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C Weber

Publications and source records attributed to C Weber.

At least 271 records · Page 15Linked to original sources
PubMed1991

Selective 13C-labelling of cyclosporin A.

Cyclosporin A is biosynthetically labelled with 13C by growing an overproducing strain of Tolypocladium inflatum on minimal media containing either [1-13C]-, [2-13C]-, [3-13C]- or [6-13C]glucose as the only carbon source. NMR analysis of the 13C-labelled peptide showed a labelling pattern in which 13C occurs at specific sites. These can be predicted by consideration of the relevant biosynthetic pathways. Quantitation of the site-specific enrichments revealed that the 13C-label incorporation is efficient and selective. Metabolic fluxes through alternative pathways can also be estimated from these results. Isotopically labelled peptides will be a very useful tool for the study of molecular interactions with their receptors.

Amino Acid Sequence↗
PubMed1991

The NMR structure of cyclosporin A bound to cyclophilin in aqueous solution.

Cyclosporin A bound to the presumed receptor protein cyclophilin was studied in aqueous solution at pH 6.0 by nuclear magnetic resonance spectroscopy using uniform 15N- or 13C-labeling of cyclosporin A and heteronuclear spectral editing techniques. Sequence-specific assignments were obtained for all but one of the cyclosporin A proton resonances. With an input of 108 intramolecular NOEs and four vicinal 3JHN alpha coupling constants, the three-dimensional structure of cyclosporin A bound to cyclophilin was calculated with the distance geometry program DISMAN, and the structures resulting from 181 converged calculations were energy refined with the program FANTOM. A group of 120 conformers was selected on the basis of the residual constraint violations and energy criteria to represent the solution structure. The average of the pairwise root-mean-square distances calculated for the backbone atoms of the 120 structures was 0.58 A. The structure represents a novel conformation of cyclosporin A, for which the backbone conformation is significantly different from the previously reported structures in single crystals and in chloroform solution. The structure has all peptide bonds in the trans form, contains no elements of regular secondary structure and no intramolecular hydrogen bonds, and exposes nearly all polar groups to its environment. The root-mean-square distance between the backbone atoms of the crystal structure of cyclosporin A and the mean of the 120 conformers representing the NMR structure of cyclosporin A bound to cyclophilin is 2.5 A.

Amino Acid Isomerases↗
PubMed1991

Two-dimensional 1H nuclear magnetic resonance study of AaH IT, an anti-insect toxin from the scorpion Androctonus australis Hector. Sequential resonance assignments and folding of the polypeptide chain.

Sequence-specific nuclear magnetic resonance assignments for the polypeptide backbone and for most of the amino acid side-chain protons, as well as the general folding of AaH IT, are described. AaH IT is a neurotoxin purified from the venom of the scorpion Androctonus australis Hector and is specifically active on the insect nervous system. The secondary structure and the hydrogen-bonding patterns in the regular secondary structure elements are deduced from nuclear Overhauser effects and the sequence locations of the slowly exchanging amide protons. The backbone folding is determined by distance geometry calculations with the DISMAN program. The regular secondary structure includes two and a half turns of alpha-helix running from residues 21 to 30 and a three-stranded antiparallel beta-sheet including peptides 3-5, 34-38, and 41-46. Two tight turns are present, one connecting the end of the alpha-helix to an external strand of the beta-sheet, i.e., turn 31-34, and another connecting this same strand to the central one, i.e., turn 38-41. These structure elements are very similar to the secondary structure reported in single crystals for either variant 3 from the scorpion Centruroides sculpturatus Ewing (CsE V3) or toxin II from the scorpion A. australis Hector (AaH II). The differences in the specificity of these related proteins, which are able to discriminate between mammalian and insect voltage-dependent sodium channels of excitable tissues, are most probably brought about by the position of the C-terminal peptide with regard to a hydrophobic surface common to all scorpion toxins examined thus far. This surface is made of an aromatic cluster that is surrounded by long hydrophobic side-chain residues, as well as the loops protruding out of it. Thus, the interaction of a given scorpion toxin with its receptor might well be governed by the presence of this solvent-exposed hydrophobic surface, whereas adjacent areas modulate the specificity of the interaction.

Amino Acid Sequence↗
PubMed1991

Determination of 6-keto-PGF1 alpha, 2,3-dinor-6-keto-PGF1 alpha, thromboxane B2, 2,3-dinor-thromboxane B2, PGE2, PGD2 and PGF2 alpha in human urine by gas chromatography-negative ion chemical ionization mass spectrometry.

