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Biomedical subjects

G Johnson

Publications and source records attributed to G Johnson.

At least 253 records · Page 14Linked to original sources
PubMed1993

Synthesis and pharmacological evaluation of 4a-phenanthrenamine derivatives acting at the phencyclidine binding site of the N-methyl-D-aspartate receptor complex.

A novel series of octahydrophenanthrenamines and their heterocyclic analogues have been synthesized as potential noncompetitive antagonists of the N-methyl-D-aspartate (NMDA) receptor complex. The compounds were evaluated for their affinity at the phencyclidine (PCP) binding site by determining their ability to displace [3H]TCP from crude rat brain synaptic membranes. A wide range of affinities were observed, with the most potent analogs possessing IC50's equivalent to that of the reference agent MK-801 (3, dizocilpine). NMDA antagonist activity was demonstrated by prevention of glutamate-induced accumulation of [45Ca2+] in cultured rat cortical neurons. Selected compounds were also studied in vivo to determine their ability to prevent the lethal effects of systemically injected NMDA in the mouse. In general, the SAR of the phenanthrenamine series may be summarized as follows: (a) for the amino group at C4a, NHMe > NH2 > NHEt >> NC5H10; (b) for the B-ring substitution, X = CH2 > S > O; (c) unsaturation of the C ring decreases receptor affinity; (d) cis-ring fusion between the B and C rings is desirable; (e) 6-hydroxy or 6-methoxy substitution of the phenanthrenamine system identified an additional hydrogen bonding interaction that substantially increased receptor affinity; (f) spiro analogues (such as 55, IC50 = 3400 nM), which altered the point of attachment of the C ring, caused a substantial reduction in PCP-site affinity. Molecules from this series were useful for refining a pharmacophore model consistent with previous models of the PCP site. In this model, the (R)-(+)-phenanthrenamine 13 superimposes closely onto MK-801 (3), and the angular 4a-amino group is believed to hydrogen bond with a putative receptor site atom. In the phenanthrenamine and thiaphenanthrenamine series, the (R)-(+)-enantiomers (9, 13, and 44) are more potent by approximately 5-10-fold than their corresponding (S)-(-)-enantiomers with respect to their affinity for the PCP site, their ability to prevent accumulation of [45Ca2+] in cultured neuronal cells, and their protection against the lethal effects of NMDA in mice. In general, there was no separation between the dose that prevented NMDA lethality and the dose that produced ataxia in mice, except in the case of the thiaphenanthrenamines 41 and 43. We have not yet obtained evidence that this small separation in activity offers a therapeutic advantage in the treatment of cerebral ischemia or other neurodegenerative disorders.

Animals↗
PubMed1993

Synthesis and pharmacological evaluation of hexahydrofluorenamines as noncompetitive antagonists at the N-methyl-D-aspartate receptor.

The noncompetitive (PCP) site of the N-methyl-D-aspartate (NMDA) receptor complex has been implicated in a number of pathologies, including the etiology of ischemic stroke. Recent testing has shown that cis-1,2,3,4,9,9a-hexahydro-N-methyl-4aH-fluoren-4a-amine (1), a rigid analog of PCP, is a potent antagonist at this site (IC50 = 30 nM for displacement of [3H]TCP). On the basis of this finding, a number of derivatives encompassing variations in stereochemistry, amine substitution and position, aromatic and aliphatic ring substitution, and heteroatom ring substitution have been prepared to explore the structure-activity relationships around this ring system. All compounds were evaluated for their PCP receptor affinity; potent compounds were also tested in vitro (cultured neurons) and in vivo (prevention of NMDA-induced lethality in mice). The present hexahydrofluorenamines demonstrated a wide range of potencies, with optimal affinity concentrated in analogs containing a heteroatom (sulfur) in the B ring (IC50 of 11 nM versus [3H]TCP for 16b), methyl substitution on the amine, and R stereochemistry at the 4a position. No significant improvement in affinity was seen with aromatic ring substitution. Aliphatic ring substitution, large amine substituents, and alterations in the position of amine substitution on the ring system resulted in a loss of potency. To explore the effect of simultaneous hydrogen bonding with a putative receptor atom from two directions, the 2-hydroxymethyl derivatives were prepared. This substitution resulted in a loss in receptor binding affinity. Molecular modeling, X-ray, and NMR studies have been used to determine an optimal conformation of the hexahydrofluoreneamines at the receptor site.

Animals↗
PubMed1993

Polystyrene chemistry affects vitronectin activity: an explanation for cell attachment to tissue culture polystyrene but not to unmodified polystyrene.

