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J K Johnson

Publications and source records attributed to J K Johnson.

18 recordsLinked to original sources
PubMed1992

Mechanistic investigation of medium-chain fatty acyl-CoA dehydrogenase utilizing 3-indolepropionyl/acryloyl-CoA as chromophoric substrate analogues.

The CoA derivative 3-indolepropionyl-CoA (IPCoA) serves as a competent pseudosubstrate for the medium-chain fatty acyl-CoA dehydrogenase (MCAD)-catalyzed reaction. The reaction product trans-3-indoleacryloyl-CoA (IACoA) exhibits a characteristic UV-vis absorption spectrum with lambda max = 367 nm and epsilon 367 = 26,500 M-1 cm-1. The chromophoric nature of IACoA allows us to measure the direct conversion of substrate to product (at 367 nm) without recourse to absorption signals for either the enzyme-bound flavin or the coupling electron acceptors, as well as probe the enzyme site environment. The interaction of IACoA with medium chain fatty acyl-CoA dehydrogenase (MCAD)-FAD is characterized by resultant (spectra of the mixture minus the individual components) absorption peaks at 490, 417, and 355 nm. These absorption peaks increase in magnitude as the pH of the buffer media decreases. Transient kinetic analysis for the interaction of MCAD-FAD with IACoA suggests that the formation of the enzyme-IACoA complex proceeds in two steps. The first (fast) step involves the formation of an E-IACoA collision complex, which [formula: see text] is isomerized (concomitant with changes in the protein structure) to an E*-IACoA complex in the second (slow) step. We have studied the effect of pH on Kc, k2, and k-2. While Kc shows practically no dependence on pH (within a 2-fold variation between pH 6.0 and 9.5), k2 and k-2 show a strong dependence on pH. Both k2 and k-2 exhibit a sigmoidal dependence on the pH of the buffer media, with pKa's of 7.53 and 8.30, respectively. In accordance with the model presented herein, the pKa of 7.53 represents an enzyme site group which is involved in the interaction with IACoA within the E-IACoA collision complex. This pKa is perturbed to 8.30 upon isomerization of the collision complex. The pH-dependent changes in k2 and k-2 are such that the equilibrium distribution between E-IACoA and E*-IACoA is favored to the latter complex (by about 20-fold) at lower pH than at higher pH. A cumulative account of the spectral, kinetic, and thermodynamic properties of the enzyme-IACoA complexes has allowed us delineate the microscopic pathway by which the E-IACoA isomerization (presumably via protein conformational changes) is coupled to the proton equilibration steps.

Acyl Coenzyme A↗
PubMed1992

Using serial observations to identify predictors of progression to AIDS in the Toronto Sexual Contact Study.

The Toronto Sexual Contact Study comprises a cohort of 249 male sexual contacts of men with HIV disease which has been followed every 3 months for almost 5 years. On enrollment 143 were seropositive and 16 seroconverted during the follow-up period. By 31 December 1989, 41 of the 159 seropositive cohort members had developed AIDS. Using Cox relative risk regression models, we investigated the association of a number of laboratory and clinical variables and progression to AIDS. Fixed covariate models examined laboratory variables from the enrollment visit of cohort members, with time calculated from this date. In models assessing time dependent covariates, time was calculated from the estimated date of HIV infection. In the univariate models of either fixed or time dependent covariates, many variables were significantly associated with risk of progression to AIDS (T4 cell count, T4/T8 ratio, blastogenic responses to phytohemagglutinin, concanavalin A, and pokeweed mitogen, serum IgA, appearance of p24 antigen, and the development of oral hairy leukoplakia, thrush, or herpes zoster). Appearance of persistent generalized lymphadenopathy was not associated with increased risk of progression. In the multivariate model which evaluated fixed laboratory covariates, T4/T8 ratio, IgA level, and PHA response at enrollment were significantly associated with elevated risk.(ABSTRACT TRUNCATED AT 250 WORDS)

AIDS Serodiagnosis↗
PubMed1991

Labeling of human retinohypothalamic tract with the carbocyanine dye, DiI.

The carbocyanine dye, DiI, was used to demonstrate human retinohypothalamic tract (RHT) projections in 6 normal human postmortem brains. In 5 of 6 brains, labeling was seen extending from the site of implantation in the distal optic nerve to both the ipsilateral and contralateral suprachiasmatic nuclei. This study confirms the presence of RHT projections in humans, and demonstrates the usefulness of DiI for neuronal tracing in human postmortem tissue.

