Experimenting with Lipoprotein(a)

lpa

[Part 1 of 2]

In the late 1980s into early 1990, Linus Pauling and a colleague, Matthias Rath, worked intensively on the health benefits of Vitamin C and Lipoprotein(a) binding inhibitors. In 1990 they applied for two patents related to that research. The first, applied for in April, was titled “Use of ascorbate and tranexamic acid solution for organ and blood vessel treatment prior to transplantation.” The second, submitted in July, was titled “Prevention and treatment of occlusive cardiovascular disease with ascorbate and substances that inhibit the binding of lipoprotein (A).”

The technique that Pauling and Rath were attempting to patent in April was both a method and a pharmaceutical agent designed to prevent and treat fatty plaque buildup in arteries and organs and also prevent blood loss during surgery by introducing into a patient (or organ) a mixture of ascorbate and lipoprotein(a) [Lp(a)] binding inhibitors, such as tranexamic acid.

Tranexamic acid is a synthetic version of Lysine, and ascorbate is the shortened name for L-ascorbic acid, or more commonly, Vitamin C. Lp(a) is a biochemical compound of lipids and proteins which binds to fibrin and fibrogen in the walls of arteries and other organs, which causes plaque buildup, which in turn often results in atherosclerosis – the thickening and embrittling of arterial walls – and cardiovascular disease (CVD), one of the most common causes of death in the United States. The second patent described effectively the same method, but focused more on CVD and less on surgery.

Pauling and Rath noticed that humans and a select few other animals are the only creatures that suffer from heart attacks and other issues associated with the buildup of plaque in the circulatory system. One common link between all of these creatures is the fact that they do not naturally produce Vitamin C, and therefore must obtain it solely through diet. The duo hypothesized that the cause of Lp(a) buildup was due to a lack of Vitamin C, and that if Vitamin C intake was increased, it would help the body filter out Lp(a) and therefore decrease the amount of Lp(a) in the bloodstream. They decided to run tests on Hartley guinea pigs, since they are one of the few other animals that don’t synthesize their own Vitamin C.

rath

The first test was run on three female guinea pigs, each about a year old and weighing 800 grams. The animals were all fed a diet devoid of ascorbate (e.g., a hypoascorbate diet), and given an injection daily of ascorbate so that Pauling and Rath could easily monitor and control their intake. The first pig was given ascorbate at a ratio equivalent to 1 mg per kilogram of body weight (1 mg/kg BW). The second pig was given 4 mg/kg BW, and the third was given 40 mg/kg BW.

The experiment only lasted three weeks, because Pauling and Rath didn’t want to inflict scurvy upon the guinea pigs. Creatures deprived of Vitamin C for prolonged periods develop scurvy, an incredibly painful condition where the victim becomes lethargic and begins to suffer skin color and texture changes, easy bruising, brittle and painful bones, poor wound healing, neuropathy, fever and eventually death.

The guinea pigs had their blood drawn at the start of the test, then once again after ten days. At the end of three weeks, the animals were anesthetized and euthanized, then dissected. Their results showed that the hypoascorbate guinea pigs had noticeably higher plaque buildup and general amounts of Lp(a) in their bloodstream. Upon closer analysis of the organs and the arterial wall, the researchers discovered that the guinea pigs had also developed lesions along the walls of their arteries, to which Lp(a) was binding even more than normal.

Pauling and Rath then ran a more expansive second test, with a test time of seven weeks and a test group of thirty-three male Hartley guinea pigs, each approximately five months old and weighing 550g. At the outset, the subjects were split into multiple groups. Group A consisted of eight guinea pigs and was given 40 mg/kg BW of ascorbate daily, while Group B consisted of 16 guinea pigs given 2 mg/kg BW daily. At five weeks all of Group A was euthanized and studied, as was half of Group B. The second half of Group B then had their daily dosage increased to 1.3 g/kg BW for two weeks before being euthanized.

Once again, it was observed that the hypoascorbate guinea pigs had developed lesions in their arterial walls and organs, as well as increased plaque buildup and Lp(a) levels. On the same token, the second half of Group B showed decreased levels of Lp(a) in their blood and decreased amounts of plaque after their ascorbate intake was dramatically increased.

Pauling and Rath felt that their research was confirming their hypothesis, and wanted to see how it would function on humans. Their method here was to obtain post-mortem pieces of human arterial wall. They cut the pieces into smaller sections, and for one minute placed a piece weighing 100 mg into a glass potter containing 2.5 ml of a mixture of ascorbate and tranexamic acid. Compared to the other pieces, the portions in the mixture released sizable amount of Lp(a).

