Control of globin gene transcription by erythropoietin in erythroblasts from friend virus-infected mice.

Splenic erythroblasts of mice infected with the anemia-inducing strain of Friend virus can be isolated in large numbers with less than 5% contamination with other cell types. In short-term culture, the isolated cells will initiate globin synthesis and undergo other aspects of terminal differentiation only if erythropoietin (EP) is added to the medium. An early effect of the hormone on these cells is stimulation of total RNA synthesis. EP also causes initiation of transcription of the beta-globin genes after a lag period of 4 to 6 h. By 6 h, the transcription rate of beta-globin RNA is enhanced threefold, and by 12 h, it is nearly maximal at ca. 20 times the level of control cells which received no EP. Transcription rates of alpha and beta-globin genes are approximately equal to each other throughout the period of terminal differentiation. In the splenic erythroblasts, the chromatin structure in the vicinity of the beta-major globin gene was analyzed with two nucleases during these transcription rate changes. No S1 nuclease-hypersensitive site is detectable near the gene. The beta-major gene is quite sensitive to DNase I in comparison with the albumin gene; however, the level of sensitivity is the same before EP addition as it is during maximal gene transcription after EP addition. Also, a hypersensitive site near the 5' cap site of the beta-major gene is quantitatively equivalent both before and after EP addition. Analysis of cytosine methylation at two sites upstream from the gene showed no changes upon induction of beta-globin gene transcription by EP. Thus, the initiation of beta-globin transcription by EP appears to be at some step after chromatin structural alteration such as synthesis, release, or activation of a specific transcription initiation factor.

Thrombocytopenia in a prospective, randomized, double-blind trial of bovine and porcine heparin.

A prospective, randomized, double-blind clinical trial of bovine and porcine heparin was conducted. One hundred forty-one patients were randomized, of whom 89 received heparin treatment for six or more days (mean, ten days). Two developed severe thrombocytopenia (platelet count less than 20,000 per microliter); both were randomized to the bovine heparin group, but one inadvertently received two doses of porcine heparin. Laboratory investigation suggested that the thrombocytopenia in these two patients was immunologically mediated, and platelet reactivity to both bovine and porcine heparin was demonstrated. Twenty patients had a decline in platelet count of greater than 50,000 from baseline, although the total count remained above 150,000 per microliter. In seven of these subjects, the platelet count returned to its original level while heparin therapy was continued. Of the 13 patients with a persistent decrease in platelet count, ten had received bovine heparin and their counts decreased by an average of 88,000 per microliter; the reduction in the three porcine-treated patients was 68,000; this difference was not statistically significant. In the remaining patients, the post-treatment platelet counts in both groups were significantly higher than pretreatment values (p less than .005), perhaps indicating a cessation of the platelet consumption that accompanied the original thrombotic event.(ABSTRACT TRUNCATED AT 250 WORDS)