Irwin Stone: An Influential Man

Irwin Stone. (Image by Oscar Falconi)

Irwin Stone. (Image by Oscar Falconi)

[Part 1 of 2]

Dr. Irwin Stone was a biochemist and chemical engineer who maintained a particular interest in and enthusiasm for vitamin C. Stone was the person who first raised Linus Pauling’s interest in vitamin C, leading to Pauling’s extensive program of research on vitamin C and its uses for the prevention and treatment of disease. Pauling’s contributions to the field are one of the big reasons why many people believe in taking vitamin C for the prevention and treatment of colds today.  But for Pauling, it all started with Irwin Stone.

Stone was born in 1907 and grew up in New York City. He attended the College of the City of New York and then worked at the Pease Laboratories, a well-known biological and chemical consulting lab, from 1924 to 1934. Stone started out as a bacteriologist, but was promoted to Assistant to the Chief Chemist and then to Chief Chemist.

In 1934 the Wallerstein Company, a large manufacturer of industrial enzymes, recruited Stone to set up and direct an enzyme and fermentation research laboratory. Ascorbic acid, or vitamin C, had just been identified and synthesized by a Hungarian research team led by Albert Szent-Györgyi, who later won the 1937 Nobel Prize in Medicine for his work. Stone pioneered processes for implementing the antioxidant properties of ascorbic acid in industrial settings. One specific application that Stone developed was the use of ascorbic acid as a preservative for food – an innovation that landed him three patents.

Stone’s interest in vitamin C lasted throughout his life. He began to study scurvy intensely and by the late 1950s he had formulated a hypothesis that scurvy was not merely a dietary issue, but a flaw in human genetics. (He called it “a universal, potentially-fatal human birth defect for the liver enzyme GLO.”) Stone considered the amount of vitamin C that nutritionists recommended in a healthy diet – the Recommended Daily Allowance (RDA) - to be far from sufficient. In 1968 that recommendation was 55 mg for women and 60 mg for men. The current standard is slightly increased at 75 mg for women and 90 mg for men, with higher recommendations for pregnant and lactating women. But none of these figures are anywhere near Stone’s recommendations.

Stone believed that humans suffer from “hypoascorbemia,” a severe deficiency of vitamin C, caused by our inability to synthesize the substance the way that virtually all other mammals do. Most other mammals synthesize vitamin C in large quantities relative to body weight; proportionately, humans theoretically should be taking between 10-20 grams daily. Stone suggested that about 25 million years ago the primate ancestors of human beings lived in an environment in which they were able to consume relatively massive amounts of ascorbic acid, compared with what we get from our diets today. These material circumstances created an environment in which a genetic mutation occurred that allowed these human ancestors to stop synthesizing the substance. In present day, Stone noted, these amounts of ascorbic acid are not readily available in our diets, so humans may only be getting 1-2% of what they need.

This hypothesis initially led Stone to propose a vitamin C intake of 3 grams for optimal health, 50 times the RDA, and as he further researched ascorbic acid, he recommended increasingly higher doses. He was convinced that taking less than the amount that he recommended would cause “chronic subclinical scurvy,” a state of lowered immunity that increased susceptibility to a variety of illnesses. He felt that large doses of ascorbic acid should be used to prevent and treat infectious and cardiovascular diseases, collagen breakdown, cancer, SIDS, birth defects, AIDS, and health problems normally associated with aging.

Practicing what he preached, Stone and his wife began taking megadoses of vitamin C and they found that it greatly improved their overall health. When the couple both incurred injuries from a serious car accident, they treated themselves in part with large doses of vitamin C and reported a swift recovery. Stone attributed their rapid healing to the large doses of vitamin C.

Letter from Irwin Stone to Linus Pauling, April 4, 1966.  This is the communication that spurred Pauling's interest in vitamin C.

Letter from Irwin Stone to Linus Pauling, April 4, 1966. This is the communication that spurred Pauling’s interest in vitamin C.

In March 1966, Linus Pauling gave a speech on the occasion of his receiving the Carl Neuberg Medal for his work in integrating new medical and biological knowledge. In the speech, Pauling – who was 65 years old at the time – mentioned that he hoped to live for another fifteen years so that he might see several advances of science in medicine that he anticipated to be emerging during that time period.

Irwin Stone was in the audience at this lecture and, on April 4, 1966, he wrote Pauling a fateful letter in which he noted

You expressed the desire, during the talk, that you would like to survive for the next 15 or so years….I am taking the liberty of sending you my High Level Ascorbic Acid Regimen, because I would like to see you remain in good health for the next 50 years.

Pauling was initially skeptical of Stone’s advice, but he had recently learned about other uses of megavitamin therapy and their successes, so he decided to give the regimen a try. It was at that point that Linus and Ava Helen Pauling began taking 3 grams of vitamin C a day.

In July Pauling wrote back to Stone: “I have enjoyed reading your paper and manuscript about hypoascorbemia. I have decided to try your high level ascorbic acid regimen, and to see if it helps me to keep from catching colds.”

Pauling, as it turned out, was impressed by the results. For most of his adult life, he had suffered from severe colds several times a year and had taken a daily dose of penicillin off and on from 1948 to the early 1960s. Pauling thought that the penicillin doses were his primary defense against colds but, in all likelihood, he was probably just killing off his good bacteria and making himself more susceptible to colds through his overuse of antibiotics. Once the Paulings started taking vitamin C, they reporting a noticeable uptick in their physical and emotional energy, and seemed to suffer from fewer colds.

Two years after their initial communications, Stone noticed that Pauling had cited him in a recently published journal article. Stone described his difficulties in getting his research published and the backlash that he was experiencing from physicians. He also asked about Pauling’s health.

