PubMed HealthSearch

Biomedical subjects

R L Kaspar

Publications and source records attributed to R L Kaspar.

12 recordsLinked to original sources

A regulatory cis element and a specific binding factor involved in the mitogenic control of murine ribosomal protein L32 translation.

The mRNA encoding ribosomal protein L32 redistributes from untranslated subribosomal particles into polysomes after mitogenic activation of quiescent T-lymphocytes and fibroblasts. To identify the regions of the L32 mRNA which are important in regulating its cytoplasmic location we constructed a plasmid containing the murine L32 cDNA under the control of the Rous sarcoma virus (RSV) long terminal repeat promoter and introduced this construct into murine 3T3 fibroblasts. The mRNA transcribed from the RSV-L32 construct redistributed from subribosomal particles into polysomes in response to mitogenic activation in a manner similar to endogenous L32 mRNA. A conserved polypyrimidine region present at the 5' terminus of all ribosomal protein mRNAs is required for translational regulation of L32 mRNA since deletion of this sequence resulted in a mRNA that was not sequestered in subribosomal particles in quiescent cells. A radioactive RNA probe containing the first 34 nucleotides of the L32 5'-untranslated region, including the polypyrimidine region, specifically interacted with a protein of about 56 kDa. This protein did not bind detectably to RNA probes lacking the polypyrimidine sequence. Binding activity was similar in protein extracts made from resting and activated cells, suggesting that binding of the 56-kDa protein as measured in this assay is not regulated. This protein is a member of what may be an emerging family of polyribopyrimidine-binding proteins with diverse biochemical functions.

3T3 Cells

Simultaneous cytoplasmic redistribution of ribosomal protein L32 mRNA and phosphorylation of eukaryotic initiation factor 4E after mitogenic stimulation of Swiss 3T3 cells.

Ribosomal protein L32 mRNA moved from messenger ribonucleoprotein particles into polysomes following serum activation of quiescent Swiss 3T3 cells. This redistribution of the mRNA into a translationally active state began by 1 h and was complete by 3 h after activation. In contrast, actin mRNA showed no translational control, being found predominantly in polysomes in both quiescent and activated cultures. The phosphorylation state of eukaryotic initiation factor (eIF) 4E, which binds mRNA caps, was examined in parallel. eIF-4E phosphorylation was elevated by 1 h following serum activation and reached a peak by 3-5 h. Treatment of resting cells with phorbol ester also simultaneously stimulated eIF-4E phosphorylation and the movement of L32 mRNA into polysomes. These results are consistent with a model in which mitogen-induced phosphorylation increases the pool of active eIF-4E molecules, which in turn cause the recruitment of translationally controlled mRNAs to actively synthesizing ribosomes.

Actins

Purification and physical analysis of Bacillus anthracis plasmids pXO1 and pXO2.

Virulent strains of Bacillus anthracis contain two large plasmids. pXO1 encodes the three component protein exotoxin and pXO2 is necessary for synthesis of the poly-D-glutamic acid capsule. A procedure for the isolation of these plasmids which yields high quantities of pure DNA is described. Restriction endonuclease analysis of these plasmids shows that they are not related. pXO1 is 174 kilobase pairs and pXO2 is 95 kilobase pairs. From their bouyant densities and melting temperatures we also determined their GC contents. pXO1 contains 31.1% GC base pairs and pXO2 is 31.4% GC. Both of these values are close to the GC content of B. anthracis genomic DNA which is 32.2%.

Bacillus anthracis

Amikacin therapy of patients with multiply antibiotic-resistant Serratia marcescens infections: development of increasing resistance during therapy.

Over a recent 22 month period, 222 patients in two adjacent hospitals became infected with a multiply antibiotic-resistant strain of Serratia marcescens; 13 were bacteremic. Nineteen patients with clinically significant infections received amikacin. Nine of 11 patients with urinary tract infections were cured. In contrast, only one of eight patients with pneumonia or other deep tissue infections was cured and four died. These eight patients were severely ill; many had infections with multiple microorganisms. In four of five patients in whom the infection failed to clear promptly. Serratia strains became increasingly resistant to amikacin during therapy and these strains contributed to the death of two of these patients. Amikacin proved useful in treating patients with infections due to gentamicin-resistant S. marcescens organisms, especially urinary tract infections. However, the capacity of some strains of S. marcescens to develop resistance to amikacin may limit the usefulness of this antibiotic in the treatment of deep tissue infections which involve this microorganism.

Adult

Perihepatitis and hepatitis as complications of experimental endocarditis due to Neisseria gonorrhoeae in the rabbit.

