Search PubMed⌕ Search

Biomedical subjects

G Arthur

Publications and source records attributed to G Arthur.

At least 19 recordsLinked to original sources
PubMed1996

1-O-octadecyl-2-O-methyl-glycerophosphocholine inhibits the transduction of growth signals via the MAPK cascade in cultured MCF-7 cells.

1-O-Octadecyl-2-O-methyl-glycerophosphocholine (ET18-OCH3) is an ether lipid with selective antiproliferative properties whose mechanism of action is still unresolved. We hypothesized that since ET18-OCH3 affects a wide variety of cells, its mechanism of action was likely to involve the inhibition of a common widely used pathway for transducing growth signals such as the mitogen-activated protein kinase (MAPK) cascade. To test this, we established conditions whereby quiescent MCF-7 cells took up ET18-OCH3 in sufficient quantities that inhibited cell proliferation subsequent to the addition of growth medium and examined the activation of components of the MAPK cascade under these conditions. ET18-OCH3 inhibited the sustained phosphorylation of MAPK resulting in a decrease in the magnitude and duration of activation of MAPK in cells stimulated with serum or EGF. ET18-OCH3 had no effect on the binding of EGF to its receptors, their activation, or p21ras activation. However, an interference in the association of Raf-1 with membranes and a resultant decrease in Raf-1 kinase activity in membranes of ET18-OCH3-treated cells was observed. ET18-OCH3 had no direct effect on MAPK or Raf-1 kinase activity. A direct correlation between ET18-OCH3 accumulation, inhibition of cell proliferation, Raf association with the membrane, and MAPK activation was also established. These results suggest that inhibition of the MAPK cascade by ET18-OCH3 as a result of its effect on Raf-1 activation may be an important mechanism by which ET18-OCH3 inhibits cell proliferation.

Calcium-Calmodulin-Dependent Protein Kinases↗
PubMed1996

Synthesis and evaluation of the antiproliferative effects of 1-O-hexadecyl-2-O-methyl-3-O-(2'-acetamido-2'-deoxy-beta-D- glucopyranosyl)-sn-glycerol and 1-O-hexadecyl-2-O-methyl-3-0- (2'-amino-2'-deoxy-beta-D-glucopyranosyl)-sn-glycerol on epithelial cancer cell growth.

Two ether glucosyl diglyceride analogs were synthesized, and their antiproliferative activity against four epithelial cancer cell lines was evaluated. 1-O-Hexadecyl-2-O-methyl-3-O-(2'-acetamido-2'-deoxy-beta-D- glucopyranosyl)-sn-glycerol (4) was synthesized by reaction of 2-acetamido-2-deoxy-3,4,6-tri-O-acetyl-alpha-D-glucopyranosyl chloride with 1-O-hexadecyl-2-O-methyl-sn-glycerol followed by deacetylation by methanolic hydrolysis. The N-acetyl group of 4 was removed by hydrolysis with ethanolic potassium hydroxide to form 1-O-hexadecyl-2-O-methyl-3-O-(2'-amino-2'-deoxy-beta-D-glucopyranosyl)- sn-glycerol (5). Compounds 4 and 5 inhibited the proliferation of MCF-7, A549, A427, and T84 cancer cell lines. The IC(50) values for 5 ranged from 6.5 to 12.2 microM, whereas 4 was more effective against A549 cells (IC(50) 9 microM) than against MCF-7 (IC(50) 17 microM) and A427 (IC(50) 25 microM) cells and was inactive against T84 cells. Under identical incubation conditions, compounds 4 and 5 were potent inhibitors of the proliferation of OVCAR-3 cells with IC(50) values of 12 and 4 microM, respectively, whereas ET-18-OCH(3), hexadecylphosphocholine, and erucylphosphocholine had IC(50) values of 24, >30, and >30 microM, respectively. The cell-inhibitory profile of these ether-linked glucosyl diglycerides strengthens the hypothesis that such glycolipids represent a distinct group of antitumor ether lipids, having antineoplastic activities that differ from the well-known alkylphosphocholines and alkyllysophospholipids.

