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Biomedical subjects

R B Clark

Publications and source records attributed to R B Clark.

At least 145 records · Page 8Linked to original sources
PubMed1987

New causes of pneumonia, meningitis, and disseminated infections associated with immersion.

Immersion in water increases the risk of infection by certain microorganisms, including bacteria, fungi, and parasites. Physicians should be aware of this relationship because many of these infections are not normally encountered except in association with water exposure. The morbidity and mortality associated with these infections may be substantial. Clinical features of pneumonia, disseminated infection, and central nervous system infections are detailed.

Humans↗
PubMed1986

T cell lines derived from the spinal cords of mice with experimental allergic encephalomyelitis are self reactive.

Experimental allergic encephalomyelitis (EAE) is an animal model of T cell-mediated, central nervous system neuropathology that may be a relevant animal model for multiple sclerosis. EAE is usually induced by sensitization of animals with a xenogeneic myelin basic protein (MBP). Recently, MBP-reactive T cell lines and clones derived from lymphoid tissue of animals with EAE have proved very useful in elucidating certain aspects of the pathogenesis in EAE. However, questions relating to how T cells actually mediate the pathologic changes seen in EAE remain unresolved. We now report for the first time the derivation of long-term, interleukin 2-dependent T cell lines and sublines from a site of pathology in murine EAE--the spinal cord. All of the spinal cord-derived T cell lines and sublines were found to be "autoreactive" in that they responded to self (murine) MBP as well as to the xenogeneic immunogen, porcine MBP. The ability to derive T cell lines and sublines from the spinal cords of mice with EAE should now aid in the elucidation of pathogenetic mechanisms in EAE by allowing for a characterization of those T cells found at the site of pathology.

Animals↗
PubMed1986

Homologous desensitization of beta-adrenergic receptors in lymphoma cells is not altered by the inactivation of Ni (Gi), the inhibitory guanine nucleotide regulatory protein.

We had previously demonstrated that the cyc- mutant of S49 wild-type lymphoma cells both desensitizes and undergoes a sequestration-internalization of the beta-receptor in response to short-term treatment with adrenaline. The cyc- mutant of S49 wild-type lymphoma cells lacks the alpha s subunit of the stimulatory coupling protein Ns, but has fully functional Ni, the inhibitory component of the regulatory complex. This suggested that functional Ns was not required for desensitization. To examine the role of Ni in desensitization, both S49 wild-type and cyc- cells were treated with islet-activating protein under conditions that led to over 85% attenuation of Ni function in S49 wild-type cells and approx. 50% attenuation of Ni function in cyc- cells. This treatment had no effect on the adrenaline-induced desensitization of adenylate cyclase or the sequestration event measured by the apparent movement of beta-adrenergic receptors to a light-vesicle fraction. Further, the desensitization event, which occurs before the sequestration event, observable only in intact cells, was also not altered by islet-activating-protein pretreatment of S49 wild-type cells. The data suggest that a functional Ni is not required for desensitization in the S49 lymphoma cells.

Adenosine Diphosphate↗
PubMed1986

Comparison of steady-state electrophysiological properties of isolated cells from bullfrog atrium and sinus venosus.

Single electrode whole cell voltage-clamp experiments and frequency domain analyses have been used to study and compare the K+ currents in enzymatically dispersed single cells from the atrium and the sinus venosus (pacemaker region) of the bullfrog heart. Admittance measurements made near the "resting' or zero-current potential yield data from which the equivalent circuit of each cell type may be obtained. Data from both atrial and pacemaker cells are well-fitted by a model consisting only of parallel resistance-capacitative elements, as predicted from their micro-anatomy. Neither of these amphibian cardiac cells contain a transverse tubule system (TT) and both have very little sarcoplasmic reticulum (SR). These results complement and extend two earlier investigations: (i) Moore, Schmid and Isenberg (J. Membrane Biol. 81:29-40, 1984) have reported that in guinea pig ventricle cells (which do contain an internal membrane system consisting of transverse tubules and a substantial SR) the SR may be electrically coupled to the sarcolemma; (ii) Shibata and Giles (Biophys. J. 45:136a, 1984) have shown that although bullfrog atrial cells have an inwardly rectifying background K+ current, IK1, pacemaker cells from the immediately adjacent sinus venosus do not. Data from admittance measurements also provide evidence that a TTX-insensitive inward Ca2+ current is activated in the pacemaker range of potentials.

Action Potentials↗
PubMed1986

A1 and A2 adenosine receptors regulate adenylate cyclase in cultured human lung fibroblasts.