The last time I wrote you in 1966, you mentioned that you were going to try my high level ascorbic acid regimen to see if it would help prevent your catching colds. How did it work? At the time you also had a broken leg. I know from personal experience [a reference to his car accident] that it is excellent in bone healing.

Pauling replied

I can report that both my wife and I have been less troubled by colds during the last two years, during which we have been taking 3 to 5 grams of ascorbic a day, than we had been before beginning your regimen.

He also asked about Stone’s research on ascorbic acid and leukemia.

During the late 1960s, Pauling did not make a point of promoting vitamin C megadoses, though he did support the use of megavitamin therapy for the treatment of schizophrenia. But by 1969, he was finally fully convinced of Irwin Stone’s arguments as well as his own personal successes with vitamin C, and he began to promote vitamin C publicly.

Alejandro Zaffaroni, 1923-2014

Alejandro Zaffaroni. (Life Sciences Foundation image)

Alejandro Zaffaroni. (Life Sciences Foundation image)

In a 1997 interview with Jill Wolfson and Tejinder Singh, Alejandro Zaffaroni shared what it was like for him growing up in Montevideo, Uruguay, where he was born on February 27, 1923. He described himself as “kind of a sick child” with asthma severe enough to keep him isolated from his peers. As a boy he spent his time alone outdoors playing, exploring, and thinking about what he found. And when not playing outside, he attended a Jesuit school where he rebelled against the strict disciplinary regime. Zaffaroni described this as an opportunity to “think a lot about all kinds of strategies,” a skill that came in handy in his adult life.

As Zaffaroni transitioned into a public school at the age of twelve, his mother passed away. His father, a banker, began to spend more time with the boy, exposing him to the cultural life of Montevideo through the symphony, opera, and other outings. Encouraged by his father to develop his own interests, Zaffaroni found chemistry and, with the help of a friend who had a much better grasp of the content than the rest of the class, found a subject he could excel in. Unfortunately, just five years after his mother had done so, Zaffaroni’s father also passed away. Yet his example continued to inspire his son.

As a pre-med student at the University of Montevideo, Zaffaroni encountered biochemistry, a subject he pursued further as the first Uruguayan to enroll in a biochemistry Ph.D. program in the United States. In July 1945 he headed to Rochester University which Zaffaroni chose over other options, including Harvard, because of the promise of freedom to follow his own research path in endocrinology, with a focus on steroids. After finishing his doctorate in 1949, he continued his work with steroids with the support of a grant from the National Institutes of Health and published his first article, “Adrenal Cortical Hormones,” in Science with Robert B. Burton and E. Henry Keutmann.

Finished with his education and ready for the next step, Zaffaroni had his pick of offers from several universities and private labs. Harvard again appeared to be a possibility, one that briefly brought him to the attention of Linus Pauling. In 1953, George. B. Kistiakowsky of Harvard wrote to Pauling for advice on a list of candidates for a new biochemistry professorship that included “Alessandro Zaffaroni.” Pauling underlined two names, Frank H. Westheimer, which he annotated with “best,” and Zaffaroni, which he annotated with “never heard of him.” Zaffaroni chose another path as Westheimer ultimately got the position, delaying further contact between Zaffaroni and Pauling for another fifteen years.

Continuing to follow his father’s earlier directive that he seek out what interested him most, Zaffaroni also turned down a position at the newly established Sloan-Kettering Institute which, according to a 2012 article in Life Sciences Foundation Magazine, had “one of the world’s top steroid labs.” Expanding on an established relationship with George Rosenkranz at Syntex, Zaffaroni chose instead to head to Mexico City to work for the smaller company, where he felt he would have fewer restrictions on his own research. At this time, in 1953, Zaffaroni also received his first patent, which concerned the extraction of adrenal hormones from bovine and porcine adrenal glands.

At Syntex, Zaffaroni worked on synthesizing steroids using a phytoestrogen extracted from yams. When he noticed that the quality of the yams interfered with the process, he went to the supplying yam farm himself and reorganized harvesting and transportation while also increasing its worker’s wages. Zaffaroni’s efforts were noticed by Charles Allen, who bought Syntex in 1956 and gave the young researcher a promotion. Zaffaroni and Rosenkranz quickly built the company into a major supplier of topical corticosteroids.

In 1962 Zaffaroni was named president of Syntex’s subsidiary in Palo Alto, California, and given the specific charge to gain access to the US pharmaceutical market. The company entered the nascent birth control pill market in 1964 and started creating offshoot companies to take advantage of Syntex’s various lines of research. This led to the creation, in 1966, of the Syva Corporation, which produced diagnostic equipment, as well as the pest control company, Zoecon, incorporated in 1968.

In addition to his executive duties, Zaffaroni also began thinking about new methods of drug delivery, but the culture at Syntex was not supportive of this work. So, funded by $3 million of his own money, Zaffaroni started ALZA in 1968 to focus on this new path of research. ALZA’s main products included an ocular insert designed to administer glaucoma medication, an intrauterine device for birth control medication, and transdermal patches, each of which incorporated timed release mechanisms. Though ALZA’s products were innovative, the pharmaceutical market was hard to move as eye drops, pills, and injections maintained their dominance.

The creation of ALZA, Inc., based in Palo Alto, coincided with Linus Pauling’s move to Stanford University in 1969. In March of that year, Zaffaroni introduced himself to Pauling, writing that he was “extremely pleased” that Pauling would soon be nearby and noting his eagerness to talk with Pauling in person about the burgeoning field of orthomolecular psychiatry. By enclosing some company literature, Zaffaroni also got Pauling interested in ALZA. The two met in April as Linus and Ava Helen were in the midst of their house hunting. The meeting was especially fruitful for Pauling as Zaffaroni provided him with a $100,000 grant to be divided over his next four years at Stanford.

