Research Completed at LPISM in 1988 – Reproduced and Extended in 2014

The author in his laboratory at the Linus Pauling Institute of Science and Medicine. Originally published in Science Digest, June 1986.

The author in his laboratory at the Linus Pauling Institute of Science and Medicine. Originally published in Science Digest, June 1986.

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

In 1987, my colleagues at the Pauling Institute in Palo Alto, colleagues at Stanford and I published a paper that clearly demonstrated that expression of a charge-altered mutant human beta-actin (glycine to aspartic acid substitution at amino acid 245; G245D) caused non-tumorigenic, immortalized human fibroblasts to form aggressive tumors in nude mice (Leavitt et al, 1987a). When these tumor-derived cells were examined, we discovered that they exhibited further elevated expression of the mutant beta-actin and these tumor-derived cells formed tumors even more rapidly – observations that were consistent with the role of this mutation in the tumorigenic phenotype. Furthermore, over-expression of mutant beta-actin was associated with down-regulation of three abundant tropomyosin isoforms in a well-documented transformation-sensitive manner (Leavitt et al, 1986; Leavitt et al, 1987a and Ng et al, 1988). These final papers were the culmination of research conducted at the Linus Pauling Institute of Science and Medicine (LPISM) from December 1981 to March 1988.

Normally when a scientific observation is never repeated it is usually not worth remembering. In this case, twenty-six years after our 1987 publication, a study was published by Schoenenberger et al. at the Biozentrum in Basel, Switzerland, that reproduced our findings in a different cell system, a rat fibroblast model (provided to them by LPISM in 1986). Furthermore, these investigators extended our findings by characterizing new aspects of abnormal behavior of the mutant beta-actin and cells that express this aberrant protein, which help to explain its potential role in cancer such as enhancement of tumor cell motility and invasiveness.

In addition to enhancement of tumor growth and alteration of cell shape, the Swiss investigators presented the following findings to clarify and support the oncogenicity of this mutation:

  1. The mutant actin stimulated formation of ruffles at the cell periphery as shown by staining of cells with an antibody that bound specifically to the mutant epitope of the mutant beta-actin (left image below)
  2. The mutant actin concentrated primarily in these ruffles (palloidin staining reveals the location of filamentous actin in stress fibers; right image below)
  3. The expression of mutant actin inhibited the tropomysin binding to filamentous actin and tropomysin did not accumulate in the ruffles
  4. Mutant actin colocalized with Rac1 (a GTPase mediator of membrane ruffling) and beta1-integrin (adhesion protein) in the ruffles

ruffles

Back-tracking several years, the discovery of this actin mutation was made in a mutagenized cell line isolated by Takeo Kakunaga at the National Cancer Institute (NCI) in 1978. During the month that his paper was published, I walked over to NCI from my lab across the street at the Bureau of Biologics (FDA) to have a chat with Takeo about using his in vitro transformed Syrian Hamster cells as a model system to identify changes in protein expression that correlated with neoplastic transformation. After describing what I wanted to do, he seemed agreeable but then casually mentioned that he had succeed in transforming human fibroblasts into tumor forming cells. I nearly fell off my chair because human cells had never been transformed in vitro before, a major problem for cancer researchers at that time.

I blurted out that we should do the work that I had proposed in his human cell model system, comparing protein expression by the transformed neoplastic cells with their normal precursor cells. My hypothesis was that this comparison would allow identification of proteins that were turned on or turned off in expression by comparing protein profiles of the most abundant 1,000 proteins expressed in these cells and resolved by high-resolution 2-D gel separation (protein profiling). My plan was to look for charge-altering mutations in proteins that might govern neoplastic transformation and tumorigenesis. A fall-back goal was to define the pattern of qualitative and quantitative changes in protein synthesis to try and get a handle on the mysterious mechanism of human cancer development. A summary of the global changes in gene expression of neoplastic human fibroblasts was published from LPISM in 1982 (Leavitt et al, 1982).

Within two weeks, in May of 1978, I was metabolically labeling the total cellular proteins (with the amino acid S-35 methionine) of the normal fibroblasts and three strains of cell lines derived from the normal culture which were immortalized, only one of which formed subcutaneous tumors in nude mice. After four hours of labeling, I prepared extracts of S-35 methionine labeled proteins from each of the four cultures and loaded 25-microliter aliquots of each sample onto the top of clear noodle-like isoelectric focusing gels (7-inch long urea-polyacrylamide gels with the thickness of thin spaghetti) which separated the complex mixture of total cellular proteins by their net charge (isoelectric point). These gels were subjected to isoelectric electrofocusing of the proteins overnight. The next morning I harvested the spaghetti-like gels, and incubated them in a detergent that would bind to the proteins to help separate them by their molecular weights in a second dimension. So, these proteins were first denatured and separated by their net charge and then, in a second dimension, separated by their size on a thin rectangular slab gel.

