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Biomedical subjects

R Schafer

Publications and source records attributed to R Schafer.

At least 19 recordsLinked to original sources
PubMed1992

Induction of a cellular immune response to a foreign antigen by a recombinant Listeria monocytogenes vaccine.

A recombinant strain of Listeria monocytogenes that stably and constitutively expresses Escherichia coli beta-galactosidase was used as a live vaccine vector. BALB/c mice were immunized orally or parenterally with the recombinant L. monocytogenes, and their cellular and humoral immune responses to beta-galactosidase were measured. Spleen cells taken 1 week after oral inoculation or 5 weeks after oral or parenteral inoculation (with a boost at 4 weeks) showed beta-galactosidase-specific CTL responses. The CTL line derived from mice immunized i.p. was also shown to be class I restricted and Thy-1.2+, CD8+, and TCR alpha beta+. All mice immunized with the recombinant L. monocytogenes had positive delayed-type hypersensitivity responses to heat-killed L. monocytogenes, but only 15% had a positive delayed-type hypersensitivity reaction to beta-galactosidase. Individual serum samples from mice immunized i.p. or i.v. were tested for antibody to beta-galactosidase. Approximately 11% had low positive titers for beta-galactosidase antibodies. These results demonstrate that both oral and parenteral immunization with recombinant L. monocytogenes results in a cellular immune response to the foreign protein, which is primarily a specific CD8+ CTL response.

Animals↗
PubMed1992

Forcing expression of a soybean root glutamine synthetase gene in tobacco leaves induces a native gene encoding cytosolic enzyme.

Glutamine synthetase (GS; EC 6.3.1.2) is present in different subcellular compartments in plants. It is located in the cytoplasm in root and root nodules while generally present in the chloroplasts in leaves. The expression of GS gene(s) is enhanced in root nodules and in soybean roots treated with ammonia. We have isolated four genes encoding subunits of cytosolic GS from soybean (Glycine max L. cv. Prize). Promoter analysis of one of these genes (GS15) showed that it is expressed in a root-specific manner in transgenic tobacco and Lotus corniculatus, but is induced by ammonia only in the legume background. Making the GS15 gene expression constitutive by fusion with the CaMV-35S promoter led to the expression of GS in the leaves of transgenic tobacco plants. The soybean GS was functional and was located in the cytoplasm in tobacco leaves where this enzyme is not normally present. Forcing this change in the location of GS caused concomitant induction of the mRNA for a native cytosolic GS in the leaves of transgenic tobacco. Shifting the subcellular location of GS in transgenic plants apparently altered the nitrogen metabolism and forced the induction in leaves of a native GS gene encoding a cytosolic enzyme. The latter is normally expressed only in the root tissue of tobacco. This phenomenon may suggest a hitherto uncharacterized metabolic control on the expression of certain genes in plants.

Base Sequence↗
PubMed1992

Natural killer cells mediate protection induced by a Salmonella aroA mutant.

We have previously shown that an avirulent strain of Salmonella typhimurium, SL3235, blocked in aromatic synthesis, confers high levels of resistance to challenge with virulent Salmonella as early as 3 days postvaccination. In the present studies, it was found that immunization with SL3235 resulted in high levels of natural killer (NK) cell activity in the spleens and peritoneal cavities of C3H/HeJ mice, as measured by cytotoxicity against YAC-1 targets. NK cell activity was at its maximum 2 to 4 days after immunization and was ablated by in vivo or in vitro treatment with anti-asialo GM1. In vivo treatment with anti-asialo GM1 during the first week after immunization with SL3235 depleted NK cell activity and markedly increased mortality in mice challenged with a virulent Salmonella strain. These results are compatible with a role for NK cells as one important component in the resistance against virulent Salmonella infection induced by a live, attenuated vaccine.

Animals↗
PubMed1989

Abnormal typical pattern of platelet function and thromboxane generation in unstable angina.