Cadaver↗
PubMed1991

Interaction of ligands with pig heart citrate synthase: conformational changes and catalysis.

The fluorescence polarization of 8-hydroxypyrene (1,3,6)trisulfonate (HPT) increases upon interaction with pig heart citrate synthase. Titration of HPT with increasing concentrations of citrate synthase exhibits a hyperbolic saturation behavior, from which the dissociation constant of the enzyme-HPT complex (3.64 +/- 0.3 microM) was determined. The enzyme-HPT interaction is competitively inhibited by oxaloacetate (but not affected by acetyl CoA) with a Ki of 4.3 +/- 1.8 microM. This value is similar to the dissociation constant (Kd = 4.5 +/- 1.6 microM) for the enzyme-oxalocetate complex (determined in the absence of any effector ligand), as well as to the Km for oxaloacetate (3.9 +/- 0.7 microM) in a steady-state citrate synthase catalyzed reaction at a saturating concentration of acetyl CoA. However, the dissociation constant for the citrate synthase-oxaloacetate complex determined by the urea denaturation method is at least 25-fold lower than those determined by the other methods. This suggests an effector role of urea in strengthening the enzyme-oxaloacetate interaction. At low nondenaturing concentrations, urea inhibits the citrate synthase catalyzed reaction in an uncompetitive manner with respect to oxaloacetate, i.e., the Km for oxaloacetate decreases with an increase in urea concentration. This further suggests that urea stabilizes the interaction between citrate synthase and oxaloacetate. The effect of urea is specific for the substrate oxaloacetate, and not for the substrate analogue, HPT, although both these ligands bind citrate synthase with equal affinities, and protect the enzyme against thermal denaturation with equal magnitudes. The results presented herein are discussed in the light of known conformational states of the enzyme.

Acetyl Coenzyme A↗
PubMed1991

In vitro activity of the human neutrophil cathepsin G on Eimeria tenella sporozoites.

The role of human neutrophil cathepsin G (Cat G) on Eimeria tenella sporozoites was studied in vitro. Sporozoites were incubated for 2 hr at 37 C in PO4 buffer, 0.9% NaCl (PBS), pH 7.6 in the presence of Cat G (50 micrograms/ml), diisopropyl fluorophosphate-inhibited Cat G (DFP-Cat G) (50 micrograms/ml) or PBS alone, prior to being inoculated into embryonated eggs. As judged by oocyst production on day 7 postinoculation, embryo mortality and the hemorrhage scores, both Cat G and DFP-Cat G demonstrated anticoccidial activity; greater activity was obtained with the DFP-Cat G. Sporozoites were exposed also to increasing concentrations of native and trypsin-digested DFP-Cat G (0-100 micrograms/ml) under the same conditions. Significant protection (37% and 49% for native and digested DFP-Cat G, respectively) was obtained with a low concentration (5 mu/ml), and higher concentrations resulted in 70% and 84% protection, respectively. The primary bactericidal domain of Cat G, the HPQYNQR peptide, at 3 concentrations (25, 50, and 100 micrograms/ml), reduced the oocyst production by 46%, 16%, and 15%, respectively. The anticoccidial activity of Cat G may involve a peptide fragment different from the antimicrobial domain of the enzyme.

Amino Acid Sequence↗
PubMed1990

Studies on the stage of action of lasalocid against Eimeria tenella and Eimeria acervulina in the chicken.

Broiler chickens in battery pens were either fed a diet containing 100 ppm lasalocid or no drug for 24 h prior to inoculation with sporulated oocysts of Eimeria tenella or Eimeria acervulina. Different groups of birds remained on medicated feed for 24, 48, 72, 96, 120 or 144 h after inoculation. Conversely, other groups started on an unmedicated diet, were given medicated feed at different times after oocyst inoculation. Starting lasalocid medication 24 h (E. tenella) or 48 h (E. acervulina) after inoculation reduced the lesions and improved the weight gain. There was no significant difference in performance of birds after withdrawal of the drug at 48 h (E. tenella) or 72 h (E. acervulina) and thereafter. Starting lasalocid medication at 96 or 120 h did not suppress but rather reduced oocyst production.

Animals↗
PubMed1990

Cofactors of progression to acquired immunodeficiency syndrome in a cohort of male sexual contacts of men with human immunodeficiency virus disease.