Antineoplastic Agents↗
PubMed1995

Evidence for receptor and G-protein regulation of a phosphatidylethanolamine-hydrolysing phospholipase A1 in guinea-pig heart microsomes: stimulation of phospholipase A1 activity by DL-isoprenaline and guanine nucleotides.

While evidence has been presented for the receptor-mediated activation of phospholipases A2, C and D, the activation of phospholipase A1 subsequent to receptor activation has not been established. Phospholipase A1-catalysed hydrolysis of 1-palmitoyl-2-linoleoyl-glycerophosphoethanolamine (GPE) by guinea-pig heart microsomes was stimulated 40-60% by isoprenaline. This isoprenaline-mediated increase in activity was blocked by propranolol and butoxamine, a specific beta 2-adrenergic antagonist, but not by atenolol, a specific beta 1-adrenergic antagonist. Neither clonidine nor phenylephrine, alpha 1- and alpha 2-adrenergic agonists respectively, had a stimulatory effect on the hydrolysis of the PE substrate. Guanosine 5'(-)[gamma-thio]triphosphate (GTP[S]) and guanosine 5'(-)[beta,gamma-imido]triphosphate, but not guanosine 5'(-)[beta-thio]diphosphate (GDP[S]) or adenosine 5'(-)[gamma-thio]triphosphate, stimulated the hydrolysis of 1-palmitoyl-2-linoleoyl-GPE by phospholipase A1. GDP[S] inhibited the isoprenaline-mediated stimulation of phospholipase A1 activity. Phospholipase A1 hydrolysis of 1-palmitoyl-2-linoleoyl-GPE was not dependent on cations; however, the stimulatory effects of isoprenaline and GTP[S] on the hydrolytic activity were abolished by cation chelators. The above data suggest that phospholipase A1 activity in guinea-pig heart microsomes is activated by the binding of isoprenaline to beta 2-adrenergic receptors. Furthermore the stimulation of phospholipase A1 activity by the agonist may be mediated via activation of G-proteins.

Adrenergic beta-Agonists↗
PubMed1995

Effect of 1-O-octadecyl-2-O-methyl-glycerophosphocholine on phosphatidylcholine and phosphatidylethanolamine synthesis in MCF-7 and A549 cells and its relationship to inhibition of cell proliferation.

The role of perturbation of lipid synthesis in the inhibition of cell proliferation by OctMeGroPCho was investigated with sensitive (MCF-7) and resilient (A549) cell lines. It inhibited de novo synthesis of phosphatidylcholine in both cells but increased triacylglycerol synthesis in A549 cells and phosphatidylethanolamine, phosphatidic acid and diacylglycerol synthesis in MCF-7 cells. The inhibition of synthesis of CDP-choline metabolites in MCF-7 cells and phosphatidylcholine biosynthetic enzyme activities in vitro by OctMeGroPCho suggests that direct inhibition of phosphocholine cytidylyltransferase may contribute to the observed inhibition of phosphatidylcholine synthesis. The activation of phosphoethanolamine cytidylyltransferase and ethanolamine phosphotransferase activities by OctMeGroPCho in vitro and increased production of CDP-ethanolamine suggest that stimulation of the above enzymes by OctMeGroPCho in the cells is responsible for the increased phosphatidylethanolamine synthesis. The apparent effect of OctMeGroPCho on intracellular lipid-metabolising enzymes is a strong indication that it may be widely distributed intracellularly and not just confined to the plasma membrane. The decrease in phosphatidylcholine synthesis by OctMeGroPCho in MCF-7 cells was prevented by co-incubation with oleic acid without any effect on the inhibition of cell growth. Although OctMeGroPCho resulted in similar decreases in phosphatidylcholine content in both cells, this did not affect the proliferation of A549 cells. The above results indicate that, although OctMeGroPCho has profound effects on lipid metabolism, these changes are not responsible for the inhibition of proliferation observed in MCF-7 cells.

Antineoplastic Agents↗
PubMed1994

Calcium-supported calpain degradation rates for cardiac myofibrils in diabetes. Sulfhydryl and hydrophobic interactions.

