THE INCORPORATION OF PALMITATE-1-14C BY RAT LUNG IN VITRO

1967 ◽  
Vol 45 (4) ◽  
pp. 597-607 ◽  
Author(s):  
A. Naimark ◽  
D. Klass
Keyword(s):  
Specific Activity ◽  
Specific Radioactivity ◽  
Phosphatidyl Inositol ◽  
Phosphatidyl Serine ◽  
Rat Lung ◽  
Exchange Reactions ◽  
Ethanolamine Phosphatidyl ◽  
Exogenous Glucose ◽  
Lipid Fractions

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.

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
Keyword(s):  
Microsomal Fraction ◽  
Phosphatidyl Ethanolamine ◽  
Lipid Fraction ◽  
Specific Radioactivity ◽  
Phosphatidyl Inositol ◽  
Phosphatidyl Serine ◽  
Ethanolamine Phosphatidyl ◽  
Ehrlich Ascites ◽  
Ethanolamine Plasmalogen

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.

scholarly journals Selective cytotoxicity of 125I-labeled monoclonal antibody T101 in human malignant T cell lines

Blood ◽  
1986 ◽  
Vol 67 (2) ◽  
pp. 429-435
Author(s):  
E Boven ◽  
T Lindmo ◽  
JB Mitchell ◽  
PA Jr Bunn
Keyword(s):  
T Cell ◽  
Cell Lines ◽  
Specific Activity ◽  
Specific Radioactivity ◽  
High Specific Activity ◽  
Tumor Localization ◽  
Cytotoxic Effects ◽  
High Specific Radioactivity ◽  
T Cell Lines

The radiolabeled anti-T cell antibody T101 can be used for specific tumor localization, but unlabeled T101 produces limited cytotoxicity in patients. We thus studied the in vitro cytotoxic effects of T101 labeled with 125I, a radionuclide known for its short-range, high- linear-energy electrons. We showed that 125I-T101 could be readily prepared at high specific activity with high immunoreactivity. Human malignant T cell lines HUT 102, MOLT-4, and HUT 78 were found to differ in the number of T65 determinants (the antigen recognized by T101) and the sensitivity to external x-ray radiation, which were of significance for the cytotoxicity of 125I-T101 in vitro. The cytotoxic effects of 125I-T101 were also found to be dose dependent and increased with exposure time under frozen conditions. As controls, unlabeled T101 had no cytotoxic effect, while free Na 125I or the 125I-labeled irrelevant antibody 9.2.27 exerted minor cytotoxicity. In HUT 102 and MOLT-4, more than 3 logs' cell killing was achieved within four weeks. Because considerable cytotoxicity was demonstrated in vitro by 125I-T101 on T65- positive malignant cells, and because low-dose 111In-T101 can be used successfully for tumor localization, future trials using 125I-T101 at high specific radioactivity may improve therapeutic results in patients with T65-positive malignancies.

scholarly journals Screening Tests of Reproductive Immunology in Systemic Lupus Erythematosus

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Zdenka Ulcova-Gallova ◽  
Alice Mockova ◽  
Miroslava Cedikova
Keyword(s):  
Systemic Lupus Erythematosus ◽  
Lupus Erythematosus ◽  
Phosphatidyl Ethanolamine ◽  
Phosphatidyl Inositol ◽  
Phosphatidyl Serine ◽  
Sperm Cells ◽  
Systemic Lupus ◽  
Ethanolamine Phosphatidyl ◽  
Glycoprotein I ◽  
Beta 2

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.

scholarly journals Therapy of cervical cancer using 131I-labeled nanoparticles

2018 ◽  
Vol 46 (6) ◽  
pp. 2359-2370 ◽  
Author(s):  
Wei Li ◽  
Danyang Sun ◽  
Ning Li ◽  
Yiming Shen ◽  
Yiming Hu ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Specific Activity ◽  
Specific Radioactivity ◽  
Xenograft Model ◽  
Uv Spectrophotometry ◽  
Cytotoxicity Test ◽  
Mouse Xenograft Model ◽  
Late Apoptosis

