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L M McManus

Publications and source records attributed to L M McManus.

At least 55 records · Page 3Linked to original sources
PubMed1984

A facile route to semi-synthesis of acetyl glycerylether phosphoethanolamine and its choline analogue.

A facile route to the semi-synthesis of acetyl glycerylether phosphoethanolamine and, subsequently, its choline analogue (platelet-activating factor) has been developed. In essence, this technique takes advantage of the fact that the phosphatidylethanolamine fraction of bovine erythrocytes contains 75-80% of a 1-O-alkyl-2 fatty acyl derivative. Isolation of the latter by silicic acid chromatography followed by base-catalyzed methanolysis allowed good recovery (60-70%) of the 1-O-alkyl-(lyso)-sn-glyceryl-3-phosphoethanolamine, which contained a mixture of long chain alkyl ethers. This compound was treated with acetic anhydride in the presence of trace amounts of perchloric acid for 45 sec to give, in excellent yield, 1-O-alkyl-2-acetyl-sn-glyceryl-3-phosphoethanolamine (AGEPE). This procedure gave a 70% yield of purified AGEPE, based on the starting component, 1-O-alkyl-(lyso)-sn-glyceryl-3-phosphoethanolamine. Separation of AGEPE into fractions individually enriched in the 16:0, 18:0, and 18:1 alkylether substituents was accomplished by silica gel G combined with silver nitrate-impregnated silica gel H thin-layer chromatography. The AGEPE or its individual molecular species can be converted in high yields to the corresponding 1-O-alkyl-2-acetyl-sn-glyceryl-3-phosphocholine (AGEPC) analogues by reaction with methyl iodide in the presence of a crown ether. Characterization of the derivatives was achieved through thin-layer chromatography, infrared spectroscopy, gas-liquid chromatography, and combined gas-liquid chromatography-mass spectrometry. The ability of these analogues to induce irreversible aggregation and secretion of serotonin from washed rabbit platelets was evaluated.

Acetylation↗
PubMed1983

Thromboxane B2 (TxB2) release following acetyl glyceryl ether phosphorylcholine (AGEPC) infusion in the rabbit.

Intravenous infusion of acetyl glyceryl ether phosphorylcholine (AGEPC) into rabbits resulted in an AGEPC dose-dependent elevation of plasma thromboxane B2 (TxB2) levels within 30 seconds. In contrast to AGEPC, the deacetylated derivative, lyso-GEPC (36.8 micrograms), did not increase intravascular TxB2 levels when similarly infused into rabbits. Intravascular TxB2 levels were maximal at 60 seconds after the infusion of 0.61 micrograms AGEPC whereas at higher doses of AGEPC (1.21-2.4 micrograms), plasma TxB2 levels were maximally elevated within 30 seconds after initiation of AGEPC infusion. These acute increases in the intravascular levels of TxB2 were accompanied by the development of thrombocytopenia, neutropenia, and basopenia which occurred concomitantly with AGEPC dose-dependent elevations in the plasma levels of platelet factor 4. Elevations in the plasma TxB2 levels returned to pre-infusion levels within 10-20 minutes after the initiation of AGEPC infusion. Thus, in vivo, one potent phospholipid, AGEPC, stimulates the production of another class of potent lipid mediators, the thromboxanes.

Animals↗
PubMed1983

Complement localization in ischemic baboon myocardium.

