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Biomedical subjects

M S Johnson

Publications and source records attributed to M S Johnson.

At least 19 recordsLinked to original sources
PubMed1997

The three-dimensional structure at 2.4 A resolution of glycosylated proteinase A from the lysosome-like vacuole of Saccharomyces cerevisiae.

The crystal structures of glycosylated native proteinase A, an aspartic proteinase found in the vacuole of Saccharomyces cerevisiae, and its complex with a difluorostatone-containing tripeptide have been determined by molecular replacement to 3.5 A and 2.4 A resolutions, respectively. Superposition of the bound and native forms gave an r.m.s. difference of 0.6 A largely reflecting the poor resolution of the native crystal structure. The secondary and tertiary structures are highly similar to those found in porcine pepsin and lysosomal cathepsin D; superposition of the structure of proteinase A bound to the difluorostatone inhibitor on those of pepsin and cathepsin D gave pairwise r.m.s. differences for C(alpha) atoms of 1.36 A and 0.88 A. Most differences occur in loop regions. Comparison of the structure of the proteinase A-difluorostatone complex with that of endothiapepsin bound with the same inhibitor shows that the conformation and hydrogen bond interactions of the inhibitor in the active site are very similar, even though the enzymes have only 27% sequence identity. Electron density for the crystal structure of the proteinase A complex reveals five residues of the oligosaccharide structure attached to Asn67: Man-(1 --> 2)-alpha-Man-(1 --> 3)-beta-Man-(1 --> 4)-beta-GlcNAc-(1 --> 4)-beta-GlcNAc-Asn-67. The first three residues of the oligosaccharide cover the same region of the protein surface as those of the oligosaccharide attached to the equivalent position in cathepsin D. The second carbohydrate attachment site is disordered beyond the first carbohydrate residue in both enzymes.

Aspartic Acid Endopeptidases↗
PubMed1997

Acute oral toxicity study of pyridostigmine bromide, permethrin, and DEET in the laboratory rat.

This study investigated the lethal interaction of pyridostigmine bromide (PB), permethrin, and DEET when given to adult male rats by gavage and was separated into two phases. Phase I determined the acute oral lethal dose-response relationship of each compound with the vehicle, propylene glycol. Phase II was divided into two portions: a dose-response study using probit units obtained from phase I [lethal dose (LD) 16, 30, 50, 70, and 84], and an interaction study that contained low levels (calculated LD16, additive LD32) of the two compounds while the concentration of the third compound was varied. Rats were fasted overnight, dosed, and observed for 14 d. A significant increase in lethality occurred when PB, permethrin, and DEET were given concurrently when compared to expected additive values. Furthermore, solutions containing PB and permethrin or PB and DEET also caused a significant increase in lethality when compared to expected additive values. This information suggests that lethality in this study was more than an additive effect.

Animals↗
PubMed1997

Fluoride-induced lesions in the teeth of the short-tailed field vole (Microtus agrestis): a description of the dental pathology.

The effect of fluoride on the appearance of the teeth of the short-tailed field vole, Microtus agrestis, was investigated in both wild animals collected from field sites affected by different levels of industrial fluoride contamination and laboratory-reared animals consuming experimental grass diets of known fluoride concentration or with known fluoride concentrations in drinking water. The extent and severity of lesions on the surface and structure of both incisors and molars are described as six lesion types and related to the amount of biologically available orally ingested fluoride. In the incisors of voles consuming relatively low fluoride diets, lesions are mainly confined to those resulting from disruption of enamel pigmentation expressing itself as concentric bands of pigmentation-free areas on incisor surfaces. The visible effects on molars at low fluoride levels are confined to minor alterations in surface appearance. At higher levels of available dietary fluoride, effects on enamel pigmentation are superseded by alterations in the formation, composition, and strength of both enamel and dentine. The incisors exhibit a marked to severe increase in the cutting tip erosion rates with comparable increases in the extent of abnormal surface changes (enamel hypoplasia) and the loss of enamel pigmentation. The grinding surfaces of molars from animals exposed to high levels of dietary fluoride exhibit increasingly severe erosion of outer enamel regions, combined with cavitation and staining of the exposed central dentine. The mechanisms through which fluoride elicits increasingly visible and pathological alterations to the surface and subsurfaces of rodent teeth are discussed.

