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Biomedical subjects

R A Proctor

Publications and source records attributed to R A Proctor.

At least 73 records · Page 4Linked to original sources
PubMed1990

Lipid A precursors protect against endotoxin challenge.

These studies provide exciting prospects for the future treatment of gram-negative infections. The anti-endotoxin activity of lipid X, a monosaccharide precursor of lipid A, may be a prototypic compound for agents that can block the toxic effects of endotoxin that is being actively released. In contrast, monophosphoryl lipid A, a disaccharide derivative of lipid A, stimulates host defenses against infections and tumors. With further understanding of the mechanisms by which these compounds exert these effects, we can anticipate that new and more active compounds will be developed and that further activities of existing compounds will be found.

Animals↗
PubMed1990

Fibronectin-staphylococcal interactions in endovascular infections.

Fibronectin is a large molecular weight protein that is found coating the surfaces of sites where endovascular damage has occurred. These are also the sites most commonly infected by bacteremic strains of staphylococci. Epidemiologic studies show a correlation between expression of fibronectin receptors and development of invasive infections. In vitro studies using cultured cells, artificial matrices containing fibronectin, blood clots, natural inflammatory matrices, and anti-fibronectin antibodies implicate fibronectin as an important ligand for staphylococcal attachment to host tissues and prosthetic devices. In the rat endocarditis model, S. aureus strains that lack the fibronectin receptor due to site-directed mutagenesis were unable to colonize the traumatized heart valves. These data suggest that the fibronectin receptor on staphylococci is important in the pathogenesis of endovascular infections. Because the fibronectin receptor is widely expressed on pathogenic of staphylococci, a broadly protective vaccine against S. aureus might be possible.

Animals↗
PubMed1989

Immunoelectron microscopic localization of fibronectin in adherence of Staphylococcus aureus to cultured bovine endothelial cells.

Fibronectin binds to Staphylococcus aureus and may have a role in mediating its adherence to host tissue. Fibronectin was localized ultrastructurally on S. aureus in suspension and in interactions with cultured bovine endothelial cells. Probes were constructed by adsorbing fibronectin or its antibodies to colloidal gold beads (FN-Au or aFN-Au, respectively). Sites of fibronectin binding to S. aureus were demonstrated by reacting FN-Au with S. aureus in suspension. Transmission electron microscopy showed that FN-Au localized uniformly over the surface of S. aureus in suspension; most localized within 65 nm of the cell wall. The distribution of aFN-Au on S. aureus adherent to endothelial cells was concentrated in areas between S. aureus and endothelial cells. Areas of S. aureus not facing endothelial cells bound few aFN-Au. This suggests that the fibronectin labeled by aFN-Au in areas between S. aureus and endothelial cells was involved in adherence of the S. aureus, consistent with a role for fibronectin in endocarditis.

Animals↗
PubMed1989

Reduced adherence to traumatized rat heart valves by a low-fibronectin-binding mutant of Staphylococcus aureus.

Isogenic strains of Staphylococcus aureus, differing in fibronectin binding, were constructed for studies of the contribution of fibronectin binding to the in vivo pathogenesis of staphylococcal disease. Mutagenesis of S. aureus 879R4S was accomplished by mating with Enterococcus faecalis FA378 that carried the transposon Tn918. Four low-fibronectin-binding mutants were identified that bound 24- to 35-fold less fibronectin than the parent strain did. A spectinomycin-resistant strain, R4SSp, was transduced by a bacteriophage (80 alpha) lysate propagated on a low-binding mutant of 879R4S to produce R4SSp/1536, which bound less fibronectin, contained a single copy of the transposon, and grew on spectinomycin-containing medium. Using a rat model of endocarditis, we determined the distribution of S. aureus R4SSp and its transductant in normal and cardiac catheterized rats. Cultures of heart tissue showed that catheterized rats challenged with the fibronectin-binding parent strain had over 250-fold more organisms in the left heart than did rats challenged with the low-binding transductant. The ability to bind fibronectin had no effect on the number of S. aureus cells cultured from other tissues. These data suggest that the ability of S. aureus to bind fibronectin is an important factor in establishing adherence to damaged heart valves in vivo.

Animals↗
PubMed1988

Cytotoxic effects of ingested Staphylococcus aureus on bovine endothelial cells: role of S. aureus alpha-hemolysin.