Complement localization was examined by direct immunoperoxidase procedures on frozen sections of baboon myocardium obtained 24 hours after ligation of the left anterior descending coronary artery. There was extensive localization of C3, C4, and C5 in most infarcted myocardial fibers; however, in these infarcted areas of myocardium, complement components were not found in myocytes immediately adjacent to either the endocardium or epicardium. Although C3, C4, and C5 were all present within the same myocardial fibers as assessed in adjacent serial sections, the light microscopic distribution of these components was dissimilar, i.e., C3 and C5 were present in both a granular and a diffuse pattern within myocytes, whereas C4 was always localized in a diffuse pattern. Complement components C3 and C5, but not C4, were also localized in the walls of small muscular arteries in infarcted myocardium. No complement was observed in myocardial fibers or blood vessels in normal baboon myocardium. Electron microscopic evaluation of C3 localization within infarcted myocardium indicated that C3 was associated with contractile elements of myocytes, as well as with membranes of myocyte nuclei, mitochondria, and sarcoplasmic reticulum. Within vascular smooth muscular cells, C3 was associated with myofilaments and mitochondrial membranes. Thus, the results of this study provide new information regarding the cellular and subcellular distribution of complement components in infarcted baboon myocardium. If this localization of C3, C4, and C5 is a result of their in situ activation within the ischemic myocardium, a variety of complement-derived phlogistic products would be expected to have been produced and to have effected, in part, the subsequent inflammatory response.

Animals↗
PubMed1982

Vasoactive properties of acetyl glyceryl ether phosphorylcholine and analogues.

Intradermal injection of 0.1 pmole (52 pg.) of acetyl glyceryl ether phosphorylcholine (AGEPC) in guinea pigs induced increased vascular permeability as assessed by a skin-blueing model. Rabbits and rats showed increased vascular permeability with 1.0 pmole (520 pg.) of AGEPC. On a molar basis, from 1,000 to 10,000 times more histamine was required to induce skin blueing in the same animals. In rabbits and guinea pigs, the increased vascular permeability induced by AGEPC occurred as an early, transient phase and as a delayed, prolonged phase. The early, transient phase was not inhibited by chlorpheniramine doses which markedly reduced or abrogated histamine-induced blueing. AGEPC was 100 to 1000 times more potent than histamine on a molar basis for the induction of vasoconstriction in a blued skin assay in guinea pigs. These studies demonstrate that AGEPC has potent vasoactive properties and provide additional information that implicates this acetylated alkyl phosphoglyceride as a mediator of the acute inflammatory process.

Animals↗
PubMed1982

Isolation of human platelet membrane microparticles from plasma and serum.

Methods have been developed to isolate human platelet membrane fragments from plasma and serum. Rabbit antibody produced against the human platelet membrane glycoprotein complex, IIb/IIIa, was utilized in an immunoelectrophoretic assay to evaluate the amount of this antigen in various microparticle preparations. The serum concentration of platelet microparticles was more than tenfold greater than that observed for plasma (65 micrograms/ml versus 4.4 micrograms/ml, respectively). Ultrastructural evaluation of either plasma or serum-derived microparticles disclosed a variety of membrane fragments and membrane-bound vesicles with occasional fragments of red blood cells, white blood cells, and platelets. In contrast, microparticle preparations derived from isolated washed platelets after thrombin stimulation contained a heterogeneous array of membrane fragments, vesicles, and granules but no identifiable red cell, white cell, or platelet fragments. Thus, these studies demonstrate that normal human plasma and serum contain platelet membrane fragments that are produced during cell activation. If a similar loss of platelet membranes occurs in vivo following reversible platelet activation, it is possible that the resulting membrane modifications may be of importance in both the structural and functional changes that develop during platelet senescence.

Antibody Specificity↗
PubMed1981

Modification of the polar head group of acetyl glyceryl ether phosphorylcholine and subsequent effects upon platelet activation.

