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Biomedical subjects

G Beck

Publications and source records attributed to G Beck.

At least 163 records · Page 9Linked to original sources
PubMed1985

Chemical and biologic characterization of a lipopolysaccharide extracted from the Lyme disease spirochete (Borrelia burgdorferi).

A lipopolysaccharide (LPS) was isolated from the Lyme disease spirochete by a modification of the hot phenol-water method. The material was composed of 45% carbohydrate, 8% protein, 44% lipid A, and 1% 3-deoxy-D-mannooctulosonic acid and accounted for approximately 1.5% of the cellular dry weight. The isolated LPS possessed several biologic activities characteristic of endotoxins. The LPS was pyrogenic for rabbits, mitogenic for human mononuclear cells and murine splenocytes, capable of clotting limulus lysate, and cytotoxic for murine macrophages. LPS extracted from Borrelia burgdorferi by the petroleum-ether:chloroform:liquid-phenol procedure was also characterized. The results show that the Lyme disease spirochete contains a hitherto unknown LPS that is biologically active in vitro, and the expression of such activities in vivo may play an important role in the pathogenesis of Lyme disease. Some of the clinical manifestations of other spirochetal disease may be explained by similar endotoxins in those organisms. To our knowledge this is the first report of an LPS extracted from a spirochete that is known to be a human pathogen.

Animals↗
PubMed1985

Rheological and transport properties of airway secretions in cystic fibrosis--relationships with the degree of infection and severity of the disease.

It has recently been suggested that in cystic fibrosis (CF), there is no rheological abnormality of airway secretions other than that associated with purulence, and that the apparent inhibition in the mucociliary transport rate might be partly due to a ciliary inhibitor present in these secretions. In order to ascertain this assumption, expectorated airway secretions were collected without salivary contamination in twenty-four CF patients and the rheological properties were measured. Using a photometric method, the effects of CF sputum samples were analysed on the ciliary beat frequency (Fm) of the frog palate, and we measured their mucociliary transport rate (TR). In all but one CF sputum, TR and Fm were lower than that of the control frog mucus (median TR: 18.7 and 11.6 mm min-1; median Fm: 12.3 and 11.3 Hz, respectively). In the eighteen patients in whom the rheological properties were outside the range for optimal mucociliary transport, the clinical Shwachman score was significantly (P less than 0.05) lower (median score: 66.2 points) than in the six patients with optimal rheologic properties (median score: 73 points). In the eleven CF patients with superinfection, the apparent viscosity (eta o) was significantly higher (P less than 0.01; median eta o: 24.4 Pa. s) and TR, expressed as a percentage of the reference value, was significantly lower (P less than 0.05; median Tr: 54.5%) in comparison with the values obtained for the thirteen non-superinfected CF patients (median eta o: 15 Pa. s and median TR: 66% respectively). The CF patients with markedly hyperviscous sputum (eta o higher than 30 Pa. s) exhibited a low Shwachman score.

Adolescent↗
PubMed1985

Effect of the cytotoxin of Clostridium difficile on cultured hepatoma cells.

Clostridium difficile is the major etiologic agent of human pseudomembranous colitis. It produces two toxins: an enterotoxin and a cytotoxin. In cultured hepatoma cells, at very low doses, the cytotoxin inhibits the incorporation of precursors into biological macromolecules. Protein synthesis is more affected than RNA and DNA synthesis. The toxin also induces severe alterations of the cell morphology consisting in damages to the cytoskeleton and to the cell shape.

Animals↗
PubMed1985

Skin reactivity of unsensitized monkeys upon challenge with staphylococcal enterotoxin B: a new approach for investigating the site of toxin action.

