METABOLISM OF EHRLICH ASCITES TUMOR IN VITRO: SUBCELLULAR DISTRIBUTION OF PHOSPHOLIPIDS NEWLY FORMED FROM14C-LABELED PRECURSORS

1966 ◽  
Vol 44 (11) ◽  
pp. 1461-1468 ◽  
Author(s):  
V. Donisch ◽  
R. J. Rossiter

When Ehrlich ascites cells were incubated in a suitable medium containing choline-1,2-14C, ethanolamine-1,2-14C, L-serine-14C, or glycerol-1-14C, radioactivity was recovered from the lipid fraction. With choline-1,2-14C, radioactivity was incorporated into the three choline-containing phospholipids, lecithin, choline plasmalogen, and sphingomyelin. Radioactivity from ethanolamine-1,2-14C was incorporated into phosphatidyl ethanolamine, ethanolamine plasmalogen, choline plasmalogen, and lecithin. Radioactivity from L-serine-14C was incorporated into phosphatidyl serine, serine plasmalogen, and phosphatide acid, with lesser amounts into phosphatidyl ethanolamine, lecithin, ethanolamine plasmalogen, choline plasmalogen, and sphingomyelin. Radioactivity from glycerol-1-14C was incorporated into the glycerophosphatides, phosphatidic acid, lecithin, phosphatidyl ethanolamine, phosphatidyl serine, phosphatidyl inositol, and choline plasmalogen. Radioactivity from this precursor was also incorporated into sphingomyelin.In all instances, radioactivity was recovered from the phosphatides in the nuclear, mitochondrial, and microsomal fractions of the tumor. Usually, the specific radioactivity of the phosphatides in the microsomal fraction exceeded that in the other two subcellular fractions.

1964 ◽  
Vol 42 (3) ◽  
pp. 299-308 ◽  
Author(s):  
H. David ◽  
R. J. Rossiter

The following phosphatides (in approximate order of concentration) were studied in cells of the Ehrlich ascites carcinoma incubated in a medium containing inorganic P32: lecithin > sphingomyelin > phosphatidyl ethanolamine = phosphatidyl inositol = phosphatidic acid > choline plasmalogen = phosphatidyl serine > ethanolamine plasmalogen. The specific radioactivity of the diacyl-glycerophosphatide fraction exceeded that of both the plasmalogen and the sphingomyelin – glycerol ether phosphatide fraction, the specific radioactivity of the individual phosphatides being as follows: phosphatidic acid > phosphatidyl inositol > ethanolamine plasmalogen > phosphatidyl ethanolamine = choline plasmalogen = lecithin > sphingomyelin. The microsomal fraction contained more phospholipid, followed by the mitochondrial and nuclear fractions, in that order. The specific radioactivities of the phospholipids of the microsomes and nuclei were greater than that of the mitochondria, chiefly because of the high specific radioactivity of the diacylglycerophosphatide fraction. The high specific radioactivity of the diacylglycerophosphatides was largely the result of a very active incorporation of inorganic P32into phosphatidic acid, particularly in the microsomal fraction. The significance of these findings is discussed.


1967 ◽  
Vol 45 (4) ◽  
pp. 597-607 ◽  
Author(s):  
A. Naimark ◽  
D. Klass

The incorporation of palmitate-1-14C into various lipid fractions was studied in rat lung in vitro. Most of the radioactivity was found in phospholipid, although in terms of decreasing specific activity the lipids were ranked: free fatty acid (FFA), glycerides, phospholipid. In addition to the usual glycerophosphatides, rat lung contained a substance with some of the chromatographic characteristics of phosphatidyl dimethylethanolamine. In terms of decreasing specific activities the phospholipids were ranked: phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl dimethylethanolamine, phosphatidyl serine plus phosphatidyl inositol, sphingomyelin plus lysophosphatidyl choline. Inhibition of oxidative energy production by hypoxia, cyanide, or low temperature markedly depressed the esterification of palmitate-1-14C. Less marked depression was observed in the absence of exogenous glucose. In all cases the decreased incorporation was associated with an increase in the total and specific radioactivity in tissue FFA. It is concluded that energy-independent exchange reactions are probably of little importance in the incorporation of FFA into esterified lipids of lung tissue. Under conditions of metabolic inhibition the penetration of labelled FFA into the tissue FFA pool does not appear to limit esterification.


