scholarly journals Influence of chain length of pyrene fatty acids on their uptake and metabolism by Epstein–Barr-virus-transformed lymphoid cell lines from a patient with multisystemic lipid storage myopathy and from control subjects

1990 ◽  
Vol 269 (1) ◽  
pp. 107-113 ◽  
Author(s):  
J Radom ◽  
R Salvayre ◽  
T Levade ◽  
L Douste-Blazy

The uptake and intracellular metabolism of 4-(1-pyrene)butanoic acid (P4), 10-(1-pyrene)decanoic acid (P10) and 12-(1-pyrene)dodecanoic acid (P12) were investigated in cultured lymphoid cell lines from normal individuals and from a patient with multisystemic lipid storage myopathy (MLSM). The cellular uptake was shown to be dependent on the fatty-acid chain length, but no significant difference in the uptake of pyrene fatty acids was observed between MLSM and control lymphoid cells. After incubation for 1 h the distribution of fluorescent fatty acids taken up by the lymphoid cell lines also differed with the chain length, most of the fluorescence being associated with phospholipid and triacylglycerols. In contrast with P10 and P12, P4 was not incorporated into neutral lipids. When the cells were incubated for 24 h with the pyrene fatty acids, the amount of fluorescent lipids synthesized by the cells was proportional to the fatty acid concentration in the culture medium. After a 24 h incubation in the presence of P10 or P12, at any concentration, the fluorescent triacylglycerol content of MLSM cells was 2-5-fold higher than that of control cells. Concentrations of pyrene fatty acids higher than 40 microM seemed to be more toxic for mutant cells than for control cells. This cytotoxicity was dependent on the fluorescent-fatty-acid chain length (P12 greater than P10 greater than P4). Pulse-chase experiments permitted one to demonstrate the defect in the degradation of endogenously biosynthesized triacylglycerols in MLSM cells (residual activity was around 10-25% of controls on the basis of half-lives and initial rates of P10- or P12-labelled-triacylglycerol catabolism); MLSM lymphoid cells exhibited a mild phenotypic expression of the lipid storage (less severe than that observed in fibroblasts). P4 was not utilized in the synthesis of triacylglycerols, and thus did not accumulate in MLSM cells: this suggests that natural short-chain fatty acids might induce a lesser lipid storage in this disease.

1986 ◽  
Vol 6 (2) ◽  
pp. 703-706
Author(s):  
F Toneguzzo ◽  
A C Hayday ◽  
A Keating

The technique of DNA transfer by electroporation was investigated in an effort to evaluate its utility for the identification of developmentally controlled regulatory sequences. Transient and stable gene expression was detected in a variety of lymphoid cell lines subjected to electroporation. No correlation existed between the levels of chloramphenicol acetyltransferase (acetyl-CoA; chloramphenicol 3-O-acetyltransferase, EC 2.3.1.28) expression and stable transfection frequency. In all lymphoid cell lines tested, the simian virus 40 early region was a better promoter than was the Rous sarcoma virus long terminal repeat.


1991 ◽  
Vol 260 (2) ◽  
pp. L44-L51 ◽  
Author(s):  
K. J. Longmuir ◽  
S. Haynes

This study was undertaken to determine those structural features of phospholipid molecules which influence their enrichment in type II cell lamellar body material. Cultured fetal rabbit lung tissue was labeled with [1-14C]acetate, type II cells were isolated, and extracellular lamellar body and microsomal fractions were prepared. Radiolabeled molecular species of phosphatidylcholine (PC) and phosphatidylethanolamine were analyzed by high-performance liquid chromatography (HPLC), followed by silver nitrate thin-layer chromatography of HPLC peak fractions that overlapped. Compared with microsomes, lamellar body PC was selectively enriched with molecular species containing 14- and 16-carbon fatty acids and depleted of species containing 18-carbon fatty acids. Palmitoleic acid and an ether linkage positively influenced the enrichment of PC molecular species in the lamellar body material, but these structural features were secondary to the predominant influence of fatty acid chain length. In vivo, lung tissue normally contains low levels of palmitoleic acid; hence most unsaturated fatty acids are 18-carbons or longer. A cellular lipid-sorting mechanism that selects PCs by recognition of 14- and 16-carbon fatty acid chains (and not by recognition of fatty acid saturation) should serve to enrich the resulting pulmonary surfactant with disaturated molecular species of PC.


Blood ◽  
2011 ◽  
Vol 118 (18) ◽  
pp. 4872-4881 ◽  
Author(s):  
Husheng Ding ◽  
Jennifer Hackbarth ◽  
Paula A. Schneider ◽  
Kevin L. Peterson ◽  
X. Wei Meng ◽  
...  