In a cohort of 249 male sexual contacts of men with acquired immunodeficiency syndrome (AIDS) or an AIDS-related condition in Toronto, Ontario, Canada, 143 cohort members were seropositive on enrollment and 16 seroconverted between initial recruitment in July 1984 to July 1985 and December 1988. Data on age, smoking and drinking status, recreational drug use, and history of sexually transmitted diseases and other diseases were obtained from interviews at induction and during follow-up on the cohort members every 3 months. Cox relative risk regression models, in which time was calculated from estimated date of human immunodeficiency virus (HIV) infection for seroprevalent cohort members and from 90 days prior to the first positive test for seroconverters, examined the potential effect of use of a variety of recreational drugs and the occurrence of selected infections on the risk of development of AIDS. Thirty-five cohort members developed AIDS while under study. No significant association with risk of progression to AIDS was noted for use of various recreational drugs (singly or in combination), history of specific infections, age at enrollment, or smoking and drinking status at enrollment. Only estimated duration of HIV infection appeared to be associated with increasing risk of development of AIDS.

Acquired Immunodeficiency Syndrome↗
PubMed1989

Blunt arterial injuries associated with multiple trauma.

In this retrospective study, we reviewed the records of 62 patients who were treated at a tertiary care community teaching hospital between 1977 and 1987 for major arterial injury caused by blunt trauma. Nearly half (45%, 28/62) of the injuries were to the thoracic aorta, 24% (15/62) were to arteries of the head and neck, 21% (13/62) were to the arteries of the extremities, and 10% (6/62) were to abdominal arteries. Twenty-eight thoracic aortic transections were repaired, 25 with Dacron grafts and three by primary repair. One patient developed an adventitial hematoma in the thoracic aorta, which was evacuated. Two patients required nephrectomies secondary to renal artery injury. Ten patients with internal carotid artery dissection were successfully treated with anticoagulation therapy. Eight (13%) of the 62 patients died: four from exsanguination, one from cardiac tamponade, one from renal failure, one from pulmonary emboli, and one from cerebral infarction secondary to intracerebral edema. Such injuries are amenable to treatment, with patient and end-organ viability, if recognized and treated promptly by the trauma surgeon.

Abdomen↗
PubMed1979

Floor pen studies on the anticoccidial efficacy of arprinocid in turkeys.

Arprinocid was tested against coccidiosis in turkeys reared in floor pens. Contamination of pens with Eimeria meleagrimitis, E. adenoeides, and E. gallopavonis was accomplished by the infected seeder bird technique. Performance improved with increasing levels of arprinocid through 120 ppm, but weight gains were depressed at 180 ppm. Lesion scores were significantly and progressively reduced by treatment with 60, 90, 120, and 180 ppm of arprinocid. Control of lesion scores and performance of birds treated with 120 ppm of arprinocid were comparable to the amprolium (125 ppm) treatment. In a separate experiment, the oocyst passage of E. meleagrimitis and of E. adenoeides was progressively reduced by increasing levels of arprinocid, and sporulation rate was decreased. Sportulation was not obtained in oocysts from birds fed arprinocid at 120 ppm or higher.

Adenine↗
PubMed1979

Anticoccidial activity of narasin in battery raised broiler chickens.

The anticoccidial activity of the ionophorus antibiotic narasin was tested against six species of coccidia (Eimeria acervulina, Eimeria mivati, Eimeria maxima, Eimeria necatrix, Eimeria brunetti, and Eimeria tenella) in battery-raised broilers. Feeding ration medicated with 60, 80, or 100 ppm narasin significantly improved weight gains during the periods of D 0 to D7 and D 0 to D 14 (D 0 = day of inoculation with sporulated oocysts), compared with the weight gains in corresponding inoculated groups fed unmedicated feed. A similar protective effect of the medication was seen with feed conversion ratios (feed consumed/bird weight) and coccidiosis-induced mortality. With most species studied, 40 and 60 ppm narasin was not as efficacious as 80 or 100 ppm. The maximum numerial improvement in weight gain and feed conversion ratio was with 80 ppm narasin. Gross intestinal lesion scores were reduced by medication compared with the scores in birds fed unmedicated feed. The overall trend was for a larger reduction in lesion score with higher drug levels. Narasin at 80 or 100 ppm was generally more effective in controlling individual or mixed species infections of coccidia than 99 ppm monensin.

Animals↗
PubMed1976

Evaluation and restoration of endodontically treated posterior teeth.

The increasing acceptance and utilization of endodontic therapy for the preservation of broken-down posterior teeth presents the practitioner with many difficult restoration problems. If 25% or more of the coronal tooth structure is lost, some form of artificial dentin augmentation is required to provide adequate support and retention for the final restoration. Criteria to be used in the prerestorative evaluation of endodontically treated teeth are presented. Various methods and techniques to protect the remaining toth structure from fracture and to provide support and retention for the final restoration are described.