Adenosine stimulates and inhibits adenylate cyclase activity and cAMP levels in WI-38 and VA13 fibroblasts. The inhibitory effects appear to be mediated by both A1 receptors and the P-site. Results supporting these conclusions are as follows: Adenosine by itself increased cAMP accumulation in these cells. PGE1-stimulated cAMP accumulation was inhibited by adenosine in a concentration-dependent fashion. IAP treatment blocked adenosine inhibition of cAMP accumulation and adenylate cyclase activity and enhanced adenosine stimulation of cAMP accumulation in VA13 cells. Theophylline and MIX attenuated adenosine inhibition of cAMP accumulation. Adenosine analogs with substitutions in the purine ring inhibited PGE1-stimulated cAMP accumulation and adenylate cyclase activity. PGE1-stimulated cAMP accumulation was inhibited by the P-site agonist 2'5'-dideoxyadenosine, but this inhibition was not attenuated by MIX or IAP treatment. These data support the idea that adenosine may inhibit cAMP accumulation in VA13 or WI-38 cells by acting at an A1 receptor of the P-site. The decrease in cAMP accumulation mediated by the A1 receptor appeared to be due at least in part to an Ni-mediated inhibition of adenylate cyclase.

1-Methyl-3-isobutylxanthine↗
PubMed1986

Cell-mediated immunity in rheumatic disease.

Rheumatoid arthritis, systemic lupus erythematosus, and systemic sclerosis are the three most common systemic rheumatic diseases in which disordered immune function is thought to play a pathogenetic role. Each disease has different and characteristic abnormalities of the cellular immune system. In rheumatoid arthritis the identified abnormalities of immunoregulation are largely limited to specific antigens: Epstein-Barr virus and collagen. Systemic lupus erythematosus is characterized by exuberant B-cell activity with exaggerated humoral response, a diversity of autoantibodies, non-antigen-specific loss of suppressor cell function, and general suppression of cell-mediated immunity. In systemic sclerosis systemic defects of cellular and humoral immune function are mild, but the release of lymphokines and monokines at sites of inflammatory lesions is thought to be important in the pathogenesis of the disease. Similar immune cell-connetive tissue cell interactions are probably important in the propagation of rheumatoid synovitis. Thus, despite the many shared clinical and serologic features of these diseases as well as the presence of many patients who have clinically overlapping features of more than one of these entities, the immune defects and the immunopathogenesis of these disorders appear to be distinct.

Animals↗
PubMed1986

Phorbol ester-induced augmentation and inhibition of epinephrine-stimulated adenylate cyclase in S49 lymphoma cells.

The effects of 4-beta phorbol 12-myristate 13-acetate (PMA) on hormone and forskolin-stimulated adenylate cyclase were evaluated in S49 lymphoma cells. Treatment of wild type (WT) S49 cells with PMA caused stimulation, inhibition or had no effect on epinephrine stimulation of cAMP accumulation. The effect observed was dependent on the length of PMA treatment, the concentration of PMA and the concentration of hormone (or forskolin) used to stimulate cAMP accumulation. Longer treatment times with PMA and higher PMA concentrations favored the inhibitory effects. Pretreating WT with 0.5 microM PMA for 18 min caused an increase in the EC50 and maximal levels for epinephrine stimulation of cAMP accumulation. Thus inhibition was seen at relatively low epinephrine concentrations and augmentation with high concentrations. The inhibitory effects of PMA on epinephrine-stimulated adenylate cyclase activity were observed only at low free Mg++ concentrations (0.75 mM). The effects of PMA on PGE1-stimulated cAMP accumulation were similar to those observed for epinephrine. In S49 WT cells 100 nM PMA augmented 5 microM forskolin-stimulated cAMP accumulation; however with 100 microM forskolin, PMA effects were minimal. PMA also attenuated Gi-mediated Gpp(NH)p inhibition of forskolin-stimulated adenylate cyclase in both WT and cyc- membranes, resembling the effects of pertussis toxin. The effects of various phorbol analogues on epinephrine-stimulated cAMP accumulation were as follows: 4 beta-phorbol 12,13-didecanoate had similar effects to PMA, 4 alpha-phorbol 12,13-didecanoate had no effects and 1-oleoyl, 2-acetylglycerol augmented epinephrine-stimulated cAMP accumulation at concentrations greater than or equal to 5 microM. Our results are consistent with a dual mechanism of PMA action on adenylate cyclase involving protein kinase C-mediated phosphorylation of Gi and of the beta-adrenergic receptor, the former leading to augmentation and the latter to inhibition of hormone-stimulated adenylate cyclase.

Adenylate Cyclase Toxin↗
PubMed1985

Compartmentalized immune responses: antigen-specificity of cerebrospinal fluid T-cell lines maintained in the absence of antigen.