OBJECTIVE: The purpose was to investigate the calcium required for calpain-mediated degradation of selected cardiac myofibril proteins modified by diabetes, sulfhydryl (SH) and hydrophobic reagents. METHODS: After 20 weeks of streptozotocin-induced (55 mg.kg-1) diabetes, calcium sensitive calpain (1.5 U.ml-1) degradation rates of purified cardiac myofibrillar proteins (1 mg.ml-1) were measured, in vitro, and compared to degradation rates for N-ethylmaleimide (NEM) and 2-p-toluidinylnapthalene-6-sulfonate (TNS) treated samples. RESULTS: Diabetes (blood glucose of 550 +/- 32 mg.dl-1) reduced the yield of purified myofibrillar protein with minimal change in fibril protein composition. Total SH group reactivities (nmol.mg-1.30min) were 220 +/- 21, 163 +/- 17 and 156 +/- 24 for control, diabetic and NEM-treated (0.5 mM) myofibrils (p < or = 0.05). Calpain degradation rates were faster for all diabetic and SH modified myofibrillar proteins (p < or = 0.05), with a 45 and 35% reduction in the pCa50 for a 37 kDa protein of diabetic and NEM-treated fibril complexes. For control myofibrils, both 100 and 200 uM TNS, reduced calpain degradation rates to a similar extent for all substrate proteins. In contrast, diabetic and NEM-treated samples showed a further reduction in calpain degradation rates with increasing TNS from 100 to 200 uM. CONCLUSION: Our results support the hypothesis that in diabetes the calcium requirements for calpain degradation rates are reduced and dependent upon sulfhydryl group status and Ca(2+)-induced hydrophobic interactions, implicating a 37 kDa myofbillar-complexed protein.

Animals↗
PubMed1994

Krypton fluoride excimer laser ablation of tooth tissues: precision tissue machining.

A variety of lasers using different wavelengths have been used to remove dental hard tissue. The infrared lasers produce their effects photothermally whereas ultraviolet excimer lasers remove tissue in a controlled and precise manner by photoablation. This study investigates the use of 248 nm laser radiation in the precision removal of both enamel and dentine using diffraction limited ultraviolet optics. The data showed that enamel and dentine were machined to a high level of precision (1-2 micron tolerances). The rate of removal was greater in dentine than enamel at a range of energy densities between 1.15 and 2.2 J/cm2. The method of removal of both tissues appears to be by the preferential ablation of the organic phases of each, exposing the anatomical details of their structure. An explanation of the possible method of ablation is proposed for these tissues.

Dental Enamel↗
PubMed1994

Wedge factors for rectangular fields.

The variation of wedge factor with field size was measured for a range of square and rectangular fields for 45 degrees and 60 degrees wedges. Measurements were performed on accelerators with both externally mounted wedges, the Varian 600C of nominal energy 6MV and internally mounted wedges, the Philips SL75/5 of nominal energy 6MV and the Philips SL25 of nominal energy 25MV. Analysis of the results confirm previous investigations reported in the literature of the variation of wedge factor with field size and in particular the significantly greater variation for internally mounted wedges. A wedge factor for a rectangular field based on the wedge factor of the open field equivalent square gives a maximum error of 1.5%. A wedge factor for a rectangular field based on the wedge factor of the square field of equal area reduces the maximum error to 0.5% for all three accelerators. Analysis of results reported in the literature show a similar reduction.

Particle Accelerators↗
PubMed1993

The ethanolamine requirement of keratinocytes for growth is not due to defective synthesis of ethanolamine phosphoacylglycerols by the decarboxylation pathway.