Seven strains of Neisseria gonorrhoeae (colony type 1 or 2) were tested for their ability to produce endocarditis in rabbits with transaortic valve catheters. The gonococci exhibited three auxotype patterns and a broad range of susceptibility to penicillin and to complement-mediated serum bactericidal activity. Only four strains produced endocarditis; infectivity appeared to be related to serum resistance. Neither arthritis nor skin lesions were observed in infected animals, but 40% had hepatitis, often with fibrinous perihepatitis. Hepatic lesions could be induced by the continuous infusion of gonococci without a transvalvular catheter. Suppurative hepatitis in the rabbit endocarditis model is of particular interest in light of the unusual hepatic involvement reported in association with gonococcal endocarditis in humans. Since perihepatitis also occurs as a complication of experimental gonococcal bacteremia, perhaps the Fitz-Hugh-Curtis (gonococcal perihepatitis) syndrome appearing in women occurs more often as a function of undiagnosed bacteremia than has heretofore been suspected.

Animals

Rapid, simple bioassay for 5-fluorocytosine in the presence of amphotericin B.

It is important that serum levels of 5-fluorocytosine (5FC) be measured to insure therapeutic levels while avoiding toxicity. This is particularly true in patients with renal insufficiency. The concurrent use of amphotericin B and 5FC complicates the measurement of 5FC in the serum, since fungi used in conventional bioassay systems are uniformly susceptible to amphotericin B. This paper describes the development of a simple, reliable, 6-h bioassay for 5FC in the presence of amphotericin B. The assay is based upon the fact that 5FC diffuses readily through yeast nitrogen base agar, whereas amphotericin B apparently does not. This assay allows rapid adjustments in therapy of patients receiving both 5FC and amphotericin B and has permitted us to maintain 5FC serum levels between limits of 25 and 120 mug/ml in patients with altered renal function.

Amphotericin B

Specificity of autonomic influences on cardiac responses during myocardial ischemia.

A possible role of the autonomic nervous system in the left ventricular response to acute regional myocardial ischemia was sought in conscious dogs instrumented for measurement of left ventricular pressure, internal diameter, and aortic flow. Ischemia produced by occluding the left circumflex coronary artery caused tachycardia and reduced contractility. Changes during control occlusions were compared with those during occlusion.s after beta-adrenergic blockade, parasympathetic blockade, and combined sympathetic and parasymphatetic blockade. Beta-blockade did reduce the tachycardia and slightly reduced left ventricular diameter changes in response to coronary occlusion. Results obtained in animals following surgical cardiac sympathectomy indicated reduced tachycardia and no effects on other parameters. The principal effect of parasympathetic blockade was to augment the increase in end diastolic diameter during occlusion Right atrial pacing indicated this change was due to higher initial heart rates. Combined parasympathetic and sympathetic blockade did not alter inotropic responses to coronary occlusion. Results indicated that inotropic support due to changes in activity in autonomic nerves is not increased during acute occlusion of the left circumflex coronary artery.

Animals

The maximum derivatives of left ventricular pressure and transverse internal diameter as indices of the inotropic state of the left ventricle in conscious dogs.

1. In normal, conscious dogs, I.V. infusion of isoprenaline caused inases in heart rate and the maximal derivatives of left ventricular pressure (dP/dt max) and left ventricular internal diameter (dD/dt max). The changes in both derivatives were linearly related to the increase in heart rate.2. Increments in heart rate produced by right atrial pacing caused only minimally significant increases in both dP/dt max and dD/dt max at heart rates of 180 beats/min. Increases in heart rate, with end diastolic diameter maintained constant, resulted in small but significant increases in dP/dt max and no significant increase in dD/dt max.3. Increasing preload by volume infusion had little effect on either derivative, while increasing afterload by phenylephrine administration produced a small but significant increase in dP/dt max and no change in dD/dt max.4. Both dP/dt max and dD/dt max are equally reliable as indices of the inotropic state of the myocardium and are minimally influenced by changes in preload, afterload or heart rate.

Animals

A simple method for diagnosing pneumonia in intubated or tracheostomized patients.

A safe, simple, reliable technique for sampling uncontaminated peripheral bronchial secretions in patients with endotracheal tubes has been developed. It is easily performed and requires no special training or equipment. This report summarizes the techniques's usefulness in providing diagnostic samples of peripheral airway secretions from 20 patients with pneumonic infiltrates. In eight of the patients, a single organism was recovered from the peripheral airway despite polymicrobial colonization of the upper airways. An additional patient had two anaerobes. The recovered organism's role as a pathogen was verified by an appropriate response to specific antibiotic therapy in two patients, isolation from blood or empyema fluid in five, confirmation by bronchoscopy in one, and transtracheal aspiration after extubation in one. Peripheral bronchial secretions were sterile in the remaining 11 patients, even though multiple organsims were isolated from usual tracheal suctionings.

Bacteria