Animals↗
PubMed1997

Isolation and characterization of five actin cDNAs from the cestode Diphyllobothrium dendriticum: a phylogenetic study of the multigene family.

Five cDNAs (pDidact2-pDidact6), representing different actin genes, were isolated from a Diphyllobothrium dendriticum cDNA library, and the DNA as well as the putative amino acid sequences were determined. The corresponding Didact2 and Didact4 genes code for peptides 376 amino acids long, with molecular weights 41,772 and 41,744 Da, respectively, while the deduced Didact3 protein is 377 amino acids long and weighs 41,912 Da. The pDidact5 and -6 cDNAs lack nucleotides corresponding to three to six amino acids at the amino-terminus. Two of the five cDNAs contain the conventional AATAAA as the putative polyadenylation signal, one has the common variant ATTAAA, whereas the hexanucleotide AATAGA is found 15 and 18 nucleotides, respectively, upstream of the poly(A) site in two of the cDNAs. Phylogenetic studies including 102 actin protein sequences revealed that there are at least four different types of cestode actins. In this study three of these types were found to be expressed in the adult D. dendriticum tapeworm. Structurally the cestode actin groupings differ from each other to an extent seen only among the metazoan actins between the vertebrate muscle and cytoplasmic isoforms. In the phylogenetic trees constructed, cestode actins were seen to map to two different regions, one on the border of the metazoan actins and the other within this group. It is, however, difficult to say whether the cestode actins branched off early in the metazoan evolution or if this position in the phylogenetic tree only reflects upon differences in evolutionary rate.

Actins↗
PubMed1997

Successful treatment of an iatrogenic infrarenal aortic dissection with serial Wallstents.

We report a patient who presented with acute lower extremity ischemia 13 days following removal of an intraaortic balloon pump. Dissection of the infrarenal aorta, extending into the left iliac artery, was found at operation. The patient's recent myocardial infarction, prolonged ischemia with compartment syndrome, otherwise normal aortoiliac and infrainguinal arteries and the necessity for suprarenal aortic clamping for surgical repair, were all factors in the decision for endovascular repair of the dissection. Infrarenal aortic and left iliac Wallstents, placed intraoperatively, successfully occluded the false lumen of the dissection. The patient was discharged after an uncomplicated recovery and is asymptomatic with palpable pedal pulses and triphasic pedal Doppler waveforms 1 year following the procedure.

Aortic Dissection↗
PubMed1997

Glycerol elicits energy taxis of Escherichia coli and Salmonella typhimurium.

Escherichia coli and Salmonella typhimurium show positive chemotaxis to glycerol, a chemical previously reported to be a repellent for E. coli. The threshold of the attractant response in both species was 10(-6) M glycerol. Glycerol chemotaxis was energy dependent and coincident with an increase in membrane potential. Metabolism of glycerol was required for chemotaxis, and when lactate was present to maintain energy production in the absence of glycerol, the increases in membrane potential and chemotactic response upon addition of glycerol were abolished. Methylation of a chemotaxis receptor was not required for positive glycerol chemotaxis in E. coli or S. typhimurium but is involved in the negative chemotaxis of E. coli to high concentrations of glycerol. We propose that positive chemotaxis to glycerol in E. coli and S. typhimurium is an example of energy taxis mediated via a signal transduction pathway that responds to changes in the cellular energy level.

Bacterial Proteins↗
PubMed1997

Outcome of tunneled hemodialysis catheters placed via the right internal jugular vein by interventional radiologists.