We have investigated whether the growth requirement of keratinocytes for ethanolamine is due to defective synthesis of ethanolamine phosphoacylglycerols (EPG) via decarboxylation of serine phosphoacylglycerols. Proliferating keratinocytes readily incorporated [3H]ethanolamine into phosphatidylethanolamine (PE) and [3H]serine into phosphatidylserine (PS) and PE. Non-proliferating keratinocytes in ethanolamine-free medium incorporated [3H]glycerol into phosphatidylcholine (PC), PS and PE in decreasing order of label incorporated. The order of decreasing incorporation of glycerol after addition of ethanolamine to the medium was PC > PE > PS. Incubation of non-proliferating keratinocytes with [3H]serine resulted in incorporation of label into PS and PE. The extent of incorporation of [3H]serine into PS in non-proliferating keratinocytes was not less than that in proliferating cells. Addition of ethanolamine to the medium of non-proliferating keratinocytes did not change the quantity of label incorporated into PS, but resulted in a decrease of label incorporated into PE. When cells were prelabelled overnight with [3H]serine and subsequently incubated in medium containing ethanolamine, the loss of label from PS was inhibited relative to that of control cells incubated in medium without ethanolamine. The activity of PS decarboxylase activity in keratinocyte mitochondria was inhibited by phosphoethanolamine and PE, but not by ethanolamine or CDP-ethanolamine. Both proliferating and non-proliferating keratinocytes incorporated [3H]serine into ether-linked ethanolamine phospholipids. Taken together, the above results suggest that (1) both proliferating and non-proliferating keratinocytes are able to synthesize PE and ether-linked ethanolamine phospholipids from serine, and therefore the ethanolamine-requirement of the cells is not due to a defective decarboxylase pathway; (2) any inability of the decarboxylase pathway to meet cellular EPG requirement is not due to decreased synthesis of serine phospholipids; (3) synthesis of PE via decarboxylation, the major route in nonproliferating keratinocytes, appears to decrease when ethanolamine is made available and the CDP-ethanolamine pathway is functioning; (4) phosphoethanolamine and increased PE produced from the CDP-ethanolamine pathway may inhibit PS decarboxylase activity in the cells and provide a means of coordinating the synthesis of PE by the two pathways to prevent excess production.

Cell Division↗
PubMed1993

Effect of delta 9-tetrahydrocannabinol and merthiolate on acyltransferase activities in guinea pig liver microsomes.

delta 9-Tetrahydrocannabinol (THC) and merthiolate have been utilized as lysophospholipid acyltransferase inhibitors in metabolic studies. However, their effects on acyltransferases other than lysophosphatidylcholine:acyl-CoA acyltransferase (LPCAT) are not known. We have therefore investigated the effectiveness of THC and merthiolate in inhibiting the acylation of lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylserine, lysophosphatidylinositol (LPI) and lysophosphatidic acid (LPA) in guinea pig liver microsomes using oleoyl-CoA and arachidonoyl-CoA as acyl donors. THC inhibited LPCAT and lysophosphatidylethanolamine:acyl-CoA acyltransferase (LPEAT) by 40-50%, but had no effect or only slightly increased the activities of the other acyltransferases when assayed with oleoyl-CoA as the acyl donor. The results obtained with arachidonoyl-CoA were similar to those with oleoyl-CoA, with the exception of a 40% inhibition of lysophosphatidylserine:acyl-CoA acyltransferase (LPSAT) at concentrations of 50 microM or higher. At similar concentrations, merthiolate was more effective than THC in inhibiting the acyltransferases examined. Selective effects on the acyltransferases were observed at low concentrations of merthiolate (20 microM or less). Thus, LPCAT was most susceptible, followed by LPI acyltransferases, LPSAT, LPEAT and lysophosphatidic acid:acyl-CoA acyltransferases (LPAAT). The presence of LPA did not affect the inhibition of LPCAT by merthiolate. Thus the resilience of LPAAT to merthiolate inhibition was not due to chelation of the compound by the acidic lysolipid. Thiol reagents including N-ethyl-maleiamide, 5,5'-dithio-bis-nitrobenzoic acid, iodoacetate, beta-mercaptoethanol and dithiothreitol had little or no effect on the acyltransferases relative to equimolar concentrations of merthiolate.(ABSTRACT TRUNCATED AT 250 WORDS)

Acyl Coenzyme A↗
PubMed1993

3-Deazaadenosine and MDL29350 differentially affect the methylation of serine-and ethanolamine-derived phosphatidylethanolamine in Hep G2 cells.

The effect of 3-deazaadenosine (DZA) and the hypolipidemic drug MDL29350 (2-[3,5-di(t-butyl-4-hydroxyphenyl)thio]hexanoic acid) on the synthesis and methylation of phosphatidylethanolamine (PE) originating from the cytidine diphosphate (CDP) ethanolamine pathway and PE originating from decarboxylation of phosphatidylserine (PS) was investigated. DZA and MDL29350 did not affect the synthesis of PE by either pathway; however, methylation of ethanolamine-derived PE was inhibited by 80% and methylation of serine-derived PE was inhibited by 36% by 20 mumol/LDZA or MDL29350. The differential inhibition of the methylation of PE synthesized via serine or ethanolamine suggests that in Hep G2 cells PE-N-methyltransferase (PENMT) may be segregated into distinct compartments that are differentially accessible to the drugs.