For the next few years the two maintained an informal relationship by visiting each other, sharing ideas, and extending invitations to social gatherings. In 1974 Pauling brought a formal element to their relationship by asking Zaffaroni to become a member of the Board of Associates of the Linus Pauling Institute of Science and Medicine, at that point still in its infancy. Zaffaroni told Pauling in November, “I have been approached by many groups to participate in directorships of various worthwhile organizations, and have been forced to decline. But because of my great admiration for you and for your accomplishments, I am prepared to accept.” He warned Pauling that his busy schedule might interfere at times and in reply Pauling promised that he would not make too many demands. Zaffaroni’s fundraising experience quickly became central to his activities as an associate and he began a correspondence with Art Robinson – at that time LPISM’s Assistant Director – concerning the development of a prospectus for potential donors.

In the summer of 1975, Pauling asked Zaffaroni to extend his relationship with the Institute by joining its Board of Trustees. Again, Pauling promised that Zaffaroni’s duties would be minimal. Zaffaroni made one stipulation in his acceptance: that the Institute revisit the operation of its fledgling medical clinic. Zaffaroni told Robinson that the psychiatric research going on at the clinic needed a better review system. Robinson responded by suspending all outpatient services at the clinic. By November, with the changes in place, Zaffaroni agreed to join the Board.

In his new capacity, Zaffaroni continued his involvement in helping Robinson with fundraising. Their first outreach effort involved “two popular appeals,” one in Prevention and the other in Executive Health. The Institute published an article in both publications and placed a request for donations at the end of each one. Robinson reported to Zaffaroni in March 1976 that the Prevention article had generated 183 donations worth $5,101.50 while the Executive Health article received 103 donations worth $8,031.50.

(Life Sciences Foundation image)

(Life Sciences Foundation image)

Besides running ALZA and serving on the Board at LPISM, Zaffaroni continued his own scientific work. This included “Special requirements for hormone releasing intrauterine devices,” published in Acta Endocrinologica in 1974, and “Contraception by intrauterine release of steroids,” which appeared in the Journal of Steroid Biochemistry in 1975. He also received patents for his transdermal bandages in 1974 and 1976, and patents for controlled release tablets in 1976 and 1977. The following year he published an article, “Therapeutic Systems: The Key to Rational Drug Therapy,” in Drug Metabolism Reviews, that described some of his new developments.

Pauling tried to tie in his own interests to these drug delivery systems. In March 1979, Pauling wrote to Zaffaroni about a study on the improvement of patients with anorectal cancer who were given time released capsules of ascorbic acid. Pauling saw the improvement as being due to topical effects and shared his idea for a similar “slow release” capsule for stomach cancer, which was then afflicting Ava Helen. Pauling’s capsule would “spring into the shape of a sphere, which would cause it to be retained in the stomach” as the ascorbic acid was released into the stomach and afterwards broken down and digested. Zaffaroni responded that ALZA was already at work on something similar to what Pauling described and that he was eager to talk more about it. By 1986 ALZA had released a “once-a-day” Vitamin C supplement utilizing its controlled-release technology.

Amidst the biotech boom of the early 1980s, Zaffaroni’s business practices came under closer examination. In 1982 Time highlighted Zaffaroni’s recent start-up, DNAX, as one of the genetic engineering companies “having trouble living up to their early billing” as one of the “hottest companies on Wall Street.” The article pointed out that to start the company Zaffaroni had “easily raised $5.5 million,” but was now “spending nearly $4 million annually on research, and…does not expect to see any profits for at least another six or seven years.” Other sources in the popular press were often more critical, focusing on Zaffaroni’s commercial, rather than scientific, performance.

In August 1985, Zaffaroni decided to resign from LPISM’s board. He felt that his work was done, telling Pauling that the Institute “has now attained worldwide recognition” and that, with Pauling’s “guidance it will continue to evolve in many positive directions.” He continued

My pattern, as you know, has been to participate in the founding of various enterprises and, once they are established, to go on to new ones. That is because I believe that what is done initially counts more than anything else. Thus, my focus has generally been on contributing innovative concepts at the outset rather than remaining permanently associated with any particular endeavor. That pattern enables me to do what I do best and to keep from becoming stale. It also leaves room for others to follow with fresh insights and new concepts. Thus, I believe it serves everyone well.

I am sure that you know, without my saying it, that the main inducement to my acting as Trustee has been the opportunity it gave me to work with you. That association has brought me great personal pleasure, intellectual challenges, and a keen appreciation of your many gifts of heart and head. You may be sure that I stand ready to help you at any time in any way I can, should the need arise.

Zaffaroni did, however, remain on the Institute’s Board of Associates until 1996 and maintained his connections to LPI into the 2000s when he met with longtime administrative officer Stephen Lawson in Palo Alto to discuss the Institute’s collaborative research on ALS and peroxynitrite in Uruguay.

Congratulatory note from Pauling to Zaffaroni on the occasion of Zaffaroni being honored by the Weizmann Institute of Science, November 1989.

Congratulatory note from Pauling to Zaffaroni on the occasion of Zaffaroni being honored by the Weizmann Institute of Science, November 1989.

After leaving LPISM, Zaffaroni also stepped down as ALZA’s CEO to seek out new endeavors. In 1988 he asked Pauling to become the Honorary Scientific Advisor for the newly forming Affymax Research Institute. As part of the deal, LPISM received 25,000 shares of stock in the company. By June 1990, Zaffaroni told Pauling that Affymax was “moving from an early stage ‘start-up’ to a successful development stage pharmaceutical company.”