This promising data in hand, Pauling and Rath then began to think about patenting and marketing their work.

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John Yudkin, Linus Pauling and the Sugar Question

In my book I say you shouldn’t eat sweet desserts, but I also quote a professor who says that this doesn’t mean that if your hostess has made this wonderful dessert you should turn it down.  My wife used to say I always looked for that hostess.

-Linus Pauling, 1987.

Linus Pauling and John Yudkin shared a semisweet bond that was nearly equal parts contradiction, respect and humor, and which lasted from the mid-1970s until Pauling’s death in 1994. The two men held radically different views on a number of topics including the effects of vitamins, especially vitamin C, but shared an identical view on the dangers of sugar. Indeed, Yudkin’s claims in his 1976 book This Nutrition Business that Pauling’s beliefs about vitamin C were completely incorrect did not deter Pauling from citing Yudkin’s work on sugar in a favorable light in How to Live Longer and Feel Better, published ten years later.

John Yudkin was born in London in 1910, earned a degree in chemistry and a Ph. D in biochemistry, and later studied medicine in London. As the Chair of Physiology at London University at Queen Elizabeth College, he persuaded the university to institute a Department of Nutrition in 1954, the first department in Europe devoted to undergraduate and postgraduate teaching and research in nutrition. In 1954 Yudkin became the Chair of Nutrition for Queen Elizabeth College. In the 1960s, he grew increasingly concerned with the role of nutrition in western afflictions like obesity and diabetes, and spoke of the problem of “the malnutrition of affluence.” Yudkin retired from Queen Elizabeth College in 1971, and became Emeritus Professor of Nutrition.

Pauling first commented on Yudkin’s work – chiefly his book Sugar: Sweet and Dangerous – in a 1972 article for the newsletter Executive Health. In it, Pauling summarized Yudkin’s belief that sugar is an important cause of coronary heart disease, and that saturated fat and cholesterol are not. He also described a study carried out by Yudkin in 1957, in which the death rate from coronary disease in fifteen countries was correlated in relation to the average intake of sugar. The study concluded that men consuming relatively large amounts of sucrose faced far greater odds of developing heart disease in the age range of 45 to 65, than did those who did not ingest as much sucrose. Pauling agreed with Yudkin’s findings that sugar not only provided “empty calories,” but also contributed to various diseases.

In 1976 Pauling received a copy of Chapter 12 of Yudkin’s book, This Nutrition Business, in the mail.  In this chapter, titled “What You Can Expect from Vitamins,” Yudkin stated that Pauling’s claims about vitamin C were untrue. Yudkin suggested that the human body needs a certain amount of vitamins and no more, and that to ingest more vitamins than are required is a waste – thinking that was common at the time. He added that he knew Pauling personally and thought of him as warm and friendly, but also that “I think sincerely that he is wrong in most of what he says about vitamin C and about the use in large amounts of this and other vitamins in the preservation of health and in the treatment of disease.”

He then proceeded to find fault in Pauling’s argument that the best diet is one of raw fruits and vegetables; a diet that would provide roughly the same amounts of vitamin C that humans consumed millions of years ago. Yudkin instead argued that humans have subsisted on an omnivorous diet for at least two million years, and that if they really weren’t ingesting enough vitamin C they would have died off long ago.

The year after Yudkin wrote about Pauling in his book, Pauling – in what may have been a retaliation of sorts – singled out Yudkin as an example of subjective reporting on nutrition.  Pauling mailed his editorial “Needed: More Responsibility, More Objectivity, Less Bias,” to Yudkin along with a short note telling him that he was sorry to have to use him as an example, and that he hoped Yudkin would “get around to examining the evidence about nutrition in relation to disease in an unbiased and responsible way sooner or later.” Yudkin answered Pauling with a terse note informing him that his views were simply different, and that Pauling should not accuse people of being biased and irresponsible just because they had differences of opinion.

It was clear by 1986 that all was forgiven, when Pauling cited Yudkin extensively in his book How to Live Longer and Feel Better.  In Chapter 6, Pauling discussed Yudkin’s book Sugar: Sweet and Dangerous, in which Yudkin demonstrated that ingesting sucrose leads to coronary disease. According to Pauling,

Against the general public acceptance of the proposition that coronary heart disease is caused by a high intake of animal fat (saturated fat) and the eating of foods containing cholesterol, Yudkin himself has shown that for some countries the correlation of coronary disease with intake of sugar is much better than that with intake of fat.

