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D H Albert

Publications and source records attributed to D H Albert.

31 records · Page 2Linked to original sources
PubMed1992

Synthesis and structure-activity relationships of a series of novel benzopyran-containing platelet activating factor antagonists.

A class of N-substituted tetrahydrobenzopyrano[3,4-c]pyridines, I, have been identified as antagonists of platelet activating factor (PAF). The structural features essential for PAF binding were determined by systematic modification of three sites in the molecule. While O-alkyl analogues had little effect on binding potency, N-alkyl analogues exhibited a wide range of activity. Structural changes in the core ring system generally resulted in a loss of binding activity. Optimization of the N- and O-substituents resulted in the analogues 25-27 which exhibited Ki values ranging between 131 and 167 nM in a [3H]PAF binding assay. Compound 23 was also active in a model of PAF-induced shock in the mouse following intravenous administration.

Alkylation↗
PubMed1991

5-lipoxygenase inhibitory activity of zileuton.

Zileuton [N-(1-benzo[b]thien-2-ylethyl)-N-hydroxyure] inhibited 5-hydroxyeicosatetraenoic acid synthesis by rat basophilic leukemia cell 20,000 x g supernatant and rat polymorphonuclear leukocytes (PMNL) (IC50 = 0.5 and 0.3 microM) respectively. It also inhibited leukotriene (LT)B4 biosynthesis by rat PMNL (IC50 = 0.4 microM), human PMNL (IC50 = 0.4 microM) and human whole blood (IC50 = 0.9 microM). Inhibition of human PMNL LTB4 biosynthesis was removed readily by a simple wash procedure. At concentrations up to 100 microM, the compound produced little or no inhibition of several related enzymes, such as platelet 12-lipoxygenase, soybean and rabbit reticulocyte 15-lipoxygenase and sheep seminal vesicle cyclooxygenase. At p.o. doses from 0.5 to 5 mg/kg in the dog, zileuton produced a rapid and sustained inhibition of ex vivo blood LTB4 biosynthesis which correlated with the pharmacokinetic behavior of the compound. In a similar ex vivo study in the rat, the compound displayed an p.o. ED50 of 2 mg/kg. Zileuton was highly effective in preventing 6-sulfidopeptide LT formation in the rat peritoneal cavity triggered by an antigen-antibody reaction with an ED50 of 3 mg/kg. In experimental models of inflammation, zileuton significantly reduced arachidonic-acid induced mouse ear edema (ED50 = 31 mg/kg) and also attenuated inflammatory cell accumulation in the rat pleural Arthus reaction. The effectiveness of this compound for preventing LT formation in vitro, ex vivo and in vivo suggests its utility for preventing the pathophysiological effects of the LTs and other 5-lipoxygenase products in animals and in humans.

Administration, Oral↗
PubMed1984

Release of arachidonic acid from 1-alkyl-2-acyl-sn-glycero-3-phosphocholine, a precursor of platelet-activating factor, in rat alveolar macrophages.

Platelet activating factor and the bioactive metabolites of arachidonic acid are secreted by alveolar macrophages in response to stimulation by phagocytic agents or calcium ionophore. We have previously shown a deacylation-acetylation sequence in the formation of 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine (PAF) from alkylacyl-(long chain)-GPC (Albert, D.H. and Snyder, F. (1983) J. Biol. Chem. 258, 97-102). This sequence may be an important source of 20:4 during inflammatory reactions since, in alveolar macrophages, the ether lipid precursor of PAF represents 35% of the choline glycerophospholipids and has a much higher content (35%) of 20:4 in the sn-2 position than does diacyl-GPC (17%). Alveolar macrophages prelabeled with 14C-labeled fatty acids (16:0, 18:1, 18:2 and 20:4) and [1-3H]alkyllyso-GPC were used to study the release of fatty acids from ether-linked and diacyl phospholipids. Each of these fatty acids was incorporated primarily into the choline glycerophospholipids of alveolar macrophages. The release of 20:4 from macrophage phospholipids was increased by treatment of the labeled cells with the calcium ionophore A23187 (2 microM) or zymosan (1 mg/ml), whereas the release of 16:0, 18:1 and 18:2 was not increased above control levels by either stimuli. Although more of the labeled 20:4 is released from the diacyl-GPC (50% of the total released), substantial amounts (44%) of 20:4 are derived from alkylacyl-GPC after incubating the stimulated cells for 60 min. The loss of 20:4 continued from the diacyl species throughout the incubation period studied, whereas a slower net release of 20:4 lost from the alkylacyl-GPC fraction was evident after 2 h. We conclude that the deacylation-reacylation cycle is an important aspect of the metabolism of 20:4 and alkylacyl-GPC during inflammatory stimulation of alveolar macrophages and that the deacylation of this ether-linked phospholipid (which is the first step in the formation of PAF) is responsible for a significant amount of the 20:4 released.