The press continued to be somewhat cautious. The following year, the New York Times still referred to Affymax as a promising start-up, but was concerned with Zaffaroni’s avoidance of investment bankers by relying on his many contacts to raise his own funds. The Times followed up with questions as to how Affymax “burned through over $20 million so fast” and by noting that its central product -VLSIPS, short for Very Large Scale Immobilized Polymer Synthesis, which were biological compounds produced on silica chips developed by the semiconductor industry - was not selling well. Zaffaroni was not concerned with Affymax’s profitability at all, informing Pauling in May 1991 that he had raised $26 million and was ready for the company to “aggressively pursue our scientific and commercial goals” as well as move into their new research facility. By the end of the year, Affymax began offering public stock.

At the end of the 1990s, Zaffaroni began attracting more praise in the press, particularly for his founding, in 1995, of Smyx, a company focusing on the applications of combinatorial chemistry in the development of drugs. But he was also running up against new criticism echoing that issued in the early 1980s about the viability of DNAX. This time those fears were directed at Affymetrix, which had been spun off from Affymax in 1992 to focus on VLSIPS. An article in Forbes reported that the chips, though potentially “a godsend to medicine,”  may not be a “godsend…to Affymetrix’s bottom line.”

Zaffaroni soon had more direct problems to deal with. In 1995 he organized the sale of Affymax to Glaxo for $533 million. Some of Zaffaroni’s friends and family members began trading shares of Affymax just before the sale was announced publicly, which drew the attention of the Securities and Exchange Commission. Charged by the SEC with insider trading, Zaffaroni and six others agreed to pay fines of $1.85 million and immediately tried to move on. According to the New York Times, Zaffaroni’s lawyer stated that his client wanted “to devote his time to science and charity rather than litigation.” And that’s just what he did as he continued to produce more patents related to drug delivery and VLSIPS.

The 2000s saw Zaffaroni open up another line of research, this time for a drug delivery system that used the cigarette as a model. In 2000 Zaffaroni started Alexza to focus on this research and, before too long, he, along with Joshua D. Rabinowitz and Dennis W. Solas, produced patents for delivering insomnia, anti-inflammatory, antipsychotic, pain relieving, and several other drugs through inhalation. Unlike other inhalation drugs, these relied on the slight heating of the drugs before delivery. Zaffaroni and others described their research in “Fast Onset Medications through Thermally Generated Aerosols” for the Journal of Pharmacology and Experimental Therapeutics in 2004. In 2012 Alexza’s first product, aimed at schizophrenia and bipolar disorder, was ready for market. That same year, at the age of 89, Zaffaroni was inducted into the National Inventors Hall of Fame.

Zaffaroni passed away this past March 1st from complications related to dementia. When asked in 1997 how he wanted to be remembered, Zaffaroni said

Well, the one thing that was always very important in my life is human relations. So in looking at candidates for any of the jobs that I had, I wanted to have people who shared my value system, in addition to being the kinds of capable individuals that I needed to have.

Perhaps drawing on his father’s support and encouragement of him as a youth, Zaffaroni continued,

It is tremendously important in building this company that I create a very, very warm caring environment, so that people have an opportunity to do the best of their work. One of the key things to success is never to worry about failing. Many people do not do a lot of the things that could be done because they do not want to have a negative result. If you don’t go for the new breakthrough, if you are going just to stay in the areas which we all know, we are stationary.

Now on the other hand, if you make a huge effort with a new idea and you don’t succeed, the big companies don’t see that as a good thing. So why take risk, if there is no opportunity to be rewarded by the effort?

In my view, the only thing attractive in life is continually to move forward, to be looking for new opportunities, and to support people and let them fail safely.

Rebecca Mertens, Resident Scholar

Rebecca Mertens

Rebecca Mertens

Rebecca Mertens of Bielefeld University, located in northwest Germany, is the latest visitor to complete a term as Resident Scholar in the Oregon State University Libraries Special Collections and Archives Research Center.  A Ph.D. candidate in the philosophy and history of science, Mertens spent a month stateside, visiting both the OSU Libraries as well as the Caltech Archives.

During her stay she braved both a major (and unusual) snow event in Corvallis as well as torrential rains in southern California.  Despite these obstacles, Mertens enjoyed a fruitful visit to the west coast as she pursued her research on Linus Pauling’s contributions to the lock-and-key model of biological specificity and the influence that this model imparted upon the sweep of modern biochemistry.

The conditions that awaited Mertens upon her arrival at OSU.

The conditions that awaited Mertens upon her arrival at OSU.

An outgrowth of his research on antibodies and antigens, Linus Pauling’s work on biological specificity comprised a major contribution to contemporary thinking on biochemical topics.  Pauling biographer Thomas Hager gives us this primer on what is meant by by the term, “biological specificity.”

Pauling demonstrated that the precise binding of antigen to antibody was accomplished not by typical chemical means – that is, through covalent or ionic bonds — but solely through shape. Antibodies recognized and bound to antigens because one fit the other, as a glove fits a hand. Their shapes were complementary. When the fit was tight, the surfaces of antibody and antigen came into very close contact, making possible the formation of many weak links that operated at close quarters and were considered relatively unimportant in traditional chemistry — van der Waals’ forces, hydrogen bonds, and so forth. To work, the fit had to be incredibly precise. Even a single atom out of place could significantly affect the binding.

In her Resident Scholar presentation, Mertens described the thrust of her research, which focuses on how one should interpret the contributions that Pauling made in this particular arena.