2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Zdenka Ulcova-Gallova ◽  
Alice Mockova ◽  
Miroslava Cedikova

Female patients in reproductive age with systemic lupus erythematosus and fertility complications together are observed by rheumatologists, gynecologists, and reproductive immunologists. The paper notes the presence of autoantibodies to zona pellucida, to phospholipids (phosphatidyl serine, phosphatidyl ethanolamine, phosphatidyl inositol, phosphatidyl glycerol, phosphatidic acid, annexin V, beta-2 glycoprotein I, and cardiolipin) and of isoantibodies to sperm cells. Isoantibodies to sperm cells are not significantly predominant, but autoimmunity is well expressed in IgG positivity against phosphatidyl inositol, phosphatidyl ethanolamine, phosphatidyl serine, cardiolipin, and beta-2 glycoprotein I, as well as antizona pellucida antibodies in IgG isotype. According to the levels of autoantibodies we have to choose preventive treatment to protect mother and her foetus.


1974 ◽  
Vol 32 (02/03) ◽  
pp. 382-390 ◽  
Author(s):  
P Gautheron ◽  
E Dumont ◽  
S Renaud

SummaryThe clotting activity of man and rat platelet phospholipid fractions separated by bi-dimensional TLC, resuspended in Tyrode by sonication, was studied in the recalcification (manual) and in the Stypven (recalcification plus Russell’s viper venom) clotting time (determined in a coagulometer). Phosphatidyl serine was the most active fraction to shorten the two clotting tests utilized, in both rat and man, but it was much more effective in the Stypven time. The phosphatidyl ethanolamine was the second most active fraction, in the Stypven time; this fraction was almost as active as phosphatidyl serine in both animal species. The other fractions studied (phosphatidyl inositol, phosphatidyl choline and sphingomyelin) were sligthly active or not active, depending on the experimental conditions.The clotting activity of platelet phosphatidyl serine from rat, at concentrations corresponding to platelet counts from 1 to 10 ( X105), was much smaller than this of the disrupted (sonicated) platelets from which it originated. However, the clotting activity of sonicated platelets could be completely reproduced, either at each concentration studied (Stypven time) or at a concentration corresponding to lOxlO5 platelets (recalcification time), by adding to phosphatidyl serine the other four phospholipid fractions (phosphatidyl inositol, phosphatidyl ethanolamine, phosphatidyl choline, sphingomyelin) dispersed in a homogeneous way by sonication.The feeding of a butter-rich diet to rats considerably increased the activity of each of the platelet phospholipid fractions in the two clotting tests carried out.


1972 ◽  
Vol 18 (7) ◽  
pp. 1059-1063 ◽  
Author(s):  
J. A. Brushaber ◽  
J. J. Child ◽  
R. H. Haskins

Addition of exogenous cholesterol to Pythium initially increases the growth rate, but the final dry weight yield is reduced. Cholesterol induces an overall increase in lipid synthesis after the initial period of rapid growth. The lipid content of cholesterol-grown mycelium becomes about double that of mycelium grown without cholesterol. The proportion of phosphatidyl serine relative to other phospholipids is reduced by half in mycelia grown with cholesterol. The major phospholipids are phosphatidyl ethanolamine, phosphatidyl serine, and phosphatidyl choline. Minor phospholipids identified are phosphatidyl inositol, lysophosphatidyl choline, lysophosphatidyl ethanolamine, phosphatidyl glycerol, and cardiolipin. No significant differences were noted in fatty acid composition.


1966 ◽  
Vol 44 (6) ◽  
pp. 763-774 ◽  
Author(s):  
Demetrios Sgoutas

The livers of male chicks which had been fed a soybean and corn type ration for 8 weeks were exhaustively extracted with chloroform–methanol. The total (4.47%) lipid extract was separated into neutral and phospholipid fractions on a Unisil column and the mixed fatty acids in each fraction determined with the aid of gas chromatography. The neutral lipid fraction contained 3.16% hydrocarbons, 4.73% cholesterol esters, 74.99% triglycerides, 0.28% free fatty acids, 11.57% cholesterol, 3.37% diglycerides, and 1.54% monoglycerides. The phospholipid fraction contained 5.7% cardiolipin, 32.3% phosphatidyl ethanolamine, 7.3% phosphatidyl inositol, 3.0% phosphatidyl serine, 47.9% phosphatidyl choline, and 3.8% sphingomyelin and lysophosphatidyl choline. The cholesterol ester and diglyceride fractions contained twice as much linoleic acid as the other neutral lipid fractions. Phospholipids were characterized by a high content of polyunsaturated fatty acids.


