The vaccinia virus bifunctional gene J3 (nucleoside-2'-O-)-methyltransferase and poly(A) polymerase stimulatory factor is implicated as a positive transcription elongation factor by two genetic approaches.

Vaccinia virus genes A18 and G2 affect the elongation and termination of postreplicative viral gene transcription in opposite ways. Viruses with mutations in gene A18 produce abnormally long transcripts, indicating that A18 is a negative transcription elongation factor. Viruses containing mutations in gene G2 produce transcripts that are abnormally short, truncated specifically from their 3' ends, indicating that G2 is a positive transcription elongation factor. Despite the fact that both A18 and G2 are essential genes, A18-G2 double-mutant viruses are viable, presumably because the effects of the mutations are mutually compensatory. In addition, the anti-poxviral drug isatin-beta-thiosemicarbazone (IBT) seems to enhance elongation during a vaccinia infection: IBT treatment of a wildtype vaccinia infection induces a phenotype identical to an A18 mutant infection, and G2 mutant viruses are dependent on IBT for growth, presumably because IBT restores the G2 mutant truncated transcripts to a normal length. These observations inspire two independent genetic selections that have now been used to identify an additional vaccinia gene, J3, that regulates postreplicative transcription elongation. In the first selection, a single virus that contains an extragenic suppressor of the A18 temperature-sensitive mutant, Cts23, was isolated. In the second selection, several spontaneous IBT-dependent (IBT(d)) mutant viruses were isolated and characterized genetically. Marker rescue mapping and DNA sequence analysis show that the extragenic suppressor of Cts23 contains a point mutation in the J3 gene, while each of seven new IBT(d) mutants contains null mutations in the J3 gene. The J3 protein has previously been identified as a (nucleoside-2'-O-)-methyltransferase and as a processivity subunit for the heterodimeric viral poly(A) polymerase. The nature of the two independent selections used to isolate the J3 mutants strongly suggests that the J3 protein serves as a positive postreplicative transcription elongation factor during a normal virus infection.

Chronic pancreatitis progressing to duodenal obstruction in the absence of classic symptoms.

We report the case of a 34-year-old alcoholic who was initially seen in March 1985 because of acute pancreatitis. A mass was demonstrated in the head of the pancreas. Serial sonogram and computed tomography scans over 4 1/2 years revealed progressive encroachment of the duodenum without symptoms attributable to obstruction. In 1989, three separate endoscopies with multiple biopsies showed chronic inflammation and strictures. Hypotonic duodenography confirmed stricture and obstructed duodenum. Surgical intervention is being considered. Duodenal obstruction secondary to chronic pancreatitis is rare. It may proceed subclinically for several years independent of continued alcohol use. Only when obstruction became severe in our patient did the classic symptoms of postprandial nausea, emesis, and weight loss become manifest. Obstructive jaundice from chronic pancreatitis due to stricture in the pancreatic portion of the common bile duct is uncommon.

Myosin phosphorylation, agonist concentration and contraction of tracheal smooth muscle.

Myosin phosphorylation plays an important part in excitation--contraction coupling in smooth muscle. Phosphorylation by a Ca2+, calmodulin-dependent kinase stimulates the actin-activated Mg2+-ATPase activity of smooth muscle myosin, suggesting that myosin phosphorylation regulates smooth muscle contraction. This hypothesis is supported by evidence that myosin is phosphorylated during contraction and dephosphorylated during relaxation of intact smooth muscles stimulated with a single agonist concentration. However, there is little information regarding the response to stimulation with various agonist concentrations. As the dose-response relationships for phosphorylation and tension should be similar if myosin phosphorylation does, in fact, regulate smooth muscle contraction, we studied myosin phosphorylation in tracheal smooth muscle stimulated with a broad range of concentrations of the cholinergic agonist, methacholine. The results of these experiments are consistent with the hypothesis that myosin phosphorylation regulates smooth muscle contraction but they indicate a relatively complex relationship between myosin phosphorylation and the generation of isometric tension.

Long-term hemodynamic assessment of the porcine heterograft in the mitral position. Late development of valvular stenosis.

We undertook a study of patients who had porcine mitral valves in place for more than 5 years and who had no clinical signs or symptoms suggestive of valve dysfunction. Of the first 54 patients who had porcine valves implanted in the mitral position, 18 were available for catheterization; all had a routine hemodynamic study postoperatively (mean 7 months) for comparison. Mean follow-up was 85 months (range 61-111 months). Compared with the early postoperative data, there was a significant increase in mean mitral valve gradient, from 5.9 +/- 0.7 to 8.6 +/- 0.7 mm Hg (p less than 0.01), and a significant decrease in calculated mitral valve area, from 2.2 +/- 0.2 to 1.7 +/- 0.2 cm2 (p less than 0.01). Moreover, seven patients showed a decrease in valve area greater than 1.0 cm2, five with valves in place for more than 80 months and only two of 11 patients with valves in place for 80 months or less (p less than 0.05). We conclude that there is a significant incidence of hemodynamic deterioration of porcine heterografts in the mitral position for greater than 5 years, even in patients who are clinically stable.

Verapamil therapy: a new approach to pharmacologic treatment of hypertrophic cardiomyopathy.

The hemodynamic and clinical effects of verapamil administration were examined in patients with hypertrophic cardiomyopathy. Infusion of 0.021 mg/kg/min of verapamil decreased the basal left ventricular outflow tract gradient from 94 +/- 14 to 49 +/- 14 mm Hg, while cardiac index increased from 2.5 +/- 0.2 to 2.8 +/- 0.2 L/min/sq m. Orally administered, 80 or 120 mg four times daily (QID), increased exercise capacity relative to placebo by 26% (1.6 +/- 0.5 minute; p less than 0.005) and propranolol, 40 or 80 mg QID, increased it by 21 +/- 8% (1.3 +/- 0.5 minute, p less than 0.025). Nine patients considered their symptomatic status "best" while receiving verapamil, seven while receiving placebo, and only three while receiving propranolol. Seventy-three patients have been discharged from the hospital receiving long-term verapamil therapy, and 49 have reported continuing symptomatic benefit with many showing improved exercise capacity up to two years later. Thus, verapamil can improve exercise capacity and symptomatic status in certain patients with hypertrophic cardiomyopathy, thereby providing physicians with a new therapeutic agent for the treatment of this disorder.