In the course of his research on antibodies, Linus Pauling postulated that the complementary structure of two molecules or two parts of a molecule determined the specificity of reactions in the living organism. However, the idea that molecular complementarity and biological specificity are deeply connected was already mentioned by Emil Fischer at the end of the 19th century. Thus, Pauling’s novel contribution was not the initial articulation of the model, but rather his emphasis on the importance of molecular complementarity for all biological phenomena.

Through examination of the Ava Helen and Linus Pauling Papers, as well as the institutional records held at Caltech, Mertens is pursuing the idea that “Pauling’s interdisciplinary reputation, his public presence and his engagement in the organization of scientific institutions led to the popularity of the lock-and-key model and to its standardization in the second half of the twentieth century.”  These forces of Pauling’s status and personality in turn made an impact on questions of “financial support, networking and science popularization within the administration of scientific projects.”


Beyond uncovering and detailing the history of Pauling’s role in the development of the lock-and-key model, Mertens is also using her research to “suggest an approach to the study of analogical models that considers social and political factors on successful model usage…[and] the formation and consolidation of model-based research programs.” Mertens returned to Germany with a large volume of content to sift through and absorb as she continues to develop her thinking on these issues.

Now entering its seventh year, the Resident Scholar Program at OSU Libraries provides research stipends of up to $2,500 to support work conducted in the Special Collections and Archives Research Center.  Applications for the 2014 class of scholars are being accepted now – the deadline for entry is April 30, 2014.  For more details, please see the program homepage.

An Interview with Zia Mian

Dr. Zia Mian, who will be traveling to Oregon in April to accept the 2014 Linus Pauling Legacy Award, was kind enough to give us a bit of his time not long ago for an interview.  In it he discussed a whole range of topics including the development of his socio-political consciousness, his admiration for Pauling and his thoughts on healing old wounds in South Asia.  The transcript of our conversation is presented below.

For a more technical perspective on Mian’s thinking with particular respect to the Comprehensive Test Ban Treaty, see the embedded video above.  An excellent profile of Mian, published by his home institution, Princeton University, is likewise available here.

Pauling Blog: You studied physics in graduate school. Were you already interested in socio-political issues? Or did you experience an awakening of sorts, as happened to Pauling with Hiroshima and Nagasaki?

Zia Mian: I’m of a generation of people that were growing up during the period of the late 1970s and the early 1980s, what has come to be called the Second Cold War, where President Reagan and the United States, and I believe it was Western Europe, moved new nuclear missiles into Western Europe as a response to new Soviet missiles that had been developed. And so there was a great risk of nuclear war again and peace movements across Europe and in the United States became very active. We had some of the largest demonstrations by these groups that had ever been seen in New York and London and other cities. And the presence of such a large and determined and active social movement raises questions for all kinds of people, such as “what do I think about this issue? What does this mean? How does this impact society and what is my role in what’s going on?”

And so as a young physics student it became obvious that nuclear weapons were something that I had to think about and to try and understand what I thought about them and what they might mean. And so as a consequence I think that it wasn’t so much like a calling of having a Hiroshima or Nagasaki type moment, but the existence of a large and determined peace movement raising the issue to people across the world, that this is an issue you have to take seriously and come to a position on. That led me to think about what nuclear weapons meant and how I felt about them.

PB: With Pauling and several other scientists at the beginning of the nuclear age, they could understand the science behind nuclear weapons as well, and that seemed to lend itself toward their activism, in the sense that they could understand how they worked and the amounts of energy they could release. Did that play in for you as well?

ZM: At the beginning of the nuclear age certainly many scientists, including ones who had worked on the Manhattan Project, realized that the public and policy makers needed to understand the new dangers that nuclear weapons and nuclear materials posed to the world. And having a technical background made it easier to understand some of the things that nuclear weapons mean, without having to know secrets. Because the science was sufficiently clear that you could make this understanding of what was going on. What you have to remember is that lots of other people came to a similar understanding about nuclear dangers without being scientists. One thinks of Mahatma Gandhi writing about the danger of nuclear weapons soon after Hiroshima and Nagasaki, or the French writer and philosopher Albert Camus or the English writer George Orwell or the American writer Lewis Mumford. All of them, within months or the first year or so after Hiroshima, tried to explain to people that these nuclear weapons posed a profound and unimaginable new danger, without being scientists themselves.

But the scientists—being experts gives you a somewhat privileged position to debate, because people have a tendency to look to scientists as being people who can understand and explain some of the more detailed factual and technical basis of what nuclear weapons and their production and use mean, rather than just talking about the politics of what nuclear weapons mean or the ethics and morality of what nuclear weapons mean. But I can’t emphasize strongly enough that many of the early scientists like Pauling and others, as well as writers like Mumford and Bertrand Russell and Albert Camus and George Orwell who wrote about nuclear weapons, combined both a technical understanding and a political understanding and a moral and ethical sensibility about what these weapons would mean. And it was only by taking them all together that one can see what kind of intervention they made in helping people understand the nuclear danger.

Continue reading

Dr. Pauling’s Chiral Aliens

[A guest post expanding on Pauling's idea for a science fiction novel. Post authored by the blog's East Coast Bureau Chief, Dr. John LeavittNerac, Inc., Tolland, CT.]

Pauling lecturing with the "fish model" (foreground) that he used to demonstrate chirality, ca. 1960s.

Pauling lecturing with the “fish model” (foreground) that he used to demonstrate chirality, ca. 1960s.