After about five hours of separation in the second dimension, I was soon to learn that I had separated more than 1,000 denatured protein subunits (polypeptides) by their differing charges and molecular weights. The final step before autoradiography, which revealed the full protein profile, was to fix and stain the gels to get a glimpse of the resolution of these peptide patterns. The staining of these rectangular gels revealed only the most abundant architectural cellular proteins, the largest number of which were cytoplasmic beta- and gamma-actin, at a ratio of about 2:1 in abundance, respectively. The figure below shows what quickly appeared as the gels were de-stained. In the one tumorigenic cell line, instead of seeing a 2:1 ratio of beta- to gamma-actin, a new abundant protein at about one unit charge more negatively charged (more acidic), and half of the normal beta-actin was lost. The pixilation of these three radioactive “spots” immediately suggested to me that one of the two functional genes (alleles) encoding beta-actin had mutated, possibly due to the replacement of a neutral amino acid with a negatively charged amino acid. This prediction was no mystery to me as I had demonstrated this type of electrophoretic shift in another protein a year earlier at Johns Hopkins.

mutant actin further annotated

A number of experiments were done to build the case for the beta-actin mutation, and then I wrote a letter to Klaus Weber at the Max-Planck Institute in Goettingen, Germany, asking for his help in sequencing these actins. His lab was the only one in the world sequencing actins, e.g. the four muscle forms of actins. It only took Klaus two weeks to respond affirmatively, an indication to me that he was eager. I provided him with the actin proteins from this cell line and it took a postdoctoral fellow, Joel Vandekerhkove, and Klaus a little over a year to determine the complete amino acid sequences of the mutant beta-actin and both the wildtype beta- and gamma-actins, to define the mutation that had occurred. We published the result shown above in the top journal Cell in December 1980. Four years later, my colleagues at Stanford and I published my cloning of the mutant and wildtype human beta-actin gene, and the experiments that formally proved the mutation at the level of the gene (Leavitt et al, 1984). Three years after that, we published the experiments that demonstrated the tumorigenic effect of this mutation in immortalized human fibroblasts.

The dramatic nature of this discovery was never fully appreciated, perhaps, because no other actin mutations had been reported and it took Scheonenberger, et al. twenty-six years to complete the work published in September 2013. In another recent related development, Lohr et al. reported reoccurring beta-actin mutations in a panel of tumor cell samples from patients with diffuse large B-cell lymphoma.

One interesting piece of information that came out of our initial sequencing of these actins was the degree of evolutionary conservation of human beta- and gamma-actin. These two actins differ by only four amino acids at the N-terminus, whereas the four muscle-specific human isoforms are more divergent. Comparing the sequence of actin cloned from Saccharomyces cerevisiae (yeast) with these human sequences (sequences stored at the National Center for Biotechnology Information; NCBI) reveals that yeast and human cytoplasmic actins are 92% identical in their sequences (differing by only 31 amino acids out of 375) and most of these amino acid exchanges are conservative replacements both structurally and thermodynamically. This makes these actins the most highly conserved proteins (on a par with histones H3 and H4) among the 20,000 or so known human protein sequences. This fact presents an argument for the fundamental importance of non-muscle cytoplasmic actins in eukaryotic life. It turns out that among actin sequences of all species, no replacement of the Glycine 245 has ever been documented as a result of species divergence or mammalian isoform divergence.

When we introduced the mutant beta-actin gene into a non-tumorigenic immortalized fibroblast strain by gene transfer (Leavitt et al, 1987a), we isolated a transfected clone in which the ratio of exogenous mutant beta-actin to wildtype beta- + gamma-actin was 0.88 – a 76% higher level of expression than the mutant actin in the original mutated cell line in which the mutation arose (0.5 ratio). When we isolated and cultured the cells from a tumor formed by this cell line, the ratio of exogenous mutant beta-actin to wildtype beta- + gamma-actin had increased to 1.95, indicating that about 64% of the total cytoplasmic actin was the mutated beta-actin. Whereas the initial transfectant cell line produced visible tumors at about six weeks, the tumor-derived transfectant cells expressing 64% mutant actin formed visible tumors at about 1.5 weeks. Thus, expression of this mutation was not inhibitory to cell growth.

The other surprising finding was that cell lines expressing the transfected mutant actin gene did not have higher levels of cytoplasmic actins in them because the two endogenous wildtype beta- and gamma-actin genes were coordinately down-regulated (auto-regulated) so that the relative rates of total actin synthesis remained around 30% compared to S-35 methionine incorporation into 600 surrounding non-actin polypeptides in the protein profile (Leavitt et al, 1987b). This auto-regulation phenomenon was reproduced by Minamide et al. (1997) ten years later.

Cytoskeletal rearrangement of actin microfilaments, as well as changes in composition of tropomyosin isoforms and other actin-binding proteins, have long been associated with neoplastic transformation. However, before our study, causal mutations in a cytoplasmic actin had apparently not been considered. It is perhaps consistent then that Ning et al. (2014) have recently described genetically inherited polymorphisms in the actin-bundling protein, plastin (also discovered and cloned at LPISM), that significantly affect the time of tumor recurrence in colorectal cancer after resection and chemotherapy.

During my tenure at the Pauling Institute, I felt that Dr. Pauling understood and appreciated this work and its relevance to the fundamental nature of cancer development. Progress can be slow, but ultimately true understanding of cancer will emerge from this type of research…and I predict that cytoplasmic actins and actin-binding proteins that regulate actin organization and function in the cytoskeleton will be understood to play a central role in the manifestation of the tumorigenic phenotype.

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Some Personal Thoughts on Vitamin C in the 1980s and Now

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

The author in his laboratory at the Linus Pauling Institute of Science and Medicine. Originally published in Science Digest, June 1986.

The author in his laboratory at the Linus Pauling Institute of Science and Medicine. Originally published in Science Digest, June 1986.

During my daily work for pharmaceutical and biotech clients, I am continuously learning about developments resulting from my research at the Linus Pauling Institute of Science and Medicine in Palo Alto, CA in the 1980s. Likewise, I am regularly coming into contact with new medically related developments focusing on vitamin C, an interest of Linus Pauling in those years.

