The Rapeutation of Walter Nelson-Rees, An Excerpt From "A Co

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The Rapeutation of Walter Nelson-Rees, An Excerpt From "A Co

Postby admin » Wed Mar 09, 2016 7:55 am

The Rapeutation of Walter Nelson-Rees
Excerpt From: "A Conspiracy of Cells: One Woman's Immortal Legacy and the Medical Scandal It Caused"
by Michael Gold
© 1986 Michael Gold

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Image
Henrietta Lacks

8: Spreading the Word

The daily press knew how to handle this story. So what if Science buried Nelson-Rees's report in the back pages under the stodgy title "Banded Marker Chromosomes as Indicators of Intraspecies Cellular Contamination." The newspapers, properly horrified, played it on page one with headlines more to the point:

CANCER WAR SET BACK
GOOF COSTS 20 YEARS OF RESEARCH

A line of human tumor cells used by laboratories around the world for more than 20 years may have invalidated millions of dollars worth of cancer research, according to a scientist's report .... As a result, says the author, Dr. Walter A. Nelson-Rees, checks are in order for dozens of laboratories engaged in cancer research. -- Los Angeles Herald Examiner


DEAD WOMAN'S CANCER CELLS SPREADING

Dr. Walter Nelson-Rees, one of the most experienced cell biologists in the world ... has reported that many cell lines are by no means what they are thought to be by the laboratories handling them. -- Miami Herald


A SHOCKER FOR SCIENTISTS

"The main situation has probably existed for years," said the main author of the report, Walter A. Ne1son-Rees, a highly respected researcher. ... Ne1son-Reessaid the contaminating potential of the HeLa cells is well known, but that sufficient precautions against it have apparently not been taken. -- San Francisco Chronicle


All this publicity made no sense to a number of scientists. Why was Nelson-Rees taking bows now when Stan Gartler had dropped the original bomb in 1966?

Part of the reason was that Nelson-Rees's paper was printed in Science, one of the few technical journals that nonscientists, particularly reporters, find accessible. One section, prepared by the journal's news staff, was actually written in English, and in the 7 June 1974 issue, the section carried a story that translated Nelson-Rees's article beautifully. "If Nelson-Rees is right," wrote Barbara Culliton, "a lot of people may have been spending a lot of time and money on misguided research. If, for example, you are studying the properties of human breast tumor cells, hoping to find features that distinguish breast cells from others, and are, all the while, dealing unknowingly with cervical tumor cells, you've got a problem." That was plain enough even for a newspaper reporter to understand, and to embellish and bang out for the morning edition.

But what really made Nelson-Rees a media star was the dramatic background of his shocking results: "The War." In Gartler's day, HeLa contamination had been the dirty little family secret of the tissue culture crowd. Its broader impact was not obvious. In 1974, however, "The War" had been officially declared and raging for several years. Everybody knew that the nation's most brilliant medical experts were at this very moment working feverishly against the scourge of cancer. It was a national priority.

Nelson-Rees's message made this large and serious effort seem a little silly. Sure, the institute was spending millions of dollars sending its brave recruits over the top against the enemy. But it turns out our boys were shooting with blanks! It was a scandal, and there's nothing the press likes better than a scandal. Besides, this story came with a bonus -- the awkward adventure of the Russian HeLa cells, until then unpublicized. The reporters loved sprinkling that one in: we've not only screwed up cancer research, folks, we almost blew detente on account of these crazy cells.

***

Nelson-Rees returned from his week in Miami like the local boy come home from battle. Reporters were still pursuing him. Friends showered him with congratulatory phone calls and letters. And the laboratory's resident bard immortalized his accomplishments in a limerick that appeared on one of the office bulletin boards:

A perceptive young Nelson named Rees,
Dumbfounded genetic police
When HeLa he found
To abound all around,
In cell lines from West and from East.


In addition to all the excitement it generated, the publication of their report brought great relief to Nelson-Rees, Flandermeyer, and the rest of the crew. It had been frustrating to know what they knew without having a means of broadcasting it. Now that the news was out, it might be easier to spread the word about future screwups.