Caproates↗
PubMed1993

Binding of dog immunoglobulins G, A, M, and E to concanavalin A.

The binding of dog immunoglobulins G, A, M and E to concanavalin A (Con A) has been investigated. A passive cutaneous anaphylaxis test was used for measurement of dog IgE, and enzyme-linked immunosorbent assay was used for measurement of dog IgG, IgA and IgM. After the dog serum fraction was applied to a Con A-Sepharose column, sequential elution with different buffers was performed; 100% of IgE and IgM, 60% of IgG and 58% of IgA bound to the Con A-Sepharose. IgE was eluted by mannose, methylglucose, and methylmannoside. IgG was eluted by glucose, mannose, methylglucose, and methylmannoside. IgA and IgM were eluted by methylmannoside only. This provides a useful technique in the purification of dog immunoglobulins, especially dog IgE.

Animals↗
PubMed1993

Purification and identification of polyclonal IgE antibodies from ragweed-sensitized dog sera.

We have purified and characterized polyclonal dog IgE. Serum IgE was precipitated by (NH4)2SO4 and then purified by two different procedures. Ion exchange on DEAE-Sephacel, followed by HPLC using Tonen hydroxylapatite and then Protein G-Sepharose, produced a highly purified IgE fraction (No. 1) free of IgG, IgA and IgM as measured by ELISA, but recovery of IgE as measured by passive cutaneous anaphylaxis was low. Gel filtration on Sephacryl S-300, Con A-Sepharose and Protein G-Sepharose recovered 18% of initial IgE, 0.02% IgG, 0.4% IgM and 0.3% IgA. This IgE fraction (No. 2) was used to induce antibody production in rabbits. Western blot analysis was then performed for dog IgE fractions No. 1 and 2. Using the rabbit anti-dog IgE, a prominent IgE band with an apparent molecular mass of 226 kD was identified in fractions No. 1 and 2 subjected to nonreducing SDS-PAGE. This band also reacted with anti-human IgE, but not with anti-dog IgG or anti-dog IgA. Under reducing conditions the approximate molecular mass for the IgE & chain, estimated by Western blot using rabbit anti-dog IgE, was 73 kD, providing a molecular mass of 196 kD for dog IgE.

Allergens↗
PubMed1992

Evidence for the regulation of guinea-pig heart microsomal phosphatidylcholine-hydrolysing phospholipase A1 by guanosine 5'-[gamma-thio]triphosphate.

We have recently characterized lysophospholipase A2 activities in guinea-pig heart microsomes and postulated that these enzymes act sequentially with phospholipases A1 to release fatty acids selectively from phosphatidylcholine (PC) and phosphatidylethanolamine, thus providing an alternative route to the phospholipase A2 mode of release. In a further investigation of the postulated pathway, we have characterized the PC-hydrolysing phospholipase A1 in guinea-pig heart microsomes. Our results show that the enzyme may have a preference for substrates with C16:0 over C18:0 at the sn-1 position. In addition, although the enzyme cleaves the sn-1 fatty acid, the rate of hydrolysis of PC substrates with C16:0 at the sn-1 position was influenced by the nature of the fatty acid at the sn-2 position. The order of decreasing preference was C18:2 > C20:4 = C18:1 > C16:0. The hydrolyses of the molecular species were differentially affected by heating at 60 degrees C. An investigation into the effect of nucleotides on the activity of the enzyme showed that guanosine 5'-[gamma-thio]triphosphate (GTP[S]) inhibited the hydrolysis of PC by phospholipase A1 activity, whereas GTP, guanosine 5'-[beta-thio]diphosphate (GDP[S]), GDP, ATP and adenosine 5'-[gamma-thio]triphosphate (ATP[S]) did not affect the activity. The inhibitory effect of GTP[S] on phospholipase A1 activity was blocked by preincubation with GDP[S]. A differential effect of GTP[S] on the hydrolysis of different molecular species was also observed. Taken together, the results of this study suggest the presence of more than one phospholipase A1 in the microsomes with different substrate specificities, which act sequentially with lysophospholipase A2 to release linoleic or arachidonic acid selectively from PC under resting conditions. Upon stimulation and activation of the G-protein, the release of fatty acids would be inhibited.