Animals↗
PubMed1983

Biosynthesis of 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine (platelet-activating factor) from 1-alkyl-2-acyl-sn-glycero-3-phosphocholine by rat alveolar macrophages. Phospholipase A2 and acetyltransferase activities during phagocytosis and ionophore stimulation.

1-Alkyl-2-acyl-sn-glycero-3-phosphocholine (alkyl-acyl-GPC) comprises 11% of the total phospholipids of rat alveolar macrophages. This endogenous pool of alkylacyl-GPC was prelabeled by incubating the macrophages with [1,2-3H]alkyllyso-GPC (54 Ci/mmol), which enters the cells and is acylated. The effect of various stimuli on the synthesis and release into the media of labeled alkylacetyl-GPC (platelet-activating factor) from the cells was used to establish the role of inactive alkylacyl-GPC as a precursor of the biologically active derivative. A phagocytic agent (zymosan, 100 micrograms/ml) and an ionophore (A23187, 2 microM) stimulated the release of both alkylacetyl-GPC and alkyllyso-GPC into the media at the expense of cellular alkylacyl-GPC. Phospholipase A2 activity (at pH 4.5 and in 1 mM EDTA) was also increased in the media. The stimulatory effect of zymosan and the ionophore on alkylacetyl-GPC release was prevented by mepacrine (0.1 mM), an agent that inhibits the release of fatty acids from phospholipids. These data indicate that phospholipase activity is required for the biosynthesis of alkylacetyl-GPC. However, since the inhibitory effect of mepacrine was not apparent when acetate was present, it appears that the acetylation step is rate limiting. Exposure of alveolar macrophages in culture to zymosan or A23187 stimulated acetyltransferase activity 250-300%. In contrast, phorbol myristate acetate (1.6 microM), which stimulated the accumulation of lysophospholipids but not the level of alkylacetyl-GPC in the media, did not substantially increase acetyltransferase activity. We conclude that alkylacyl-GPC serves as a precursor of alkylacetyl-GPC and that the production of this potent mediator by rat alveolar macrophages can be stimulated by agents that affect phospholipase A2 and acetyltransferase activities. The latter enzyme appears to have a regulatory function in the biosynthesis of alkylacetyl-GPC.

Acetyltransferases↗
PubMed1982

Cholesterol metabolism in human monocyte-derived macrophages: stimulation of cholesteryl ester formation and cholesterol excretion by serum lipoproteins.

The role of lipoproteins and serum in the formation and accumulation of cholesteryl esters in human monocyte-derived macrophages (HMD macrophages) was investigated; studies were also carried out with IC21 cells (a cell line derived from mouse peritoneal macrophages). Following preincubation of HMD macrophages with lipoprotein-depleted serum (LPDS), both native and acetylated low density lipoprotein (LDL and AcLDL, respectively) stimulated the formation of cholesteryl esters with a resultant increase in cellular cholesteryl ester content. Cholesteryl ester formation and accumulation was also stimulated in macrophages exposed continuously to 25-hydroxycholesterol. However, the stimulation of cholesterol esterification by either lipoproteins or 25-hydroxycholesterol was not inhibited by progesterone in HMD macrophages, but was in the IC21 cells. Cholesterol efflux and the hydrolysis of cellular cholesterol ester, promoted by serum components, were studied in HMD macrophages preloaded with cholesteryl ester by incubation with 25-hydroxy cholesterol. Replacement of the medium with one devoid of 25-hydroxycholesterol resulted within 24 hr in at least a 30% decrease in the cholesteryl ester content of the HMD macrophages; replacement with a medium high in cholesterol acceptor content (LPDS or high density lipoprotein) and incubation for three days led to the most marked decreases in cellular cholesterol content. Thus, hydrolysis of the cholesteryl esters by HMD macrophages was not dependent on the presence of cholesterol acceptors in the medium, but cellular cholesterol content was.