Tissue culture polystyrene (TCPS) supports good attachment of adherent cells whereas unmodified polystyrene (PS) does not, but the mechanism of this difference is not well characterized. We have compared TCPS and PS for the amounts of vitronectin (Vn) and fibronectin (Fn) which adsorb from the fetal bovine serum (FBS) component of the culture medium. The significance of the amounts of Vn and Fn which adsorbed onto TCPS and PS was determined by reference to the concentration dependence of the cell attachment activity of Vn and Fn when adsorbed onto TCPS and PS, assayed using human vein endothelial cells and BHK-21 fibroblasts. The amount of Vn which adsorbed onto TCPS from medium containing 3-30% (v/v) FBS was supraoptimal for the attachment of endothelial cells and fibroblasts. On PS, the amount of Vn which adsorbed from this medium was less than for TCPS and was suboptimal for cell attachment. Higher levels of Fn adsorbed onto TCPS than to PS, but even the amounts of Fn which adsorbed onto TCPS were suboptimal for cell attachment. We propose that the principal mechanistic difference between TCPS and PS for the initial attachment and spreading of cells is that more Vn adsorbs onto TCPS from the serum component of the culture medium.

Adsorption↗
PubMed1993

Self-correction of proton spectroscopic images for gradient eddy current distortions and static field inhomogeneities.

A postprocessing method of correcting for gradient eddy current distortions and inter-voxel static field inhomogeneity in spectroscopic imaging is presented. Data is acquired normally and all spatial processing is performed. The FID in each voxel is then digitally filtered to extract the signal from a single reference line. Phase multiplying the original FID by the phase of this reference signal corrects for gradient eddy currents and static field offsets. Computer simulations show that the method is robust with respect to noise, filter bandwidth and the presence of small lines close to the reference line. The method is demonstrated on proton spectroscopic images of phantoms.

Computer Simulation↗
PubMed1993

Length distribution of CDRH3 in antibodies.

Sequences of the third complementarity determining region of antibody heavy chains (CDRH3s) are listed according to their length. Human sequences vary from 2 to 26 amino acids residues, but less extensively in other species. When combined with the other five complementarity determining regions, this enormous length variation of CDRH3, together with amino acid substitutions in their sequences, can provide a very large number of antibody specificities and can influence the shape of antibody combining sites.

Amino Acid Sequence↗
PubMed1993

Progression of occlusive disease following femorofemoral crossover bypass graft.

Progression of distal disease is considered the most common cause of femorofemoral artery crossover bypass graft (FFBPG) failure. Twenty-seven patients with patient grafts (mean 53 months) were evaluated with segmental Doppler and duplex scan arterial studies for evidence of disease progression. In the early postoperative period (compared with preoperative levels), 26 patients (95.3%) showed a significant improvement (> 0.1) in the recipient limb ankle-brachial index (ABI) (mean increase of 0.38; SD = 0.24) and/or ankle spectral arterial waveform. However, there was a statistically significant decrease (p = 0.0001) in the donor limb ABI, and 12 patients (44.4%) had a > 0.1 deterioration. On long-term follow-up (compared with preoperative levels) this difference was no longer significant (p = 0.49); only seven donor limbs remained with a > 0.1 decrease in ABI. The recipient limbs maintained a significant improvement (> 0.1) in the ABI compared to preoperative levels (p < 0.0001; mean of 0.39; SD = 0.16) except for three limbs that had decreased by 0.1. However, eight patients (29.6%) developed an increase in their donor common femoral artery acceleration time > 133 msec and/or increased blood flow velocity without a simultaneous significant decrease in their recipient limb ABI. In the latter group the preoperative donor limb common femoral artery acceleration time and ABI and the immediate postoperative change in donor limb ABI were not significantly different (p > 0.05) than in the remaining patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Arterial Occlusive Diseases↗
PubMed1993

Altered expression and phosphorylation of amyloid precursor protein in heat shocked neuronal PC12 cells.

The pathology of the Alzheimer's disease (AD) brain, including amyloid plaques, neurofibrillary tangles and neuronal degeneration, indicates that neurons affected by AD exist under conditions of stress. In fact, the brains of AD patients undergo many changes classically associated with the heat shock response, which is one form of a stress response. These changes include reduced protein synthesis, disrupted cytoskeleton, increased number of proteins associated with ubiquitin, and the induction of heat shock proteins. To investigate the response of neurons to stress, we examined neuronal PC12 cells incubated at either 37 degrees C (control cells) or 45 degrees C (heat-shocked cells). After a 30 min exposure at 45 degrees C, the heat-shocked cells exhibited several features characteristic of the classical heat shock response including a 45% reduction in total protein synthesis, the induction of heat shock protein 72, and an increased phosphorylation of the protein synthesis initiation factor eIF-2 alpha. We used this cellular model system to study the neuronal response to stress specifically focusing on protein synthesis elongation factor 2 (EF-2) and the Alzheimer's amyloid precursor protein (APP), the precursor form of beta-amyloid peptide. Hyperphosphorylation of EF-2 has been observed in the neocortex and hippocampus of AD brain. However, in our system, we find no hyperphosphorylation of EF-2 in response to heat shock. Heat-shocked neuronal PC12 cells exhibited two additional APP-like polypeptides not present in controls. We also found a significant decrease in the phosphorylation state of APP in response to heat shock.(ABSTRACT TRUNCATED AT 250 WORDS)

Amyloid beta-Protein Precursor↗
PubMed1993

Reversible phosphorylation of tau to form A68 in heat-shocked neuronal PC12 cells.

