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Biomedical subjects

R G Johnson

Publications and source records attributed to R G Johnson.

At least 181 records · Page 10Linked to original sources
PubMed1985

Reoperation in congenital aortic stenosis.

Over a 22-year period, 81 patients underwent initial operations for critical aortic stenosis at our institution. Their ages ranged from 3 days to 20 years (mean, 7.3 +/- 5.9 years). Fourteen (17%) were infants less than 1 year old. Three children died perioperatively (3.7%). We have followed the survivors and 3 children who underwent initial operations elsewhere for a mean of 9.0 +/- 6.8 years (range, 2 to 23 years). To the present, 27 patients have undergone one reoperation (24 of our initial survivors) at a mean interval of 7.3 years, with 2 perioperative deaths. Ten of these patients have required a second reoperation at a mean interval of 3.7 years, with 2 deaths perioperatively. There were 3 late cardiac deaths after the initial procedure and 1 after a third operation. Actuarial reoperation-free survival is 56.7% at 10 years. While overall survival is 88.6% at 10 years, we find a significantly poorer survival among those patients with valvular stenosis compared with those with subvalvular lesions (p = 0.03). We believe that for children with all levels of aortic stenosis, good functional results and survival can best be obtained by follow-up, recatheterization, and reoperation.

Actuarial Analysis↗
PubMed1985

The effect of adding mannitol or albumin to a crystalloid cardioplegic solution: a prospective, randomized clinical study.

To determine if the myocardial protection afforded by a cold crystalloid potassium cardioplegic solution could be improved by the addition of either mannitol or albumin, a prospective clinical study was undertaken in which 58 patients undergoing elective aortocoronary bypass were randomized to one of three groups. Each group featured a different cardioplegic solution. The solutions were a standard potassium crystalloid solution, a solution containing mannitol sufficient to raise the osmolality by 20 to 30 mOsm, and a solution containing 5% albumin. Preoperative, intraoperative, and postoperative evaluation included serial measurements of ejection fraction, myocardial-specific isoenzyme, and hemodynamic indexes of performance. Electrocardiographic evaluation for perioperative myocardial infarction and the need for postoperative inotropic and mechanical support were also included. No differences were found among the groups. Therefore, although the use of mannitol or albumin has been shown to be beneficial in an experimental setting, superiority of either additive could not be demonstrated clinically.

Albumins↗
PubMed1985

Junctions between lens fiber cells are labeled with a monoclonal antibody shown to be specific for MP26.

A monoclonal antibody (mcAb) that recognizes an intracellular domain of the major lens membrane protein in both chicken and bovine lenses is described. Mice were immunized with chicken lens fiber cell membranes that had been washed with 7 M urea. Hybridomas were screened by means of enzyme-linked immunosorbent assays and the molecular specificities of the mcAbs were determined using electrophoretic transfer procedures, "Westerns." One of these mcAbs, an IgG designated B2, reacted with a single band of 28,000 Mr from the chicken embryo lens (MP28) and the analogous 26,000 Mr protein in the bovine lens (MP26). Monoclonal B2 was shown to be specific for these proteins, since (a) heating in SDS caused MP26 to aggregate and reduced B2 binding to the protein band at an Mr of 26,000 in Western transfer analysis; (b) apparent dimers were bound by B2 in Western transfers; (c) soluble protein fractions from the lens contained no detectable B2 antigens; and (d) a cyanogen bromide fragment of MP26 was bound by B2. Studies with several proteases indicated that the antigenic site for B2 resides on a 2-kd, protease-sensitive region at the C-terminal end of MP26 and MP28. Evidence for B2 binding on the cytoplasmic side of the membrane comes from labeling studies done at the ultrastructural level. These studies, utilizing indirect methods with peroxidase and colloidal gold markers, clearly demonstrated that B2 labels two types of junctional profiles. In our calf lens membrane preparations after tannic acid staining, the predominant type (80%) measured 16-18 nn thick, with the second type measuring only 12-14 nm. Chick embryo lens cells that had differentiated in vitro and formed groups of lens fiber-like cells (termed lentoids), fluoresced brightly only when they had been permeabilized before labeling with B2 and a fluorochrome-conjugated antibody. This binding was concentrated at the plasma membranes of cells within the lentoids, even outside areas of cell-cell contact. Surrounding epithelioid cells were not stained. Solubilized lens cultures, examined by Westerns, displayed a single immunoreactive band, which co-migrated with MP28.

Animals↗
PubMed1985

Lens junctional protein: analyzing MP26 with monoclonal antibodies.