Previously we observed that Staphylococcus aureus phagocytized by cultured bovine endothelial cells do not proliferate intracellularly, but are cytotoxic to bovine endothelial cells. To investigate S. aureus virulence factors which may be produced intracellularly and cause lysis of endothelial cells, we tested S. aureus mutants defective in production of one or more potential virulence factors and corresponding parent strains for cytotoxicity to endothelial cell monolayers subsequent to being ingested. Following incubation of endothelial cell monolayers with S. aureus for 3.5 h, cultures were supplemented with lysostaphin to destroy extracellular but not intracellular S. aureus. At subsequent times, viability of endothelial cells was assayed by retention of 3H-adenine and the number of intracellular S. aureus was measured. The cytotoxic activity of S. aureus culture supernatants was also characterized. The results indicate that S. aureus alpha-hemolysin is cytotoxic to bovine endothelial cells and plays an important role in the damage suffered by bovine endothelial cell monolayers following ingestion of S. aureus. Ingestion of alpha-hemolysin-producing S. aureus by endothelial cells in vivo might be expected to result in destruction of endothelium followed by development of platelet-fibrin vegetations. This possible sequence of events is compatible with the frequently fulminant course of S. aureus endocarditis.

Animals↗
PubMed1988

Elimination and tissue distribution of the monosaccharide lipid A precursor, lipid X, in mice and sheep.

Lipid X (2,3-diacylglucosamine 1-phosphate) is a novel monosaccharide precursor of lipid A (the active moiety of gram-negative endotoxin) and has been found to be protective against endotoxin administered to mice and sheep and against life-threatening gram-negative infections in mice. Because of the need to design optimal dosing regimens in experimental models of ovine and murine septicemia, the pharmacokinetic profile of lipid X was investigated in sheep and in two strains of mice by using 32P-labeled lipid X. In sheep, peak whole blood lipid X levels after a bolus injection of 100 micrograms of lipid X per kg were 900 ng/ml. An initial rapid distribution phase of 7.98 +/- 0.1 min was observed, followed by a prolonged elimination phase of 3.0 +/- 0.5 h; the area under the curve from time zero to infinity was 428 +/- 27 ng.h/ml. The serum half-lives of lipid X were slightly shorter than whole blood half-lives, suggesting that lipid X associates with cellular elements. Metabolites of lipid X could not be detected in serum over a 4-h period. Lipid X appears to accumulate mainly in the liver, and the tissue distribution of lipid X resembles that of lipopolysaccharide. The elimination rate of lipid X in mice was approximately four times as rapid as that seen in sheep. Lipid X pharmacokinetics in lipopolysaccharide-sensitive DBA/2J mice were virtually identical with those seen in endotoxin-resistant C3H/HeJ mice. The pharmacokinetics described here should greatly aid in the design and interpretation of animal studies investigating the therapeutic applications of lipid X in gram-negative septicemia.

Animals↗
PubMed1988

Lipid X protects mice against fatal Escherichia coli infection.

Lipid X, the major monosaccharide precursor of lipid A, is nontoxic and has previously been shown to protect mice and sheep from the harmful effects of endotoxin. To test the hypothesis that lipid X could be therapeutic against infections with gram-negative organisms, neutropenic ICR mice were infected by intramuscular inoculation of Escherichia coli and subsequently treated with lipid X alone or in combination with the antibiotic ticarcillin. Lipid X slightly prolonged survival; treated mice had a significantly improved rate of survival 18 h after intramuscular inoculation as compared with controls (P less than 0.025). By 24 h, however, this difference disappeared. When lipid X was combined with ticarcillin, survival differences were both significant and prolonged. Treatment of mice with one to two doses of lipid X for a total dose of 1 mg intravenously and with 1,200 mg of ticarcillin per kg every 6 h improved survival over a 48-h treatment period from 5 to 23% (P less than 0.0025). Treatment with lipid X and ticarcillin over a broad range of antibiotic dosages in 362 mice demonstrated improved survival of two- to fourfold (P less than 0.0001 at 24 h after inoculation, P less than or equal to 0.0005 at 48 h, and P less than or equal to 0.0001 at 5 days). Lipid X enabled the dose of ticarcillin necessary to protect 50% of mice from death to be reduced by two- to fivefold. Pretreatment with lipid X was not necessary to improve survival: 16 of 17 (94%) infected and visibly ill animals that received lipid X and ticarcillin 6 h after thigh inoculation survived versus 30 of 44 (68%) control animals treated with ticarcillin alone (P less than 0.0001). Lipid X had no antimicrobial activity in vitro. Lipid X is a novel agent that enhances survival in an animal model of severe infection with gram-negative organisms.

