Search PubMed⌕ Search

Biomedical subjects

W J Boyle

Publications and source records attributed to W J Boyle.

42 records · Page 3Linked to original sources

Serum testosterone measurements.

Serum testosterone and especially free testosterone is one of the parameters commonly used to evaluate androgen excess or deficiency. The authors equilibrated serum samples with 14C-labeled testosterone followed by an ammonium sulfate precipitation to compare the "apparent free testosterone concentration" with "total" serum testosterone concentration in the following populations: normal males and females; females presenting with gynecologic problems, particularly hirsutism and/or virilization; and males and females on maintenance hemodialysis. Total serum testosterone for each specimen was assayed with five different commercially available RIA kits encompassing a variety of technics: direct assay technics, assays utilizing extraction procedures prior to RIA; tritium-labeled tracer as well as iodine-labeled tracers. Clinical correlations improve strikingly when apparent free testosterone concentrations rather than total serum testosterone concentrations are used.

Female↗

Identification of the leukemogenic protein of avian myeloblastosis virus and of its normal cellular homologue.

The genome of the replication-defective avian myeloblastosis virus (AMV) contains an inserted cellular sequence (amv) that is part of the oncogene responsible for acute myeloblastic leukemia in chickens infected with AMV. Three antisera raised against distinct synthetic peptides predicted from the long open reading frame of amv specifically precipitated the same 48-kilodalton protein (p48amv) from leukemic myeloblasts but not from normal hematopoietic tissue, fibroblasts, or from fibroblasts infected with the AMV helper virus, MAV-1 (myeloblastosis-associated virus type 1). p48amv is not glycosylated or phosphorylated and does not appear to act as a protein kinase in vitro. The same three antisera that recognized p48amv also specifically precipitated a common 110-kilodalton protein from normal uninfected hematopoietic tissue. This normal cellular homologue of the AMV leukemogenic protein, p110proto-amv, was not present in normal fibroblasts, MAV-1 infected fibroblasts, or, interestingly, in some leukemic myeloblasts. We conclude that p48amv is the leukemogenic product of an altered, transduced, partial protooncogene. Short helper-virus sequences provide its carboxyl terminus and also may provide the amino terminus.

Animals↗

Isolation of poly A(+) RNA from Schistosoma mansoni and immunoprecipitation of its in vitro translation products.

Total RNA was extracted from S. mansoni by homogenization in 4 M guanidine thiocyanate followed by centrifugation through cesium chloride. Poly A(+) RNA was isolated by oligo(dT)-cellulose chromatography. The recovered poly A(+) RNA was subsequently shown to stimulate protein synthesis in a cell-free, rabbit reticulocyte system. Among the translation products were polypeptides with molecular weights of 18, 20, 26, 39, 42, and 50 kilodaltons that were recognized by rabbit anti-schistosome-denuded body IgG. Human infection serum IgG precipitated polypeptides with molecular weights of 16, 18, 30, 35, 37, and 42 kilodaltons. These results identify several mRNA's that may provide useful templates for preparation of cDNA for eventual cloning or as probes in various related experiments.

Animals↗

Regulation of osteoclastogenesis and RANK expression by TGF-beta1.

Transforming growth factor-beta (TGF-beta) has been shown to both inhibit and to stimulate bone resorption and osteoclastogenesis. This may be due, in part, to differential effects on bone marrow stromal cells that support osteoclastogenesis vs. direct effects on osteoclastic precursor cells. In the present study, we used the murine monocytic cell line, RAW 264.7, to define direct effects of TGF-beta on pre-osteoclastic cells. In the presence of macrophage-colony stimulating factor (M-CSF) (20 ng/ml) and receptor activator of NF-kappaB ligand (RANK-L) (50 ng/ml), TGF-beta1 (0.01-5 ng/ml) dose-dependently stimulated (by up to 120-fold) osteoclast formation (assessed by the presence of tartrate-resistant acid phosphatase (TRAP) positive multinucleated cells and expression of calcitonin and vitronectin receptors). In addition, TGF-beta1 also increased steady state RANK mRNA levels in a time- (by up to 3.5-fold at 48 h) and dose-dependent manner (by up to 2.2-fold at 10 ng/ml). TGF-beta1 induction of RANK mRNA levels was present both in undifferentiated RAW cells as well as in cells that had been induced to differentiate into osteoclasts by a 7-day treatment with M-CSF and RANK-L. Using a fluorescence-labeled RANK-L probe, we also demonstrated by flow cytometry that TGF-beta1 resulted in a significant increase in the percentage of RANK+ RAW cells (P < 0.05), as well as an increase in the fluorescence intensity per cell (P < 0.05), the latter consistent with an increase in RANK protein expression per cell. These data thus indicate that TGF-beta directly stimulates osteoclastic differentiation, and this is accompanied by increased RANK mRNA and protein expression.

Acid Phosphatase↗