Nitrogen mustard (NH2) and Nor-nitrogen mustard (Nor-HN2) both inhibit the polymerization of deoxyhemoglobin S in solution and in intact erythrocytes. Metabolic studies were undertaken to determine the feasability of an extracorporeal treatment with these or related agents. Glucose utilization, hexose monophosphate shunt activity, methemoglobin reduction, and incubation with acetylphenylhydrazine for Heinz body formation were performed, as well as specific assays for hexokinase, pyruvate kinase, glucose-6-phosphate dehydrogenase, glutathione reductase, ATP, reduced glutathione (GSH), and survival of autologous mustard-treated cells in rabbits. HN2 was found to enter red cells rapidly and bind to intracellular contents. Metabolic studies revealed no significant inhibition or alteration of function by Nor-HN2 at 10 mg/ml of whole blood. Rabbit red cell survival was also normal. HN2, however, inhibited glutathione reductase and blocked the free sulfhydryl group of GSH by forming serveral addition products of alkylated GSH. Heinz body test with acetylphenylhydrazine became positive in HN2-treated cells, and rabbit red cell survival was shortened considerably in the concentration range used to inhibit sickling. Ascorbic acid stimulation of the hexose shunt pathway was inhibited by HN2, but methylene blue stimulation remained unaffected. 14-C-HN2 remains bound to red cells in vivo, and the disappearance of radioactivity is similar to that found with 14-C-DFP (disopropylfluorophosphate). Oxygen affinity of both HN2 and Nor-HN2 treated human red cells remains virtually the same as that found in control samples. It is concluded that Nor-HN2 may be a suitable agent for an extracorporeal therapy, and that each mustard needs to be evaluated individually for its antisickling effects and its suitability for extracorporeal use.

Annals of the New York Academy of SciencesVolume 241, Issue 1 p. 334-346 REGULATORY ROLE OF HEME * Barry H. Kaplan, Barry H. Kaplan Departments of Medicine and Biochemistry Albert Einstein College of Medicine Bronx, New York 10461Search for more papers by this authorMarianela Tricoche, Marianela Tricoche Departments of Medicine and Biochemistry Albert Einstein College of Medicine Bronx, New York 10461Search for more papers by this authorGrace Vanderhoff, Grace Vanderhoff Departments of Medicine and Biochemistry Albert Einstein College of Medicine Bronx, New York 10461Search for more papers by this author Barry H. Kaplan, Barry H. Kaplan Departments of Medicine and Biochemistry Albert Einstein College of Medicine Bronx, New York 10461Search for more papers by this authorMarianela Tricoche, Marianela Tricoche Departments of Medicine and Biochemistry Albert Einstein College of Medicine Bronx, New York 10461Search for more papers by this authorGrace Vanderhoff, Grace Vanderhoff Departments of Medicine and Biochemistry Albert Einstein College of Medicine Bronx, New York 10461Search for more papers by this author First published: November 1974 https://doi.org/10.1111/j.1749-6632.1974.tb21891.xCitations: 7 * This research is supported by a Grant-in-Aid from the American Heart Association and grants from the National Institutes of Health, AM-13147 and AM-13698. B. Kaplan is an Established Investigator of the American Heart Association. AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Citing Literature Volume241, Issue1Hemoglobins: Comparative Molecular Biology Models for the Study of DiseaseNovember 1974Pages 334-346 RelatedInformation

Double-stranded RNA from HeLa cell nuclei was isolated and purified using chromatography on CF-11 cellulose columns. Analysis by polyacrylamide gel electrophoresis indicated that the [3H]uridine-labeled RNA had a molecular size slightly greater than that of transfer RNA. The double-stranded RNA was a potent inhibitor of initiation of protein synthesis, as measured in rabbit reticulocyte lysates in vitro.

1.1. δ-Aminolevulinic acid synthetase, the rate-limiting enzyme for haem biosynthesis, has been purified 11.2-fold in 26% yield from the particulate fraction of liver of porphyric rats.2.2. Enzymatic activity is stable indefinitely at −150° and can be stabilized during purification with 0.1 mM pyridoxal phosphate or in the presence of 0.2 M potassium phosphate (pH 7.4).3.3. The enzyme is inhibited 48% by 5·10−5 M haemin. The inhibition is non-competitive with succinyl-CoA, but haemin is a mixed inhibitor with respect to glycine.4.4. Despite the observation that 58% of the enzymatic activity does not precipitate upon centrifugation at 140 000 × g for 2 h, studies with gel filtration, polyacrylamide gel electrophoresis, and electronmicroscopy indicate that the enzymatic activity is contained in a large protein aggregate. The implications of this observation are discussed.