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Biomedical subjects

M L Pearson

Publications and source records attributed to M L Pearson.

67 records · Page 4Linked to original sources
PubMed1975

Isolation and characterization of an alpha-amanitin-resistant rat myoblast mutant cell line possessing alpha-amanitin-resistant RNA polymerase II.

Cultures of the rat skeletal muscle myoblast cell line, L6, were treated with the mutagen ethylmethanesulfonate and grown in the presence of alpha-amanitin, an inhibitor of RNA polymerase II in vitro. One clonal cell line, Ama102, resistant tc the cytotoxic action of 2 mu-g/ml of alpha-amanitin was isolated and extensively characterized. Ama102 cells were about 30-fold more resistant to alpha-amanitin than their Ama+ parent cells based on a comparison of the concentration of alpha-amanitin required to reduce their plating efficiencies to similar extents. The RNA polymerase activities from Ama+ and Ama102 cells were solubilized and separated by DEAE-Sephadex chromatography. Whereas all of the Ama+ RNA polymerase II activity was inhibited by 0.1 mu-g/ml of alpha-amanitin, about 30% of the activity in the Ama102 RNA polymerase II peak was resistant to this concentration of alpha-amanitin and was inhibited only by much higher concentrations (25 mu-g/ml) of alpha-amanitin. This alpha-amanitin-resistant activity in Ama102 cells was identified as a bona fide RNA polymerase II by its chromatographic behavior on DEAE-Sephadex, salt optimum, preference for denatured DNA as template, insensitivity to inhibition by potassium phosphate, thermal inactivation kinetics, and inactivation by anti-RNA polymerase II antiserum. Both RNA polymerase IIa and IIb from Ama102 cells exhibited the partial alpha-amanitin resistance, as did this activity when purified further on phosphocellusose. Unlike the parental Ama+ cells, Ama102 cells neither fused at confluence nor showed an increase in the specific activity of creatine kinase. The altered sensitivity of the Ama102 RNA polymerase II to alpha-amanitin appears to account for the drug-resistant phenotype of these cells.

Amanitins↗
PubMed1975

Repression and autogenous stimulation in vitro by bacteriophage lambda repressor.

Purified lambda repressor protein is shown to reduce the lambda DNA-directed synthesis of proteins in vitro as determined both by net amino-acid incorporation and by analysis of specific lambda-coded proteins resolved by sodium dodecyl sulfate/polyacrylamide slab gel electrophoresis. By means of different lambda DNA templates carrying deletion and point mutations in the operators o-L or o-R, it has been possible to demonstrate repression of the synthesis of two classes of lambda proteins. The synthesis of one, class c, appears to be controlled from the operator o-L and is more efficiently repressed at low concentrations of the repressor than that of the other class of repressible lambda proteins, class d, which is controlled from the operator o-R. Several other proteins synthesized in vitro are not repressible. Some of these are coded by the J-att region. In addition, the repressor appears to have another activity, that of stimulating the synthesis of a protein identified as the repressor itself. Lambda repressor appears to stimulate its own synthesis by acting at prm, a site defined by the cis-acting mutation prm 116.

Chromosome Mapping↗
PubMed1973

Regulation by N gene protein of phage lambda of anthranilate synthetase synthesis in vitro.

The N protein of bacteriophage lambda is a positive regulator of early lambda gene expression. In a lambdatrp transducing phage, lambdatrp46Nam, the synthesis of trp enzymes in vivo is also dependent on the presence of active N protein. The DNA of this phage has been used in a protein-synthesizing system in vitro to develop a biochemical assay for the activity of the N protein. From the following observations it appears that it is the N protein itself that stimulates trp enzyme synthesis in vitro. An activity that stimulates trp enzyme synthesis can be made in vitro by lambdaN(+) DNA but not by lambdaNam DNA. This activity can also be detected in extracts of induced lambdaN(+) lysogens but not of lambdaNam lysogens. Furthermore, the activity is temperature sensitive in similar extracts of lambdaNts lysogens. No such activity is detectable in extracts of induced lysogens of lambdaimm(21). This phage makes all lambda-coded gene products outside the immunity region, but lacks an N protein activity able to substitute for N(lambda) protein in vivo. Our experiments also show that the action of N protein is inhibited by the cI repressor protein in vivo as it is in vivo.

Chromosome Mapping↗
PubMedDate not supplied

Evidence-based care for depression in managed primary care practices.

This paper evaluates whether externally designed, evidence-based interventions for improving care for depression can be locally implemented in managed care organizations. The interventions were carried out as part of a randomized trial involving forty-six practices within six diverse, nonacademic managed care plans. Based on evaluation of adherence to the intervention protocol, we determined that local practice leaders are able to implement predesigned interventions for improving depression care. Adherence rates for most key intervention activities were above 70 percent, and many were near 100 percent. Three intervention activities fell short of the goal of 70 percent implementation and should be targets for future improvement.

Depressive Disorder↗
PubMedDate not supplied

Clinical efficacy of the Abbott Tacrolimus II assay for the IMx.

Monitoring tacrolimus is essential to maintain therapeutic concentrations. Performance of the new Abbott Tacrolimus assay (FK II) was evaluated and compared to the original tacrolimus assay (FK I). 189 trough whole blood samples from transplant cases were included in the study. Samples (n = 117) with FK I concentrations > 5 ng/mL were reanalyzed with the FK II assay. Patient samples (n = 43) that had FK I concentration < 5 ng/mL with apparent mean and range of 3.1 ng/mL and 0.7 to 4.5 ng/mL, respectively, were also reanalyzed with FK II to yield a mean of 5.9 ng/mL with a range of 2.9 to 10.8 ng/mL. Checking for patient compliance, samples (n = 10) with a FK I concentration of 0 ng/mL were re-analyzed. With one exception of a mislabeled cyclosporine sample, all samples (n = 9) showed FK506 levels greater than 2 ng/mL with the FK II assay. The FK II assay was shown to be a clinically efficacious assay, with improved sensitivity and acceptable precision versus the previous FK I assay.

Adult↗