"When I retired from active involvement in science in 1995 some of my colleagues with whom I worked closely over the years generously penned the following description of my career in science. My thanks to each of them, and to many others." Paul Hochstein, 2006

Kelvin j. A. Davies*, Joanna M.S.. Davies*, Enrique Cadenas**, Lester Packer`, Alex Sevanian**, Henry Jay Forman**, Timothy M. Chan**, Joseph R. Landolph**, and Fulvio Ursini


We are delighted to have the opportunity to dedicate this volume to our friend, teacher, collaborator, and colleague, Paul Hochstein. Paul is an inspiration to all who have worked with him and our lives are the richer for his influence.

We began this article by stating that we were all friends, students, collaborators, and/or colleagues of Paul Hochstein. Thus it is with great pride that we write this abbreviated scientific history and tribute. Several of the articles in this Festschrift book (a "livingtribute") are based on oral presentations made at the meeting "Oxidants & Antioxidants in Biology, A Festschrift in Honor of Paul Hochstein" which took place from February 4-5, 1994 in Pasadena, California. The meeting was organized by the Oxygen Club of California, and sponsored by the University of California at Berkeley, the University of Southern California, and the Oxygen Society. Over 200 of Paul's friends and colleagues travelled to Pasadena for this 2 day tribute to his work and scientific influence on the young field of free radical biology. During the Festschrift letters of congratulations and gratitude from US President Bill Clinton, California Governor Pete Wilson, and Los Angeles Mayor Tom Bradley were read to the assembly, and speeches were made by University of Southern California Vice President for Health Affairs, Joseph P. Van Der Meulen and School of Pharmacy Dean John A. Biles. Several of us also took the opportunity to publicly thank Paul for his leadership, example, and friendship.

Articles based on talks at the Paul Hochstein Festschrift have been combined with papers based on lectures given at the first Oxygen Society Meeting held in Charleston, South Carolina from November 12-17, 1993 (Oxygen `93). Approximately 500 delegates attended Oxygen `93 and made a great success of this first major meeting of the Oxygen Society. The Oxygen Society, which Paul Hochstein helped to establish in 1987, now boasts over 1,000 members and is a constituent member of the International Society for Free Radical Research (SFRR International), representing North, Central, and South America. This book, thus, reflects both a tribute to Paul Hochstein's work and a synthesis of the latest free radical research.

We are delighted to have been involved with this project to produce a living tribute to our friend Paul Hochstein. We are particularly pleased that Paul will be able to read this book. Paul is a man of many talents and his "retirement" from the scientific arena has simply served to give him the time he needs to pursue his remarkable talent for painting and sculpting. An often heard "Paul Hochstein-ism" is that, "Suffering is good for you". Paul we thank you for sharing an important part of your life with us and we hope to continue "suffering" your company for a long time to come.

Paul Hochstein was born on February 7, 1926 in New York City, the son of Ida and Samuel Hochstein. He attended the famous High School of Science in New York (1940-1944) and served in the US Army from 1944-1946. Paul went on to earn a B.S. degree (1950) in Biology from Rutgers University in New Jersey. For his graduate work Paul went to the University of Maryland where he earned an M.S. degree in Plant Pathology (1952), and completed his Ph.D. degree in Biology (1954) in the laboratory of Carrol Cox. From 1954-1957 Paul was a US Public Health Service Fellow in the Laboratory of Dean Burk at the National Cancer Institute. From 1957-1962 Paul was an Associate in Biochemistry at the College of Physicians and Surgeons of Columbia University in New York. For the period 1962-1963 Paul was a senior National Science Foundation Fellow (USA) in the laboratory of Lars Ernster, at the WennerGren Institute in Stockholm, Sweden. This interaction with Lars Ernster was to develop into a lifelong friendship and collaboration (see next article by Lars Ernster).

Returning to the USA Paul spent the next six years (1963-1969) at Duke University in North Carolina, first as an Assistant Professor (1963) and then as an Associate Professor (1965) of Pharmacology, and Chief of the Laboratory of Cellular Pharmacology, in the Department of Physiology & Pharmacology. While at Duke, Paul regularly interacted with Irwin Fridovich and Phil Handler. During the summer of 1965 Paul was Visiting Scientist at the Institute for Comparative Biochemistry in La Jolla, California.