A68, the primary protein constituent of Alzheimer's disease-associated neurofibrillary tangles, is an abnormally phosphorylated form of the microtubule-associated protein tau. We find that A68 is formed in neuronal PC12 cells when the cells are subjected to a heat shock (45 degrees C for 30 min). A68 was identified by immunoprecipitation with two different anti-tau antibodies (tau-2 and Alz50). Upon separation by SDS-polyacrylamide gel electrophoresis, the tau immunoprecipitates from heat-shocked cells exhibited an additional polypeptide of reduced electrophoretic mobility (approximately 68 kDa) when compared to control cells. A68 was formed with heat shock in the presence of cycloheximide, suggesting that its production occurred by post-translational modification of existing polypeptides. The tau/A68 polypeptides were identified as phosphoproteins by incorporation of 32P into the immunoprecipitates. The phosphorylation of tau to form A68 was reversed with recovery of the intact cells from the heat shock. Finally, immunoprecipitation of lysates from heat-shocked cells with antibodies to heat shock protein (hsp) 72/73 resulted in co-precipitation of tau with hsp 72, which indicates a stable complex formation between these two proteins. On the other hand, A68 remained unassociated with hsp during the heat shock. These results suggest that tau is reversibly phosphorylated to form A68 in neuronal PC12 cells under conditions of stress.

Animals↗
PubMed1993

Occupational experience and mortality among a cohort of metal components manufacturing workers.

We extended a previous cohort study of workers at a metal components manufacturing facility, which found elevated rats of lung cancer and unspecified nonmalignant respiratory diseases, by evaluating mortality rates from 1950 to 1987 for 3,630 workers categorized according to potential exposures to metal dusts, cutting oils/fluids, metal fumes, and solvents. Lung cancer rates were elevated for workers with exposure to metal dusts and cutting oils/fluids [standardized mortality ratio (SMR) = 1.8; 95% confidence interval (CI) = 1.2-2.6]. We found that elevated mortality was restricted to those workers first exposed during 1950-1959 (SMR = 2.5; 95% CI = 1.6-3.7) and that mortality rates were similar for workers with exposure durations of < 2 years, 2-9 years, and > or = 10 years, respectively. The elevated mortality for workers with < 2 years exposure and the lack of a trend with exposure duration raise the possibilities of either a short-lived occupational carcinogen or an unspecified confounding factor. Mortality rates for stomach or pancreatic cancers--cancers frequently reported to be in excess for metalworking populations--or for nonmalignant respiratory diseases were consistent with expected values for all exposure subgroups.

Cohort Studies↗
PubMed1993

Implantable transvenous cardioverter-defibrillators.

BACKGROUND: Implantable transvenous cardioverter-defibrillators offer a significant opportunity to decrease procedural morbidity and medical costs in the care of patients with life-threatening ventricular arrhythmias who otherwise would have required a sternotomy or thoracotomy for device insertion. The purpose of this study was to examine prospectively the safety, efficacy, and limitations associated with the use of a transvenously implanted, tiered-therapy cardioverter-defibrillator with antitachycardia pacing function in a consecutive population of 84 ventricular fibrillation (VF) and sustained ventricular tachycardia (VT) survivors. METHODS AND RESULTS: The index arrhythmia promoting transvenous cardioverter-defibrillator implantation was VF in 41 patients, VT in 27, and both VF and VT in 16. In each patient, transvenous defibrillation via a coronary sinus, a right ventricular, a superior vena caval, and/or a subcutaneous chest patch lead system was attempted. The pulsing methods used include two-electrode single-pathway pulsing or three-electrode dual-pathway simultaneous or sequential pulsing. A transvenous cardioverter-defibrillator was inserted if the defibrillation threshold (DFT) was < or = 20 J. Successful implantation of a transvenous cardioverter-defibrillator was possible in 80 of 84 (95%) patients. The mean implant DFT was 10.9 +/- 4.8 J. After cardioverter-defibrillator implantation, all patients were extubated in the operating room and sent to a standard telemetry ward for monitoring. No patient suffered a postoperative pulmonary complication or perioperative flurry of cardiac arrhythmias. Postoperative complications included lead dislodgments in eight, transient long thoracic nerve injury in one, asymptomatic left subclavian vein occlusion in two, asymptomatic small pericardial effusion in one, subcutaneous patch pocket hematomas in four, pulse generator pocket infection in one, and lead fracture in one. As experience was gained with the procedure, it was routine to discharge patients 3 days after surgery. The mean hospital stay was 6.0 +/- 2.4 days. Upon discharge, all patients returned to their prehospital activities including those with complications except for the patient with a pocket infection, who required intravenous antibiotic therapy. Patient survival using an intention-to-treat analysis was 98% over an 11 +/- 7-month follow-up period. During this time period, 31 of the 80 patients (39%) with transvenous lead systems were successfully treated by their device for sustained VT or VF. Antitachycardia pacing was used in 424 episodes of monomorphic VT and was successful in 371 (88%). All episodes of VF were aborted by the device. Antiarrhythmic drugs were used after device implantation in only eight of 80 patients (10%). CONCLUSIONS: Transvenous cardioverter-defibrillator implantation is practical in most candidates. Implant DFTs are usually low, surgical morbidity and postoperative complications are modest, therapy of VT and VF is efficient, and survival is excellent.

