ABSTRACT
OBJECTIVE: Noninvasive assessment of the immediate and delayed cardiopulmonary response to a 2% aminophylline-based topical thigh reducing cream. DESIGN: Prospective, double-blind, randomized, counterbalanced study with application of: no cream (NC), placebo cream (PC) or 2% aminophylline cream (AC). SUBJECTS: Nine healthy women (aged: 23+/-3 y; weight: 58+/-3 kg; height: 165+/-7 cm; body fat: 19+/-6%; estimated maximal aerobic fitness VO2max): 40+/-4 ml/kg/min). MEASUREMENTS: Medical history, skin patch test, skinfolds, YMCA submaximal cycle ergometry test, psychological evaluations (POMS and Speilberger STAI-1). Pulmonary function and spectral analysis on heart rate variability, measured immediately post-and 4 h post-treatment, on three separate days within a three-week period. RESULTS: Pulmonary function did not change. The averaged R-R interval (ms) was significantly lower for the immediate post AC treatment, but returned to baseline in 4 h. CONCLUSION: Application of a 2% aminophylline-based thigh cream does not affect pulmonary function, however, it may cause a temporary, transient reduction in the averaged R-R interval.
Subject(s)
Aminophylline/pharmacology , Anti-Obesity Agents/pharmacology , Heart Rate/drug effects , Respiration/drug effects , Administration, Cutaneous , Adult , Aminophylline/administration & dosage , Anti-Obesity Agents/administration & dosage , Double-Blind Method , Female , Humans , Patch Tests , Prospective Studies , Reference Values , ThighABSTRACT
Although the glycoprotein (GP) Ib-IX-V complex and FcgammaRIIA are distinct platelet membrane receptors, previous studies have suggested that these structures may be co-localized. To determine more directly the proximity of GP Ib-IX-V and FcgammaRIIA, we assessed the effects of anti-GP Ibalpha monoclonal antibodies on FcgammaRIIA-mediated platelet aggregation and on the direct binding of polymeric IgG to human platelets. In addition, we directly examined the proximity of FcgammaRII and GP Ib-IX-V using flow cytometric fluorescence energy transfer and immunoprecipitation studies. Preincubation of platelets with either of two monoclonal antibodies (AN51 or SZ2) directed against GP Ibalpha completely blocked platelet aggregation by polymeric IgG. Similarly, these antibodies totally inhibited platelet aggregation by two strains of viridans group streptococci known to induce aggregation via FcgammaRIIA. In addition, AN51 and SZ2 significantly reduced the binding of polymeric IgG to washed fixed platelets. When assessed by flow cytometry, significant levels of bidirectional energy transfer were detected between FcgammaRIIA and GP Ibalpha, indicating a physical proximity of less than 10 nm between these receptors. This energy transfer was not due to high receptor density, because no homoassociative energy transfer was seen. Moreover, immunoprecipitation of FcgammaRIIA from platelet lysates also co-precipitated GP Ibalpha. These results indicate that GP Ibalpha and FcgammaRIIA are co-localized on the platelet membrane and that this association is not random.
Subject(s)
Antigens, CD/isolation & purification , Blood Coagulation Factors/isolation & purification , Blood Platelets/ultrastructure , Platelet Glycoprotein GPIb-IX Complex/isolation & purification , Receptors, IgG/isolation & purification , Antibodies, Monoclonal , Antigens, CD/metabolism , Blood Coagulation Factors/metabolism , Blood Platelets/drug effects , Blood Platelets/immunology , Blood Platelets/metabolism , Cell Membrane/drug effects , Cell Membrane/immunology , Cell Membrane/metabolism , Energy Transfer , Fluorescent Dyes , Humans , Immunoglobulin G/pharmacology , Platelet Aggregation , Platelet Glycoprotein GPIb-IX Complex/metabolism , Precipitin Tests , Protein Binding , Receptors, IgG/metabolismABSTRACT
The direct binding of platelets by bacteria is a postulated central mechanism in the pathogenesis of endocarditis. To address the role of binding more definitively, we employed Tn551 insertional mutagenesis of Staphylococcus aureus parental strain ISP479 to generate an isogenic variant (strain PS12) that bound platelets minimally. As compared with the binding of ISP479, the binding of PS12 to platelet monolayers was reduced by 67.2%. Similarly, the binding of PS12 to platelets in suspension was reduced by 71.3%, as measured by flow cytometry. The low-binding phenotype was transducible into both ISP479 and S. aureus Newman. Southern blotting indicated that a single copy of Tn551 was inserted within the chromosomes of PS12 and the transductants. When tested in a rabbit model, animals inoculated with PS12 were significantly less likely to develop endocarditis and had lower densities of organisms (CFU per gram) within vegetations and a decreased incidence of renal abscess formation, as compared with animals inoculated with the parental strain. The diminished virulence of PS12 was not attributable to a reduction in the initial attachment of organisms to the damaged endocardium, since 30 min after inoculation, PS12-infected animals had microbial densities on the valve surface comparable to those seen with the parental strain. These results indicate that the direct binding of Staphylococcus aureus to platelets is a major determinant of virulence in the pathogenesis of endocarditis. Staphylococcus-platelet binding appears to be critical for pathogenetic events occurring after the initial colonization of the valve surface, such as vegetation formation and septic embolization.