Animals↗
PubMed1992

Perturbations of cellular acylation processes by the synthetic alkyl-lysophospholipid 1-O-octadecyl-2-O-methylglycero-3-phosphocholine do not correlate with inhibition of proliferation of MCF7 and T84 cell lines.

We have investigated the hypothesis that the antiproliferative effect of 1-O-octadecyl-2-O-methylglycero-3-phosphocholine (ET-18-OCH3) is mediated through the inhibition of cellular acylation processes that control the unsaturated fatty acid complement of phospholipids. The effect of ET-18-OCH3 on the incorporation of radiolabeled oleic, linoleic, and arachidonic acids into MCF7 and T84 phospholipids was investigated. Incubation of MCF7 cells with fatty acids and 2.75 micrograms/ml ET-18-OCH3, which inhibited the proliferation of the cells after 8 h, resulted in decreased incorporation of fatty acids into a number of phospholipids, notably phosphatidylcholine; however, increased incorporation of fatty acids into other phospholipids was also observed. After 12 h incubation with the alkyl-lysophospholipid, differences in the distribution of newly incorporated fatty acids into the phospholipid classes were observed without any effect on the total amount of fatty acid incorporated. Incubation of MCF7 cells with 5 micrograms/ml ET-18-OCH3, which caused a cessation in proliferation, had a similar effect on the incorporation of the fatty acids into the phospholipids, but the redistribution of newly incorporated fatty acids in the phospholipids was accompanied by a decrease in the amount of associated radiolabeled fatty acid. Incubation of T84 cells with the labeled fatty acids and 3.5 micrograms/ml ET-18-OCH3, which significantly decreased proliferation after 8 h, resulted in decreased incorporation of oleic acid into phosphatidylcholine and increased incorporation of oleic, linoleic, and arachidonic acids into phosphatidylethanolamine, prior to the decrease in proliferation. After 12 h incubation with alkyl-lysophospholipid, significant increases in the total amount of labeled oleic and arachidonic acids incorporated in the phospholipid fraction were observed. These results clearly indicate that the antiproliferative effect of ET-18-OCH3 in MCF7 and T84 cells is not dependent on inhibition of acylation processes and the above hypothesis may not be applicable to all alkyl-lysophospholipid-sensitive cells.

Acylation↗
PubMed1992

The differential susceptibility of A427 and A549 cell lines to the growth-inhibitory effects of ET-18-OCH3 does not correlate with the relative effects of the alkyl-lysophospholipid on the incorporation of fatty acids into cellular phospholipids.

Proliferation of A427, a lung cancer cell line, was significantly decreased 10 h after incubation with 5 micrograms/ml 1-O-octadecyl-2-O-methylglycero-3-phosphocholine (ET-18-OCH3) while the proliferation of A549, another lung cancer cell line, was unaffected until 15 h after incubation with the alkyl-lysophospholipid (ALP). The relative sensitivity of cells to the antiproliferative effect of ET-18-OCH3 has been postulated to be due to the degree of inhibition of cellular acylation processes. We therefore investigated the effect of 5 micrograms/ml ET-18-OCH3 on the incorporation of fatty acids for up to 12 h, into A427 and A549 phospholipids. Significant changes observed in the incorporation of fatty acids into A427 phospholipids by the ALP were a decreased incorporation of oleic acid into PC after 8 h, an increased incorporation of linoleic acid into PE after 12 h, decreased incorporation of arachidonate into PE after 3 h, and increased incorporation into PA after 5 h. Although the above changes affected the distribution of newly esterified fatty acids in the phospholipids, there was no effect on the total quantity of label incorporated in the phospholipid fraction between the experimental and control cells after 12 h. Incubation of A549 cells with ET-18-OCH3 resulted in decreased esterification of oleic acid into PC, SM, and LPC after 5 h; decreased incorporation of linoleic into PE after 12 h; and a decreased incorporation of arachidonate into SM after 1.5 h. After 12 h incubation with ET-18-OCH3, changes in the distribution of radiolabeled fatty acids were observed in the quantitatively minor phospholipids, SM and LPC. A 20% decrease in the quantity of oleic acid incorporated into the phospholipids was observed in cells incubated with the ALP; however, no differences were observed in the quantity of linoleic or arachidonic acid incorporated into the phospholipids. The lack of common effects of the ALP on the incorporation of fatty acids into A427 and A549 phospholipids, coupled with the absence of changes that were more severe or manifested earlier in the more sensitive A427 cell line, suggests that the effect of ET-18-OCH3 on the acylation processes depends on the cell type and the fatty acid species and is unlikely to be responsible for the relative sensitivities of the cells to the compound. Radiolabeled ET-18-OCH3 was used to examine the correlation between the amount of the compound accumulated in A427, A549, MCF7, T84, and LS174T cells and the relative susceptibilities of the cells to the ALP.(ABSTRACT TRUNCATED AT 400 WORDS)