Animals↗
PubMed1987

In-vitro studies of antibiotic combinations for multiply-resistant coagulase-negative staphylococci.

Coagulase-negative staphylococci are being implicated as pathogens with increasing frequency and this may in part be due to their development of resistance to a wide variety of antibiotics. Because vancomycin is the only drug generally available for treating these multiply resistant organisms, we have tested several combinations of antibiotics for activity in vitro against these organisms. These in-vitro studies suggested that rifampicin combined with either gentamicin or cefamandole might occasionally provide an efficacious alternative to vancomycin, while the novobiocin plus rifampicin combination might be an effective oral regimen against methicillin-resistant coagulase negative staphylococci. Resistance to co-trimoxazole was found in nearly all isolates tested.

Anti-Bacterial Agents↗
PubMed1987

Haemophilus parainfluenzae vertebral osteomyelitis.

Haemophilus parainfluenzae is a normal inhabitant of the nose and oropharynx. The authors report a case of adult vertebral osteomyelitis due to this organism that occurred 3 months after nasal septoplasty. Successful therapy was accomplished by a combination of inpatient and outpatient therapy.

Haemophilus Infections↗
PubMed1987

Lipid X ameliorates pulmonary hypertension and protects sheep from death due to endotoxin.

Lipid X (2,3-diacylglucosamine-1-phosphate) is a novel monosaccharide precursor of lipid A that has some of the physiologic activities of endotoxin but little toxicity. To determine whether lipid X would interfere with the toxic effects of endotoxin, we pretreated sheep with either 100 or 200 micrograms of lipid X per kg of body weight and then challenged them with a potentially fatal dose of Escherichia coli endotoxin (20 micrograms/kg). Twenty-one sheep underwent pulmonary artery catheterization and were monitored for changes in pulmonary artery pressure, temperature, pH, partial O2 pressure, partial CO2 pressure, blood pressure, and cell counts over 7 h. Overall mortality for control animals was 37% versus 5.3% for pretreated animals. None of the 13 animals pretreated with 100 micrograms of lipid X per kg died. These differences in survival were significant (P less than 0.05). Animals pretreated with 100 micrograms of lipid X per kg had significantly lower pulmonary artery pressure during both phases 1 and 2 of endotoxin-induced pulmonary artery hypertension. A higher dose of lipid X, 200 micrograms/kg, produced pulmonary hypertension. Perhaps because lipid X is a subunit of lipid A, lipid X shows a partial pyrogenic effect while also decreasing the pyrogenic activity of complete lipopolysaccharide (LPS). Lipid X did not prevent endotoxin-induced neutropenia or moderate hypotension in response to LPS. Lipid X is a potential prototype compound for a new type of chemotherapy directed at blocking the harmful effects of LPS during bacterial septicemia.

Animals↗
PubMed1987

Ingestion of Staphylococcus aureus by bovine endothelial cells results in time- and inoculum-dependent damage to endothelial cell monolayers.

Cultured endothelial cells phagocytize Staphylococcus aureus, but the resultant effects are unknown. Monolayers of cultured bovine endothelial cells with or without [3H]adenine label were exposed to 100, 10, or 1 S. aureus organism per endothelial cell for 3.5 h. Lysostaphin was then applied to all cultures to destroy extracellular but not phagocytized S. aureus. In cultures treated for only 20 min with lysostaphin, S. aureus multiplied exponentially after a 9- to 12-h lag period. In cultures treated continuously with lysostaphin, numbers of S. aureus remained constant or decreased. These results indicate that S. aureus became extracellular and multiplied but did not multiply intracellularly. In parallel experiments, the release of 3H-adenine from prelabeled endothelial cell monolayers was assayed to indicate cytotoxicity. Results indicated that the loss of 3H-adenine from endothelial cell monolayers depended on the following: (i) the size of the S. aureus inoculum, (ii) the strain of S. aureus, and (iii) the length of time after exposure to S. aureus. S. aureus endocarditis and persistent septicemia could arise, at least in part, from ingestion of S. aureus by host endothelium. The intracellular location would afford S. aureus protection from host defenses and antibiotics. Eventual damage to endothelial cells could expose collagen, thus resulting in platelet adherence and vegetation formation. Intracellular S. aureus would be continuously released into the circulation, possibly accounting for the persistent bacteremia that is found in S. aureus endovascular infections.