Algorithms↗
PubMed1993

A simplified, single-lead unipolar transvenous cardioversion-defibrillation system.

BACKGROUND: Transvenous implantable cardioverter-defibrillators provide significant advantages in the treatment of patients with life-threatening ventricular arrhythmias. However, present technology requires considerable electrophysiology expertise, multiple incisions, and long operative times for successful implementation. METHODS AND RESULTS: In this study, we present a prototype of a new, easy-to-insert unipolar transvenous defibrillation system that has the reliability of epicardial defibrillation but the ease of pacemaker insertion. This system incorporates a single anodal right ventricular defibrillation electrode using a 65% tilt biphasic pulse delivered to a 108-cm2 surface area pulse generator titanium alloy shell as an active cathode placed in a left infraclavicular pocket. Testing of this system was performed before implantation of a standard nonthoracotomy-transvenous defibrillation system in 40 consecutive patients with a history of ventricular tachycardia or fibrillation. The simplified unipolar single-lead system resulted in a defibrillation threshold of 9.3 +/- 6.0 J with 37 of 40 patients (93%) having a defibrillation threshold of less than 20 J. Moreover, the unipolar defibrillation system was efficiently used requiring only 3.4 +/- 0.8 ventricular fibrillation inductions to measure the defibrillation threshold and 100 +/- 28 minutes to implement. CONCLUSIONS: This new unipolar transvenous defibrillation system is as simple to insert as a pacemaker, requires few ventricular fibrillation inductions, demands less technical expertise, and provides defibrillation at energy levels comparable to that reported with epicardial lead systems. It should substantially reduce the morbidity, time, and cost of defibrillator implantation.

Adult↗
PubMed1993

Attachment of human bone cells to tissue culture polystyrene and to unmodified polystyrene: the effect of surface chemistry upon initial cell attachment.

Cell culture studies have often been used in the determination of the suitability of biomaterials as surfaces for the attachment and growth of cells. For such studies of surfaces for potential use in bone implants, cells derived from bone may be maintained in culture on tissue culture polystyrene (TCPS). We have determined the contribution that serum fibronectin (FN) or vitronectin (VN) make to the attachment and spreading of cells cultured from explanted human bone (bone-derived cells) during the first 90 min following seeding on culture surfaces. The attachment of bone-derived cells to TCPS was simulated two-fold by the addition of 10% (v/v) fetal bovine serum (FBS) to the seeding culture medium. The roles of FN and VN were determined by selective removal of the FN or VN from the FBS prior to addition to the culture medium. FBS from which the VN had been removed did not have this stimulatory activity. In contrast, the attachment of bone-derived cells onto TCPS from medium containing FN-depleted serum (which contained VN) was the same as when intact FBS was used. There was incomplete attachment of bone-derived cells (27% of cells) when seeded in medium containing FBS depleted of both VN and FN. Our results show that for human bone-derived cells, the attachment onto TCPS of cells planted in medium containing FBS during the first 90 min of culture is principally as a result of adsorption onto the surface of serum VN. As unmodified polystyrene (PS) has also been used previously as a model biomaterial surface, PS was compared to TCPS for attachment of the bone-derived cells. Attachment of bone-derived cells to TCPS was twice that onto PS, both when the medium was serum-free and when it contained FBS. Bone-derived cells attached to TCPS or PS onto which purified VN or FN had been precoated, with VN adsorbed onto PS being as effective as was VN adsorbed onto TCPS. With FN, there was an effect of the polystyrene surface chemistry which was evident in that suboptimal concentrations of FN had a slightly higher potency when adsorbed onto TCPS than did the same concentrations of FN coated onto PS. When preadsorbed onto TCPS, the potency of FN for attachment of bone-derived cells was at least equal to that of VN.

Bone and Bones↗