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Biomedical subjects

M Glaser

Publications and source records attributed to M Glaser.

At least 91 records · Page 5Linked to original sources

Rotational relaxation times of 1,6-diphenyl-1,3,5-hexatriene in phospholipids isolated from LM cell membranes. Effects of phospholipid polar head-group and fatty acid composition.

Phospholipids were isolated from mitochondrial, microsomal, and plasma membranes of LM cells and fractionated into individual phospholipid classes on silicic acid columns. The fatty acid composition and the rotational relaxation time of 1,6-diphenyl-1,3,5-hexatriene (DPH) were determined for each phospholipid class. Sphingomyelin was the only phospholipid isolated from LM cell membranes that showed a phase transition within the temperature range investigated, 5-40 degrees C. The rotational relaxation times for DPH were lowest in phosphatidylcholine in all the membrane fractions. Phosphatidylcholine isolated from the three membrane fractions of choline-supplemented cells had similar rotational relaxation times and phosphatidylcholine isolated from microsomal membranes of linoleate-supplemented cells had lower rotational relaxation times. The results indicate that the differences in the rotational relaxation times of DPH between mitochondrial, microsomal, and plasma membrane phospholipids could be explained primarily by differences in the polar head-group composition, while differences in the fatty acid composition had only a minor effect. This provides a basis for understanding how the different lipid components in these cells contribute to membrane fluidity.

Animals↗

Regulation of specific cell-mediated cytotoxic response against SV40-induced tumor associated antigens by depletion of suppressor T cells with cyclophosphamide in mice.

When cyclophosphamide was administered to mice before immunization with syngeneic SV40 transformed cells, the specific immune response elicited, as was measured by in vitro 51Cr release assay was stronger and lasted longer when compared to the response generated in noncyclophosphamide-treated mice. The augmentation effect of the drug was dependent on cyclophosphamide concentration being optimal at 100 mg/kg and on the time of drug administration in relation to antigen immunization being optimal at 2 d before antigen administration. Transfer of T cells from normal syngeneic mice to drug-treated animals abolished the cyclophosphamide-induced augmentation of immune response. These results implied that cyclophosphamide sensitive T cells suppressed the in vivo generation of specific effector T cells against SV40-induced tumor-associated antigens.

Animals↗

Effect of membrane phospholipid compositional changes on adenylate cyclase in LM cells.

Adenylate cyclase activities were examined in mouse LM cell membranes which had been supplemented with polar head groups and/or fatty acids. Basal, fluoride-, and PGE1-stimulated activities varied systematically with changes in phospholipid composition, and PGE1-stimulated activities correlated with the average degree of unsaturation of the phospholipid fatty acids or with the primary amino group character of the phospholipid polar head groups. In addition, the Km for ATP of basal adenylate cyclase was systematically changed by both polar head group and fatty acid supplementation. Alteration of the membrane lipid composition also changed the temperature dependence of the enzyme and the lag time between addition of PGE1 and the onset of a change in catalytic rate. However, none of the alterations in the enzyme activity could be correlated with the viscosities of supplemented membranes and, instead, seemed to be characteristic for a specific polar head group or fatty acid composition. The data suggest a specific interaction of the enzyme with phospholipids and indicate that structural features of phospholipids may play a role in regulating adenylate cyclase activity. It is proposed that adenylate cyclase can exist in several different conformations in the membrane depending upon the phospholipid composition.

Adenylyl Cyclases↗

Deuterium nuclear magnetic resonance investigation of the effects of proteins and polypeptides on hydrocarbon chain order in model membrane systems.

Deuterium Fourier-transform nuclear magnetic resonance spectra have been obtained of 1-myristoyl 2-(14,14,14-trideutero)myristoyl phosphatidylcholine bilayers at 34.1 MHz by using the quadrupole echo pulse technique. Thereby, we have investigated the effects upon the deuterated dimyristoyl phosphatidylcholine bilayers of the following proteins and polypeptides: gramicidin A, bacteriophage f1 coat protein, beef brain myelin proteolipid apoprotein, cytochrome b(5), and cytochrome c oxidase (ferrocytochrome c:oxygen oxidoreductase, EC 1.9.3.1). Above T(c), the transition temperature between the gel and liquid crystal phases, the quadrupole splitting of the deuterium-labeled methyl group is reduced or collapsed in the presence of protein or polypeptide. No evidence has been found for ordered "boundary lipid." Below T(c), the spectra show that the hydrocarbon chains are prevented from crystallizing by the protein (or polypeptide) incorporated in the membrane. Similar disordering effects above T(c) are also seen when an unsaturated lipid, 1-(16,16,16-trideutero)palmitoyl 2-palmitoleyl phosphatidylcholine is complexed with cytochrome oxidase.

Journal Article↗

Use of a fluorescent probe to determine the viscosity of LM cell membranes with altered phospholipid compositions.

The phospholipid compostition of LM cells grown in tissue culture was altered by substituting ethanolamine for choline in the growth medium. The plasma membrane isolated from cells grown in medium conatining ethanolamine for 83 h had a sixfold increase in the ratio of phosphatidylethanolamine to phosphatidylcholine, the two major phospholipid classes. This was accompanied by small changes in other lipid components of the membrane. There was also a sixfold increase in the amount of triacylglycerols and alkyldiacylglycerols which were not associated with the membrane fraction of the cell. No significant changes occurred in the lipid composition of cells during growth in choline containing medium. The viscosity of plasma membranes was studied in whole cells and isolated membranes using the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene. Plasma membranes isolated from ethanolamine-supplemented cells had greater viscosities than membranes isolated from choline-supplemented cells. When whole cells were labeled with the fluorescent probe, the opposite trend in the apparent membrane viscosity was observed. This was due primarily to the probe penetrating into nonmembranous neutral lipids rather than remaining localized in the surface membrane of the cells. Since the enthanolamine-supplemented cells contained more low viscosity neutral lipids, the whole cells gave an apparently lower viscosity as compared with choline-supplemented cells, thus, measurements carried out on whole cells gave an inaccurate determination of the viscosity of the surface membrane.

Adenosine Triphosphatases↗

Suppressor cells in tumor bearing mice and rats.

Suppressor macrophages in spleens of tumor bearing mice and rats were found to inhibit "in vitro" reactivity of lymphocytes to T and B cell mitogens and to allogeneic cells in the mixed lymphocyte tumor cell interaction and the "in vitro" generation of secondary cytotoxic effector cells. We propose that similar suppressor cells may play an important role in the general immunosuppression of tumor bearing hosts and in the inability of tumor bearing hosts to effectively reject the tumor.

Animals↗

Deuterium magnetic resonance spectroscopy of isotopically labeled mammalian cells.

Choline completely deuterated in the methyl groups has been incorporated into two mammalian cell systems. The first was the chemically transformed mouse fibroblast LM cell line, grown in suspension culture. The second system consisted of mitochondria from Sprague-Dawley rats which were fed upon a choline-deficient diet enriched with deuterated choline chloride. Two classes of deuterium nuclear magnetic resonance (NMR) signal were obtained from each system. The first class of signal exhibited no residual quadrupole coupling constant and was assigned to the natural abundance of 2H in water, together with free labeled choline. The second class of signal had a residual quadrupole coupling of about 1 HKz and was assigned to 2H-labeled choline headgroups of phospholipids. These results are in conflict with a previous study of deuterated rat liver mitochondrial membranes. Deuterium magnetic resonance spectra can be obtained on mammalian membranes that have incorporated specific deuterium-labeled compounds and, therefore, provides a very powerful method for studying the dynamic structure of membranes.

Animals↗