A method for quantification of 6-keto-PGF1 alpha, 2,3-dinor-6-keto-PGF1 alpha TXB2, 2,3-dinor TXB2, PGE2, PGD2 and PGF2 alpha in human urine samples, using gas chromatography-negative ion chemical ionization mass spectrometry, is described. Deuterated analogues were used as internal standards. Methoximation was carried out in urine samples which were subsequently applied to phenylboronic acid cartridges, reversed-phase cartridges and thin-layer chromatography. The eluents were further derivatized to pentafluorobenzyl ester trimethylsilyl ethers for final quantification by gas chromatography-mass spectrometry. The overall recovery was 77% for tritiated 6-keto-PGF1 alpha and 55% for tritiated TXB2. Urinary levels of prostanoids were determined in a group of six volunteers before and after intake of the thromboxane synthase inhibitor Ridogrel, and related to creatinine clearance.

6-Ketoprostaglandin F1 alpha↗
PubMed1991

Production of thromboxane and prostaglandins in human blood in the presence of thromboxane synthase inhibitors: a comparison of RIA and GC/MS determinations.

The radioimmunological determination (RIA) of primary prostaglandins (PGs) in serum and plasma was evaluated with gas chromatography/mass spectrometry (GC/MS). Human blood was stimulated in vitro in the presence or absence of the specific thromboxane synthase inhibitor ridogrel. TxB2, 6-keto-PGF1 alpha, PGE2, PGD2 and PGF2 alpha were determined with RIA and GC/MS on the same samples. For GC/MS analysis, prostanoids were extracted and derivatized to their methoximepentafluorobenzyl-esters-trimethylsilyl-ethers. An excellent correlation was observed between levels of all eicosanoids determined by RIA or GC/MS (r = 0.996-0.999) when plasma was spiked with known amounts of PGs and TxB2 (2-500 ng/ml). In stimulated blood, with or without inhibition of thromboxane synthase, the correlation between RIA and GC/MS values remained good, except for 6-keto-PGF1 alpha. RIA largely overestimated the levels of 6-keto-PGF1 alpha and no significant correlation was found with levels detected by GC/MS. The results demonstrate the importance of corroborating the reliability of RIA with GC/MS.

6-Ketoprostaglandin F1 alpha↗
PubMed1991

Detection of circulating Fc epsilon R2/CD23+ monocytes in patients with rheumatic diseases.

Recently, in vitro studies have demonstrated that expression of Fc epsilon R2/CD23 on normal monocytes can be specifically induced by IL-4. In order to investigate the interaction of IL-4 and monocytes in rheumatic diseases, flow cytometry studies were performed. Elevated numbers of circulating Fc epsilon R2/CD23+ monocytes were detected in patients with progressive systemic sclerosis (PSS) as compared with controls. In addition, supernatants derived from phytohaemagglutinin-stimulated peripheral blood mononuclear cells of PSS patients contained high activity to induce Fc epsilon R2/CD23 on CD14+ monocytes. An increased frequency of Fc epsilon R2/CD23+ monocytes was also observed in rheumatoid arthritis, and sequential studies in patients with systemic lupus erythematosus showed a close relationship between Fc epsilon R2/CD23+ monocytes and disease activity. It is suggested that IL-4 has an important role in the pathogenesis of PSS by activating monocytes, and might also contribute to monocyte activation in other rheumatic diseases.

Adult↗
PubMed1991

[T-helper cell subsets in patients with inflammatory rheumatic diseases undergoing immunosuppressive therapy].

To determine the influence of immunosuppressive therapy on T-helper-cell subsets in patients with systemic lupus erythematosus and rheumatoid arthritis flow cytometric analysis was performed. Longitudinal studies showed reduced numbers of CD45R+CD4+ lymphocytes in patients treated with cyclophosphamide or azathioprine. Patients receiving steroids had low frequencies of CD29+CD4+ cells. The data suggest that cytotoxic drugs and steroids affect T-helper cells at different activation stages.

Antineoplastic Agents↗
PubMed1990

Relationship between CD5+ B lymphocytes and the activity of systemic autoimmunity.