1963 ◽  
Vol 09 (01) ◽  
pp. 164-174 ◽  
Author(s):  
Albert R Pappenhagen ◽  
J. L Koppel ◽  
John H Olwin

SummaryData have been presented on the in vitro effects of human chylomicra, low-density human plasma lipoproteins, and partially purified preparations of various phospholipids on human plasma euglobulin lysis. Euglobulin lysis was found to be accelerated by preparations of mixed soybean phospholipids (aso-lectin), cephalin, phosphatidyl inositol, phophatidyl serine and phosphatidyl ethanolamine. In contrast, it was found to be inhibited by preparations of human chylomicra, low-density human plasma liproproteins and lecithin. Inhibition of euglobulin lysis produced by any of these three agents could be diminished or completely overcome by the simultaneous presence of suitable levels of any one of the accelerating agents. In all cases studied, both inhibitory and accelerating effects were observed to be concentration-dependent. Evidence has been obtained to suggest that in the case of the accelerating agents the observed increased rate of euglobulin lysis is not a direct effect on lysis itself, but rather is due to more complete precipitation of plasminogen in the presence of these substances. On the other hand, it appears that the inhibitory effects observed are not related to the extent of plasminogen precipitation, but are actually true inhibitions of euglobulin lysis. The possible clinical significance of some of these observations has been briefly discussed.


1969 ◽  
Vol 111 (2) ◽  
pp. 157-165 ◽  
Author(s):  
A. Sheltawy ◽  
R. M. C. Dawson

1. The distribution of individual phospholipids was determined in hen brain and compared with that in sciatic nerve obtained in a previous investigation. Sciatic nerve is more enriched in the myelinic phospholipids ethanolamine plasmalogen, phosphatidylserine and sphingomyelin, but it contains relatively less triphosphoinositide, and much less diphosphoinositide, than the brain. 2. The course of incorporation of intraperitoneally injected 32P into the acid-soluble phosphorus, phosphoinositides and total phospholipids of hen brain and sciatic nerve was followed. Although the maximum specific radioactivity in sciatic nerve of acid-soluble phosphorus is 4·5 times, and that of triphosphoinositide six times, that in the brain, the relative rate of triphosphoinositide phosphorus synthesis per gram of brain is three times that in sciatic nerve. 3. Administration of the demyelinating agent tri-o-cresyl phosphate to hens has no significant effect on the amounts or the rate of 32P incorporation into the total phospholipids of the sciatic nerve. However, the rate of incorporation of 32P into triphosphoinositide, although not its concentration, is raised from the first day after administration of the drug and remains thus 13 and 23 days later. 4. The incorporation of 32P into polyphosphoinositides of hen brain slices in vitro was studied. The recovery of triphosphoinositide from the slices is markedly increased in the presence of EDTA, although the rate of incorporation of 32P is unaffected. The incorporation of 32P is dependent on the presence of Mg2+ and Ca2+ in the medium, and is decreased when Na+ is replaced with K+ or cholinium ions.


1957 ◽  
Vol 35 (1) ◽  
pp. 945-951 ◽  
Author(s):  
R. J. Rossiter ◽  
I. M. McLeod ◽  
K. P. Strickland

The radioactivity of cytidine diphosphate choline labelled with P32(CMP-P32Ch) was incorporated in vitro into the phosphatides of both hypotonic homogenates of rat brain and mitochondria prepared from isotonic sucrose homogenates. With both preparations the incorporation was increased by addition of D-α,β-diglycerides prepared enzymatically from naturally-occurring lecithins.Hydrolysis of the labelled phosphatides, followed by separation of the hydrolysis products by two-dimensional paper chromatography, indicated that most of the radioactivity was in lecithin (phosphatidyl choline). Only traces of activity were present in phosphatidyl ethanolamine and phosphatidyl serine. There was also some radioactivity in an alkali-stable fraction that was designated 'sphingomyelin'.These findings are consistent with the view that the phosphatides of brain are formed in situ from smaller molecules and that the metabolic pathways for their formation are similar to those postulated by Kennedy and colleagues for the biosynthesis of phosphatides in chicken liver.The radioactivity of CMP-P32Ch also was incorporated into the phosphatides of homogenates of rat sciatic nerve. This incorporation was greater in homogenates prepared from nerves removed 13 days after previous section. It is suggested that these and other increases, found in the in vitro labelling of phosphatides in nerves degenerating after previous section, reported elsewhere, represent an increase in the potentiality of the nerve for remyelination. These increases are associated with the increase in the number of Schwann cells in the nerve.


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