Specific antibodies are versatile tools for analyzing cell surface proteins. This study involves the characterization of monoclonal antibodies which are specific for the junctional protein found in the lens fiber cell. This protein can be expected to include regions on the external membrane surface for junction formation, others on the cytoplasmic surface for regulation of junctional properties and, if cell-cell channels are indeed involved, transmembrane domains forming the hydrophilic connection between adjacent cytoplasms. Antibodies to these various regions would provide for an experimental analysis of the junctional protein, e.g., the identification of "active sites" for junction formation. Three monoclonal antibodies specific for the lens junctional protein in the chicken are described here. The first, termed B2, also recognizes the bovine junctional protein, MP26 (5). We have characterized the submolecular specificity of B2 and have found that it binds approximately ten amino acid residues from the C-terminus of MP26. In isolated lens junction preparations, B2 binds to the cytoplasmic surfaces of the lens junctions (both 12 nm and 16 nm thick forms). Thus, we consider MP26 a component of the lens junction. Monoclonal A4, the second antibody considered in detail here, was produced by immunization with lens membranes after treatment with low pH. We have found that lens junctional membranes are separated, or "split," by treatment at pH 2.5-3.0. It appears that A4 binds to the external surface of the junctional membrane; EM studies to confirm this are in progress. In order to map the A4 binding site within the chicken junctional protein and to explore the arrangement of this protein within the membrane, a number of procedures were used to generate fragments of MP26. These included reactions with N-chlorosuccinimide and proteases after acid treatment. Antibody binding to fragments was evaluated with immunotransfer ("Western") procedures. These studies mapped the A4 binding site to the center of the molecule and suggested that MP26 projected externally from the membrane at two different points. These results are consistent with a recent model, based on sequence data (6), for the arrangement of MP26 within the bovine lens membrane.

Animals↗
PubMed1984

Cardiovascular parameters after acute myocardial infarction and streptokinase administration in patients receiving coronary artery bypass grafts.

In 16 patients requiring coronary artery bypass grafting (10 control and 6 streptokinase patients), we compared the preoperative, operative, and postoperative cardiovascular parameters. Streptokinase patients had an acute myocardial infarction and attempted reperfusion with streptokinase before coronary artery bypass grafting. One patient failed to recannalize with streptokinase and one patient had reocclusion after withdrawal of heparin necessitating coronary artery bypass grafting. Examination of hemodynamic parameters revealed a lower preoperative mean blood pressure and an elevated pulmonary artery wedge pressure in streptokinase patients. The elevated pulmonary artery wedge pressure persisted through the postoperative period of observation. These results indicate that only minor differences exist between control and streptokinase patients. Emergency and elective coronary artery bypass grafting can be safely performed in patients treated with streptokinase for acute myocardial infarction without associated cardiogenic shock.

Blood Pressure↗
PubMed1984

Chicken embryo lens cultures mimic differentiation in the lens.

Embryonic chicken lenses, which had been disrupted by trypsin, were grown in culture. These cultures mimic lens development as it occurred in vivo, forming lens-like structures known as lentoids. Using a variety of techniques including electron microscopic analysis, autoradiography, immunofluorescence, and polyacrylamide gel electrophoresis, it was shown that the lentoid cells had many characteristics in common with the differentiated cells of the intact lens, the elongated fiber cells. These characteristics included a shut off of DNA synthesis, a loss of cell organelles, an increase in cell volume, an increase in delta-crystallin protein, and the development of extensive intercellular junctions. The cultures began as a simple epithelial monolayer but then underwent extensive morphogenesis as they differentiated. This morphogenesis involved three distinctive morphological types which appeared in sequence as an epithelial monolayer of polygonal shaped cells with pavement packing, elongated cells oriented end to end, and the multilayered, multicellular lentoids. These distinct morphological stages of differentiation in culture mimic morphogenesis as it occurs in the lens.

Animals↗
PubMed1983

Inability of seven-pinhole myocardial tomography to obtain accurate 201Tl kinetic data.