In 1969 Paul made what now appears to have become a permanent move to the University of Southern California (USC) in Los Angeles, CA. At the University of Southern California Paul was Professor of Pharmacology from 1969 to 1980. In 1976 Paul took a short leave as Visiting Professor in the Department of Biochemistry at the Royal Free Hospital of London University in England, where he worked with Tony Diplock and Catherine Rice-Evans. From 1978-1979 Paul took a full sabbatical as Visiting Professor at the Department of Biochemistry of the University of Stockholm in Sweden, where he renewed his scientific collaboration with Lars Ernster. Returning to the University of Southern California, Paul left the Department of Pharmacology to take up two new appointments. In 1980 he became the founding Director and Professor of the Institute for Toxicology, and assumed a joint appointment as Professor of Biochemistry.

Paul has a long list of honors, awards and achievements. Early in his career he won fellowships from the NIH, the NSF, and the Swedish Research Council. He was also awarded an NIH Career Development Award in 1965. In 1985 Paul was Visiting Professor of the University of Genoa in Italy and 1986 he was awarded a Doctoral Degree (honoris causae) by the University of Stockholm in Sweden. Paul was the first President of the Southern California Chapter of the Society of Toxicology (1987-90), and was awarded an endowed chair as the first Charles Krown Alumni Professor at the University of Southern California (1988). In 1990 Paul became one of the first Fellows of the Oxygen Society,and was elected an Honorary Member of the Oxygen Club of Washington, DC. Paul was a founding member of the Oxygen Society, and from 1989-1992 served as a member of the Oxygen Society Council.

A member of some 15 scientific and professional societies, Paul has also served as cochairman of the Peroxide Group (1965-1970), correspondent for the FASEB Public Affairs Committee (1970-1975), and as a member of the Oxygen Radicals Gordon Conference Organizing Committee (1983-85). Paul has been an editorial board member or reviewer for some 17 journals including Free Radical Biology & Medicine, the Journal of Biological Chemistry, Science, the Journal of Clinical Investigation, and Proceedings of the National Academy of Science. Since 1968 Paul has been a consultant to the National Science Foundation. From 1969-1973 he was a consultant to the Western Cooperative Cancer Research Group. Since 1970 Paul has been a consultant to the U.S. Veterans Administration of the National Institutes of Health. He served as a member of the NIH Toxicology Study Section from 1983-87. Always a superb teacher and mentor, Paul was recognized as Outstanding Basic Science Teacher of the Year, for three consecutive years (1973, 1974, and 1975), at the University of Southern California School of Medicine. Paul Hochstein's research contributions begin with an abstract published in 1952 and continue to this day: 43 years thus far! We shall attempt to summarize his work but please understand that the task is almost impossible. In point of fact we have had to limit ourselves to a brief review of less than 50 of Paul's publications. It should be noted that Paul so far has well over 200 publications to his name. One of the earliest investigators to explore the effects of organic fungicides on the metabolism of plant pathogenic fungi, Paul demonstrated that certain quinone antifungal agents block fungus growth by oxygen-dependent inhibition of glycolytic enzymes(1). Paul also carried out the first investigations on the fungitoxic actions of carboxamides. In this instance he demonstrated inhibition of cell growth accompanied by competitive inhibition of decarboxylation reactions requiring thiamine pyrophosphate as a coenzyme (2).

Next Paul demonstrated the high degree of association of glycolytic enzymes, in particular hexokinase, with tumor cell mitochondria(3). He also explored the regulation of glycolytic enzymes in tumor-bearing animals exposed to heat stress and demonstrated the insulinreversible nature of stress-induced inhibition of glycolytic metabolism in cellular and subcellular systems of mellanotic tumors prepared from stressed animals(4). In 1960 Paul demonstrated the high sensitivity of brain glucose metabolism to oxygen-dependent inhibition by quinones and dihydric pbenols(5). He proposed that such effects might be related to several diseases involving altered neurological functions(6).

Paul Hochstein is justifiably well known for his work on hydrogen peroxide detoxification in red blood cells. Paul discovered that glucose-6-phosphate dehydrogenase (G-6-PD) activity was essential for the detoxification of hydrogen peroxide in intact, catalase-rich, erythrocytes(7). This was the first demonstration of the operation of the glutathione peroxidase pathway for peroxide detoxification in intact cells. Paul proposed that the failure of this pathway in erythrocytes of individuals with a genetic deficiency of glucose-6phosphate dehydrogenase (favism) was the biochemical basis of the hemolytic anemia associated with the ingestion of many drugs(8,9). Paul went on to demonstrate that many such hemolytic agents caused hydrogen peroxide accumulation in erythrocytes, when administered to animals(10). Since many of these agents which produced oxidative damage were, in fact, reducing substances it was proposed that they reduced molecular oxygen to hydrogen peroxide and that peroxide sensitivity was the basis for this disorder. These concepts were later summarized in a 1988 review in Free Radical Biology & Medicine(11).