We studied the relationship between CD5+ B cells and the activity of the disease process in patients with autoimmune diseases. In rheumatoid arthritis (RA), levels of CD5+ B cells were associated with autoantibody production as determined by serum rheumatoid factor and antinuclear antibodies. In addition, CD5+ B cells were significantly correlated with C-reactive protein, and data from longitudinal studies showed a marked influence of corticosteroid treatment on numbers of CD5+ B cells. Patients with systemic lupus erythematosus (SLE) had slightly elevated levels of CD5+ B cells as compared with normals, but a close association with measures of an active disease was not observed. In a group of patients with type I diabetes mellitus, CD5+ B cells were detected in patients with anti-islet cell antibodies. Our results suggest that CD5+ B cells are related to the activity of the autoimmune process and can be modulated by therapy in patients with RA. Although CD5+ B cells do not seem to have a major role in SLE, polyclonal activation might affect this B cell subset as well in this disease. Further studies are needed to define the precise role of CD5+ B cells in organ-specific autoimmunity.

Adrenal Cortex Hormones↗
PubMed1990

Treatment of pacemaker patients with extracorporeal shock wave lithotripsy: experience from 2 continents.

Patients with cardiac pacemakers currently are treated by extracorporeal shock wave lithotripsy only by agreement between individual physicians and patients. We present the results of a survey of patterns of extracorporeal shock wave lithotripsy treatment (or nontreatment) of patients with pacemakers in the United States and Europe. Letters of inquiry regarding treatment of pacemaker patients were sent to 141 United States extracorporeal shock wave lithotripsy sites and 55 sites in Europe. Responses were received from 81 (57%) and 17 (31%) sites, respectively. A total of 131 patients received 142 treatments and pacemaker-related complications occurred in 4 (none was lethal, 3 were minor and all were corrected immediately). No patient required replacement of the pacemaker. No correlation existed between complications and pacemaker type or manufacturer. Urologists who treat patients with a pacemaker required previous clearance of the patients by the cardiologist, a cardiologist in or near the extracorporeal shock wave lithotripsy suite and standby of corrective equipment or temporary pacemakers.

Europe↗
PubMed1990

Eicosanoids in rat brain during ischemia and reperfusion--correlation to DC depolarization.

The effects of complete ischemia on cerebral arachidonic acid (AA) metabolism were investigated in the isolated perfused rat brain. During 12.5 min of ischemia, AA, 5-hydroxy-6,8,11,14-eicosatetraenoic acid, and 15-hydroxy-5,8,11,13-eicosatetraenoic acid increased 129-, 4-, and 10-fold, respectively, while subsequent reperfusion for 30 min resulted in normalized levels independently of the duration of preceding ischemia. Prostaglandin (PG) F2 alpha, PGE2, PGD2, 6-keto-PGF1 alpha, and thromboxane (Tx) B2 remained at preischemic levels during 12.5 min of complete ischemia. However, at the end of subsequent reperfusion for 30 min, the levels of the prostanoids PGF2 alpha, PGE2, PGD2, 6-keto-PGF1 alpha, and TxB2 increased according to the preceding ischemic time. The levels reached a maximum after 7.5 min of ischemia and were elevated by 7-, 14-, 48-, 3-, and 30-fold, respectively. A prolongation of ischemia of up to 12.5 min was not associated with further increases of prostanoids at the end of reperfusion. The mechanisms underlying the metabolism of eicosanoids are discussed in relation to the changes of cortical direct current potential.

Animals↗
PubMed1990

Microvascular changes in rabbit kidneys after extracorporeal shock wave treatment.

Concerns about the safety of extracorporeal shock wave lithotripsy include the risk of vascular damage and the genesis of hypertension. We evaluated gross specimens and barium microangiograms of the right kidneys of 14 rabbits exposed to shock waves; the untreated left kidneys served as controls. Six rabbits were assessed immediately after shock wave treatment, whereas the other eight rabbits had the same procedure 3 months later. All six acute phase animals showed microangiographic changes indicative of shock wave trauma, including cortical, subcapsular, and medullary extravasation of barium and patchy loss of efferent vessels. No significant vascular abnormalities were seen in the control kidneys. All kidneys treated from the animals that had delayed microangiography showed macroscopic evidence of fibrotic capsular thickening and parenchymal scarring. Microscopically, there was crowding of glomeruli and areas of avascularity. In this model, shock waves damaged the cortical circulation and resulted in persistent alterations of the microvasculature that potentially could be responsible for the abnormal secretion of renin.

Animals↗
PubMed1990

Gaining control in a changing environment.

The rapid evolution of home care has frequently produced chaotic and unmanageable situations for home care personnel. This article will assist clinical supervisors with daily operations as they teach their staff how to effectively gain control and stabilize the chaos--how to manage, rather than be managed by, home care.

Community Health Nursing↗