Seven-pinhole myocardial tomography has been reported to enhance the accuracy of thallium-201 (201Tl) studies in detecting patients with coronary artery disease. To determine if this approach can accurately assess regional 201Tl kinetics, 12 dogs with temporary occlusion (mean 15 min) of either the left anterior descending (LAD) (n = 6) or left circumflex (LCX) (n = 6) coronary artery were studied. Thallium-201 was injected and serial 7-pinhole images were acquired during occlusion and following reflow (mean duration 175 min). Time-activity analysis was obtained from normal and ischemic regions of interest in the central pinhole image and the reconstructed tomographic images (TOMO-ROI). Time-activity data from corresponding normal and ischemic regions were also obtained using a circumferential graph program (TOMO-MAX). In addition, regional myocardial 201Tl activity was recorded continuously with a cadmium telluride radiation probe sutured directly to the posterior myocardial wall. Defects in 201Tl distribution with subsequent partial or complete redistribution were present in 11 of 12 central pinhole images and tomographic studies. In the normal and ischemic anterior myocardial segments the percentage change in 201Tl activity in the central pinhole image from occlusion to the end of reflow correlated well with the percentage change in activity for both TOMO-ROI (r = 0.93) and TOMO-MAX (r = 0.85). In the normal posterior segments the percentage change in 201Tl activity in the central pinhole image correlated well with TOMO-ROI (r = 0.98) and TOMO-MAX (r = 0.87).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1983

The efficacy of postoperative autotransfusion in patients undergoing cardiac operations.

The efficacy of postoperative autotransfusion in lowering the requirement for banked-blood transfusion was studied in two groups, each having 168 patients, who underwent cardiac operations between April, 1979, and May, 1980. A Sorenson autotransfusion system was available for use in the autotransfusion group, whereas the control group received routine closed mediastinal drainage. Of the autotransfusion group, 81% met the criterion for autotransfusion (mediastinal losses of 450 ml or more during 4 hours), but only 61% of the autotransfusion group actually received autologous blood (mean autotransfusion volume, 399 +/- 25 ml). The patients receiving autologous blood required significantly less banked blood than their matched controls (447 +/- 60 ml and 744 +/- 83 ml, respectively; p less than 0.001). In the subgroup of patients with large mediastinal losses (more than 1,250 ml), this difference was even greater (autotransfusion, 642 ml compared with control, 1,145 ml; p less than 0.01). Postoperative autotransfusion is a simple, safe, and cost-effective method to reduce dependence on banked blood, especially when mediastinal losses are large. Obtaining maximum benefit requires familiarity of staff with the system and use of a consistent protocol.

Blood Transfusion↗
PubMed1983

Fracture-dislocation of the ankle with anterior dislocation of the fibula.

A fracture-dislocation of the ankle with fixed anterior displacement of the proximal fibular fragment is reported. The mechanism of this injury was likely eversion of the foot which produced rupture of the deltoid ligament and distal tibiofibular diastasis, and lateral dislocation of the talus. The proximal fibular fragment was probably displaced laterally and returned to a position anterior to the fibular notch of the tibia. This injury was extremely unstable and required open reduction with internal fixation, not only to reduce the anterior displacement of the fibula but also to repair the associated disruption of the syndesmotic ligament and anterior capsule. The key radiographic finding was anterior displacement of the distal fibula seen on the lateral view.

Ankle Injuries↗
PubMed1983

The response of articular cartilage to the in vivo replacement of synovial fluid with saline.

Experiments on rabbits were performed to determine whether continuous irrigation of a synovial joint produced irreversible degenerative changes in the articular cartilage. Continuous irrigation of a synovial joint permits observations of the effect on articular cartilage of altering the intra-articular environment. The right knee joints were irrigated with normal saline for varying periods, up to 14 days, and the left knees served as nonirrigated controls. One group of animals (short-term) was killed immediately following irrigation, while another group (long-term) was assessed after six months of cage activity following the irrigation period. Within each group, half were skeletally immature and half were mature. No differences in cell density, nuclei-to-lacunae ratio, or the ratio of calcified layer to total cartilage thickness were found in any group. Irrigation did produce a significant reduction in matrix staining with safranin-O. This was first detectable following three days of irrigation and was severe by seven to 14 days. Staining properties returned to normal after six months of cage activity. No obvious difference was found between immature and mature animals.

Age Factors↗
PubMed1982

H+ ATPase of chromaffin granules. Kinetics, regulation, and stoichiometry.