Yes, spreading the word, that was the goal here. The publicity was fun, of course. It was nice for an "outhouse project" to steal the show for a moment. But getting the news out to those who needed to hear it -- the scientific community -- that was the main point of all this. And the publication of their paper was really just the beginning.

Frantic scientists had been calling and writing from all over the United States and several other countries to request copies of the article. In a few weeks Nelson-Rees's secretary had mailed out all 400 reprints and had to order another batch from Science. The author himself was in equally high demand. Would he give a lecture at the Stanford School of Medicine? Could he address a meeting of viral cancer researchers in Hershey, Pennsylvania? Would he brief a group at the Argonne National Laboratory in Illinois?

***

Nelson-Rees hit the road, preaching and proselytizing like biology's Billy Graham. At one stop he would tell the stories of such victims as Bassin and Plata who, through lack of vigilance, fell prey to HeLa's sabotage. At another he would describe the waste and futility of trying to learn about breast or prostrate or kidney cells by studying cultures of cervical cancer. And he would always conclude with the exhortation: Never trade cells without reliable information about what they are and where they've been. And always double-check them, before and after your experiment.

In some respects Nelson-Rees's early evangelizing looked like a touring revival of Gartler's performance at the Bedford Springs Hotel. Like Gartler, Nelson-Rees was telling audiences they had torpedoed years of their own work by being sloppy and letting HeLa creep in on them. He encountered the same reactions, shock and skepticism. And he too was offering new tools, chromosome banding along with other techniques, to help set things straight.

But Nelson-Rees soon added a new message that was less scientific in tone and more philosophical, or perhaps more political. It was at a 1975 meeting of cell culturists and cancer researchers in Lake Placid, New York, that he began to talk about two different reasons for cell mixups. One was simple sloppiness. "It can be combatted in individual laboratories by adherence to increasingly stricter techniques," he explained. The other effect was "more lasting and insidious." It had to do with researchers' attitudes, "frailties of the human ego ... exigencies of profit margins ... the threat of cuts of support in contractual arrangements."

Most members of the audience had gone into his talk thinking they had a pretty good idea what Nelson-Rees was going to say. But when he hit this stuff about scientists' attitudes and frailties of the human ego, they didn't know quite what to make of it.

It was curious, he told his fellow researchers, how well they handled a problem involving bacteria, for example, or viruses. Most of them quickly faced up to it when a cell culture became spoiled by such infectious agents. Not so with cell mix-ups. They seemed to take it personally when someone claimed their cultures had been overtaken by other human cells, he said.

"Cases of cellular contamination have been known to precipitate lengthy diatribes and are the reason for lectures such as this one," he clucked. "This kind of contamination would certainly be easy to control if one could frankly and readily discuss it and eradicate it without fear of offending colleagues' feelings."

At the moment, said Nelson-Rees, there was no friendly forum for discussing cell mix-ups and no means of rapidly notifying the scientific community of contaminated cultures. The audience raised a few eyebrows in disapproval as he said, "I would now like to describe to you how difficult it was for us to publish our results on those cultures which we have vouched are HeLa derivatives."

He began with the story of the Russian cells. He recounted how reluctant institute officials had been to believe his results, how for months they merely ignored the rapidly accumulating data. He read aloud the reviewer's critique that accused him of making a "gratuitous attack" on the Russians, and he named a few names. Then he moved on to the American cells: HBT3, HBT39B, HEK, and the others. He explained that Science had originally turned down the now celebrated report, quoting sarcastically from Abelson's rejection letter: "The manuscript and the referees' comments are enclosed. I trust the comments will be helpful to you when you prepare the paper for submission elsewhere."

Stan Gartler had certainly never done this. He had presented his findings, disturbing as they were, made a couple of suggestions, and taken his leave. But here was Nelson-Rees hauling out the dirty laundry. He was pointing out the stains and explaining how they had got there, and more than a few members of the audience were starting to squirm.

To say that a few cell lines had got shuffled around, that was one thing. But to suggest that scientists were letting their egos get in the way of good science or that they avoided publishing important information because it might be controversial ... well, it just wasn't done.

