Long-term protection of hematopoiesis against the cytotoxic effects of multiple doses of nitrosourea by retrovirus-mediated expression of human O6-alkylguanine-DNA-alkyltransferase

A human O6-alkylguanine-DNA-alkyltransferase (ATase) cDNA-containing retrovirus was used to infect murine long-term primary bone marrow cultures. High levels of ATase expression were obtained, and colony- forming cells of the granulocyte-macrophage lineage from the cultures transduced with the human ATase retrovirus were three times more resistant to the alkylating agent, N-methyl-N-nitrosourea (MNU), than control cultures. Furthermore, expression of the human ATase protected long-term hematopoiesis, measured as the output of progenitor cells to the nonadherent fraction of the culture, against the cytotoxic effects of repeated exposures to MNU. These results clearly show that a human ATase cDNA-containing retrovirus can be used to infect long-term primary bone marrow cultures and that this attenuates their sensitivity to nitrosoureas.     

The surgical decision in tetralogy of Fallot: weighing risks and benefits with decision analysis

The preferred management of the symptomatic infant less than 2 years of age with tetralogy of Fallot remains unsettled. In this study decision analysis was used to assess the risks and benefits of three courses of action: (1) primary intracardiac repair; (2) palliative operation in infancy and delayed intracardiac repair; and (3) treatment with propranolol in infancy and delayed intracardiac repair. For each action the likelihood was determined of three possible outcomes for the patient: (1) death, (2) poor surgical result, or (3) good surgical result. Each outcome was associated with an estimated life expectancy. The best action was defined as that resulting in the longest life expectancy. With use of previously reported data and those from experience at one institution, a range of probabilities was estimated for death before operation, surgical mortality, success of propranolol treatment and hemodynamic result of operation. The choice of action was affected most by the mortality of primary intracardiac repair and the likelihood of a good hemodynamic result from intracardiac repair. Less important factors were the outcome of a palliative operation and the likelihood of success with propranolol therapy. It was found that in the institution studied if the mortality rate of primary intracardiac repair is 10 percent or less, intracardiac repair should be performed; otherwise propranolol treatment, which allows delay in intracardiac repair, should be selected. Both alternatives are preferable to palliative operation. With decision analysis, the clinician can use probabilities and life expectancy appropriate to a given clinical setting in determining the best management for the infant with tetralogy of Fallot.     

Enhancement of chemotactic factor-stimulated neutrophil oxidative metabolism by leukotriene B4

Leukotriene B4 (LTB4) is a potent primary stimulator of neutrophil chemotaxis, aggregation, and degranulation and induces superoxide production at higher concentrations. In order to determine whether LTB4 modulates neutrophil responses to oxidative stimuli, human neutrophils (PMNs) were incubated with LTB4 prior to stimulation with f-Met-Leu-Phe (fMLP, 10(-7) mol/L), opsonized zymosan (OZ, 250 micrograms/mL), or phorbol myristate acetate (PMA, 32 nmol/L). Superoxide (O2-) production by stimulated PMNs was assessed by the superoxide dismutase-inhibitable reduction of cytochrome c. LTB4 alone did not stimulate O2- production in concentrations below 10(-7) mol/L and had no effect on the O2- assay. In the concentration range of 10(-12) to 10(-8) mol/L, LTB4 did not alter O2- release induced by OZ or PMA. In contrast, LTB4-treated cells demonstrated enhanced O2- production following exposure to fMLP, and in the presence of 10 nmol/LLTB4, generated 180% +/- 41% of O-2 quantities produced by control cells (n = 23). Enhancement was LTB4 dose-dependent, was maximal in the range of 1 to 10 nmol/L LTB4, was not reversed by removal of the lipid from the medium prior to fMLP stimulation, and was not dependent on the presence of Ca++ or Mg++ in the suspending medium. Chemiluminescence of fMLP-stimulated neutrophils was increased to 323% of controls in neutrophils preincubated with 10 nmol/L LTB4. Unlike augmentation of oxidative responses to fMLP seen with other degranulating stimuli, enhancement by LTB4 was not correlated with an increase in 3H-fMLP receptor binding. These results indicate that, in addition to its primary effects on neutrophil function, LTB4 modulates PMN oxidative responses to the chemotactic peptide and, thus, may amplify the release of oxygen metabolites at inflammatory foci.     