Animals↗
PubMed1987

Pulmonary pressor responses in sheep to chemically defined precursors of E. coli endotoxin.

The toxicity of various monosaccharide and disaccharide endotoxin precursors has now been studied in sheep. We measured the early pulmonary arterial pressure responses after injections of the monosaccharides lipid X (2,3-diacylglucosamine 1-phosphate) and MAGP (2-monoacylglucosamine 1-phosphate), of the tetraacyl disaccharide diphosphate precursor of lipid A, IV-A (Federation Proc. 43: 1567, 1984), and of Escherichia coli bacterial endotoxin (lipopolysaccharide). We also measured the response of lipid X after prior administration of indomethacin and MAGP. Lipid X, at a total cumulative dose of 40 micrograms/kg, produced an immediate, but transient dose-dependent pulmonary arterial vasoconstrictive response. MAGP, at a total dose of 40 micrograms/kg, had no pulmonary pressure activity but did increase extravascular lung water and produce some histological changes in the lung. Disaccharide precursor IV-A, at a total dose of 40 micrograms/kg, produced an immediate dose-dependent pulmonary arterial vasoconstrictive response that was prolonged for greater than 2 h. E. coli endotoxin caused a delayed (15-min) increase in the pulmonary arterial pressure but one that also persisted for greater than 2 h. Prior administration of indomethacin blocked the pulmonary pressor activity of lipid X, whereas prior administration of MAGP increased both the magnitude and the duration of the pulmonary pressure response of lipid X. We conclude that the initial pulmonary hypertension seen after lipid X injection may involve cyclooxygenase-dependent formation of prostaglandins and that the genesis of this pulmonary pressor activity is at least in part dependent on the ester-linked hydroxymyristoyl moiety at position 3 of the lipid X molecule.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Protection of mice against lethal endotoxemia by a lipid A precursor.

Lipid X, the major biosynthetic precursor of lipid A, has recently been described. Although lipid X is a mitogen and coagulates the Limulus amebocyte lysate, we found that it is not lethal for mice, even when given in large doses (2 X 10(6) micrograms/kg). Furthermore, lipid X was found to give partial protection against a 100% lethal dose of endotoxin, even if the lipid X was given as late as 6 h after endotoxin challenge.

Animals↗
PubMed1986

Phagocytosis of Staphylococcus aureus by cultured bovine aortic endothelial cells: model for postadherence events in endovascular infections.

We examined the interaction of Staphylococcus aureus with cultured bovine aortic endothelial cells as a model for the initial events in the pathogenesis of endovascular infections. Confluent monolayers of cultured endothelial cells were incubated with S. aureus. Cell-associated bacteria were measured by washing away nonadherent organisms, disrupting the monolayers, and performing quantitative cultures. Phagocytosis was differentiated from adherence by treating the cells with lysostaphin; approximately 60% of cell-associated bacteria was found to be intracellular. Phagocytosis could be blocked by using cytochalasin B, which interferes with microfilament function. Addition of fibronectin resulted in a 63% increase in adherence of S. aureus to the endothelial cells but did not increase ingestion. Transmission electron microscopy demonstrated a sequence of events similar to that which occurs during ingestion by professional phagocytes, including: adherence of bacteria to the endothelial cell; formation and elongation of surface extensions of the endothelial cell to surround the adherent bacteria; and complete enclosure within apparent phagosomes. Phagocytosis of bacteria by endothelial cells, followed by intracellular persistence, may be an important postadherence event in the pathogenesis and pathophysiology of endovascular infections.

Animals↗
PubMed1986

Oxidative metabolism of circulating neutrophils in infantile (Philadelphia chromosome-negative) chronic myelogenous leukemia.

Mature polymorphonuclear neutrophilic leukocytes (PMNL) from a 2.5-year-old female with infantile chronic myelogenous leukemia (CML) were stimulated with phorbol myristate acetate or opsonized zymosan. The resulting enzymatic NADPH oxidation and the cellular O2- release and luminol-enhanced chemiluminescence were measured. The patient's PMNL responded normally in all respects. Thus, mature infantile CML PMNL undergo a normal respiratory burst following either soluble or particulate stimulation. Our review of the literature emphasizes the importance of studying a well-defined population of PMNL in patients with myelodysplasia.

Child, Preschool↗