Composite Resins↗
PubMed1976

Effects of high environmental temperatures on cecal coccidiosis.

Maintenance of broiler or White Leghorn cockerels in a high temperature (32 C.), high humidity (90%) environment for six days following artificial infection with Eimeria tenella significantly reduced coccidiosis severity as compared to birds similarly infected and maintained in an ambient environment averaging 24.5 C., and 62% humidity. Disease severity was determined by lesion scores, fecal scores, and hematocrit readings. The heavier broiler stock maintained body temperatures of 0.7-1.6 C. higher than those birds kept at ambient temperatures for the entire six days. Body temperatures of heat-treated Leghorn cockerels were 0.9 C. higher during the first 48 hours post-inoculation (P.I.), but similar to ambient temperature controls for the last four days. Elevated body temperatures during the first two days following inoculation impaired the survival and development of some parasites while shorter periods (24 hours) of heat treatment did not affect parasite survival.

Animals↗
PubMedDate not supplied

Evaluation of treatment with single-dose ampicillin/sulbactam with probenecid or ceftriaxone in patients with uncomplicated gonorrhea.

This study compared ampicillin/sulbactam plus probenecid with ceftriaxone for treatment of uncomplicated gonorrhea. Of the 297 men and women who were enrolled and randomized to receive either ampicillin/sulbactam (1.0 g/0.5 g) with probenecid (1 g) or ceftriaxone (0.25 g), 274 patients were evaluable. Both ampicillin/sulbactam and ceftriaxone were administered by intramuscular injections. Patients were gonococcal contacts, had positive culture results for Neisseria gonorrhoeae, or had clinical evidence of gonorrhea. Specimens for gonococcal cultures were collected from the cervix (female patients), urethra, rectum, and pharynx at pretreatment and test-of-cure visits. The presence of N. gonorrhoeae and a test-of-cure visit were required for drug efficacy analysis. Of the 274 evaluable patients, 195 (71.2%) had positive culture results for N. gonorrhoeae. Cure was achieved in 93 (94.9%) of 98 patients receiving ampicillin/sulbactam with probenecid and in 96 (99.0%) of 97 patients receiving ceftriaxone. Penicillinase-producing N. gonorrhoeae strains were found in 21 (10.8%) patients; these were eradicated by either ampicillin/sulbactam with probenecid (N = 9) or ceftriaxone (N = 12). Overall, the two drug regimens were very well tolerated and no serious adverse effects were noted. Ampicillin/sulbactam with probenecid may be useful as single-dose therapy in patients with uncomplicated genitorectal gonorrhea.

Adolescent↗
PubMedDate not supplied

Human type A botulism and treatment with 3,4-diaminopyridine.

3,4-diaminopyridine was evaluated for its ability to improve muscle strength, respiratory function and electromyographic compound muscle action potentials in human botulism. In a double blind, placebo controlled study, 3,4-diaminopyridine failed to improve these parameters in a 31-year old patient with severe food-borne type A botulism. The addition of an anti-cholinesterase medication to the 3,4-diaminopyridine did not add any benefit. Lack of clinical improvement from 3,4-diaminopyridine in this patient differed from some reports of benefit in animals experimentally poisoned with type A botulinum toxin.

4-Aminopyridine↗
PubMedDate not supplied

Efficacy of arprinocid against coccidiosis of broilers in battery and floor-pen trials.

Medication of broilers with arprinocid [9-(2-chloro-6-fluorophenylmethyl)-9H-purine-6-amine] gave protection against the effects of coccidiosis in both battery and floor-pen trials. In battery trials, efficacy was tested on single-species inoculations of Eimeria acervulina, E. mivati, E. necatrix, E. maxima, E. Brunetti, and E. tenella. Two strains of each species recently recovered from the field, were tested separately. In floor-pen trials, all six species, both field and laboratory strains, were used as a mixed infection. In batteries, 60 and 70 ppm essentially eliminated coccidiosis-induced mortality and weight depression. Effects of 50 ppm on weight gain were variable. The effectiveness of different medication levels varied between strains within a species. In floor-pen trials, 40, 60, or 80 ppm was effective in controlling mortality and weight depression, and increasing feel-conversion ratios. All levels were significantly as effective as monensin in protecting against coccidiosis. With severe exposure to coccidia, 60 and 80 ppm gave significantly lower lesion socres than did 40 ppm of arprinocid or 120 ppm monensin.

Adenine↗