Platelet aggregation (PA), platelet thromboxane B2 (TXB2) generation and 14C 5-hydroxytryptamine (5HT) release were studied in 13 patients with unstable angina, and compared to 14 patients with stable angina and 16 healthy controls. A typical pattern, distinct in 4 aspects from stable angina patients or controls, was observed in the unstable angina patients. ADP or collagen induced shape change was 3-4 times greater, the extent of epinephrine induced PA was nil or very low, the extent of collagen induced 14C 5HT release was also reduced while collagen induced platelet TXB2 generation was increased in spite of a reduced extent of PA. The extent of ADP or collagen induced PA was also significantly reduced. These results indicate a platelet membrane abnormality occurring presumably during contact of the circulating platelets with a non-occlusive thrombus observed at sites of ruptured plaques in unstable angina patients. Since also the pattern (20-30% overlap with control values) was distinct from that of stable angina patients, it might indicate an active thrombotic process. Plasma beta-thromboglobulin (beta TG) and TXB2 levels and serum TXB2 generation were also studied in the cardiac patients and controls and in another 10 patients with advanced peripheral occlusive arterial disease (POAD). Plasma beta TG and TXB2 levels were slightly elevated in the unstable angina patients and markedly elevated in the POAD patients. Serum TXB2 generation was, however, elevated in the stable angina patients (p less than 0.002) and more so in the unstable angina patients (p less than 0.001) compared to controls or to POAD patients. This was presumably mediated through enhanced thrombin generation. These results suggest that the measured plasma beta TG variable in the unstable angina patients is not useful in the assessment of in vivo platelet activation. It is presumably reflecting the sum of local enhanced platelet activation (at sites of ruptured plaques) and of reduced function of the "defective" circulating platelets. The ability of the platelets of unstable angina patients to generate large amounts of TXB2 if occurring in vivo might induce an intense coronary vasospasm.

Adult↗
PubMed1988

Induction of activated macrophages in C3H/HeJ mice by avirulent Salmonella.

A single injection of viable Salmonella typhimurium SL3235, an avirulent organism blocked in the aromatic pathway, induced the generation of activated peritoneal macrophages in three different C3H mouse strains, including macrophage-defective C3H/HeJ mice. Macrophages obtained from immunized mice were cytotoxic for B16 melanoma cells, P815 mastocytoma cells, and TU-5 fibrosarcoma cells and microbicidal in vitro for the obligate, intracellular, protozoan parasite Leishmania major. The capacity of live SL3235 to activate C3H/HeJ macrophages contrasts with the failure of live Bacillus Calmette-Guérin to induce activated macrophages in this mouse strain. Although viable SL3235 were capable of fully activating cells of both normal and defective mice, a dose-dependent difference was observed in the number of organisms necessary for induction of tumoricidal macrophages in C3HeB/FeJ (normal) and C3H/HeJ (defective) animals. As few as 80 viable SL3235 were capable of activating C3HeB/FeJ macrophages whereas 5 X 10(4) organisms were required to activate C3H/HeJ macrophages. Maximal macrophage activation occurred 7 to 10 days after SL3235 inoculation in C3H/HeJ and C3HeB/FeJ mice. Acetone-killed cells of SL3235 had some but not all of the activity of the living Salmonella. A single in vivo injection of the nonviable preparation resulted in the induction of tumoricidal macrophages in C3HeB/FeJ but not in C3H/HeJ mice, even when tested over a wide dosage range. Injection of acetone-killed cells of SL3235 did, however, result in a population of primed macrophages in C3H/HeJ mice, as explanted cells could be induced to express activated macrophage effector activities after additional treatment in vitro with either LPS or IFN-gamma. Thus, in vivo administration of viable SL3235 is, by itself, capable of eliciting the full series of steps required for activation of C3H/HeJ macrophages, whereas killed SL3235 only provides signals sufficient to prime these defective macrophages for further activation in vitro. AI 15613

Animals↗
PubMed1988

Calcium and olfactory transduction.