A series of phosphoglycerides containing the same structural unit, 1-O-alkyl-2-acetyl-sn-glycerol, was prepared with the following polar head groups: -3-phosphoric acid (AGEPA), -3-phosphorylethanol (AGEPEt), -3-phosphorylethanolamine (AGEPE), -3-phosphoryl-N-monomethylethanolamine (AGEPMME), -3-phosphoryl-N,N-dimethylethanolamine (AGEPDME), and -3-phosphorylcholine (AGEPC). These compounds were synthesized primarily by phospholipase D modification of AGEPC. The characterization of these derivatives was achieved through thin layer chromatography, infrared spectrometry, gas-liquid chromatography, and combined gas-liquid chromatograph-mass spectrometry. Their ability to induce irreversible aggregation and 50% secretion of serotonin from washed rabbit platelets was investigated. The latter secretory activity, expressed on a molar basis (60 s following platelet stimulation) was shown to be particularly sensitive to the nature of the polar head group as follows: AGEPA, 8.3 X 10(-7) M; AGEPE, 6.8 X 10(-7) M; AGEPEt, 4.0 x 10(-7) M; AGEPMME, 3.7 X 10(-9) M; AGEPDME, 4.5 X 10(-10) M; and AGEPC, 1.8 X 10(-10) M. It was concluded that the polar head group of the various acetyl glyceryl ether phosphoglycerides had an important role in expressing the biological activity of this unique type of phosphoglyceride towards platelets. It appears likely that the polar head group of the various acetyl glyceryl ether phosphoglycerides had an important role in expressing the biological activity of this unique type of phosphoglyceride towards platelets. It appears likely that the recognition site (receptor site) on platelets is more reactive to the dimethylethanolamine or teh quaternary ammonium base, choline, than to any of the other polar head groups under consideration.

Animals↗
PubMed1981

Differential effects of platelet depletion on the physiologic alterations of IgE anaphylaxis and acetyl glyceryl ether phosphorylcholine infusion in the rabbit.

Intravenously administered acetyl glyceryl ether phosphorylcholine (AGEPC) induced all of the respiratory and circulatory alterations observed during IgE anaphylaxis in the rabbit. Prior platelet depletion, however, had differential effects on these two physiologic responses. The AGEPC-induced increase in total pulmonary resistance and decrease in dynamic compliance were abrogated by prior platelet depletion, whereas these lung mechanical changes occurring as part of an IgE anaphylactic response after intravenous antigen challenge were unaffected by platelet depletion. The apneic episode observed in both the AGEPC and antigen-induced response was unaffected by platelet depletion, and the brief period of rapid shallow breathing of the AGEPC response was diminished to that characteristically seen in the anaphylactic response of both platelet-intact and platelet-depleted rabbits. Prior platelet depletion had little effect on right ventricular hypertension, bradycardia, and systemic hypotension of either the AGEPC or the anaphylactic responses. Thus, AGEPC induced lung mechanical alterations via platelet-dependent mechanisms and ventilatory and circulatory alterations by mechanisms largely independent of circulating platelets. These findings were consistent with the possibility that AGEPC released into the blood stream during IgE anaphylaxis may mediate the circulatory and ventilatory alterations but not the lung mechanical alterations of the anaphylactic response.

Anaphylaxis↗
PubMed1981

Human platelet stimulation by acetyl glyceryl ether phosphorylcholine.

Acetyl glyceryl ether phosphorylcholine (AGEPC) induced dose-dependent platelet aggregation and release of [3H]serotonin and platelet factor 4 in citrated human platelet-rich plasma. ADP scavengers or indomethacin prevented irreversible platelet aggregation responses induced by 0.2 microM AGEPC but had no effect upon platelet secretion; prostacyclin inhibited AGEPC-induced aggregation and secretion. EDTA or EGTA inhibited AGEPC-induced aggregation but had no effect on platelet secretion.

Blood Coagulation Factors↗
PubMed1981

Activation of human neutrophils with 1-O-hexadecyl/octadecyl-2-acetyl-sn-glycerol-3-phosphorylcholine (platelet activating factor).