The correlation between skin tests and emetic responses in unsensitized monkeys was used to elucidate the cellular site of action of staphylococcal enterotoxin B (SEB). Evidence is presented that SEB administered intradermally provoked immediate-type skin reactions associated with mild degranulation of cutaneous mast cells. The cytoplasma showed signs of synthetic and metabolic activity, with formation of vesicles and increased prominence of mitochondria. Carboxymethylation of histidine residues of SEB altered the molecule (cSEB) from more alkaline components to more acidic species with increased microheterogeneity. This modification caused a loss in toxicity and completely abrogated the skin-sensitizing activity without changing the immunological specificity. cSEB, however, could compete with SEB for binding sites on the target cell surface. Previously, compound 48/80-treated skin sites behaved refractively to challenge with SEB, indicating that mediators from cutaneous mast cells are required for SEB-induced skin reactions. Skin reactions as well as emetic responses challenged with SEB were completely inhibited by H2 receptor antagonists and calcium channel blockers but not by H1 antihistamine or competitive antagonists of serotonin. This new approach provides a model for investigating the mechanisms of SEB action.

Animals↗
PubMed1985

Lyme disease spirochetes induce human and murine interleukin 1 production.

IL 1 is a major immunoregulatory molecule produced by macrophages, and it appears to be the molecular orchestrator of nonspecific host defense mechanisms against a variety of environmental insults. Many investigators have used artificial agents to stimulate macrophages to produce IL 1. We now report production of large quantities of IL 1 after a physiologic stimulus. The Lyme disease spirochete, recently isolated and adapted for growth in vitro, was used to stimulate P388D1 cells or human peripheral blood monocytes. Spirochetes were added to confluent macrophage cultures in serum-free RPMI at a ratio of 10:1. The release of IL 1 was dose-dependent. The 24-hr supernatant IL 1 activity was determined by using the thymocyte Con A co-mitogenesis assay. Activity was not due to an endotoxin on, or produced by, the spirochete. A polymyxin B affinity column failed to remove activity, and polymyxin B in the spirochete-macrophage culture had no effect on IL 1 production. Supernatants were collected, were concentrated, and were subjected to size exclusion HPLC. Three areas of activity were found in P388D1 cell supernatants (Mr greater than 60,000, 40,000, and 20,000), whereas two peaks (Mr 23,000 and 13,000) were found in human monocyte supernatants. The Mr 20,000 and 13,000 peaks from murine and human cell supernatants, respectively, were subjected to SDS-PAGE and were shown to be single bands (Mr 12,400 for the mouse IL 1 and Mr 13,500 for the human IL 1). Isoelectric focusing of column-purified IL 1 preparations showed two different pI in both human (pI 7.25 and 4.4 to 5) and murine (pI 7.25 and 5.55) IL 1. Fibroblasts cultured with murine or human IL 1 preparations demonstrated both an increase in secreted collagenase and increased cell proliferation. Thus, a physiologic stimulus and simple biochemical techniques produce large amounts of very pure mouse or human IL 1. That this IL 1 is produced by Lyme disease spirochete-stimulated macrophages may explain some of the clinical manifestations of Lyme disease.

Animals↗
PubMed1985

[Efficacy of the paramunity inducers PIND-AVI and PIND-ORF as radioprotective agents].

A significant reduction of mortality after lethal irradiation (7,8 and 9 Gy X-ray total-body irradiation) was achieved by continuous therapeutic subcutaneous application of the biologic inducers PIND-AVI and PIND-ORF. This was obtained by a stimulation of the investigated spleen parameters and a stimulation of leucocytes and phagocytosis. The inducers had no significant influence on the radiogenic reduction of blood cells and bone marrow cells, of the relative spleen and thymus weight and of the DNA and protein level of spleen and thymus. The regeneration of blood leucocytes (preponderantly by PIND-AVI) and of the spleen (preponderantly by PIND-ORF) was accelerated, but not the thymus regeneration which was already rather low. The leucocyte phagocytosis which increased generally after irradiation was markedly stimulated by paramunization. This stimulation was also observed after a latent time in animals submitted to sham irradiation. Its temporal appearance and its quantitative and qualitative properties corresponded to the effects of inducers after X-ray irradiation. Both inducers were found to be harmless in all experiments.