Cholesterol↗
PubMed1982

The conversion of progesterone into 5 alpha-pregnane-3,20-dione, 3 beta-hydroxy-5 alpha-pregnan-20-one, and its fatty acid esters by preparations of bovine corpora lutea.

Homogenates obtained from bovine corpora luteal tissue were found to catalyze the synthesis of 3 beta-hydroxy-5 alpha-pregnan-20-one (allopregnanolone) from progesterone but not from pregnenolone. The major metabolites of progesterone included allopregnanolone, 5 alpha-pregnane-3,20-dione, and fatty acid esters of allopregnanolone. Incubation with labeled pregnenolone resulted in the formation of pregnenolone esters; however, neither allopregnanolone nor esterified derivatives of it were detected. The esterifying enzyme(s) leading to the formation of allopregnanolone esters was associated primarily with the microsome-enriched subcellular fraction and was stimulated by the addition of ATP and coenzyme A. With these added cofactors, the pH optimum was 6.0-6.5. The rate of steroid ester formation was enhanced by the addition to the homogenate fraction of oleic acid, which was incorporated into the steroid ester fraction. Thus, enzymatic activity, with characteristics similar to either cholesteryl ester hydrolase (EC 3.1.1.13) or acyl cholesterol acyltransferase (EC 2.3.1.26), catalyzed the esterification of allopregnanolone. The data suggest that the allopregnanolone esters found in vivo are derived from progesterone rather than from pregnenolone.

5-alpha-Dihydroprogesterone↗
PubMed1980

Identification of fatty acid esters of pregnenolone and allopregnanolone from bovine corpora lutea.

The lipoidal derivatives of steroids present in bovine corpora lutea have been identified and quantified. Nonpolar steroid derivatives were extracted from the tissue with a mixture of chloroform and methanol. The extract was subjected to column and high performance liquid chromatography in order to remove phospholipids, cholesterol, triglycerides, cholesteryl esters, and unesterified steroids. A portion of the purified nonpolar material was subjected to methanolysis. By gas-liquid chromatographic analysis, the products were shown to consist of the steroids 3 beta-hydroxy-5 alpha-pregnan-20-one (80%) and pregnenolone (20%), and the methyl esters of palmitic (34%), stearic (27%), oleic (22%), linoleic (7%), arachidonic (7%), palmitoleic (3%), and eicosatrienoic (1%) acids. Mass spectrometric analysis of the intact lipoidal derivatives and of their methoximes confirmed the presence of seven esters of allopregnanolone and pregnenolone: allopregnanolone palmitate, pregnenolone palmitate, allopregnanolone stearate, allopregnanolone oleate, allopregnanolone linoleate, allopregnanolone eicosatrienoate, and pregnenolone arachidonate. The mass spectral data suggested that four other esters were also present: pregnenolone stearate, pregnenolone oleate, pregnenolone linoleate, and allopregnanolone arachidonate.

Animals↗
PubMed1980

Lipid composition and gonadotropin-mediated lipid metabolism of the M5480 murine Leydig cell tumor.

The effects of human choriogonadotropin (HCG) stimulation on lipid composition in the murine Leydig cell tumor M5480 grown subcutaneously were determined. The main lipids of the Leydig cell tumor were found to be largely triacylglycerols and phospholipids. Daily in vivo administration of human choriogonadotropin to tumor-bearing mice for 3 days increased the phospholipid content and altered the phospholipid composition of the tumors. There was no demonstrable change in the levels of triacylglycerols, cholesterol, and cholesteryl esters. HCG had no major effect on the fatty acid patterns of the major lipid fractions with the exception of cholesteryl esters, which had a decreased amount of arachidonic acid following HCG-treatment. Results of in vitro incubations of tumor cells prelabeled with [1-14C]arachidonate showed that the label was lost more rapidly from cholesteryl esters of HCG-treated cells than from control cells during (the 12-hour) incubation. Moreover, less [1-14C]acetate was incorporated into the cholesteryl ester fraction of hormone-treated cells than in control cells. HCG stimulated the activity of cholesteryl ester hydrolase in dispersed cells within 3 hours. These results demonstrate that an acute effect of HCG on tumor Leydig cell metabolism is to increase the metabolism of cholesteryl esters, probably by stimulating cholesteryl ester hydrolase activity. The long term effect is an accumulation of phospholipids which may be utilized for membrane synthesis.