The generation of long-term interleukin 2-dependent T-cell lines from anatomically compartmentalized sites of pathology offers a unique approach to the investigation of certain autoimmune diseases. However, it is generally believed that antigen-specific T-cell lines and clones lose antigen reactivity and specificity when propagated in the absence of antigen. Therefore, the optimal application of this approach to such diseases in which the pathogenetic antigens are unknown may be difficult. In approaching this problem, we have recently demonstrated that a proportion of antigen-specific T-cell lines derived from the peripheral circulation can maintain antigen specificity if propagated with antigen-presenting cells alone or with antigen-presenting cells together with OKT3 antibody, but in either case in the absence of antigen. In this report we describe the use of this approach to maintain the antigen specificity of T cells obtained from an anatomically compartmentalized site of pathology--the cerebrospinal fluid from a patient with tuberculous meningitis. We report here that a proportion of the T-cell lines generated from such cerebrospinal fluid lymphocytes can be maintained as antigen specific in the absence of antigen if propagated with either antigen-presenting cells alone or with antigen-presenting cells and OKT3 antibody. The approach illustrated in this report should now find broad applicability in the investigation of a number of autoimmune disease.

Antibodies, Monoclonal↗
PubMed1985

Isolation of Serratia plymuthica from a human burn site.

The saprophytic bacterium Serratia plymuthica was recovered from a facial wound (burn) site of a pediatric patient. The clinical significance of the organism was undetermined due to its apparent eradication from this location by therapy with topical 1% silver sulfadiazine. Seeding of the burn with S. plymuthica may have occurred from contaminated moisture sometimes found on and around steam radiators.

Burns↗
PubMed1985

Maintenance of antigen specificity by human interleukin-2-dependent T cell lines. Use of antigen-presenting cells and OKT3 antibody in the absence of antigen.

The in vitro growth of T cells obtained from localized anatomic sites of pathology may offer a new approach to the investigation of certain human autoimmune diseases. However, if interleukin-2-dependent T cell cloning is to be useful in helping to elucidate putative pathogenetic antigens in these diseases, the expansion of the small number of T cells obtainable from localized anatomic sites of pathology will often have to be accomplished in the absence of these, as yet undetermined, antigens. At present, it is a generally held belief that antigen-responsive, interleukin-2-dependent T cell lines and clones will lose antigen responsiveness if propagated in the absence of specific antigen. Thus, the use of T cell cloning might be viewed as being of limited usefulness in the investigation of certain human autoimmune diseases. In this report we demonstrate that, when propagated in the absence of antigen, human tetanus toxoid-specific, interleukin-2-dependent T cell lines will indeed lose antigen reactivity. However, if propagated in the absence of antigen but in the presence of antigen-presenting cells, the tetanus toxoid reactivity of a subset of such lines can be maintained. Moreover, the propagation with OKT3 antibody, in addition to antigen-presenting cells, may be even more effective in maintaining antigen reactivity. These results may suggest a new approach to the use of T cell cloning technology in the investigation of certain autoimmune diseases.

Antibodies, Monoclonal↗
PubMed1985

Epinephrine-induced sequestration of the beta-adrenergic receptor in cultured S49 WT and cyc- lymphoma cells.

Pretreatment of either intact wild type S49 lymphoma cells (WT) or the uncoupled variants, cyc-, H21a, or UNC with epinephrine results in the redistribution of 20-30% of the beta-adrenergic receptors into a light vesicle fraction in sucrose gradients. Since the variants are uncoupled with respect to hormonal stimulation of adenylate cyclase, it appears that productive interaction with Gs is not required for the sequestration of beta-adrenergic receptors. Characterization of the epinephrine-induced redistribution of the beta-adrenergic receptor has revealed the following: The EC50 for the redistribution in WT cells was between 100 and 200 nM. Pretreatment of WT cells with 50 nM epinephrine for 30 min induced only a slight redistribution of receptors in sucrose gradients but produced a significant desensitization of adenylate cyclase. The desensitization was characterized by an increase in the Kact of epinephrine stimulation of adenylate cyclase while the Vmax was unaltered. Pretreatment with 10 microM epinephrine resulted in a significant decrease in the Vmax (50%) of epinephrine stimulation of adenylate cyclase and a 3-fold increase in Kact in the heavy vesicles. The beta-receptors in the light vesicle fraction of WT were uncoupled from adenylate cyclase and displayed low affinity for epinephrine binding, comparable to the cyc-. The "desensitized" receptor in the light vesicle fractions of cyc- was capable of stimulating adenylate following reconstitution with cholate extracts of WT membranes containing Gs. The molecular weight of the photolabeled beta-receptor in the light vesicle fractions (65,000 +/- 2,000) was not significantly different from the Mr 65,000 polypeptide photolabeled in the heavy fractions. The Mr 55,000 beta-receptor polypeptide was not detected in the light vesicles. Our results suggest first that the redistribution of the beta-receptor into light vesicles may follow an earlier stage of desensitization, and second that the beta-receptor in light vesicles while sequestered from Gs is capable of activating adenylate cyclase.

Adenylyl Cyclase Inhibitors↗