Animals↗
PubMed1985

Closed locked intramedullary nailing. Its application to comminuted fractures of the femur.

For many fractures of the femoral shaft, closed intramedullary nailing will not control rotation or telescoping of the fragments. Locked intramedullary nailing combines closed nailing with the percutaneous insertion of screws that interlock the bone and nail. This method permits static locking that controls rotation and telescoping and subsequently conversion to dynamic locking when weight-bearing is started after approximately twelve weeks. By providing greater stability, this method extends the indications for intramedullary nailing to severely comminuted, oblique, and spiral fractures as well as to fractures complicated by loss of bone and fractures in the proximal and distal ends of the femoral shaft. Of fifty-two patients with forty-nine severely comminuted fractures of the femoral shaft and three fractures that were complicated by loss of bone, forty-seven patients had uneventful consolidation of the fracture, with a mean time of 4.5 months for the severely comminuted fractures and seven months for the fractures that had a loss of bone. At follow-up, all forty-seven patients had normal motion of the hip, and forty-five had normal motion of the knee. Of the remaining five patients, four had a non-union that eventually healed (three after a second locked nailing and one after a third) and one had a septic non-union that eventually healed after removal of the nail and screws, débridement, and immobilization with an external fixator. Based on this experience, we concluded that this form of treatment has many advantages. The risk of infection and non-union is low, the incidence and severity of malunion are reduced, the hospital stay is short, and early mobilization of the patient is possible.

Adult↗
PubMed1985

Steroid metabolism in normal and transformed liver cells. Relation with glucocorticoid antagonism.

The metabolism of the potent antagonist RU38486 has been studied in cultured liver cells and in two hepatoma cell lines. In the liver cells, this steroid undergoes a rapid degradation, whereas in hepatoma cells grown in similar conditions only a minor degradation occurs. Moreover the rate of degradation is much higher for the antagonist steroid than for the agonist steroid tested, dexamethasone. The high antiglucocorticoid potency and the relative instability of the RU38486 molecule are very important to define its different effects and its mechanism of action in liver and in liver derived tumor cells. RU38486 may represent a useful drug in cancerotherapy.

Animals↗
PubMed1984

Antagonism of glucocorticoid action in cultured hepatoma cells.

We report here our recent data on the effects of antiglucocorticoids in two established cell lines (HTC, FAZA). These steroid hormone target tissues were designed to consider the problem of differential antagonism and lack of correlation between antiglucocorticoid activity and competition for the glucocorticoid receptor. In the systems chosen, several responses were considered which may be differentially antagonized: induction of tyrosine aminotransferase (TAT), alanine aminotransferase (AAT) and tryptophan oxygenase (TPO). By testing the anti-inducing capacities of a number of steroids, we found a few synthetic compounds like promegestone and R 25055 which exert a stronger antagonism against TAT and AAT induction than natural steroids like progesterone. The availability of highly radioactive antagonists (promegestone, progesterone) has greatly facilitated our "whole cell" study and allowed us to detect the antagonists in isolated nuclei whose purity and morphological integrity were controlled by specific criteria; our results suggest that the binding of the antagonists to the nucleus proceeds via the glucocorticoid receptor. The appearance of promegestone and progesterone in the nucleus suggests that the nucleus may be involved in the mechanism of action of anti-glucocorticoids.

Animals↗
PubMed1984

An approach to the mechanism of the potent antiglucocorticoid: 17 beta-hydroxy-11 beta-4-dimethylaminophenyl-17 alpha-propynyl-estra-4,9-dien-3-one.