PURPOSE: To assess the outcome of interventional radiologic placement of tunneled hemodialysis catheters via the right internal jugular vein. MATERIALS AND METHODS: In 194 patients, the catheter was placed via the right internal jugular vein unless thrombosis was present. Real-time ultrasound-guided puncture and fluoroscopic guidance were used. Patients were followed up until catheter removal or death. Outcomes evaluated included infection, thrombosis, and catheter malfunction. RESULTS: In 175 patients, 250 consecutive catheters were placed via the right internal jugular vein with 100% success. All catheters functioned immediately after placement. Procedural complications were limited to clinically unimportant air embolus (n = 2). No instances of pneumothorax, hemothorax, or substantial bleeding complications occurred. Follow-up was available in 173 (99%) patients. Mean and median "catheter duration" were 87 and 56 days, respectively. Catheter-related symptomatic venous thrombosis or stenosis was not observed. The rate of infection was 0.08 per 100 catheter days, and the rate of malfunction that necessitated removal was 0.22 per 100 catheter days. Definite or possible catheter thrombosis that necessitated removal occurred at a rate of 0.16 per 100 catheter days. CONCLUSION: Interventional radiologic placement of tunneled hemodialysis catheters via the right internal jugular vein showed equal or better long-term results than those reported for surgical placement. Interventional radiologic placement should be the method of choice.

Catheterization↗
PubMed1996

Effect of phorbol 12, 13-dibutyrate on ligand binding, enzyme activity and translocation of protein kinase C isoforms in the alpha T3-1 gonadotrope-derived cell line.

The effect of incubating alpha T3-1 cells with phorbol 12,13-dibutyrate (PDBu) on the protein kinase C (PKC) isoform content (predominantly alpha, epsilon and zeta isoforms) was assessed by immunoblotting, enzyme activity assay and [3H]PDBu binding. After exposure to PDBu for 17 h the immunoreactivity detected for both PKC alpha and PKC epsilon had disappeared from cytosol and had increased slightly in membranes. Immunoreactivity for PKC zeta was present as two bands in cytosol; after PDBu treatment both bands decreased in intensity, the higher molecular weight band more than the lower. The lower molecular weight band corresponded with a component of constitutive PKC activity eluting from DEAE cellulose that was defined by inhibition of basal activity with GF 109203X or H7. Investigation of very short treatment times with PDBu using binding, immunoblot and activity measurements (in the presence/absence of Ca2+) indicated that translocation of PKC alpha and epsilon was very rapid-detectable by 10 sec, maximal within minutes. Reduction of these isoforms in membranes took much longer, and was not apparent up to 150 min. The immunoblot data for PKC zeta in cytosol showed no detectable effect of PDBu treatment on the low molecular weight band up to 150 min although it was reduced at 17 h. Translocation of the upper band was detectable at 10 sec but this band may have resulted from cross-reaction with other PKC isoforms. The constitutive activity and low molecular weight ("authentic') PKC zeta immunoreactivity were partially affected after long exposure only, suggesting an action of PDBu on PKC zeta secondary to activation of the other PKC isoforms. An endogenous receptor agonist, luteinising hormone-releasing hormone (LHRH), was also used to assess by immunoblotting, translocation of the PKC isoforms. Although all the isoforms did translocate from cytosol to membrane fractions, they did so with distinctly different time courses: PKC epsilon moved more rapidly than PKC zeta which appeared to translocate more quickly than PKC alpha. After downregulation of the responsive PKC isoforms with PDBu, the remaining PKC zeta was not translocated by LHRH.

Animals↗
PubMed1996

Structure of cytokine hydrophobic cores.

Diagrams were constructed that describe, using a common frame of reference, the interactions that occur within the hydrophobic cores of the left-handed four-helix bundles of interleukin-2, -4, -5, granulocyte/macrophage-, granulocyte-, macrophage-colony-stimulating factor, beta- and gamma-interferon, growth hormone, and leukemia inhibitory factor. Based on the assumption that the topologies of the ligand-receptor complexes are similar, analogous patterns were obtained from these diagrams and lead to the detailed comparison of the hydrophobic cores. These patterns were then used to obtain pairwise rigid-body superpositions among all of the four-helix bundles; values of the root-mean-square deviations (C alpha-atoms) over the four alpha-helices of the bundles range from 1.14 A to 3.22 A. Two distinct groups are formed after clustering based on structural superposition of the ten cytokines.