Paul discovered the ADP-activated, NADPH-dependent peroxidation of endogenous lipids in microsomes prepared from rat liver(12). He showed that lipid peroxidation resulted in microsomal damage (protein release) and enzyme inhibition (glucose6-phosphatase). Paul further demonstrated that lipid peroxidation in microsomes results in the formation of short chain hydrocarbons such as ethane and ethylene(13) and he described the propagation of lipid peroxidation in microsomes to other (erythrocyte) cellular plasma membranes(14). Paul next demonstrated that lipid peroxidation induced in erythrocyte membranes by oxidative hemolytic agents was accompanied by compensatory lipid repair and the incorporation of free fatty acids into membrane phospholipids(15). He identified palmitylcarnitine transferase as an intrinsic component of erythrocyte membranes(l6). He also described morphological changes and the energy-dependent(l7) internal vacuolization of the plasma membrane of erythrocytes induced by peroxide-generating agents(18).

Paul initiated important studies of futile redox cycling and peroxide formation as a mechanism for the cytotoxicity of antitumor agents such as streptonigrin(19). These studies were later extended to include anthracycline antibiotics such as adriamycin(20,21). Paul demonstrated that cupric-cuprous transitions at the surface of membranes, and at the expense of membrane sulfhydryl groups, resulted in the formation of superoxide(22). He suggested that consequent lipid peroxidation was the basis for the cytotoxicity of copper and the hemolytic anemia associated with Wilson's disease(23,24).

Extensive experiments were carried out by Paul on the alterations of membrane proteins associated with aging and oxidative damage in erythrocytes. The polymerization of membrane spectrin in aging ce11s(25) was found to be similar to that which takes place during oxidative damage(26,27). These alterations were found to result in changes in the physicochemical properties of cells(28), as well as in the fluidity of isolated membranes(29), which presumably lead to decreased deformability of affected cells, splenic entrapment and anemia.
Uric acid was demonstrated to be an effective antioxidant defense in human beings(30). Its surprising ability to protect membranes and DNA from free radical damage was described in detail for the first time by Paul and his collaborators(31), as was as its ability to complex iron(32) and to conserve vitamin C in blood(33). These, and related studies, have now caused most scientists and nutritionists to add uric acid to their lists of important physiological antioxidants.

Paul's investigations on the function of DT diaphorase were initiated in 1962 and continue up to the present time. It was found in 1962, and reported in a Methods in Enzymology paper(34), that the enzyme was a detoxifying quinone reductase in the sense that it converted naphthoquinones to derivatives that were conjugated through their hydroquinone forms. This observation was confirmed many years later when it was found that the redox cycling of these quinones was dependent on one electron reduction and limited by two electron reduction through DT Diaphorase(35). These findings were preshadowed by experiments with antitumor quinones(36,37) and extended by later experiments with C3H/10T 1/2 cells in culture(38).

Investigation of the interaction of myoglobin with cardiotoxic agents such as adriamycin(39) and experiments on the role of hemoglobin in catalyzing oxidative damage in plasma membranes(40) led to the proposal that under conditions (e.g., ischemic-reperfusion states) where hydrogen peroxide might be produced, the resultant formation of high oxidation states (ferryl) of myoglobin might be involved in causing damage(41). This proposal has been further extended by numerous experiments on the detection(42) and biochemical interactions of ferrylmyoglobin with various cellular components(43). It has even been shown that ascorbate, and other endogenous reductants of ferrylmyoglobin may play important roles in protecting the human heart against damage during ischemia-reperfusion and/or coronary bypass surgery(44-47). A full understanding of the role of myoglobin oxidation/ reduction reactions in cardiac ischemia/reperfusion damage awaits future investigations. Some of Paul's most recent papers have returned to some of his earliest and abiding interests in hemoglobin and red blood cell redox reactions. These studies have included an investigation of the mechanism by which uric acid and ergothioneine may inhibit the oxidation of oxyhemoglobin by nitrite(41) and a study of H2O2 production and H2O2 steadystate levels in erythrocytes (49).

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REFERENCES

1. Hochstein P. & Cox C.E. (1952) The effect of tetrachloro-p-benzoquinone on certain fermentative enzymes in fungi. Phyopath. 42:11.

2. Hochstein P. & Cox C.E. (1956) Studies on the fungicidal action of N(trichloromethylthio)-4cyclohexene-1,2-dicarboximide (Captan). Am. J. Botany 43:437-441.

3. Hochstein P. (1957) Glycolysis by tumor mitochondria and the action of insulin. Science 125:496-498.

4. Hochstein P. (1959) Hormonal regulation of subcellular glycolysis in the S91 mouse melanoma. In: Pigment Cell Biology (Gordon, M., ed.) Academic Press, New York, pp 445-459.