Pauling later commented that “It has been shown in a trustworthy clinical study that the ingestion of sucrose leads to an increase in the cholesterol concentration in the blood.” The trustworthy study of which he speaks was reported by Milton Winitz along with his associates in 1964 and 1970. This investigation studied eighteen prisoners who had volunteered to be locked into an institution for about six months and have their cholesterol levels recorded as they were fed a specific diet. After a preliminary period, the group was placed on a small-molecule diet made up of seventeen amino acids, a little fat, vitamins, essential minerals, and glucose.  From there, more sucrose was added back into the diet.  During the length of the study, the group’s cholesterol levels were closely monitored.

The average cholesterol concentration during the initial period, during which the subjects had been fed a standard Western diet, had been 227 milligrams per deciliter. After two weeks on the glucose diet, the average concentration dropped to 173 and, two weeks later, to 160. After that point, a quarter of the glucose in the subjects’ diet was replaced by sucrose. In a week the average cholesterol concentration was 178, and two weeks later it had risen to 208. The glucose was then added back into the diet, replacing the sucrose, and results were evident in one week, when the average cholesterol concentration dropped to 175, and kept dropping afterward to points even lower than the 160 initially recorded. In his book, Pauling stated that this study “shows conclusively that an increased intake of sucrose leads to an increased level of blood cholesterol.”

At the end of Chapter 6, Pauling concurs with Yudkin and gives advice to the reader regarding sugar. His first admonition is to keep away from the sugar bowl – to keep it out of your coffee or tea. He also warns against prepared, frosted breakfast cereals, and to keep away from any regular intake of sweet desserts. His last piece of advice is to avoid soft drinks. In a different section of the book, Pauling advises, as part of a regimen for better health, to “keep your intake of ordinary sugar (sucrose, raw sugar, brown sugar, honey) to 50 pounds per year, which is half the present U.S. average.” (By 2003, Americans were consuming 142 pounds per year, on average.)

In December 1987, Pauling was interviewed for the magazine Outside for an article that focused specifically on his views on sugar. In it, Pauling is quoted as saying, “the increasing incidence of [coronary] disease closely parallels the increasing consumption of sugar. It is not at all correlated with the consumption of animal fat (saturated fat) or of total fat.” With this, Pauling reaffirmed his support for Yudkin’s viewpoint that sucrose is the primary culprit behind cardiovascular disease.

In 1989 Yudkin visited Pauling in person and, shortly thereafter, sent to Pauling a copy of his latest book Pure, White and Deadly. In thanking Yudkin for the book, Pauling asked if he would be willing to serves as a member of the Board of Associates of the Linus Pauling Institute of Science and Medicine. He also asked for a biographical sketch to be run in the Institute’s newsletter, and likewise asked for permission to reprint parts of the book in the publication.

Yudkin eventually agreed to join the Board of Associates (once assured by Pauling that his joining would not involve any work, since he had too much already) and provided a biographical sketch along with a letter in which he joked that he had “excluded such interesting aspects of my life as what clothes I wear, what I have for breakfast…”  Little bits of humor such as these dot the correspondence between the two men, who maintained a friendly relationship despite their occasional public disputes.

In a memo relaying news of Yudkin’s appointment to the Board, Pauling noted that “[i]t was Yudkin’s work that caused me to make my strong recommendations about decreasing the intake of sucrose.” Pauling also obtained copies of Pure, White and Deadly to distribute to members of the Institute, and continued to promote the book in the LPISM newsletter. Clearly, although Yudkin contradicted Pauling’s strong arguments in support of vitamin C, Pauling saw the logic in Yudkin’s case against sugar and stood firmly behind it.

Formulas, Pictures and Sports Drinks: The Pauling Chalkboard, Part III

Linus Pauling, 1985.

(Part 3 of 3)

While much of the real estate on Linus Pauling’s chalkboard is consumed by lists of names, a number of additional annotations, when examined, prove to be of keen interest.

Metabolic Profiling

On the right side of the board, below the last column of names, is the following text:

NSF – Mol. Str. 21 Mar.

Library 3000 21 Mar.

Aging – NIH Nutrition

American Cancer Society – Dr. Neville

Sample Bank

Mass Spectrometer

Muscular Dystrophy

Aging Patterns in mice

This particular sample of notes relates to the metabolic profiling program carried out for some time at the Linus Pauling Institute of Science and Medicine. As mentioned in part II of this series, a large number of names on the board were involved with the metabolic profiling program, and this particular column of text ties many of the names together. Pauling was working with numerous people from diverse backgrounds and professions. He was in contact with researchers at, among other organizations, the Institute on Aging and the American Cancer Society.