With regard to our research on human plastins, a gene family of proteins that we discovered, cloned, and characterized at the Pauling Institute, it has recently been reported that plastin (PLS3) is a marker of carcinoma cells circulating in the blood (for example Yokobori, et al.). Our hypothesis was that when this protein was inappropriately expressed in cells from solid tissues, as it is in many tumor types, (e.g. carcinomas, fibrosarcomas, etc.) these potential tumor cells become more like blood cells in that they are able to live and replicate in an anchorage-independent state, an essential property of metastatic tumor cells. It is metastasis that kills us when we get cancer. Thus plastins, discovered and characterized at the Pauling Institute, may turn out to be the “holy grail” of cancer research.

I often run across new information on the medical importance of vitamin C without looking for it. Back in the 1980s, we would receive an annual shipment of loose vitamin C from Hoffmann-La Roche, Inc. as a way of saying thank you to Dr. Pauling for his advocacy of the merits of vitamin C. We received no funding from Hoffmann-La Roche though. One year I recall that two dignitaries from the company visited us. Dr. Pauling, with me and several others, walked our visitors to lunch a few blocks down El Camino Real in Palo Alto to my favorite restaurant, the Captain’s Cabin.

Afterward, while walking back to the Institute, one of the guests asked Dr. Pauling if he thought the perceived benefits of vitamin C were due to the placebo effect. I was amused because I too had said something ill-advised like that to Dr. Pauling in my first few months at the Institute. I mentioned to him that I had a vitamin C-resistant cold to which he replied, “You’re not taking enough!” and told me that he takes 18 grams a day. No doubt he had calculated this number based on the amount of vitamin C that animals produce within themselves every day. He would stir 18 grams into a large glass of water and imbibe the glass with no great rush.


A few months ago I heard a physician state in the national media that taking supplemental vitamins is a waste of money. This bold assertion reminded me of the announcement of the discovery of cold fusion and another premature announcement of the discovery of a cure for AIDS. The progress of science is slow but relentless, like the new developments with plastins fifteen years after I left LPISM’s labs.

On October 31, 2013, Kim, et al. at Seoul National University in South Korea published their findings on a new strain of experimental mice. The researchers knocked out the mouse gene encoding the enzyme L-gulono-γ-lactone oxidase, known as gulo for short. This is the gene that is missing in humans and that keeps us from synthesizing our own vitamin C, unlike nearly all other animals. An extreme lack of vitamin C in our diet can lead to scurvy, caused by aberrant expression of collagen in our connective tissues because of starvation of vitamin C in our diet. In these mice the lack of this gene caused “vitamin C insufficiency” in an animal model – a model that can now be used to learn more about the importance of vitamin C.

As these mice matured they expressed known blood markers of liver damage. This damage, called fibrosis, is basically the scarring of the liver, sort of like the scarring of the skin that is caused by certain types of skin damage. Concomitantly, as the mice aged, reactive oxygen species (ROS) and lipid peroxides increased in the liver, as did activated hepatic stellate cells, which deposited abnormal collagen fibriles on the basement membrane of functional liver cells. There is a wealth of evidence that elevated ROS in the lungs, liver, and kidneys is associated with pulmonary, hepatic, and renal fibrosis. Elevated vitamin C in these tissues will quench ROS.

Currently in the United States, there are no drugs approved to treat any of these forms of fibrosis. Fortunately, Intermune’s drug, pirfenidone, is close to approval for treatment of pulmonary fibrosis and has already been approved in Canada, Europe, and Japan. This drug reduces ROS and inhibits other key targets that are suspected of playing a role in the development of fibrosis. So who is to say that supplementing your diet with vitamin C is of no consequence? It is certainly not toxic in any way. Oh, by the way, pulmonary fibrosis is worse than cancer – it kills you in three to five years once diagnosed. You basically die of asphyxiation.


In the last week I stumbled upon another interesting paper on the effects of vitamin C on humans. A 2011 paper by Juraschek, et al. at Johns Hopkins University Medical School reported the results of a significant meta-analysis (a systematic review of multiple clinical trials) of 13 randomized clinical trials involving 556 patients who took a median dose of 500 mg of vitamin C per day. (I take a full gram)

The purpose of the study was to examine the effects of vitamin C supplementation on uric acid levels in the blood. Elevated uric acid levels in the blood causes gout, because saturation of blood with uric acid causes urate crystals to form in the synovial fluids of joints (e.g. crystal arthritis). Drugs that lower uric acid in the blood are used to treat gout because lowering uric acid causes the urate crystals to dissolve to ameliorate the arthritic pain.

Admittedly gout is not as bad as cancer. But another systematic clinical review of multiple trials on humans published in 2012 by Lottmann, et al. at the IGES Institut GmBH in Germany has shown clearly that having gout is associated with both all-cause mortality and, in particular, cardiovascular mortality. So what could be worse than death by gout?

I think I will keep taking vitamin C.

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.

Pioneering the Field of Proteomics

John Leavitt, 1982.

John Leavitt, 1982.

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

In the fall of 1985, I went to a small meeting in Heidelberg, Germany, with Steve Burbeck from the Linus Pauling Institute of Science and Medicine, who had helped me by developing computerized microdensitometry to analyze two-dimensional protein profiles. At this meeting I described our protein profiling work and the discovery of the mutant beta-actins and another interesting protein which I named “plastin.”