5. Hochstein P. & Cohen G. (1960) The inhibitory effect of quinones and dihydric phenols on glucose metabolism in subcellular systems of brain. J. Neurochem. 5:370-378.

6. Cohen G. & Hochstein P. (1963) Enzymatic mechanisms of drug sensitivity in brain. Diseases of the Nervous System 24:1-4.

7. Cohen G. & Hochstein P. (1961) Glucose-6-phosphate dehydrogenase and the detoxification of hydrogen peroxide in human erythrocytes Science 134:1574-1575.

8. Cohen G. & Hochstein P. (1963) Glutathione peroxidase: The major pathway of peroxide detoxification of erythrocytes. Biochemistry 2:1420-1428.

9. Cohen G. & Hochstein P. (1964) Generation of hydrogen peroxide in erythrocytes by hemolytic agents. Biochemistry 3:895-900.

10. Cohen G. & Hochstein P. (1965) In vivo generation of hydrogen peroxide in mouse erythrocytes by hemolytic agents. J. Pharmacol. Exp. Ther. 147:139-143.

11. Hochstein P. (1988) Perspectives on hydrogen peroxide and drug-induced hemolytic anemia in glucose-6-phosphate dehydrogenase deficiency. Free Radical Biol. Med. 5:387-392.

12. Hochstein P., Nordenbrand K. & Ernster L. (1964) Evidence for the involvement of iron in the ADP-activated peroxidation of lipids in microsomes and mitochondria. Biochem. Biophys. Res. Commun. 14:323-328.

13. Lieberman M. & Hochstein P. (1966) Ethylene formation in rat liver microsomes. Science 152:213-214.

14. Hochstein P. (1966) Antioxidant mechanisms associated with lipid peroxidation. In: Proceedings III International Conference on Hyperbaric Oxygen, Publ.1404, National Academy of Science, pp 6164.

15. Wittels B. & Hochstein P. (1966) The effect of primaquine of lecithin metabolism in human erythrocytes. Biochim. Biophys. Acta 125:594-597.

16. Wittels B. & Hochstein P. (1967) The identification of palmitylcarnitine transferase in erythrocyte membranes. J. Biol. Chem. 242:126-130.

17. Berry D.H. & Hochstein P. (1970) Primaquine-induced hemolysis of normal erythrocytes in vitro: The requirement for energy. Biochem. Med 4:317-326.

18. Ginn F.L., Hochstein P. & Trump B.F. (1969) Membrane alterations in hemolysis: Internalization of plasmalemma induced by primaquine. Science 164:843-845.

19. Miller D.S., Laszlo J., McCarty K.S., Guild W.E. & Hochstein P. (1967) Mechanisms of streptonigrin action in leukemic cells. Cancer Res. 27:632-638.

20. Goodman J. & Hochstein P. (1977) Generation of free radicals and lipid peroxidation by redox cycling of adriamycin and daunomycin. Biochem. Biophys. Res. Commun. 77:797-803.

21. Davies K.J.A., Doroshow J.H. & Hochstein P. (1983) Mitochondrial NADH dehydrogenase-catalyzed oxygen radical production by adriamycin, and the relative inactivity of 5-iminodaunorubiein. FEBS Lett. 153:227-230.

22. Kumar K.S., Rowse C. & Hochstein P. (1978) Copper-induced generation of superoxide in human red cell membranes. Biochem. Biophys. Res. Commun. 83:587-592.

23. Hochstein P., Kumar K.S. & Forman S.J. (1980) Lipid peroxidation and the cytotoxicity of copper. Ann. NY Acad. Sci. 355:240-248.

24. Forman S.J., Kumar K.S., Redeker A.G. & Hochstein P. (1980) Hemolytic anemia in Wilson's disease: Clinical findings and biochemical mechanisms. Am. J. Hematol. 9:269-275.

25. Jain S.K. & Hochstein P. (1980) Polymerization of membrane components in aging and blood cells. Biochem. Biophys. Res. Commun. 92:247-254.

26. Jain S.K. & Hochstein P. (1979) Generation of superoxide radicals by hydrazine: Its role in phenylhydrazine-induced hemolytic anemia. Biochim. Biophys. Acta 586:128-136.

27. Jain S.K. & Hochstein P. (1980) Membrane alterations in phenylhydrazine-induced reticulocytes. Arch. Biochem. Biophys. 201:683-687.

28. Corry W.D., Meiselman H.J. & Hochstein P. (1980) t-Butyl hydroperoxide-induced changes in the physicochemical properties of human erythrocytes. Biochim. Biophys. Acta 597:224-234.