Mexiletine: an effective antiarrhythmic drug for treatment of ventricular arrhythmias in congenital heart disease

The use of antiarrhythmic drugs to suppress ventricular arrhythmias in pediatric patients with a structurally or hemodynamically abnormal heart appears to improve long-term prognosis. The previously successful use of phenytoin to treat serious ventricular arrhythmias led to the investigation for an alternative antiarrhythmic agent, in the same antiarrhythmic drug class, for those patients who develop side effects or become intolerant to phenytoin's antiarrhythmic effect. Forty-two children and young adults (age range 5 months to 34 years, mean 15.5 years) were treated with mexiletine. Arrhythmias treated were ventricular tachycardia (25), ventricular couplets (8), multiform ventricular premature beats (4) and frequent uniform ventricular premature beats (5). Anatomic diagnoses included congenital heart disease (postoperative in 26, unoperated in 2), cardiomyopathy (7), no heart disease (4) and other (3). Thirty-three patients had been previously treated with 1 to 5 (mean 1.6) antiarrhythmic drugs. In the short term, ventricular arrhythmias were effectively suppressed in 30 (71%) of all 42 patients treated. During follow-up (ranging to 42 months, median 10.6), 18 (60%) of the 30 acute responders continued to have excellent control. Early suppression of ventricular arrhythmias was more effective in patients with congenital heart disease (89%) than in those with cardiomyopathy (29%) or no heart disease (43%) (p less than 0.01). Initial complexity of ventricular ectopic activity had no effect on treatment results. Of 25 patients previously treated with phenytoin, in whom alternative antiarrhythmic therapy was required, 40% had long-term arrhythmia control when treated with mexiletine. Mexiletine therapy was terminated for side effects in only five patients (12%). Mexiletine is recommended for young patients with congenital heart disease and serious ventricular arrhythmias.     

Normal synthesis and expression of endothelial IIb/IIIa in Glanzmann's thrombasthenia

Glanzmann's thrombasthenia is a bleeding disorder, inherited in an autosomal recessive way and characterized by an absence or deficiency of the platelet glycoprotein (GP) IIb/IIIa complex. Recently, we and others demonstrated that cultured human umbilical vein endothelial cells synthesized a membrane protein complex similar to the platelet GP IIb/IIIa complex. In this article, we demonstrate that endothelial cells isolated from the umbilical vein of a newborn with Glanzmann's thrombasthenia, as compared with normal endothelial cells, show no difference in their ability to synthesize and express this GP IIb/IIIa complex. Our results indicate that Glanzmann's thrombasthenia is not accompanied by an "endotheliopathy."     

Prognosis for the newborn with transposition of the great arteries

To determine the prognosis for the newborn with transposition of the great arteries, the clinical course of 112 consecutive neonates with dextro-transposition was reviewed. Patients were managed with balloon atrial septostomy at initial cardiac catheterization, palliative operation if needed in the 1st year of life and Mustard's intraatrial baffle repair.The 1st month of life was the period of greatest risk (8 percent mortality rate). Between balloon septostomy and baffle repair, 14 of 103 patients at risk (14 percent) either died or had a cerebrovascular accident. The mortality rate at baffle repair was 14 percent (10 deaths in 71 patients), and there were 3 late postoperative deaths. Actuarial analysis of the data indicates that with this plan of management, approximately 50 percent of newborns with transposition of the great arteries will survive 5 years with excellent function and an additional 15 to 20 percent will survive with one or more medical handicaps.     

von Willebrand disease "Vicenza" with larger-than-normal (supranormal) von Willebrand factor multimers

When normal volunteers or patients with type I von Willebrand disease (VWD) are given desmopressin (DDAVP), a set of larger-than-normal (supranormal) von Willebrand factor (VWF) multimers, similar to those present in VWF-containing cells such as platelets megakaryocytes and endothelial cells, appear transiently in postinfusion plasma. In two kindreds with mild lifelong bleeding symptoms transmitted as an autosomal dominant trait, all ten symptomatic members (but none of the five asymptomatic members) had a supranormal multimeric structure for plasma VWF, apparently identical to that seen for postdesmopressin normal plasma. Plasma factor VIII coagulant activity (VIII:C), VWF antigen (VWF:Ag), ristocetin-induced platelet agglutination, and ristocetin cofactor (RiCof) activity were low. Platelet VWF:Ag and RiCof levels (tested for three patients only) were normal. Bleeding times were normal or slightly prolonged. The patients' platelet multimeric structure was the same as that for normal platelets. After desmopressin infusion the plasma VWF multimeric structure remained supranormal as for preinfusion plasma, with VIII:C VWF:Ag and RiCof increasing markedly over baseline values and disappearing at a normal rate. Examination of the VWF subunit composition from three of these patients indicated that proteolytic processing of their VWF did not differ from normal. This study describes the first variant of VWD with a supranormal multimeric structure.