Steve Kent, head of the protein sequencing facility in Leroy Hood’s lab at Caltech, heard my talk. We sat across from each other at dinner and he proposed a collaboration to develop methods of sequencing minute amounts of protein leached from spots in high resolution protein profiles. Lee Hood was well known for developing state-of-the-art protein and nucleic acid sequencing methods and machines, and was a founder of Applied Biosystems in Foster City, CA.

After I returned from Heidelberg, Ruedi Aebersold called me from Caltech and we began collaborating on microsequencing of pure nanomolar quantities of unknown proteins of interest eluted out of my protein profiles. In this work we essentially started the field of proteomics, which was eventually named ten years later by Jim Garrells, a protein profiler at Cold Spring Harbor. Proteomics is the search for and definition of proteins that could serve as diagnostic markers and drug targets for diagnosis and treatment of diseases, in our case cancer.

In 1987 we published a landmark paper in PNAS on the microsequencing technique that Ruedi developed. This paper would eventually be cited in references by more than 1,000 other research papers.

Ruedi

I gave a postdoctoral fellow, Mahdu Varma, the task of isolating the cancer-specific leukocyte isoform of plastin (L-plastin) from 140 protein profiles. This protein has been implicated in metastases in both melanoma and prostate cancer as well as in other aspects of cancer. The L-plastin spot was easily recognized and those spots on a nitrocellulose filter were “snipped out,” removing all the other proteins of the cell. We sent Ruedi a plastic tube containing the 140 “spots” of L-plastin. He had figured out a way to solubilize the protein from the nitrocellulose and was successful in determining the sequence of eight oligopeptides of between eight and sixteen amino acids derived by digestion of L-plastin with a proteolytic enzyme.

The peptide sequences he determined turned out to be perfectly accurate internal amino acid sequences of plastin when we decoded the sequence of the plastin gene (cDNA) clone, a reverse transcript of the messenger RNA. This was the first time that anyone had done this and it opened up the field of proteomics and led to the discovery of other diagnostic and drug targets.

plastin 1

We had chosen L-plastin, normally only expressed in white blood cells, because I had reported for years that it was a cancer marker in tumors that arose in solid tissues (identified in the image above by the two upward arrows). After we received the oligopeptide sequences from Ruedi, we made short DNA antisense probes that would hybridize to DNA sequences encoding these peptides in the human genome to fish out the full-length DNA clones that carried the sequence of the L- plastin gene.

Ching Lin and I, along with Reudi, published the sequences of the human L- and newly discovered T-plastin proteins, based upon sequencing of cDNAs, in Molecular and Cellular Biology. The discovery of a second isoform of plastin (T-plastin named for tissue plastin as opposed to L-plastin from leukocytes) was a surprise. We now had two genes to characterize at the genomic level. Today, T-plastin is a well recognized marker for cutaneous T-cell lymphoma (Sezary Lymphoma) and L-plastin, inappropriately expressed in solid tumor cells (carcinomas, fibrosarcomas, etc.), is understood to be a contributor to metastasis.

The Linus Pauling Institute was not all work and no play in the 1980s

We worked hard at the Institute and Linus Pauling was always there and visible.

We put together a softball team with Jim Fleming, Dan McQueeny, Zelek Herman, myself, and others at the Institute and played departmental teams at Stanford. I think we were called the “Pauling squeeze.” After these games we would often go dancing at the Class Reunion on El Camino Real near the corner of Page Mill Road.

We were fortunate to have on staff a first rate fundraiser in Richard (Rick) Hicks who arranged wonderful parties on Nob Hill at the Stanford Court. The most memorable of these parties occurred in late November 1986, when we honored Japanese billionaire Ryoichi Sasakawa with the annual Linus Pauling Medal. Another year Carl Sagan and Ann Druyan, who helped Carl put together the Cosmos series, likewise took part. We often saw Dr. Pauling’s sons, Linus Pauling Jr., Peter, and Crellin as well.

Here we are at the Stanford Court that night with postdoctoral fellows, Dr. Karin Sturm from Heidelberg, Germany, on the left and Dr. Madhu Varma from Madras, India, on the right. My wife, Becki, is in the middle. I recall that Dr. Pauling enjoyed this night as well.

Here we are at the Stanford Court that night with postdoctoral fellows, Dr. Karin Sturm from Heidelberg, Germany, on the left and Dr. Madhu Varma from Madras, India, on the right. My wife, Becki, is in the middle. I recall that Dr. Pauling enjoyed this night as well.

In 1988 I moved on to a new position in San Jose and then became Director of Research at Adeza Biomedical. Since we continued to live in Palo Alto, we continued to interact and party with the Linus Pauling Institute staff into the 1990s.

The 1980s at the Linus Pauling Institute – A Wonderful Place to Be

John Leavitt

John Leavitt

[Ed Note: This is part one of a two part series of guest posts written by John Leavitt, Ph.D., Nerac, Inc., Tolland, CT.]

There was an article about Linus Pauling in Time magazine in early 1981 about the fact that at the age of 80 he was still seeking a grant from the National Institutes of Health (NIH) to fund his research on ascorbic acid for treating diseases. This news caught my attention and I looked into the possibility of joining Dr. Pauling’s institute. Toward the end of the summer I was invited to visit the Pauling Institute in Palo Alto, CA to give a seminar on my research at NIH.