The effects of a new synthetic steroid, 17 beta-hydroxy-11 beta-4-dimethyl-aminophenyl-17 alpha-propynyl-estra-4,9-dien-3-one, classified under reference R38486 in the Roussel-Uclaf nomenclature [1], were investigated in two established rat hepatoma cell lines in order to gain information on the mechanism of action. The induction of tyrosine aminotransferase (TAT) and alanine aminotransferase (AAT) was totally abolished when R38486 was added with dexamethasone either on a 1-1 basis or on a 10-fold excess depending on the differentiation state of the cell. Binding studies showed a high affinity for the glucocorticoid receptor; however our "whole cell" study with [3H] R38486 indicates that only a low amount of antagonist-receptor complexes was translocated into the nucleus. Nuclear fractionation experiments showed that R38486, like the other antagonists studied, was located in the chromatin fraction where it may exert some definite role. Our observations based on whole cell experiments using physiological doses of glucocorticoid analogs indicate that the binding of activated antiglucocorticoid-receptor complexes to nuclear acceptor sites represents a physiologically significant process. Moreover the differences in the nuclear binding of antagonist-receptor- as compared to agonist-receptor-complexes may set off the machinery of antagonistic action.

Animals↗
PubMed1984

Biological activities of 6-keto-prostaglandin E1.

In this article the broad spectrum of biological properties of 6-keto-Prostaglandin E1 will be reviewed. This includes assay procedures, biosynthesis and metabolism, various aspects of pharmacological activities, disease states, therapeutic potential and analogues of 6-keto-Prostaglandin E1.

Alprostadil↗
PubMed1983

Isolation and characterization of a trypsin-like serine proteinase from the membranes of Walker 256 carcino-sarcoma cells.

A serine proteinase was isolated from Walker-256-carcino-sarcoma plasma-membrane-enriched preparations by affinity chromatography employing soya-bean trypsin inhibitor as the ligand. This enzyme was termed 'memsin' owing to its membrane location and trypsin-like substrate specificity. Analysis of this preparation by steric-exclusion high-pressure liquid chromatography (h.p.l.c.) resulted in a single peak of enzyme activity. Calculations of the rates of inactivation of memsin by peptidyl-chloromethanes and comparison with rate constants obtained with other serine proteinases indicated that memsin closely resembled trypsin and acrosin. Digestion of oxidized ribonuclease by memsin and analysis of the resulting peptides by h.p.l.c. yielded a chromatogram that was very similar to one generated by a tryptic digest of oxidized ribonuclease. This enzyme could possibly play a role in tumour-cell invasion.

Animals↗
PubMed1983

Nuclear localization of a glucocorticoid antagonist in cultured hepatoma cells.

The binding characteristics of promegestone, a typical antagonist of glucocorticoid action, has been investigated. We localized [3H] promegestone in the nuclei purified from steroid-treated HTC cells but the radioactivity specifically bound to the nucleus was much lower for the antagonist than for an inducer steroid, for instance dexamethasone. We attempted to define the nature of the "nuclear" activity and found that promegestone does not bind to the perinuclear membrane but is associated with the chromatin fraction.

Animals↗
PubMed1983

Pyridoxal 5' phosphate involvement in age-related modifications of tyrosine aminotransferase: multiple form patterns.

Multiple form patterns of tyrosine aminotransferase were studied in senescent and adult rat liver. Two main modifications were described for "old" rat enzyme: (i) appearance of a new molecular form, specific for old rats, eluted after adult form III and having other properties identical to this form; (ii) disappearance of intermediate forms II and III after enzyme induction; this result seems to be due to acceleration of the conversion process. Vitamin B6 deficiency of old rats explain this and other (previously described) post-translational modifications of "old" tyrosine aminotransferase. The influence of pyridoxal 5' phosphate and the role of protease(s) in the multiple form conversion are discussed. Moreover we show the possibility of a correlation between in vivo alterations of the enzyme molecule and modifications of tyrosine aminotransferase multiple form patterns observed in vitro.

Aging↗
PubMed1983

Direct skin test in highly sensitized guinea pigs for rapid and sensitive determination of staphylococcal enterotoxin B.

