffraction.
The complex [U{N(SiMe(3))(2)}(2){N(SiMe(3))(SiMe(2)CH(2)B(C(6)F(5))(3))}] (1) is formed in the reaction between the hydride complex [U{N(SiMe(3))(2)}(3)(H)] and B(C(6)F(5))(3), and H(2) is evolved. The X-ray [C(36)H(53)BF(15)N(3)Si(6)U.3.5C(6)D(6), triclinic, space group P&onemacr;, Z = 2, 90 K, a = 14.065(1) Å, b = 14.496(1) Å, c = 18.759(1) Å, alpha = 82.898(1) degrees, beta = 74.415(1) degrees, gamma = 62.919(1) degrees ] and neutron structure [C(36)H(53)BF(15)N(3)Si(6)U.3.5C(6)D(6), triclinic, space group P&onemacr;, Z = 2, 20 K, a = 13.993(1) Å, b = 14.484(1) Å, c = 18.720(1) Å, alpha = 82.810(1) degrees, beta = 74.200(1) degrees, gamma = 63.054(1)E] of compound 1, which crystallizes with 3.5 molecules of C(6)D(6) per asymmetric unit, show the electron deficiency of the uranium atom to be effectively compensated by the formation of multicenter bonds between U and three Si-CH(2) units of the amido ligands. The reaction of the uranium complex [U{C(Ph)(NSiMe(3))(2)}(2)(Cl)(2)] with [Na(BH(4))] gives the complex [U{C(Ph)(NSiMe(3))(2)}(2){&mgr;(3)-BH(4)}(2)] (2). The X-ray structure of 2 [C(26)H(54)B(2)N(4)Si(4)U, monoclinic, space group C2/c, Z = 4, 90 K, a = 21.613(1) Å, b = 9.233(1) Å, c = 18.132(1) Å, beta = 98.804(1) degrees ] proves unequivocally the &mgr;(3) coordination of the BH(4) moieties. In both single-crystal X-ray structure determinations, all hydrogen and deuterium atoms could be located and isotropically refined, including those which are directly coordinated to the uranium. The reliability of the refined hydrogen and deuterium positions for compound 1 is confirmed by comparison of the X-ray and neutron structure determinations. The ability to locate the hydrogen and deuterium positions in these uranium compounds by single-crystal X-ray diffraction is due to good crystal quality, the measurement of data at low temperature, and the use of image plate technology for data collection.
Journal Article↗
Development of a minimal cell-free translation system for the synthesis of presecretory and integral membrane proteins.
By combining translation and membrane integration/translocation systems, we have constructed a novel cell-free system for the production of presecretory and integral membrane proteins in vitro. A totally defined, cell-free system reconstituted from a minimal number of translation factors was supplemented with urea-washed inverted membrane vesicles (U-INVs) prepared from Escherichia coli, as well as with purified proteins mediating membrane targeting of presecretory and integral membrane proteins. Initially, efficient membrane translocation of a presecretory protein (pOmpA) was obtained simply by the addition of only SecA and SecB. Proteinase K digestion clearly showed the successful translocation of pOmpA inside the vesicles. Next, integration of an inner membrane protein (MtlA) into U-INVs was achieved in the presence of only SRP (Ffh) and SR (FtsY). Finally, a membrane protein possessing a large periplasmic region (FtsQ) and therefore requiring both factors (SRP/SR and SecA/SecB) for membrane integration/translocation was also shown to be integrated correctly in this cell-free system. Thus, our novel cell-free system provides not only an efficient strategy for the production of membrane-related proteins but also an improved platform for the biological study of protein translocation and integration mechanisms.
Adenosine Triphosphatases↗
AUDENS: a tool for automated peptide de novo sequencing.
We present AUDENS, a new platform-independent open source tool for automated de novo sequencing of peptides from MS/MS data. We implemented a dynamic programming algorithm and combined it with a flexible preprocessing module which is designed to distinguish between signal and other peaks. By applying a user-defined set of heuristics, AUDENS screens through the spectrum and assigns high relevance values to putative signal peaks. The algorithm constructs a sequence path through the MS/MS spectrum using the peak relevances to score each suggested sequence path, i.e., the corresponding amino acid sequence. At present, we consider AUDENS a prototype that unfolds its biggest potential if used in parallel with other de novo sequencing tools. AUDENS is available open source and can be downloaded with further documentation at http://www.ti.inf.ethz.ch/pw/software/audens/ .
Algorithms↗
The Tat pathway in bacteria and chloroplasts (review).
Both in prokaryotic organisms and in chloroplasts, a specialized protein transport pathway exists which is capable of translocating proteins in a fully folded conformation. Transport is mediated in both instances by signal peptides harbouring a twin-arginine consensus motif (twin-arginine translocation (Tat) pathway). The Tat translocase comprises the three functionally different membrane proteins TatA, TatB, and TatC. While TatB and TatC are involved in the specific recognition of the substrate, TatA might be the major pore-forming component. Current evidence suggests that a functional Tat translocase is assembled from separate TatBC and TatA assemblies only on demand, i.e., in the presence of transport substrate and a transmembrane H+-motive force.
Bacterial Proteins↗
Comparison of different procedures for the lipid extraction from HL-60 cells: a MALDI-TOF mass spectrometric study.
A human leukaemia cell line--HL-60--can be differentiated into neutrophils or macrophages and both differentiation processes are accompanied by changes of the lipid composition. Various methods were described for the extraction of lipids from cellular systems, but only two of them were applied to the HL-60 cell line so far. In this study we compared five selected extraction methods for the lipid extraction from HL-60 cells with regard to their qualitative analysis by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS): chloroform/methanol at volume ratios 2:1 and 1:2, isopropanol/ chloroform, isopropanol/hexane and butanol. In addition, the cholesterol and phospholipid concentrations in organic extracts were measured by colorimetric assays. Results can be summarized as follows: For the analysis of polar phospholipids obtained from HL-60 cells by MALDI-TOF MS, a chlorofom/methanol (1:2) or isopropanol/chloroform mixture or butanol can be applied as extraction systems On the other hand, if one would like to analyze changes in triacylglycerols, then chloroform/methanol (2:1) would be the method of choice.
Cholesterol↗
[Bacterially conditioned thromboembolism in dairy cows--a retrospective study of 31 necropsy cases with special consideration of the causative complex].
A study of 340 necropsied dairy cattle from northern Bavaria resulted in 31 animals (9%) showing evidence of pyemic thromboembolism. The most frequent pathomorphologic lesions consisted in endocarditis valvularis thromboticans of the tricuspid and/or mitral valve (21), embolic pneumonia (26), nephritis (13) and renal infarction (12). The most common isolate found in bacteriologic culture was Arcanobacterium pyogenes in 26 of 31 cases, followed by Staphylococcus aureus (2), Mannheimia haemolytica (2) and Streptococcus bovis (1) in rare cases. In 27 of 31 cows a possible cause of pyemic thromboembolism was found; in 23 cases claw diseases and decubital ulcera were probably responsible.
Animals↗