In late August Koloman Laki, an aging scientist at NIH, called me up and invited me over to his lab in NIH Building 10, a short walk across the campus from my lab in NIH Building 37. He was interested in talking to me about my recent discovery of mutations in human non-muscle cytoskeletal actin that was published in Cell in late 1980. This protein is the major architectural protein of all eukaryotic cells and we had shown that it was the most highly conserved protein in evolution of the species from yeast to humans. This fact made these mutations even more interesting.

Koloman was a protege of the Hungarian Nobel Prize winner Albert Szent-Györgyi who, I later learned, was much admired by Dr. Pauling because he had discovered both vitamin C and actin. Koloman described how Szent-Györgyi discovered muscle actin. When I mentioned that I was to visit the Linus Pauling Institute in late September, he told me about Emile Zuckerkandl’s and Dr. Pauling’s work on the ‘biological clock,’ which provided evidence in support of Charles Darwin’s theory on divergence of the species.

In the last week of September I flew to Oakland, CA and was picked up at the airport by Emile who was President of the Linus Pauling Institute of Science and Medicine. The next morning I stood up in front of Dr. Pauling and the institute staff to tell them about my discovery of a mutant human beta-actin and my speculation on its involvement in neoplastic transformation. The evidence suggested that I had actually discovered at least two mutations in the same gene, each of which caused a progression to a higher malignant state.

Linus Pauling was in the front row and was all smiles. He asked me if I knew who discovered actin. I was prepared to answer that question thanks to Koloman Laki. In the afternoon I met with Emile who offered me a Senior Scientist position at the Institute, which I accepted. At the time it would be me and Dr. Pauling with separate research interests. Nevertheless, Dr. Pauling could appreciate my discovery as, 32 years earlier, he had described the molecular basis for sickle cell anemia, which predicted that mutations in hemogloblin governed the sickled shape of red blood cells which caused the disease, sickle cell anemia. Likewise, human cancer cells exhibit altered shapes.

So I resigned my secure job-for-life at NIH and moved to Palo Alto to join the struggling Linus Pauling Institute. My technician, Patti Porecca, hired from Bob Gallo’s lab at NIH, would follow me to the Pauling Institute.

Cloning of the Human Beta-Actin Gene

After I arrived at the Pauling Institute, two of my colleagues at NIH and I published a comprehensive study of the changes in protein expression between normal and neoplastic cells in Carcinogenesis using high-resolution computerized microdensitometry to analyze the complex protein patterns (my first paper from the Pauling Institute). This was the first time that such a study had been published, e.g. the comparative profiling of expression of a large number of proteins in neoplastic cells. It was a study of the 1,000 most abundant proteins in normal and neoplastic human cells which revealed potential biomarkers and causative genetic events for human cancer. At the time it was staggering to view these patterns but perfect for my dyslexic brain and mind’s eye. In addition, we published another paper in Cell that described, for the first time, the progression of a neoplastic human cell to a higher malignant cell following a second mutation in the same beta-actin gene. Early in 1982, Steve Burbeck and Jerry Latter at the Institute set up the same computerized microdensitometry platform I had exploited at NIH.

Jerry Latter gave a stirring talk at Argonne Labs in Chicago demonstrating that computerized microdensitometry of protein profiles could be used to determine the identities of unknown proteins based upon determining their amino acid compositions in situ in protein profiles. This paper was published in Clinical Chemistry in 1984. At the same meeting, Steve Burbeck described a truly innovative invention that could measure beta-particles emitted from radioactive protein profiles to produce a direct image of the protein profile pattern. As a group we had entered an exciting period of discovery and innovation at the Linus Pauling Institute.

When I got to Palo Alto in December 1981, I called Professor Larry Kedes at Stanford and we embarked on a collaboration to clone the human beta-actin gene. His impressive postdoctoral fellow, Peter Gunning, taught me some basic recombinant DNA techniques, and I was off to the races. The difficulty was to identify the functional gene in a sea of actin pseudogenes (sometimes referred to as junk DNA). I used an elegant method of homologous recombination developed in Tom Maniatis’ lab at Harvard that had never been used before to clone a novel gene (In fact, cloning of human genes was just getting started at the time). This was smart because Professor Maniatis would be the chairman of the NIH study section that reviewed my first grant proposal submitted from the Pauling Institute. I did not know it at the time but within a month or two I had cloned the functional beta-actin gene a week before Christmas in 1982.

I developed a scheme to identify the correct gene among 300-400 clones of pseudogenes that Patti and I had cloned and the strategy worked. We gave Dr. Sun-Yu Ng the task of sequencing the DNA clone that we were betting on. Rather quickly we determined that we had cloned the functional human beta-actin gene because the DNA sequence that Sun-Yu determined from our candidate clone accurately encoded the amino acid sequence of human beta-actin protein that I had published in Cell in 1980 (with Klaus Weber). Quite coincidentally another lab discovered a rat oncogene that was a fusion of part of an actin gene with a tyrosine kinase gene. I sent this information off to the study section that was reviewing my grant in January 1984 as added evidence that the actin gene was in some way relevant to neoplasia.