The direct skin test in highly sensitized guinea pigs was developed as a rapid and extremely sensitive assay for detection of staphylococcal enterotoxin B (SEB) in foods. This report details the experimental conditions required to elicit optimal sensitization of guinea pigs to SEB. An intense and persistent immunoglobulin E (IgE) anti-SEB response was established in strain 13 guinea pigs pretreated with cyclophosphamide followed by four sensitizing doses of 10 micrograms of SEB 1 month apart. The conditions, however, optimal for eliciting IgE responses led to a sustained failure to produce antibody of the IgG1 subclass. With the use of highly sensitized guinea pigs, one can achieve a sensitivity ranging from 0.1 to 1.0 pg of purified SEB by the direct skin test for at least 7 months after the last challenge. For analysis of SEB in food extracts, the entire assay can be accomplished within 20 min with a sensitivity of 10 to 100 pg SEB per ml of prepared food samples, and the recovery of enterotoxin from spiked food products ranged between 75 and 89% of the amount added.

Biological Assay↗
PubMed1983

Mechanism of BCG-activated macrophage-induced tumor cell cytotoxicity: evidence for both oxygen-dependent and independent mechanisms.

Activated peritoneal macrophages have been shown to be cytotoxic to cancer cells. Bacillus Calmette-Guerin (BCG)-activated rat peritoneal macrophages have a basal cytolytic potential for 3H-thymidine-labelled Walker 256 cancer cells in vitro that can be markedly enhanced by digitonin. This stimulation of cytotoxicity can be partially inhibited by catalase and the combination of superoxide dismutase plus catalase. This suggests that digitonin stimulates activated macrophages to produce superoxide, hydrogen peroxide and possibly other free radicals which can augment macrophage-induced tumor cell cytotoxicity. After a 2-hour incubation with digitonin, macrophages are no longer stimulated by digitonin. However, after a 2-hour drug preincubation period, inhibitors of serine protease activity (DFP, TLCK, SBTI) and inhibitors of protein synthesis (cycloheximide) are potent inhibitors of basal macrophage-induced tumor cytotoxicity. We suggest that BCG-activated macrophages have two mechanisms for destroying cancer cells: one mediated by proteolytic activity, and a second mechanism dependent on the generation of oxygen-derived free radicals.

Animals↗
PubMed1983

Characterization of tumor cell membrane serine proteases by polyacrylamide gel electrophoresis.

Studies in our laboratory have indicated that tumor cell membrane-bound proteases are responsible for the ability of tumor cells to lyse normal cells in vitro. In order to evaluate the tumor cell membrane enzymes, a purified tumor cell membrane preparation was prepared and the nonionic detergent Triton X-100 was used to extract active enzymes from the cell membranes. The solubilized membrane enzymes were then studied by Triton X-100 polyacrylamide gel electrophoresis under non-denaturing conditions. Using this technique the tumor cell membranes were shown to contain esterproteases that reacted with the substrates alpha-naphthyl acetate and naphthol-AS-aminocaproate. These esterproteases were inhibited by diisopropyl fluorphosphate and tosyl lysine chloromethyl ketone but not by tosylamide phenylethyl chloromethyl ketone, soybean trypsin inhibitor p-chloromercuribenzene sulfonic acid; N-ethylmaleimide choline iodide, alpha-1-anti-trypsin. NaF, epsilon-aminocaproic acid, ethylenediamine tetraacetic acid, or eserine. SBTI affinity chromatography of the tumor cell membrane extract revealed that some of the serine esterproteases bound to the SBTI column. The proteolytic activity of the tumor cell membrane extract and a fraction eluted from the SBTI affinity column was demonstrated using casein. We conclude that the tumor cell membranes contain previously undescribed serine proteases that are identifiable by their esterase activity and inhibitor profiles in polyacrylamide gels.

Animals↗