1-O-Hexadecyl/octadecyl-2-acetyl-sn-glyceryl-3-phosphorylcholine (AGEPC), the acetylated alkyl phosphoglyceride known as platelet-activating factor, stimulated human neutrophil (PMN) exocytosis, migration, superoxide production and aggregation over a concentration range of 10(-10) to 10(-5) M. AGEPC-induced PMN exocytosis of azurophilic (myeloperoxidase and beta-glucuronidase) and specific (lactoferrin and lysozyme) lysosomal granules was rapid (T 1/2 = 20 sec), dependent on the presence of cytochalasin B, but was not associated with release of cytoplasmic LDH. As seen with the complement-derived peptide stimulus, C5a, AGEPC-initiated PMN enzyme release was dependent on temperature and cellular glycolysis but not on the presence of extracellular Ca++. When analyzed by gradient analysis, PMN migration caused by AGEPC was primarily chemotactic in nature. An unusual feature for both enzyme secretion and migration was a decrease in response between 10(-6) M and 10(-5) M AGEPC. This decreased responsiveness could be explained by rapid PMN desensitization occurring at high AGEPC concentrations, limiting the overall cellular response. Rapid desensitization for exocytosis was demonstrated in PMN stimulated with AGEPC in the absence of cytochalasin B. When cytochalasin B was added subsequently and PMN challenged with AGEPC or C5a, stimulus-specific desensitization to AGEPC but not C5a-induced lysosomal enzyme release occurred. PMN desensitized to C5a responded normally to a subsequent AGEPC challenge. Stimulation of all the PMN functions examined was markedly attenuated with removal of the 2-acetyl group from AGEPC (lyso GEPC). These results suggest that AGEPC stimulates a wide variety of human PMN responses by a receptor-like mechanism, dependent on the short chain fatty acid ester in the 2-position of the alkyl phosphoglyceride.

Animals↗
PubMed1981

Mouse c1q: light and electron microscopic immunohistochemical localization.

The localization of mouse C1q in various mouse tissues was assessed by immunohistochemistry. In all mouse strains examined, C1q was associated with cells in small focal areas of lymph nodes, Peyer's patches, and spleens often near germinal centers. Ultrastructural studies of these foci demonstrated that C1q was present in the endoplasmic reticulum of mononucleated cells that had an extensive cytoplasmic network. Occasionally, small deposits of C1q were observed in the perinuclear space of mononucleated cells that had scanty cytoplasm with few organelles. Extracellular C1q was found at the surface of interdigitating cell processes in these lymphatic foci. In addition to the presence of C1q within lympho-reticular tissues, C1q frequently was detected in renal glomeruli but not in urinary tract epithelium or intestinal epithelium of adult mice; in contrast, C1q was localized in the intestinal epithelium of neonatal mice. C1q was not detected in tissue fibroblasts nor in cultured fibroblasts; however, a subpopulation of mouse adherent peritoneal cells maintained in vitro did contain C1q. The results of these studies provide new information regarding the distribution and, thus, the possible sites of C1q biosynthesis in normal mouse tissues.

Animals↗
PubMed1981

Acetyl glyceryl ether phosphorylcholine. Intravascular alterations following intravenous infusion into the baboon.

The intravenous infusion of 1-O-hexadecyl/octadecyl-2-acetyl-sn-glyceryl-3-phosphorylcholine (AGEPC) in baboons (28 micrograms per kg.) induced acute, but reversible, thrombocytopenia and neutropenia and the intravascular release of platelet factor 4 and thromboxane B2. Maximal depression of circulating platelets and neutrophils occurred within 30 seconds after AGEPC infusion and was accompanied by significant elevations in plasma platelet factor 4 and thromboxane B2 levels (p less than 0.02). Hematocrit values increased after AGEPC infusion, but this increase was delayed relative to the other intravascular alterations, i.e., maximal hematocrit values occurred at 10 to 20 minutes after AGEPC infusion.. The thrombocytopenia induced by AGEPC was reversed within 2 to 3 minutes; in contrast, circulating neutrophils did not return to preinfusion levels until 30 minutes after AGEPC infusion. Plasma platelet factor 4 and thromboxane B2 elevations gradually decreased and returned to preinfusion levels within 30 to 60 minutes. The deacetylated derivative of AGEPC, lyso-glyceryl ether phosphorylcholine, had no effect when similarly infused into baboons. These studies demonstrate that the intravenous administration of AGEPC into baboons initiated significant but reversible intravascular alterations; thus, this unusual acetylated alkyl phosphoglyceride may be an important mediator of inflammation in primates, including man.

Animals↗
PubMed1980

Isolation and characterization of mouse C1q.