My colleagues and I at the Pauling Institute and Stanford published our successful isolation of both the mutant and wildtype human beta-actin genes in Molecular and Cellular Biology in October 1984. As shown below, we had given Armand Hammer’s name to our cancer research program because of his generosity in helping to fund the Linus Pauling Institute.

actin cloning

In January 1984 I was awarded a grant of about $110,000 a year for two years from the American Cancer Society…what a relief. Later in the spring I received word from Professor Maniatis’ NIH study section that our program would also be funded in June by a grant of about $150,000 a year for 3.5 years from the National Cancer Institute for the same work. I was able to hire Dr. Ching Lin from Iowa State University and Dr. Ng (Sun-Yu) from Kedes’ lab. By 1985 Sun-Yu finished the complete DNA sequencing of the human beta-acid gene and Ching sequenced the copy of the beta-actin gene that had two mutations to formally prove the mutations at the level of the gene. Everything that we had learned about the genetic code and amino acid sequences of proteins made our findings predictable. I had learned from my own research how Darwin’s theory of evolution and natural selection worked.

This was the year I finally successfully transferred in recombinant gene inside a cell in culture. I transferred the mutant human actin gene into a rat fibroblast cell line to show that I had cloned the functional gene which could abundantly express its protein the way the natural endogenous beta-actin gene worked (shown in a protein profile below).

mutant actin annotated

At this point I had a brief meeting arranged by Emile with Alex Zafferoni, founder and CEO of Alza Corporation, a block away on Page Mill Road. Zafferoni recommended Bert Roland as a patent attorney. I arranged a meeting with Roland, also a block away, for that afternoon to discuss patenting the human beta-actin gene promoter because of its strong constitutive nature (the engine of the gene that drives its expression). I told Bert that this was a collaboration with Peter Gunning and Larry Kedes at Stanford. Roland was famous for filing Boyer’s and Cohen’s genetic engineering patent which created Genentech and eventually funded Stanford with hundreds of millions of dollars.

We published Sun-Yu’s work on the sequence, structure, and chromosomal location (chromosome 7) of the human beta-actin gene in Molecular and Cellular Biology and we published Ching’s work locating three mutations in this gene in the Proceedings of the National Academy of Sciences, sponsored by Linus Pauling. A patent was filed on the beta-actin promoter and over the years it was licensed to about 15 biotech companies by Stanford University. This patent was prosecuted for the full 17 years (the life of a patent) but never issued. The Institute’s first royalty check was about $10,000 in 1986, but most of the royalties were earned by Stanford’s patent attorneys.

Peter, Larry and I published a paper in PNAS on the use of the human beta-actin gene promoter for expression of other genes. This vector was distributed to anyone who asked for it – and many did – and to those companies that licensed the invention. At last count this paper had more than 1,000 reference citations.

Our paper popularized the actin promoter as a strong constitutive promoter of foreign gene expression. Soon the rice actin promoter would be used to make Round-up Ready crops by DeKalb Genetics and Monsanto, and giant tilapia fish would be engineered with growth hormone under the control of the fish beta-actin promoter. There were even fluorescent mice running around in Japan created with firefly luciferase expressed by the beta-actin promoter (which I called “the cat’s meow”). Since cytoplasmic actins are the most abundant proteins in most cells you could use the promoter to abundantly express foreign genes in most cells of any animal.

In 1987 we also published the culmination of my research on the mutant beta-actin gene in Molecular and Cellular Biology. When I introduced this gene into non-tumor forming immortalized human fibroblasts they became tumorigenic. The results showed that the more abundant the expression of the mutant beta-actin, the more tumorigenic the non-tumorigenic cells became and the cells that came out of the tumors were enhanced further in the level of mutant beta-actin expression. This was a sensational finding that was the goal of research which began with the discovery of the mutant beta-actin in 1978 at NIH.

Out of Ashes, the Phoenix Rose

Linus Pauling Jr., October 14, 2011.

Linus Pauling Jr., October 14, 2011.

[Coda to our history of the Linus Pauling Institute of Science and Medicine]

Linus Pauling Science Center grand opening Keynote Address, by Linus Pauling Jr., MD. October 14, 2011.

This is a very personal account of the background that has miraculously led to this wonderful, beautiful and exciting building, I title it: OUT OF ASHES THE PHOENIX ROSE.

It was back in the spring of 1991, just over 20 years ago now, that I sat down to talk with my father at his Big Sur ranch on the rugged California coast. For many years, in fact since my mother died a decade earlier, my wife and I had made a pilgrimage to the ranch to be with my father and celebrate our three birthdays, which fortuitously fell within a two-week period.

I had been on the Board since the Palo Alto Linus Pauling Institute of Science and Medicine’s inception in 1973, so at our 1991 meeting I knew the situation had become desperate. My father, who for all his earlier life had been full of remarkable energy and ambition, now at 90 had lost that energy and was making mistakes in judgment. He was ill with the cancer that would kill him three years later.

LPISM was failing: half a million dollars of debt, laboratory research had vanished for lack of incentive and direction, donor income was being diverted to non-nutritional investigations, there were no research grants and morale was in the basement.

As his oldest son, I could not just stand by and watch this great man’s efforts of the past quarter century go down the drain, along with his reputation. If the Institute failed, all the naysayers would crow and describe him as a senile crackpot in spite of his astonishing lifetime achievements. Additionally, the thousands of donors over the years and the makers of future bequests would feel betrayed. It was obvious he needed help. As his son, I felt it was necessary to provide that help and it felt good to me to try.

lp-jr2

So we had to talk. Early in my life I realized that my father was a very special person with talents I could never hope to emulate. That was emphasized by this story which I enjoy telling. When I was about 15, my father was writing an introductory chemistry textbook for Caltech freshmen, the best and brightest college freshmen, the cream of the crop. At the end of each chapter were questions. He asked me to read a chapter and answer the questions. I tried, valiantly, but I did not understand the text and could not answer a single question. When my mother heard about this, she hurried down to the Pasadena City Hall to have my name officially changed from Linus Carl Pauling to Linus Carl Pauling Jr. so no one could possibly mistake me for him.