Abstract The mechanism of cytotoxicity of farnesyltransferase inhibitors is incompletely understood and seems to vary depending on the cell type. To identify potential determinants of sensitivity or resistance for study in the accompanying clinical trial (Witzig et al, page 4882), we examined the mechanism of cytotoxicity of tipifarnib in human lymphoid cell lines. Based on initial experiments showing that Jurkat variants lacking Fas-associated death domain or procaspase-8 undergo tipifarnib-induced apoptosis, whereas cells lacking caspase-9 or overexpressing Bcl-2 do not, we examined changes in Bcl-2 family members. Tipifarnib caused dose-dependent up-regulation of Bim in lymphoid cell lines (Jurkat, Molt3, H9, DoHH2, and RL) that undergo tipifarnib-induced apoptosis but not in lines (SKW6.4 and Hs445) that resist tipifarnib-induced apoptosis. Further analysis demonstrated that increased Bim levels reflect inhibition of signaling from c-Raf to MEK1/2 and ERK1/2. Additional experiments showed that down-regulation of the Ras guanine nucleotide exchange factor RasGRP1 diminished tipifarnib sensitivity, suggesting that H-Ras or N-Ras is a critical farnesylation target upstream of c-Raf in lymphoid cells. These results not only trace a pathway through c-Raf to Bim that contributes to tipifarnib cytotoxicity in human lymphoid cells but also identify potential determinants of sensitivity to this agent.


1973 ◽  
Vol 137 (3) ◽  
pp. 838-843 ◽  
Author(s):  
T. H. Hütteroth ◽  
H. Cleve ◽  
S. D. Litwin ◽  
M. D. Poulik

ß2-microglobulin was detected on the cell surface and in the medium of human lymphoid cells established in long-term culture. The secretion of ß2-microglobulin was relatively uniform when different cell lines were compared, whereas IgG production varied widely. κ- and µ-membrane antigens were modulated by specific antibody; ß2-microglobulin was not modulated. Anti-κ and anti-µ antisera had no effect on the expression of membrane ß2-microglobulin, nor had anti-ß2-microglobulin antiserum any effect on the expression of κ- and µ-membrane antigens.


1985 ◽  
Vol 249 (6) ◽  
pp. G745-G750 ◽  
Author(s):  
G. W. Aponte ◽  
A. S. Fink ◽  
J. H. Meyer ◽  
K. Tatemoto ◽  
I. L. Taylor

The present study examines peptide YY responses to regional intestinal perfusion of fatty acids of different chain length--dodecanoate and oleate. Six dogs with chronic gastric, duodenal, and jejunal fistulas were studied. Proximal perfusates were administered into the duodenum and diverted through an intestinal fistula placed 45 cm beyond the duodenal cannula. Distal perfusates were administered into the caudal stoma of this intestinal stoma. Peptide YY responses to proximal, distal, and whole-gut perfusion were compared. Proximal perfusion with oleate or dodecanoate failed to release peptide YY. In contrast, distal and whole-gut perfusion with either fatty acid produced significant increases that were of similar magnitude. Immunocytochemical studies demonstrated that peptide YY cells predominated in the canine ileocolonic mucosa and decreased progressively in an orad direction. We conclude that peptide YY release is not dependent on fatty acid chain length and that the duodenum does not play a dominant role in peptide YY release. As such, peptide YY release differs from that of its cousin pancreatic polypeptide and may result at least in part from direct stimulation of the peptide YY cell in the ileocolonic mucosa.


1970 ◽  
Vol 75 (1) ◽  
pp. 55-60 ◽  
Author(s):  
R. J. Andrews ◽  
D. Lewis

SUMMARYThe effect of fatty acid chain length and unsaturation on digestibility in sheep were examined using partially purified samples of lauric, myristic, palmitic, stearic, oleic and linoleic acids. The digestibility of the fatty acids was relatively constant with only a very slight decrease on increasing chain length. There was an extensive hydrogenation of the unsaturated fatty acids.The corrected digestibility coefficients for lauric acid was 91%, myristic 86%, palmitic 87% and stearic acid 81–83% whereas the corrected digestibility coefficients for oleic and linoleic acids were calculated at 87 and 93% respectively. The digestibility coefficients for the saturated fatty acids are higher than similar estimates that have been reported for non-ruminants. It is suggested that the ruminant is better able to utilize saturated fatty acids than the non-ruminant.


1972 ◽  
Vol 136 (3) ◽  
pp. 644-649 ◽  
Author(s):  
B. Shannon Danes ◽  
T. H. Hütteroth ◽  
H. Cleve ◽  
Alexander G. Bearn

Lymphoid suspension lines have been established from three patients with the Hurler syndrome and four normals. The Hurler lines can be distinguished from normals by (a) staining characteristics, (b) increase in total cellular mucopolysaccharide content, and (c) increase in dermatan sulfate. Hyaluronic acid is absent in cultured lymphoid cells from normal persons and patients with the Hurler syndrome. The availability of biochemically marked suspension cultures should prove useful for enzymatic studies as well as for further elucidation of this clinical syndrome.


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