C1q was isolated from mouse serum and ascites fluid by absorption onto human IgG-coated latex beads followed by separation on 3-10% exponential gradient sodium dodecyl sulfate (SDS) polyacrylamide gels. Mouse C1q was also purified by low ionic strength precipitation of mouse serum. The purified C1q was heat-labile (56 degrees C, 30 min) both structurally and functionally, contained 4.3% hydroxyproline, 1.38% hydroxylysine, and 18.5% glycine, had an apparent molecular weight of 380,000 daltons, and reconstituted the hemolytic complement activity of C1q-depleted mouse serum. The negatively stained ultrastructural appearance of this purified material consisted of 6 globular units connected by strands. These data demonstrate that mouse C1q structurally and functionally is similar to human and rabbit C1q. A portion of polyacrylamide gel containing mouse C1q was injected into rabbits resulting in the production of monospecific antisera against mouse C1q. Thus, this procedure is a new, rapid and simple method to obtain monospecific antisera against mouse C1q.

Adsorption↗
PubMed1980

Respiratory and circulatory alterations induced by acetyl glyceryl ether phosphorylcholine, a mediator of IgE anaphylaxis in the rabbit.

We examined the physiologic alterations induced in the rabbit by an acetyl glyceryl ether phosphorylcholine (AGEPC), a synthetic compound identical to rabbit basophil-derived platelet-activating factor. Intravenous administration of AGEPC led to a brief period of rapid, shallow breathing, a transient apnea, a decrease in dynamic compliance, and an increase in total pulmonary resistance. Circulatory alterations included brief bradycardia, increase in right ventricular pressure, and systemic hypotension. At a dose of 0.6 microgram/kg, AGEPC produced physiologic alterations both qualitatively and quantitatively indistinguishable from those which characterize IgE anaphylaxis in the rabbit. Mortality was also similar. After recovery to prechallenge baseline values, subsequent doses of AGEPC resulted in attenuated responses with respect to many of the physiologic alterations. Thrombocytopenia, leukopenia, and platelet secretion (as assessed by release of platelet factor 4 into the plasma) were also part of the response to AGEPC, further extending the similarity of this response to IgE anaphylaxis. Because AGEPC has been shown previously to be released into the circulation during IgE anaphylaxis, our results provide strong evidence that AGEPC is an important mediator of this acute allergic reaction in the rabbit.

Anaphylaxis↗
PubMed1980

Complement localization and mediation of ischemic injury in baboon myocardium.

We sought to determine whether the third component of complement (C3) is localized in ischemic baboon myocardium after coronary artery ligation. Furthermore, we assessed the effects of prior C3 depletion on myocardial necrosis. We studied seven control baboons (group I) and seven C3-depleted (group II) baboons that were killed 24 h after ligation of the anterior descending coronary artery. Multiple tissue samples were obtained from infarct, intermediate, and normal myocardial sites as defined by serial unipolar epicardial ECG mapping. In group I baboons, myocardial creatine kinase content from infarct sites was reduced as compared with normal sites (12.6+/-0.92 [SE] vs. 24.4+/-0.75 IU/mg protein, P < 0.001). The intermediate sites from group I contained more creatine kinase (19.0+/-1.25 IU/mg protein) than infarct sites (P < 0.001), but less (P < 0.025) than normal sites. In group II, intermediate sites showed no significant reduction in creatine kinase from normal sites and there was significantly less creatine kinase depletion in infarct sites when compared with group I animals (33.7+/-4.6 and 51.4+/-1.8% depletion, respectively, P < 0.001). In all seven group I baboons, uniform C3 localization was observed in infarct sites by direct immunofluorescence but appeared in mosaic patterns in intermediate sites. C3 was not demonstrated in any normal sites, nor in any site from group II baboons. Additional studies on baboons killed at earlier times after ligation indicated that C3 was localized focally on swollen myocytes in infarct sites as early as 4 h after coronary ligation. These results strongly implicate the active participation of the complement system of inflammatory proteins in the pathogenesis of myocardial tissue injury following coronary occlusion.

Animals↗