At least I had sense enough to follow a very different track from my father, one that eventually gave me skills that now could be used to help him as my thanks to him for bringing me into the world.

It was now or never, so I boldly waded in. He and I discussed the future, starting with the past. I talked about his amazing life with his multiple triumphs in so many and so very diverse arenas.

His fame was world-wide, originating with the scientific community. I pointed out that he was arguably the first, and certainly the most successful, bridge-builder between chemistry, mathematics, physics, medicine and biology, linking these disciplines to create what is now the most popular science of all, molecular biology. One result of his creativity, hard work and dedication to science, as you all know, was the Nobel Prize for Chemistry.

It was during this time period that his interest in nutrition originated, spurred by his own life-threatening kidney disease. Thanks to a rigid diet prescribed by Stanford Medical School nephrologist Dr. Thomas Addis at a time long before renal dialysis, and carefully supervised by my mother, my father not only survived a usually fatal disease but recovered completely.

lp-jr3

After World War II, prompted by my politically-liberal mother whom he certainly loved deeply and wanted to please, he embarked on a spectacularly successful two decades of humanitarian effort, educating the governments of the world and, necessarily, their peoples, about the evils of war and the dangers associated with unrestricted exposure to radiation, especially that produced by the hundreds of nuclear bomb tests being conducted. He suffered vilification by many from all parts of the world. He was hounded by the FBI and the United States government.

His crowning moment of glory, at least in my estimation, was his indomitable courage in confronting those nasty witch hunters, the United States Senate Internal Security Subcommittee, when facing imprisonment when he refused to disclose the names of his ban-the-bomb United Nations petition assistants. He knew that these conscientious people, most of them scientists, would be less able than he was to defend themselves from accusations and loss of employment. The Subcommittee, when faced by my father’s public popularity, courage, remarkable memory and command of facts, then backed off, their collective tail between their legs. His world-wide influence was so extensive and the result so positive that he was eventually awarded the Nobel Peace Prize.

So what was next for him? His old interest in nutrition as a factor in health and well-being resurfaced. Starting with vitamin C, he promoted nutrient research and encountered resistance from university, medical and government bureaucracies. He turned to the public, writing article after article and giving hundreds of talks, with the result of an explosion in popular food supplement usage. But research remained a fundamental necessity, so the private nonprofit Linus Pauling Institute of Science and Medicine was founded in 1973 and initially showed promise.

By the time of our talk in 1991, LPISM’s outlook was dismal.

At age 90, my father was tired and dispirited. Being fully occupied with his own illness, he was unwilling to devote energy to coping with his Institute’s problems. I said to him that I could not in good conscience stand by and see his eponymous Institute go down in ignominious defeat. With his incredibly illustrious past, I felt strongly that he deserved more than that. And maybe, just maybe, I could do something about it.

We decided, together, that if the Institute, and also his reputation, were to survive, the best course of action was for the Institute to affiliate with a reputable university. That would ensure the rigorous scientific attitude and protocol necessary to legitimize micronutrient research in the future. And, most important of all, we had to be ethically responsible to the thousands of past, present and future donors who believed in my father and supported the Institute. We could not let them down.

I had just retired from 35 years of the practice of psychiatry, so I had the time and energy to devote to other endeavors. After discussion with my wife, I decided to offer to take over management of the Institute. I had to have my wife’s agreement, because I was planning to spend considerable time in Palo Alto, a long way from my home in Honolulu.

lp-jr4

To his credit and with an audible sigh of relief, my father agreed. We discussed affiliation possibilities, Stanford and Caltech among them. He seemed, however, to favor Oregon State University, his undergraduate alma mater, to which he had already committed his scientific papers. If you haven’t already, you should check out the Pauling Papers at the OSU Valley Library Special Collections website. You will be impressed.

During the next years, I became President and Chairman of the Board of LPISM. We reorganized radically and survived many trials and tribulations. My essential second in command Steve Lawson and I visited many universities.

OSU, thanks to then President John Byrne, Development Director John Evey and Dean of Research Dick Scanlan, was our clear and undisputed choice.

And what a great choice it was! Here now, before us, 15 years later, is the Linus Pauling Science Center, dedicated to highest-quality research in scientific areas that would surely be of interest to my father. I’m sure, if he were here, he would have tears of joy in his eyes just as I do.

I want to thank OSU President Ed Ray, Dean Sherman Bloomer, LPI Director Balz Frei, architect Joe Collins, the many others in the system who have participated in making this possible, all the donors and the people of the great state of Oregon. I specifically thank the key major donors, Tammy Valley and Pat Reser, for allowing Linus Pauling’s name to be on this beautiful building. That is a very unusual act of generosity.

It will be a great future. Thank you all with my whole heart.

LPI Looks to the Future

lpsc-beam

[A history of the Linus Pauling Institute of Science and Medicine, Part 8 of 8]

The opening of this current decade promises to be even better for the Linus Pauling Institute than was the last. The decade got off to a great start when, in 2011, Oregon State University opened the Linus Pauling Science Center to house LPI, parts of the department of chemistry, and other lab and teaching spaces.