The words “sample bank” refer to urine and blood samples that were to be kept refrigerated for, potentially, decades, and ultimately to be analyzed by mass spectrometry. This particular undertaking was very ambitious, and could have provided a great deal of material for practical study. Unfortunately, the chronically underfunded Institute had trouble with their refrigeration units, and the project was eventually abandoned. (Despite the setbacks, some results of this program of research, headed by Pauling and Arthur Robinson, can be found in articles published at Stanford University as well as in certain of the Institute’s early news releases.)

A New Sports Drink

Another interesting bit of text can be found towards the lower right hand corner of the board:

C + glycine

dextrose

The text is likely the basic outline of a carbonated “sports drink” being worked on by the Institute in the 1980s. The drink was to be infused with vitamins, and the Institute was developing acids that would provide alternative sweeteners. Production and research eventually halted, but it is interesting to think about what may have resulted from a successfully marketed “Paulingaide.”

Vitamin C, Cancer and Heart Disease

The following words, located in the upper right portion of the column ark, have perhaps the most basic and relevant connections to Pauling’s work.

Ascorbate

stimulates

Production of Lymphocytes

The order simply implies that ascorbate, or vitamin C, stimulates the production of lymphocytes, the major cellular components of the body’s immune system. Several studies have shown that increased levels of ascorbate generally correlate with increased levels of lymphocyte production. If nothing else, this is the most centrally relevant theme of Pauling’s work with vitamin C, and the fact that it maintained such a substantial place on his overcrowded board underlines the significance that he himself placed upon it.

In the middle of the board towards its top, is the diagram of a mystery molecule that was crafted by Pauling. Mention of the molecule (given the name “2-azido-5,8-dihydroxy-1,3,4,5,7,9,9b-heptaazaphenalene”) appeared in an article titled “A Prized Collection: Pauling Memorabilia,” published in Chemical and Engineering News in August 2000.

In a 1977 interview, Pauling was asked about his chalkboard and, in particular, about the mystery molecule.  He reponded

I had an idea in the field of organic chemistry about 40 years ago. It involved this unusual compound. Benzine has a six-membered ring of carbon atoms and this compound has three six-membered rings consisting of six carbon atoms and seven nitrogen atoms and then it has these hydroxyl groups attached. It is known that the similar substance with only one ring can be made into certain derivatives that have anti-cancer activity. And I thought that this substance with only three rings might well operate in the same way and that we should study it.

In other words, Pauling was still actively contemplating an idea that had occurred to him 40 years prior – an idea that managed to stay on his chalkboard through his death in 1994. Indeed the mystery molecule exemplifies the function of Pauling’s chalkboard, not only as a mnemonic device, but as a place holder for people and ideas that span decades.

Linus Pauling, 1991.

Left of the mystery molecule towards the top of the board, one finds a series of words written one above the other. The seemingly haphazard placement of the words diverts attention from their historical significance in terms of the latter portion of Linus Pauling’s life.

LDL

Cholesterol

Lipoprotein a

The words almost certainly refer to research that Pauling began supporting in conjunction with a German physician named Matthias Rath, which investigated the possibility of a link between vitamin C and heart disease. Over the final years of his life, Pauling spoke of the relationship between vitamin C and heart disease in much the same way that he talked about vitamin C in terms of colds and cancer.

This writing was likely one of the last times that Pauling touched chalk to his board, as his collaboration with Rath did not develop until the early 1990s. The three words both acknowledge and hide the significance of the interaction between Rath and Pauling – a mercurial relationship for much of its duration.

Sandbox

Beneath an ark of name columns, adorned with the mystery molecule at its pinnacle, is a half-circle filled with pictures, figures and chemistry formulas. This area is likely where Pauling exercised the least concern for preservation, and it is supposed that this area of the board was used to aide in his discussions with visitors to his office. The space likely represents over two decades of personal interactions between Pauling and others, a spot on the board where he could explain theories and manifest abstract ideas. In essence, this half circle is where Pauling used the board in a more traditional sense – writing and erasing as suited his needs.


Linus Pauling’s chalkboard is covered in historical significance. It functioned as an important tool for a very busy man, and has preserved a telling aspect of both the history of the Linus Pauling Institute and the character of Pauling himself, in part reflecting the organization of his consciousness.

 

To be sure, the board is merely a fragment of Linus Pauling and his research, but it is unique and intriguing in a very personal sense. The names, pictures and diagrams on the board all represent important aspects of Pauling’s professional life. Not only does it make a valuable contribution to a room dedicated to the man’s work, it preserves the living memory of Pauling by displaying an intimate demonstration of his method.

Pauling's chalkboard, as preserved in the OSU Libraries Special Collections.