1. Inorganic cations, organic calcium antagonists, and calmodulin antagonists were applied to olfactory epithelia of frogs (Rana pipiens) while recording electroolfactogram (EOG) responses. 2. Inorganic cations inhibited EOGs in a rank order, reflecting their calcium channel blocking potency: La3+ greater than Zn2+ greater than Cd2+ greater than Al3+ greater than Ca2+ greater than Sr2+ greater than Co2+ greater than Ba2+ greater than Mg2+. Barium ion significantly enhanced EOGs immediately following application. 3. Diltiazem and verapamil produced dose-dependent EOG inhibition. 4. Calmodulin antagonists inhibited EOGs without correlation to their anti-calmodulin potency.

Animals↗
PubMed1987

Inhibition of protein synthesis does not block physiological responses to odors in the frog over the short term.

Frog olfactory mucosae were stimulated with an odorant every 60 sec and electrophysiological responses (electroolfactograms or EOGs) were monitored continuously--before, during, and after treatment with puromycin, a protein synthesis inhibitor. Repeated administration of puromycin at 4 hr intervals over a 36 hr period failed to suppress EOG responses. In an alternative approach, olfactory responses were inhibited with ethyl bromoacetate, a vaporous alkylating agent. EOG responses failed to reappear over a 72 hr period. We conclude that receptor turnover is not readily influenced by stimulation, and that the turnover time of frog olfactory receptor proteins is of the order of days or longer.

Animals↗
PubMed1985

Wild analysis.

Contemporary debates over psychoanalytic theory and practice warrant a reconsideration of the concept of wild analysis. Freud's initial formulation of the problem, subsequent developments in the Freudian conventions, and the work of Melanie Klein, Kohut, and Gill are compared in order to bring out different conceptions of interpretation that is wild, sound, or too tame. These different conceptions are system-bound. Moral implications of Klein's, Kohut's, and Gill's critiques and alternative systems are taken up.

Anxiety Disorders↗
PubMed1983

Effects of extracts of Trypanosoma cruzi on immune responses: induction of a nonspecific suppressor factor.

Extracts of epimastigotes of Trypanosoma cruzi were examined for the ability to augment or suppress in vivo and in vitro responses of mouse spleen cells. When administered to mice 4 days before immunization with sheep erythrocytes, the 12,000 X g pellet and the 104,000 X g pellet and supernatant fractions of extracts of T. cruzi had no effect on the subsequent plaque-forming cell responses. Testing of the two 104,000 X g fraction in vitro resulted in a slight dose-dependent modification of anti-sheep erythrocyte responses. Also, these two fractions had a moderate dose-dependent blastogenic effect on normal spleen cells. Most significantly, the 104,000 X g supernatant fraction elicited the production of a suppressor factor when incubated with spleen cells from infected mice. This suppressor factor was also apparent when spleen cells from infected mice were incubated with intact epimastigotes or trypomastigotes. Production of the suppressor factor was insensitive to indomethacin and, therefore, presumably was not a prostaglandin.

Animals↗
PubMed1981

Colchicine reversibly inhibits electrical activity in arthropod mechanoreceptors.

Colchicine and some other microtubule-active agents inhibit the electrical responses of cockroach tibial spine mechanoreceptors. Lumicolchicine, a colchicine analog which does not bind to microtubule protein, does not inhibit mechanoreceptive responses. Colchicine inhibition of peripheral mechanoreceptive responses is fully reversible and dose dependent, but colchicine has no effect on conduction in leg nerve axons. Colchicine inhibition is therefore an effect on the sensory dendrites or soma. The inhibition produced by colchicine could be produced by several effects. Colchicine may inhibit because it (1) disrupts the numerous intracellular microtubules which are a part of this sensory receptor's dendrite, (2) blocks axoplasmic transport of essential materials to the sensory dendrite, or (3) binds to tubulin or other proteins in the dendritic membrane.

Animals↗