For the Institute, the historical importance of the completion of the Linus Pauling Science Center is difficult to overstate. The building, which is the largest academic facility on the OSU campus, was a serious undertaking – it cost $62.5 million to build the four-story, 105,000 square-foot research center. The funding was acquired through donations from the Wayne and Gladys Valley Foundation ($20 million), the Al and Pat Reser family ($10.65 million), 2,600 private individuals (~$600,000), and a matching bond ($31.25 million) from the State of Oregon.  The facility is one of the cornerstone achievements of The Campaign for OSU, a capital campaign which seeks to raise $1 billion in funds by June 2014.

Constructing its own building on the OSU campus was a goal for LPI from the minute the Institute moved to Corvallis. Indeed, Linus Pauling Jr. remembers sketching potential plans on napkins while at meetings with OSU staff during the moving process and Institute Director Balz Frei has written that ever since LPI moved to OSU, building “a state-of-the-art research facility to house the Institute and serve as a high-profile working memorial for Linus Pauling” had been one of LPI’s highest priorities.

A portion of the crowd assembled for the LPSC opening ceremonies, October 14, 2011.

A portion of the crowd assembled for the LPSC opening ceremonies, October 14, 2011.

The Linus Pauling Science Center was opened on October 14, 2011. Over 250 people attended the ceremony, during which Linus Pauling Jr. and OSU President Edward J. Ray delivered the main speeches. In his remarks, Dr. Ray noted his belief that “preventive health care is the future of medicine,” and that LPI and the Linus Pauling Science Center are in strong positions to develop this in the twenty-first century.

A light painting by Stephen Knapp, Linus Pauling Science Center.

A light painting by Stephen Knapp, Linus Pauling Science Center.

The center was designed by the firm ZGF Architects LLP, based in Portland, Oregon. It is a unique building with large windows and ample natural light. In addition, each of its floors is home to several works of art, including several light paintings created by Massachusetts-based artist Stephen Knapp, and those who work in the facility enjoy an enviable lunch spot on a fourth floor balcony looking toward the Coast Range mountains.

The view from the "lunch room."

The view from the “lunch room.”

Its lab space, however, is the real highlight of the Linus Pauling Science Center. Unlike most facilities, LPSC’s labs consist mostly of open space, with only a few partial walls separating research areas. Administrator Steve Lawson commented on this decision, noting “We didn’t want a lab environment with a lot of walls… For us, it’s a way to keep the Institute coherent and increase the possibility of people communicating.” In further pursuit of this goal, most of the Institute’s noisier lab equipment is kept behind closed doors in dedicated spaces away from the work environment, thus rendering the laboratories a more pleasant place to think and interact.

Peering down the LPSC laboratory space.

Peering down the LPSC laboratory space.

In recent time, LPI has also begun working to expand the staff supporting its very popular Healthy Aging Program and Healthy Youth Program. As part of this initiative, the Institute hired Kathy Magnusson, an expert on aging, memory, and degenerative brain diseases, to fill the role of Primary Investigator and to work with the Healthy Aging Program. Likewise, Corvallis High School partnered with LPI to develop the Spartan Garden, which is primarily student-run and is linked with outdoor horticulture classes that teach students about growing and preparing healthy foods.

Currently LPI has scheduled the seventh Diet and Optimum Health Conference for May 15-18, 2013, and has established an ambitious research agenda. At the time of this writing, LPI has twelve laboratories working on:

  • Oxidative stress, lipoic acid, and essential metals in atherosclerosis
  • Vitamin E metabolism and biological functions
  • Oxidative and environmental stress in Lou Gehrig’s, Parkinson’s, and Alzheimer’s disease
  • Stress response, lipoic acid, and mitochondrial dysfunction in aging
  • Cancer chemoprotection by phytochemicals in tea and vegetables
  • Transplacental cancer chemoprotection
  • Epigenetic and epigenomic mechanisms of cancer etiology
  • Zinc and antioxidants in prostate cancer and neurodegeneration
  • Novel biological functions of vitamin C
  • Antioxidants and gene expression in diabetes
  • Dietary fats and carbohydrate and lipid metabolism
  • Vitamin D and zinc in immune function

Seventeen years after moving to Oregon with a core staff of five, LPI has regenerated its roster to 63 employees. Of particular note, Steve Lawson still works there, the only individual from the California days to remain. The Institute has remained prolific, has published three books (in addition to re-releases of two Pauling books) and continues to publish dozens of articles in various scientific and medical journals every year. The Institute also circulates a biannual research newsletter, available via the mail or through its website, lpi.oregonstate.edu.

Logo for the 2013 Diet and Optimum Health Conference.

Logo for the 2013 Diet and Optimum Health Conference.

The Institute is currently working to expand its support for its corpus of graduate student laboratory researchers, who are, as Balz Frei puts it, “the heart and soul of [LPI’s] labs at OSU.” To date, plans do not include any sort of major expansion of full-time staff, with a focus instead on further developing the staff infrastructure already in place. The Institute’s plan for 2013 and onward is to strengthen its current research projects and to acquire additional funds for scholarships, endowments, research, and educational programs. Lastly, LPI also hopes to broaden its outreach and health programs, such as the Diet and Optimum Health Conference, Healthy Aging Program, Healthy Youth Program, research newsletter, and Micronutrient Information Center.

One of LPI’s core missions is to “help people everywhere achieve a healthy and productive life, full of vitality, with minimal suffering, and free of cancer and other debilitating diseases.” As of 2013, the 40th anniversary of its founding and with many years of turbulence in its past, the Linus Pauling Institute appears to be in a better position than ever before to continue working towards this goal.