Objective To evaluate the effectiveness of two kinds of Arg-Gly-Asp (RGD)-targeted 131I-containing nanoliposomes for the treatment of cervical cancer in vitro and in vivo. Methods The nanoparticle liposomes designated RGD-131I-tyrosine peptide chain (TPC)-L and 131I-RGD-L were prepared. The emulsion solvent evaporation method was used to encapsulate the polypeptide into liposomes. The quantity of entrapped polypeptide was measured using UV spectrophotometry. The labeling rates, radiochemical purities, and total radioactivities were measured using paper chromatography. Cytotoxicity was assessed using the MTS assay and flow cytometry. Therapeutic efficacy was monitored using a mouse xenograft model of cervical cancer. Results The labeling efficiency, radiochemical purity, and specific radioactivity of RGD-131I-TPC-L were greater than those of 131I-RGD-L. The cytotoxicity test indicated that late apoptosis of cells treated with RGD-131I-TPC-L and 131I-RGD-L was higher than that of cells treated with Na131I. The therapeutic effect of RGD-131I-TPC-L was better than that of 31I-RGD-L in the mouse model. Conclusions The specific activity of liposome-encapsulated RGD-131I-TPC-L was higher than that of 131I-RGD-L, which labeled liposomes directly. Moreover, the RGD-131I-TPC-L liposomes were more effective for killing xenografted tumor cells.

scholarly journals Selective cytotoxicity of 125I-labeled monoclonal antibody T101 in human malignant T cell lines

Blood ◽  
1986 ◽  
Vol 67 (2) ◽  
pp. 429-435 ◽  
Author(s):  
E Boven ◽  
T Lindmo ◽  
JB Mitchell ◽  
PA Jr Bunn
Keyword(s):  
T Cell ◽  
Cell Lines ◽  
Specific Activity ◽  
Specific Radioactivity ◽  
High Specific Activity ◽  
Tumor Localization ◽  
Cytotoxic Effects ◽  
High Specific Radioactivity ◽  
T Cell Lines

Abstract The radiolabeled anti-T cell antibody T101 can be used for specific tumor localization, but unlabeled T101 produces limited cytotoxicity in patients. We thus studied the in vitro cytotoxic effects of T101 labeled with 125I, a radionuclide known for its short-range, high- linear-energy electrons. We showed that 125I-T101 could be readily prepared at high specific activity with high immunoreactivity. Human malignant T cell lines HUT 102, MOLT-4, and HUT 78 were found to differ in the number of T65 determinants (the antigen recognized by T101) and the sensitivity to external x-ray radiation, which were of significance for the cytotoxicity of 125I-T101 in vitro. The cytotoxic effects of 125I-T101 were also found to be dose dependent and increased with exposure time under frozen conditions. As controls, unlabeled T101 had no cytotoxic effect, while free Na 125I or the 125I-labeled irrelevant antibody 9.2.27 exerted minor cytotoxicity. In HUT 102 and MOLT-4, more than 3 logs' cell killing was achieved within four weeks. Because considerable cytotoxicity was demonstrated in vitro by 125I-T101 on T65- positive malignant cells, and because low-dose 111In-T101 can be used successfully for tumor localization, future trials using 125I-T101 at high specific radioactivity may improve therapeutic results in patients with T65-positive malignancies.

scholarly journals Schlafen 3 induction by cyclic strain regulates intestinal epithelial differentiation

2010 ◽  
Vol 298 (6) ◽  
pp. G994-G1003 ◽  
Author(s):  
Lisi Yuan ◽  
Yingjie Yu ◽  
Matthew A. Sanders ◽  
Adhip P. N. Majumdar ◽  
Marc D. Basson
Keyword(s):  
Sodium Butyrate ◽  
Transforming Growth Factor ◽  
Specific Activity ◽  
Phosphatidyl Inositol ◽  
Cyclic Strain ◽  
Epithelial Differentiation ◽  
Pi3 Kinase ◽  
Intestinal Epithelial ◽  
Gut Function