Acylation↗
PubMed1992

Improved procedures for the determination of lipid phosphorus by malachite green.

We have developed two procedures for the measurement of lipid phosphorus based on interaction between phosphomolybdenum and malachite green. One method, the "micro" assay uses 50-200 microliters of HClO4 and has a sensitivity range of 0.01-1.5 micrograms phosphorus. The second method, the "macro" assay, has a sensitivity range of 0.03-5.0 micrograms phosphorus with 100-500 microliters HClO4. Both assays are very reproducible with day to day standard deviations of less than 6% between triplicates irrespective of the HClO4 content used. At different concentrations of HClO4, each method was successfully used to determine the phosphorus content in phosphatidylethanolamine and sphingomyelin standards that covered the proposed sensitivity ranges. The increased range, sensitivity and greater volumes of HClO4 permitted in the procedures represent significant improvements over existing methods.

Molybdenum↗
PubMed1991

2-acyl-sn-glycero-3-phosphoethanolamine lysophospholipase A2 activity in guinea-pig heart microsomes.

We have recently described a lysophospholipase A2 activity in guinea-pig heart microsomes that hydrolyses 2-acyl-sn-glycero-3-phosphocholine (2-acyl-GPC). The presence of a similar activity that hydrolyses 2-acyl-sn-glycero-3-phosphoethanolamine (2-acyl-GPE) was not known. In this study, a lysophospholipase A2 activity in guinea-pig heart microsomes that hydrolyses 2-acyl-GPE has been characterized. The enzyme did not require Ca2+ for activity and exhibited a high specificity for 2-arachidonoyl-GPE and 2-linoleoyl-GPE over 2-oleoyl-GPE and 2-palmitoyl-GPE. The specificity for these unsaturated substrates was observed in the presence and absence of detergents. Selective hydrolysis of 2-arachidonoyl-GPE over 2-palmitoyl-GPE was observed when equimolar quantities of the two substrates were incubated with the enzyme. There was no preferential hydrolysis of either 2-linoleoyl- or 2-arachidonoyl-GPE when presented individually or as a mixture. Significant differences in the characteristics of 2-acyl-GPE-hydrolysing and 2-acyl-GPC-hydrolysing activities included differences in their optimum pH, the effect of Ca2+ and their acyl specificities. Taken together, these results suggest that the two activities are catalysed by different enzymes. 2-Acyl-GPE lysophospholipase activity with a preference for 2-arachidonoyl-GPE over 2-oleoyl-GPE was observed in guinea-pig brain, liver, kidney and lung microsomes. Lysophospholipase A1 activity that catalyses the hydrolysis of 1-acyl-GPE was also present in guinea-pig heart microsomes and had different characteristics from the 2-acyl-GPE-hydrolysing activity, including a preference for saturated over unsaturated substrates. The 2-acyl-GPE lysophospholipase A2 activity appeared to be distinct from Ca(2+)-independent phospholipase A2. The characteristics of the 2-acyl-GPE lysophospholipase A2 suggest it could play a role in the selective release of arachidonic and linoleic acids for further metabolism in cells.

Animals↗