In basic chemistry we have something called “chirality” which refers to a molecule with two possible non-superimposable configurations. One way to picture this is to look at your hands and place one on top of the other (not palm to palm) – your left and right hands are essentially the same shape but their shape is reversed. At the molecular level we can use one of the main building blocks of all proteins and all life – the amino acid alanine, depicted in the image below – to examine handedness.

alanine enantiomers

The diagram shows the arrangement of atoms of two alanine molecules, both of which exist in nature, arranged so that they are mirror images. They are the same molecules but if you turn the one on the right around so that it is facing in the same direction as the one on the left, the R (a single carbon atom in alanine with three bonded hydrogen atoms) on this alanine molecule faces toward the palm of the hand and the COOH moiety (a carboxyl group) and the NH2 moiety (an amino group) face outward away from the palm.

No matter how you rotate the alanine on the right, you can’t get the three moieties attached to the central carbon to line up in the same position as the alanine on the left. Likewise, you can’t get those hands to super-impose each other no matter how much you twist and turn them. So the alanine on the left is called L-alanine (levo- for the direction the molecule rotates photons) and the alanine on the right is called D-alanine (dextro- for the direction the molecule rotates photons). They are called “enantiomers,” or chiral forms, of alanine, and both exist in nature with identical chemical properties except for the way that they rotate polarized light.

There are twenty natural amino acids comprising the building blocks of all proteins. Of these twenty, only glycine is symmetrical around a central carbon atom and therefore glycine has no enantiomers. The other nineteen can exist in the L- and D-conformation.

Funny thing though, only the L-enantiomer is used to make proteins by the protein synthetic machinery of all life-forms, from single-cell organisms up to humans. It’s quite easy to understand why one enantiomer is used in life over random use of either enantiomer. In explaining this, note the pictures below, which show the three-dimensional globular structure of human beta-actin on the left and, on the right, the architectural arrangement of this actin in the cytoplasm of a cell.


The protein composed of 374 amino acids has an intricate folding pattern with coils which would not be possible if both amino acid enantiomers for the nineteen amino acids were randomly incorporated into the protein. This three-dimensional structure has to be preserved in order for actin to perform its dynamic architectural function inside living cells, as shown in the picture on the right. The coils are possible because the amino acids are all L-amino acids and glycine is neutral; otherwise the protein would behave like a wet noodle. The precise structure of the actin protein determines its function, which has been preserved and conserved since the beginning of all eukaryotic life-forms (that is, cells with a cytoplasm and a nucleus). Understanding the atomic forces that fold proteins in a unique shape is part of the reason why Linus Pauling received the Nobel Prize for Chemistry in 1954.

Aside from those who closely follow this blog, it is not well known that Linus Pauling was an avid reader of science fiction. In a 1992 interview with biographer Thomas Hager, he described his motivation to write a science fiction novel. The story line was to be the discovery of a human-like race from another planet that had evolved to use only D-amino acids (D-humans) rather than the L-isoform (L-humans). He explained that he never got around to writing this novel because the real science he was doing took all of his time.

If our L-humans met up with those D-humans, what consequences would there be? Well, what we would see in D-humans are people virtually indistinguishable from ourselves – barring, of course, the possibility that these extraterrestrials evolved out of some unearthly environmental niche. However, no mating, blood, or tissue sharing would be possible between these two races.

To explain this, consider the experience you have had when you accidently put your hand in the wrong glove. As you know, this doesn’t work well. All protein interactions and reactions catalyzed by enzymes require a direct fit to work. Substrates of enzymes have to fit precisely into the catalytic active site of the enzyme, like your hand fitting into the correct glove. Since L-humans have a different chirality from D-humans, nothing would fit or be transferrable, because of asymmetric incompatibility between L- and D- macromolecules. Even the food on our planet would not likely be nutritious for D-humans because all living things on Earth are L-organisms. In D-lifeforms, the actin coils would coil in the opposite direction and the DNA double helix would have to spiral in the opposite direction as well; otherwise the analogous D-proteins would not bind or fit on the chromosomal DNA.


It seems reasonable that D-humans might be found on other planets if you consider how life got started. By a quirk of nature on Earth, L-amino acids got a head start and self-assembled into peptides (small proteins) when this essential process for life as we know it got started. The assembly of only one enantiomer isoform into a peptide may have been favored thermodynamically over co-random assembly of L- and D-isoforms. This essential process evolved into a well-organized, membrane-protected and energy-driven protein synthetic machinery in single cell organisms like bacteria. Today, humans have essentially the same protein synthetic machinery that evolved in primordial bacteria and all life-forms on Earth have the same genetic code.

There are two essential enzymes that work together to catalyze protein synthesis in all living cells. One enzyme, called aminocacyl-tRNA synthetase, binds the amino acid to a transfer RNA molecule (there is one of these enzymes and a specific tRNA for each of the twenty amino acids). The second enzyme, peptidyl transferase, catalyzes the formation of a peptide bond linking two amino acids at the start of a chain and does this over and over again until the full length protein is synthesized and folded into its functional conformation. These two essential enzymes do not recognize the D-isoforms of the nineteen asymmetric amino acids. Thus, our chiral L-specificity has been preserved since the beginning of life.

I can’t think of any reason why the D-amino acids would not support life, but it has to be one isoform or the other, not both. Apparently Pauling felt the same way. Should it ever come to pass, D-humans will be interesting to meet and they will be equally interested to meet us, hopefully without mutual disappointment.

Dr. Pauling’s Prediction of a Mutation in Beta-Globin Which Causes Sickle Cell Anemia and How This Prediction Impacted My Research

[Guest post written by John Leavitt, Ph.D., Nerac, Inc., Tolland, CT.]