Animals↗
PubMed1979

Desaturation of eicosa-11,14-dienoic acid in human testes.

The metabolism of [1-14C]eicosa-11,14-dienoic acid was investigated in human testes using whole tissue minces and microsomal preparations. Both types of preparations catalyzed the desaturation of the labeled diene to eicosa-8,11,14-trienoic as well as eicosa-5,11,14-trienoic acid. The reported results, therefore, indicate that human testicular tissue, as well as rat testicular tissue (reported previously), is capable of utilizing eicosa-11,14-dienoic acid as a precursor of arachidonic acid. Since it is known that there is no delta 8 desaturase activity in rat liver and brain, these studies support the concept that there is a tissue variation in this enzymatic pathway.

Carbon Radioisotopes↗
PubMed1979

Accumulation, nature, and possible functions of the malachite green affinity material in ejaculated human spermatozoa.

An electron microscopic study was conducted on human sperm over an incubation period of 5 to 6 hours in Tyrode's solution at room temperature. Examination of aliquots of the cells, fixed at timed intervals, with glutaraldehyde, malachite green, and postosmication revealed that malachite green affinity material (MGA-M) was barely discernible at first but did accumulate considerably upon standing. Biochemical analysis of MGA-M, which is extractable by glutaraldehyde, revealed that MGA-M is a mixture of extractable phospholipids and of their lyso-derivatives. Some of these substances have fusogenic properties; i.e., they are able to fuse together the membranes of two different cells. The appearance and accumulation of these fusogens occurred during the incubation period of 5 to 6 hours, which was previously shown to be required to capacitate human sperm in vitro. It is probable, therefore, that human sperm, during their initial period of incubation either in vivo or in vitro, not only become capacitated and undergo the acrosome reaction but also develop the fusogenic substance(s) which are necessary for the imminent fusion of their plasma membrane to the vitelline membrane of the mature oocyte.

Cytoplasm↗
PubMed1977

Metabolism of eicosa-11,14-dienoic acid in rat testes. Evidence for delta8-desaturase activity.

The metabolism of [14C]eicosa-11,14-dienoic acid was investigated in rat testes in vivo and in vitro. Intratesticular injection of [1-14C]eicosa-11,14-dienoic acid resulted in the appearance of radioactivity (4-30% of 14C in total fatty acids) in 20-carbon trienoic fatty acids and a small amount (2-3.5%) in arachidonic acid. Analysis of the 20-carbon trienoic acid fraction by ozonolysis indicated that 15 to 34% of the 14C in this fraction was in an 8-carbon fragment originating from eicosa-8,11,14-trienoic acid. The rest (66 to 84%) was in a 5-carbon fragment, presumably originating from eicosa-5,11,14-trienoic acid. Incubation of testicular tissue minces or microsomes with [1-14C]eicosa-11,14-dienoic acid yielded labeled eicosa-8,11,14- and eicosa-5,11,14-trienoic acids in proportions similar to those obtained in vivo. Added unlabeled acetate had no effect on the formation of [14C]eicose-8,11,14-trienoic acid in vitro. Therefore, it is unlikely that the labeled eicosa-8,11,14-trienoic acid arose from elongation of octadeca-6,9,12-trienoic acid with labeled acetate derived from bio-oxidation of the labeled substrate. These results are compatible with a limited desaturation of eicosa-11,14-dienoic acid to eicosa-8,11,14-trienoic acid and provide evidence for delta8 desaturate activity in rat testis.

Acetates↗
PubMed1977

Ether-linked glycerolipids in human brain tumors.

In this investigation, the lipid composition of a number of human brain tumors was determined and compared to that of normal adult brain. Glioblastomas (11 samples), astrocytomas (4 samples), an acoustic neurinoma, an oligodendroglioma, and a meningioma were analyzed. All of the tumors had substantial levels (0.8-3.4% of total phospholipids) of choline plasmalogen which was present in only trace amounts in normal brain. With the exceptions of the acoustic neurinoma and the meningioma, the concentration of alkylacylglycerophosphorylcholine was also higher in the tumors than in normal brain. Neutral lipids of brain tumors also contained high concentrations of both alkyl (1.6-4.8% of total neutral gsults from this investigation indicate that increases in ether-linked glycerolipids may be characteristic of human brain tumors.

Astrocytoma↗