The Oxygen Paradox: A Tribute to Paul Hochstein
"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 "living tribute") 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.
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
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
It is, perhaps,
as Director of the USC Institute for Toxicology that Paul is most
widely known. In 1981 Paul took up the additional major responsibility
of being Associate Dean for Research and Graduate Affairs of the
School of Pharmacy at USC. In 1988 Paul became the first Charles
Krown Professor at USC. Paul remained as Director of the Institute
for Toxicology, Professor of Biochemistry, and Associate Dean for
Research and Graduate Affairs until 1993. Since July 1993 Paul has
been the Distinguished Emeritus Professor of Molecular Pharmacology
& Toxicology at the University of Southern California.
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).
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).
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).
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.
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
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
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.
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
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.
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.
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
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.
29. Rice-Evans C.
& Hochstein P. (1981) Alterations in erythrocyte membrane fluidity
by phenylhydrazine-induced peroxidation of lipids. Biochem. Biophys. Res.
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.
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.
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
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.
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.