The chromaffin granule ATPase mediates an inwardly directed transport of H+ against concentration gradients, thereby forming and maintaining an electrochemical transmembrane H+ gradient. The kinetics of this ATPase, its activity modulation by changes in electrochemical H+ gradients, and the stoichiometry between H+ transport and ATP hydrolysis were studied in intact bovine chromaffin granules, resealed chromaffin granule ghosts, and highly purified fragmented chromaffin granule membranes. In fragmented membranes the H+ ATPase has a KM for ATP of 69 microM, a maximum of activity at pH 7.3, and a Vmax of 111 nmol/min/mg of protein at 20 degrees C. Trimethyl tin inhibits the ATPase at much lower concentrations than dicyclohexylcarbodiimide, whereas oligomycin, reserpine, and other inhibitors were without effect. In intact chromaffin granules, the ATPase activity was stimulated up to 300% by collapsing the H+ transmembrane gradients. H+/ATP stoichiometry was measured in resealed chromaffin ghosts devoid of ATP and catecholamines under conditions where no net pH changes occur upon ATP hydrolysis. After addition of ATP, the rates of H+ accumulation in the ghosts and ATP hydrolysis were both linear for about 60-100 s, and the ratio of H+ to ATP was 1.71. These data indicate that the H+ ATPase of chromaffin granules has both kinetic similarities and dissimilarities with other known H+ ATPases. The regulation by changes in H+ gradients and the fixed H+/ATP ratio of this ATPase is further evidence of its primary role in establishing electrogenic H+ translocation and H+ gradients in chromaffin granules.

Adenosine Triphosphatases↗
PubMed1982

Catecholamine transport and energy-linked function of chromafffin granules isolated from a human pheochromocytoma.

The structure and function of chromaffin granules of human pheochromocytoma was extensively investigated in a highly purified granule fraction obtained from a single specimen of human pheochromocytoma tissue. Pheochromocytoma chromaffin granules were analyzed for catecholamine, ATP, enkephalin, phospholipid, cytochrome and ion content. Using a variety of techniques it was found that the membrane of these granules is highly impermeable to Na+, K+, and H+, and that the intragranular pH was maintained at 5.1 irrespective of suspending media. The presence of MgATP induces a transmembrane potential (delta psi) across the membrane of these granules which is positive inside and which corresponds to 90 mV. Both delta pH and delta psi are coupled to biogenic amine accumulation into the granules in a process which is reserpine sensitive. These properties are compared with those of chromaffin granules isolated from normal human tissue or from other animal species and are discussed in terms of possible explanation at a biochemical or subcellular level of the clinical manifestation of the pheochromocytoma.

Adenosine Triphosphate↗
PubMed1982

Mechanisms of accumulation of tyramine, metaraminol, and isoproterenol in isolated chromaffin granules and ghosts.

The effects of the transmembrane pH gradient (delta pH) and the transmembrane potential gradient (delta psi) on the uptake of several sympathomimetic amines were investigated, using bovine adrenal chromaffin granules isolated in isotonic sucrose. As previously described [R. Johnson and A. Scarpa, J. Biol. Chem. 254 3750 (1979)], freshly isolated chromaffin granules maintain an intragranular pH of 5.5 as measured by [14C] methylamine distribution and, in the presence of ATP, generate a delta psi of 80 mV, positive inside, as measured by [14C] methylamine distribution. When tyramine, metaraminol, and isoproterenol (1-50 mM) were added to well-buffered suspensions of granules at pH 7.0, a dose-related alkalinization of the granule interior was observed. Study of the time-resolved influx of the same amines labeled radiochemically (5-21 microM) revealed that all the amines were accumulated against an apparent concentration gradient. However, while accumulation of [14C] serotonin and [3H] isoproterenol was totally inhibited by reserpine, [14C] tryramine accumulation was inhibited by only 60% and [14C[ metaraminol uptake was unaffected. The ATP-dependent generation of a delta psi produced a stimulation of amine uptake in the order: serotonin greater than isoproterenol greater than tyramine; metaraminol accumulation was not enhanced by ATP addition. The relationship between the electrochemical proton gradient (delta micro H+) and the electrochemical gradient for each of the sympathomimetic amines (delta micro A) was investigated utilizing chromaffin ghosts devoid of endogenous matrix gradients or components. All amines were accumulated in the presence of delta pH alone. In the presence of delta psi alone, [14C] serotonin, (14C] tyramine, and [3H] isoproterenol were accumulated, but no [3H] metaraminol uptake was demonstrable. The results indicate that serotonin and isoproterenol accumulated in isolated chromaffin granules and ghosts via a reserpine-sensitive mechanism, driven by the magnitude of the electrochemical proton gradient. Conversely, metaraminol permeated the membrane of the chromaffin granule through the apolar lipid phase and distributed according to the delta pH alone. Tyramine uptake proceeded by both mechanisms. The implications of the mechanism of accumulation of these potent physiologic and pharmacologic agents for their in vivo action are discussed.

Adenosine Triphosphate↗