The intestinal epithelium is subjected to repetitive deformation during normal gut function by peristalsis and villous motility. In vitro, cyclic strain promotes intestinal epithelial proliferation and induces an absorptive phenotype characterized by increased dipeptidyl dipeptidase (DPPIV) expression. Schlafen 3 is a novel gene recently associated with cellular differentiation. We sought to evaluate whether Schlafen 3 mediates the effects of strain on the differentiation of intestinal epithelial cell (IEC)-6 in the absence or presence of cyclic strain. Strain increased Schlafen 3 mRNA and protein. In cells transfected with a control-nontargeting siRNA, strain increased DPPIV-specific activity. However, Schlafen 3 reduction by siRNA decreased basal DPPIV and prevented any stimulation of DPPIV activity by strain. Schlafen 3 reduction also prevented DPPIV induction by sodium butyrate (1 mM) or transforming growth factor (TGF)-β (0.1 ng/ml), two unrelated differentiating stimuli. However, Schlafen-3 reduction by siRNA did not prevent the mitogenic effect of strain or that of EGF. Blocking Src and phosphatidyl inositol (PI3)-kinase prevented strain induction of Schlafen 3, but Schlafen 3 induction required activation of p38 but not ERK. These results suggest that cyclic strain induces an absorptive phenotype characterized by increased DPPIV activity via Src-, p38-, and PI3-kinase-dependent induction of Schlafen 3 in rat IEC-6 cells on collagen, whereas Schlafen 3 may also be a key factor in the induction of intestinal epithelial differentiation by other stimuli such as sodium butyrate or TGF-β. The induction of Schlafen 3 or its human homologs may modulate intestinal epithelial differentiation and preserve the gut mucosa during normal gut function.

Protein metabolism by rat lung: influence of fasting, glucose, and insulin

1977 ◽  
Vol 43 (3) ◽  
pp. 463-467 ◽  
Author(s):  
L. A. Thet ◽  
M. D. Delaney ◽  
C. A. Gregorio ◽  
D. Massaro
Keyword(s):  
Protein Synthesis ◽  
Protein Content ◽  
Protein Metabolism ◽  
Specific Radioactivity ◽  
Rat Lung ◽  
Synthetic Process ◽  
In Vitro System ◽  
Other Substances

We studied protein metabolism by rat lung slices. We found that phenylalanine is not metabolized to other substances by the lung and that the rate of incorporation of L-[U-14C]phenylalanine into protein, calculated using its intracellular specific radioactivity, reached a maximum within 20 min and remained stable for the rest of a 3-h incubation. The rate of protein degradation, determined using [12C]phenylalanine as a marker, was linear over a 3-h incubation. Fasting for 3 days slowed the increase in lung protein content of fasted compared to nonfasted rats; there was also a decrease in protein synthesis and an increase in proteolysis. In fed rats, glucose, insulin, and glucose plus insulin did not alter protein synthesis. Glucose, insulin alone, and glucose plus insulin decreased proteolysis. We conclude that the in vitro system reflected changes in the in vivo protein content of the lung. Fasting decreases protein synthesis and increases proteolysis. Glucose and insulin alone modulate protein metabolism in the lung by acting on the degradative rather than the synthetic process.

INCORPORATION OF C14 OR P32 INTO THE PHOSPHATIDES OF RUNNER BEAN LEAVES

1957 ◽  
Vol 35 (6) ◽  
pp. 907-921 ◽  
Author(s):  
Frank M. Eberhardt ◽  
Morris Kates
Keyword(s):  
Fatty Acids ◽  
Specific Activity ◽  
Phosphatidyl Serine ◽  
Water Soluble ◽  
Total Lipids ◽  
Ethanolamine Phosphatidyl ◽  
Runner Bean ◽  
Free Air ◽  
Minute Period ◽  
Fraction I