Linus Pauling lecturing on sickle cell anemia, Kyoto, Japan. 1955

In September 2010, the company BlueBird Bio announced that it had cured a patient with the hemoglobinopathy, beta-thalessemia, by correcting the genetic defect in beta-globin that this patient inherited from his parents. This came 61 years after Linus Pauling and his associate, Harvey Itano, explained the cause of hemoglobinopathies such as sickle cell anemia. Beta-thalessemia like sickle cell anemia is caused by an inherited mutation in the beta-globin gene, just a different mutation. In the case of thalessemia, the defective beta-globin gene product disappears, whereas the defective beta-globin in sickle cell anemia remains stable to wreak havoc on the body. BlueBird Bio accomplished this first cure of a hemoglobinopathy by removing the blood-forming hematopoietic stem cells from the patient, engineering his cells ex vivo with a correct beta-globin gene, and then putting the cells back into the patient. The stem cell transplant sustained itself and produced red blood cells which functioned normally in the circulatory system. For the first time in this patient’s 18 year-old life, he did not have to have a monthly blood transfusion.

In late September 1981, when I gave a seminar at the Pauling Institute of Science and Medicine in Palo Alto, CA, I noticed that Dr. Pauling was smiling during the talk. He was aware of the discovery of the muscle isoform of actin by his friend Albert Szent-Györgyi and knew about the structure and function of actins (the subject of my talk). After reading Dr. Pauling’s 1949 paper on the molecular nature of the sickle cell trait, I understood that he was seeing during my talk the very same experiments in my discovery of a mutant human beta-actin that he and Harvey Itano had performed, which led to the prediction of a mutation in the hemoglobin protein that caused sickle cell anemia. His paper was the very first to describe the molecular genetic basis of a human disease. By 1981 there was plenty of conceptual evidence to suggest how I could look for mutations in proteins using electrophoretic separation of complex mixtures of cellular proteins. In 1949 though, Dr. Pauling was way ahead of his time. In his and Itano’s case, the plan was well thought out based upon years of characterization of oxygen bonding to the heme of the globin molecule. By contrast, I was very lucky to find a mutation in the most abundant structural protein of the cell, cytoskeletal actin in a human fibrosarcoma cell.

Harvey Itano.

It was probably evident in 1949 that hemoglobin amounts to about 95 percent of the total protein of a mature red blood cell; so these cells were essentially bags of hemoglobin molecules – globin polypeptides with attached heme moieties with an iron atom that bound oxygen. The heme-bound iron carries oxygen through the arterial system to cells for respiration. After delivery of oxygen to tissues, these red blood cells (RBCs) return carbon dioxide to the lungs through the venous system for expiration. In sickle cell anemia, after RBCs deliver oxygen throughout the body, the RBCs take on a sickled shape, clog the venous system and lyse, causing a wide variety of systemic problems. Pauling and Itano predicted that this change in RBC architecture was a direct consequence of “two to four” charged amino acid changes in the globin complex, which consists of two beta-globin subunits and two alpha-globin subunits (this was not known then). Because of the science that came after their discovery, we know now that the genetic mutation in the beta-globin moiety is a single amino acid exchange of glutamic acid to valine resulting from a single nucleotide transition (A to T transition) in codon 6 of the beta-globin gene encoding the 147 amino acid polypeptide. Thus two positive charges were added to the hemoglobin molecule by this mutation. Pauling and Itano concluded that these charge alterations caused RBC sickling.

Important discoveries can be quite simple. The figure below is the key experiment carried out by Pauling and Itano, an electrophoretic separation of hemoglobin based upon its isoelectric point (net charge). Because of the mutation in codon 6 present in both inherited beta-globin alleles, the hemoglobin complex migrated to the right of the normal hemoglobin by approximately “two to four” positive charges (panel B compared with panel A). At pH 6.9 the normal hemoglobin was shown to have an isoelectric point of 6.87, migrating as a negative ion, whereas the mutated hemoglobin had an isoelectric point of 7.09 migrating as a positive ion. We know now that this electrophoretic change in the hemoglobin complex described by Pauling and Itano is due to the loss of a single negative charge in a glutamic acid residue (replaced with an uncharged valine residue) near the N-terminus of the two beta-globin moieties of the hemoglobin molecule. Today, the fact remains that this is the only mutation in hemoglobin that causes sickle cell anemia, although other beta-globin mutations cause other hemoglobinopathies like beta-thalessemia. Panel C shows the electrophoretic behavior of hemoglobins in a heterozygous carrier of the disease-causing mutation (Panel D is a control mixture of the globins in panels A and B). Much more insight about these phenomena is discussed in the Pauling and Itano paper but the charge alteration in hemoglobins is the basic observation.

Pauling experiment

(click to enlarge)

Fast-forward to 1976. I decided to look for evidence of charge-altering mutations in a protein profile of about 1,000 visible proteins (polypeptides) comparing normal and neoplastic cells by looking for Pauling and Itano’s evidence of mutations, e.g. minor charge alterations in proteins in the protein profile. A technique had just been developed by Patrick O’Farrell which permitted high-resolution separation of virtually all major protein gene products of the cell.

An elegant study was performed by Greg Milman at the University of California at Berkeley who demonstrated that one could predict the occurrence of mutations in the relatively minor protein, the enzyme hypoxanthine phosphoribosyltransferase (HPRT), in HeLa cells by the positional changes in the HPRT polypeptide in high-resolution two-dimensional polyacrylamide gels within complex profile of proteins separated both by their charge (isoelectric point) and their molecular weight. When I saw Milman’s result I decided to use this technique to compare normal and neoplastic human cells to see if I could identify charge alterations similar to those demonstrated by Pauling and Itano in hemoglobin and by Milman in HPRT.