Cytokines↗
PubMed1996

Separation of H7-resistant protein kinase C isoforms from gonadotroph-derived alpha T3-1 cells.

Certain physiological events in anterior pituitary tissue are regulated by protein kinase C that is relatively resistant to the inhibitor, H7. In the present studies we have shown that two H7-resistant protein kinase C activities may be isolated from (pituitary) gonadotroph-derived alpha T3-1 cells using hydroxyapatite chromatography. These activities have the characteristics of PKC zeta and an apparently novel PKC isoform. The availability of clonal alpha T3-1 cells will facilitate investigations of the pituitary events.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1996

Physicians, pharmaceutical sales representatives, and the cost of prescribing.

OBJECTIVE: To evaluate the influence of primary care physicians' attitudes toward and use of information provided by pharmaceutical representatives on prescribing costs in ambulatory practice. DESIGN: A mailed questionnaire collected information about physician demographic and practice characteristics and attitudes toward and use of information provided by pharmaceutical representatives. PARTICIPANTS: Kentucky physicians practicing primary care adult medicine (family medicine, general practice, general medicine; n = 1603). MAIN OUTCOME MEASURE: Relative cost of prescribing, based on physician responses to treatment choices for ambulatory clinical scenarios in primary care. A multivariable regression model assessed predictive relationships between independent variables and prescription costs. RESULTS: Four hundred forty-six returned questionnaires were suitable for analysis. No significant differences were noted in age, gender, days worked per week, or years since graduation between responders and a sample of nonresponders. A significant positive correlation was found between physician cost of prescribing and perceived credibility, availability, applicability, and use of information provided by pharmaceutical representatives (P < .01, Pearson's Product-Moment Correlation Coefficient). Physicians in academic or hospital-based practice settings had significantly lower prescribing costs than physicians in nonacademic and nonhospital practices (P = .001, analysis of variance). Frequency of use of information provided by pharmaceutical representatives (P = .01, multiple linear regression) and the group practice setting (P = .02, multiple linear regression) remained significant, independent positive predictors of cost in the multivariable regression model. CONCLUSIONS: Frequency of use of information provided by pharmaceutical representatives and the group practice, nonacademic and nonhospital setting may be associated with increased primary care physician prescribing costs.

Attitude of Health Personnel↗
PubMed1996

Utilizing all opportunities in clinical practice.

Clinical practice provides an abundance of avenues through which nurses may enhance both clinical judgment and scholarly activities. Creative ways to stimulate opportunities for this knowledge exist. The article describes the approaches utilized by clinical nurse specialists in two teaching hospitals. A cardiovascular nursing unit and a neurologic nursing unit served as settings for description. Advanced practice nurses provided motivation.

Cardiovascular Diseases↗
PubMed1996

Oxygen taxis and proton motive force in Azospirillum brasilense.

The microaerophilic nitrogen-fixing bacterium Azospirillum brasilense formed a sharply defined band in a spatial gradient of oxygen. As a result of aerotaxis, the bacteria were attracted to a specific low concentration of oxygen (3 to 5 microM). Bacteria swimming away from the aerotactic band were repelled by the higher or lower concentration of oxygen that they encountered and returned to the band. This behavior was confirmed by using temporal gradients of oxygen. The cellular energy level in A. brasilense, monitored by measuring the proton motive force, was maximal at 3 to 5 microM oxygen. The proton motive force was lower at oxygen concentrations that were higher or lower than the preferred oxygen concentration. Bacteria swimming toward the aerotactic band would experience an increase in the proton motive force, and bacteria swimming away from the band would experience a decrease in the proton motive force. It is proposed that the change in the proton motive force is the signal that regulates positive and negative aerotaxis. The preferred oxygen concentration for aerotaxis was similar to the preferred oxygen concentration for nitrogen fixation. Aerotaxis is an important adaptive behavioral response that can guide these free-living diazotrophs to the optimal niche for nitrogen fixation in the rhizosphere.

Azospirillum brasilense↗