The phosphatide content of primary leaves of runner bean increased linearly with time, between the 8th and 20th day of development, at a rate proportional to the growth of the leaves.Detached leaves incorporated C14 into the total lipids (pigments, non-phosphatides, and phosphatides) when supplied with C14O2 in the light for 1 minute followed by a 30-minute period in the light or dark in tracer-free air and when supplied with pyruvate-2-C14 or acetate-1-C14 in the light. P32 was incorporated into the phosphatides after orthophosphate-P32 was supplied both in the light and dark. With each precursor, radioisotope was incorporated into the four phosphatide fractions obtained by chromatography of the total lipids: inositol–carbohydrate phosphatides (Fraction I), phosphatidyl ethanolamine – phosphatidyl serine (Fraction II), an unknown phosphatide (Fraction III), and lecithin (Fraction IV).Distribution of radioactivity among these phosphatide fractions and among the phosphatide moieties varied greatly with the precursor supplied. Thus, with C14O2 as precursor, Fraction I and III had the highest specific activities, and C14 entered both the water-soluble moieties and the fatty acids in all of the phosphatide fractions. With pyruvate-C14 or acetate-C14, lecithin was most highly labelled, and C14 was found almost exclusively in the fatty acids of the phosphatides. With orthophosphate-P32, Fraction III had the highest specific activity. The bearing of these findings on the metabolism of phosphatides in leaves is discussed.

scholarly journals Post-translational arginylation of ornithine decarboxylase from rat hepatocytes

1990 ◽  
Vol 267 (2) ◽  
pp. 343-348 ◽  
Author(s):  
J Kopitz ◽  
B Rist ◽  
P Bohley
Keyword(s):  
Ornithine Decarboxylase ◽  
Half Life ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Specific Radioactivity ◽  
Rat Hepatocytes ◽  
Nmri Mouse ◽  
Cultured Hepatocytes ◽  
Lysosomal Proteinase

Ornithine decarboxylase (ODC) was purified 6500-fold from NMRI mouse kidneys under conditions designed to inhibit degradation by proteinases. The enzyme was homogeneous by SDS/polyacrylamide-gel electrophoresis, and the specific activity was among the highest reported. The yield was 70%. A monoclonal antibody against this preparation was generated and used in studies to investigate the half-life of ODC in cultured rat hepatocytes labelled with [35S]methionine. This value was 39 +/- 4 min and was unchanged when either NH4Cl (as a lysosomotropic agent) or leupeptin (as a lysosomal proteinase inhibitor) was added to the culture medium. Thus the intracellular turnover of ODC in cultured hepatocytes occurs mainly in extra-lysosomal compartments. Arginylation of rat ODC was investigated in vitro by incubation with L-[3H]arginyl-tRNA, and the incorporation of the label was compared with that of total cytosolic proteins. Arginylated ODC had a specific radioactivity 8600 times that of the bulk of cytosolic protein. Edman degradation of this ODC showed that the post-translational arginylation occurred only at the alpha-amino end of the enzyme. The inhibitor of arginyl-tRNA:protein arginyltransferase (EC 2.3.2.8), L-glutamyl-L-valyl-L-phenylalanine, increased the half-life of ODC in cultured hepatocytes from 39 min to more than 90 min. The possible significance of the preferential post-translational arginylation of ornithine decarboxylase to its rapid turnover is discussed.

Clotting Activity of Platelet Phospholipids in Rat and Man

1974 ◽  
Vol 32 (02/03) ◽  
pp. 382-390 ◽  
Author(s):  
P Gautheron ◽  
E Dumont ◽  
S Renaud
Keyword(s):  
Phosphatidyl Choline ◽  
Phosphatidyl Ethanolamine ◽  
Phosphatidyl Inositol ◽  
Phosphatidyl Serine ◽  
The Other ◽  
Active Fraction ◽  
Platelet Phospholipid ◽  
Experimental Conditions ◽  
Ethanolamine Phosphatidyl ◽  
Recalcification Time

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.

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