29. Rice-Evans C. & Hochstein P. (1981) Alterations in erythrocyte membrane fluidity by phenylhydrazine-induced peroxidation of lipids. Biochem. Biophys. Res. Commun. 100:1537-1542.

30. Ames B.N., Cathcart R., Schwiers E. & Hochstein P. (1981) Uric acid provides an antioxidant defense in humans against oxidant- and radical-caused aging and cancer: A hypothesis. Proc. Natl. Acad. Sci (U.S.A.) 78:6858-6862.

31. Cohen A.M., Abendroth R. & Hochstein P. (1984) Inhibition of free-radical induced DNA damage by uric acid. FEBS Lett. 174:147-150.

32. Davies K.J.A., Sevanian A., Muakkassah-Kelly S.F. & Hochstein P. (1986) Uric acidiron complexes: A new aspect of the antioxidant function of uric acid. Biochem. J. 235:747-754.

33. Sevanian, A., Davies, K.J.A., and Hochstein, P. (1985) Conservation of vitamin C by uric acid in blood. J. Free Rad. Biol. Med. 1:117-124.

34. Ernster L. (1967) DT-diaphorase, Methods Enzymol. 10:309-317.

35. Lind C., Hochstein P. & Ernster L. (1982) DT-diaphorase as a quinone reductase: A cellular control device against semiquinone and superoxide radical formation. Arch. Biochem. Biophys. 216:178-185.

36. Ernster L., Atallah A.S. & Hochstein P. (1986) DT-diaphorase and cytotoxicity and mutagenicity of quinone-derived oxygen radicals. Proc. Clin. Biol. Res. 209:353-363.

37. Atallah A., Landolph J.R. & Hochstein P. (1987) DT-diaphorase and quinone detoxification. Chemica Scripta 27:141-144.

38. Atallah A.S., Landolph J.R., Ernster L. & Hochstein P. (1988) DT-diaphorase activity and the cytotoxicity of quinones in C3H/lOTl/2 mouse embryo cells. Biochem. Pharmacol. 37:2451-2459.

39. Taylor D. & Hochstein P. (1978) Inhibition by adriamycin of a metmyoglobin reductase from beef heart. Biochem. Pharmacol. 27:2079-2082.

40. Rice-Evans C., Baysal E. & Hochstein P. (1985) The role of hemoglobin in erythrocyte membrane alterations induced by t-butyl hydroperoxide. Life Chemistry Reports 3:102111.

41. Galaris D., Eddy L., Arduini A., Cadenas E. & Hochstein P. (1989) Mechanisms of reoxygenation injury in myocardial infarction: Implications of a myoglobin redox cycle. Biochem. Biophys. Res. Commun. 160:1162-1168.

42. Arduini A., Eddy L. & Hochstein P. (1990) Detection of ferryl myoglobin in the isolated ischemic rat heart. Free Radical Biol. Med. 9:551-513.

43. Galaris D., Mira D., Sevanian A., Cadenas E. & Hochstein P. (1988) Co-oxidation of salicylate and cholesterol during the oxidation of metmyoglobin by H2O2 Arch. Biochem. Biophys. 262:221-231.

44. Eddy L., Hurvitz R. & Hochstein P. (1990) A protective role for ascorbate in induced ischemic arrest associated with cardiopulmonary bypass. J. Appl. Cardiol. 5:409-414.

45. Arduini A. & Hochstein P. (1991) Myoglobin, a double-edged sword in myocardial infarction. In: Oxidative Damage & Repair (Davies, K.J.A., ed.) Pergamon Press, New York, pp. 409-414

46. Hochstein P. & Arduini A. (1992) The special role of myoglobin in cardiac ischemia-reperfusion injury. In: Biological Free Radical Oxidations and Antioxidants (Ursini F. & Cadenas E. eds.) CLEUP Press, Padova, Italy, pp. 151-158.

47. Arduini A., Mancinelli G., Radatti G.L., Damonti W., Hochstein P. & Cadenas E. (1992) Reduction of sperm whale ferrylmyoglobin by endogenous reducing agents: Potential reducible loci of ferrylmyoglobin. Free Radical Biol. Med. 13, 449-454.

48. Arduini A., Mancinelli G., Radatti G.L., Hochstein P. & Cadenas E. (1992) Possible mechanism of inhibition of nitrite-induced oxidation of oxyhemoglobin by ergothioneine and uric acid. Arch. Biochem. Biophys. 294:398-402.

49. Giulivi C., Hochstein P. & Davies K.J.A. (1994) Hydrogen peroxide production by red blood cells. Free Radical Biol. Med. 16:123-129.

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