I labeled the proteins of normal and tumor-forming human fibroblasts with S35-methionine and separated them using O’Farrell’s two-dimensional technique (isoelectric point separation is a tube gel followed by molecular weight sieving in an SDS slab gel). Then I fixed the proteins in the two-dimensional slab gel and stained these proteins with Coomassie blue dye.

mutant actin further annotated

With the dye you could only see the most abundant proteins and I was surprised to see this pattern of actins in the tumor-forming fibroblasts shown above. This image is actually the image of the radioactive protein pattern in the region of actins (pI 5.3 to pI 5.1, molecular weight Mr about 42,000) developed after a very short autoradiographic exposure to X-ray film (a digital image). Normally you only see one beta-actin spot barely separated from the gamma-actin spot. Gamma actin is a second isoform of actin encoded by a separate gene which differs by only four amino acids from beta-actin. Normally there is about twice as much beta-actin at isoelectric point 5.2 as gamma actin and both actins together amount to 5-10 percent of the total cellular protein. But half of the normal beta-actin was missing and a new more negative spot at isoelectric point 5.3 appeared. I was able to show that this was a new form of beta-actin by tryptic peptide separation and other criteria. The observation that the new variant migrated slower in the second dimension as a larger protein was later attributed to a frictional effect in the gel sieve due to an altered conformation caused by the amino acid change.

These observations and other differences in protein expression between the normal and tumor-forming fibroblasts were published in the Journal of Biological Chemistry in February 1980. A second paper was published a month later demonstrating that a T-cell leukemia also had a beta-actin anomaly which suggested loss of a beta-actin allele. It is now well established that reorganization of the actin cytoskeleton occurs when cells become cancerous, although mutations in the structural gene may be less common. These alterations can also be caused by changes in actin-binding proteins.

Later in the year, with my colleagues at the Max-Planck in Goettingen Germany, Klaus Weber and Joel Vandekerckhove, I published the sequences of the normal human beta- and gamma-actins and the mutant beta-actin in Cell. The normal and mutated sequence of human beta-actin is shown in the figure below.

The simple electrophoretic difference between the mutant and normal beta-actin was a single amino acid exchange of a neutral glycine for a negatively charged aspartic acid at amino acid residue 244 in the 374 amino acid polypeptide chain, an observation similar to Pauling and Itano’s hypothesis 32 years earlier. An amino acid exchange at this residue in the actin polypeptide chain had never been observed in any eukaryote. Two years later I cloned the mutant and wildtype human beta-actin genes at the Pauling Institute and formally proved the existence of the mutation at the level of the gene. This mutation was caused by a single nucleotide change in the gene. Several years later my colleagues and I demonstrated that acquisition of this simple mutation contributed to the tumorigenic phenotype of the cells in which it arose.

actin sequence with arrow

The sequence of human beta-actin and its amino acid 244 mutation (the most highly conserved protein in eukaryotes).

Ed Note: This week marks the sixth anniversary of the creation of the Pauling Blog.  Birthed to help promote the unveiling of a postage stamp, the blog, 461 posts later, has developed into a resource of consequence with an audience that is steadily growing.  For those who might be interested in how the project operates, please see this post that we ran one year ago.

As always, we thank you for your continued readership.  We plan to keep researching and writing, so please keep coming back!


Zia Mian is the 2014 Pauling Legacy Award Winner


Happy Linus Pauling Day!  Today marks the 113th anniversary of Pauling’s birth and, as has become tradition here at the Pauling Blog, we celebrate with an announcement: the recipient of the 2014 Linus Pauling Legacy Award is Dr. Zia Mian.

A physicist by training, Mian follows in the Pauling tradition through his deep commitment to helping solve some of the most vexing social issues confronting world society today.  Mian is a research scientist at Princeton University’s Program on Science and Global Security, directing its Project on Peace and Security in South Asia.  His research and teaching focus on nuclear weapons and nuclear energy policy, especially in Pakistan and India, and on issues of nuclear disarmament and peace.

A prolific author and engaging speaker, Mian is co-editor of Science & Global Society, an international journal of technical analysis for arms control, disarmament and nonproliferation policy. He is also a member of the International Panel on Fissile Materials and has edited a number of reports issued by the group. He has likewise helped to produce two documentary films on peace and security in South Asia – “Pakistan and India under the Nuclear Shadow,” (2001) and “Crossing the Lines: Kashmir, Pakistan, India” (2004). A native of Pakistan, Mian earned his Ph. D. in physics from the University of Newcastle-upon-Tyne.


As we continue, throughout 2014, to mark the fiftieth anniversary of Linus Pauling’s receipt of the Nobel Peace Prize, Mian’s acceptance of the Pauling Legacy Award would seem to be especially fitting.  Pauling, of course, received his award for his tireless campaign to end nuclear weapons testing.  Half a century later, Mian continues the quest to stem weapons proliferation and secure a more peaceful world.

The Linus Pauling Legacy Award medal.

The Linus Pauling Legacy Award medal.

Sponsored by the Oregon State University Libraries and Press, the Linus Pauling Legacy Award is granted every other year to an individual who has achieved in an area once of interest to Linus Pauling.  As with past recipients, Dr. Mian will deliver a public lecture in Portland, Oregon that is free of charge and open to anyone who is interested. Here are the details of this event:

    • What: “Out of the Nuclear Shadow: Scientists and the Struggle Against the Bomb.” Linus Pauling Legacy Award Lecture by Dr. Zia Mian. Free and open to the public.
    • When: Monday, April 21, 2014; 7:30 PM
    • Where: Oregon Historical Society Museum, Portland, Oregon

For more information see this page, contact the OSU Libraries and Press at 541-737-4633 or email the Special Collections & Archives Research Center at scarc[at]oregonstate[dot]edu


Get every new post delivered to your Inbox.

Join 47 other followers