***

Robert Stevenson was frequently in the audience at Nelson-Rees's presentations, though Stevenson never squirmed. Knowing Nelson-Rees as well as he did, Stevenson more or less expected him to say something startling.

Nelson-Rees and Stevenson first met in 1962. Nelson-Rees had just been hired as a research associate with U.C. Berkeley's School of Public Health. He was sent to Washington to tour the nation's central cell bank facilities at the American Type Culture Collection and to talk with Stevenson about the school's role in assisting the new program. As head of the National Cancer Institute's cell banking program, Stevenson was helping to organize and fund the network of outside support laboratories. These included Lewis Coriell's Institute for Medical Research in New Jersey; the Child Research Center of Michigan, where Cyril Stulberg and Ward Peterson were based; and Berkeley's new cell culture lab in Oakland.

Bob Stevenson was not a typical bureaucrat. For one thing, he spoke his mind. He routinely called those responsible for the confusion in cell culture rank amateurs, and he said it in print as well as in conversation. Until 1960 when the institute recruited him to develop high-quality virology research materials -- cell lines, in particular -- he had been head of a tissue culture lab at the Naval Medical Center in Bethesda. He had seen his share of accidentally contaminated cells, which gave him a firsthand appreciation of what the confusion meant. In fact, though still in his early thirties at the time, Stevenson was one of the old guard who discovered the original mixups among animal cells and called for a central cell bank in the late 1950s, years before Gartler's first discovery of HeLa contamination.

Stevenson had an honest, friendly manner that fit his cherubic face. His cousin, an illustrator for children's textbooks, had used that face as a model for Dick of the Dick and Jane elementary readers. But Stevenson also had a devilish grin and a mischievous streak. He liked to "stir up the muck," as he put it, "to get people thinking about stuff they don't usually think about, but should." When he left the lab bench for a desk at the institute, Stevenson thought of himself as being scientifically castrated. The only way to keep contributing, he figured, was through others, by encouraging them to do provocative experiments and to trumpet their own findings.

Maybe that was why he and Nelson-Rees hit it off. Nelson-Rees was a kindred spirit, a young and energetic perfectionist who was still in the lab. They quickly became allies, beginning a long partnership. Stevenson made Nelson-Rees a member of the advisory board to the cell bank, the group that judged whether a particular culture was qualified to be in the "reference library." Based on his analyses in Oakland, Nelson-Rees would tip off Stevenson about suspicious cell lines. As executive secretary of the board, Stevenson would then ensure that the proper probing questions were asked at the meetings. Nelson-Rees would report, for instance, on the nonhuman-looking chromosomes of a purportedly human culture, and the culture would be reviewed and rejected.

Later, when Stevenson moved up through the hierarchy and then left the institute in 1967, passing oversight of the Oakland lab to Jim Duff, he continued to encourage Nelson-Rees. In telephone pep talks -- usually when Nelson-Rees had uncovered something controversial as in the case of the Russian cells or the three American cultures that followed -- Stevenson would ask him, "Is your work good?"

The answer never varied.

"Well, then," Stevenson would say, telling Nelson- Rees what he wanted to hear, "it doesn't matter that the shit is going to hit the fan."

After Nelson-Rees's report appeared in Science, Stevenson kept trying to get him more involved in activities of the Tissue Culture Association, the professional group for cell culturists. Stevenson, who by then was back at the institute as manager of the Frederick Cancer Research Center, formed the association's Committee of Standardization, Collection, and Distribution of Cells and Tissues, and appointed Nelson-Rees as a member. Together they struggled to convince the group of the need for a listing of bona fide cells to be circulated among the association's members. They also launched a campaign to persuade journal editors to require complete, authentic descriptions of every cell line used in a published research report, including a summary of what tests were used to verify that the cells' species, sex, chromosomes, enzymes, and other traits matched those of the purported donor. Nelson-Rees and Stevenson never tired of saying how ludicrous it was that the association's own journal, In Vitro, had no such requirement. As chairman of the group, Stevenson also had the committee officially endorse Nelson-Rees's findings and recommend that he keep up the good work. And being a quintessential organizer, Stevenson was constantly setting up conferences to stress the need for careful monitoring of cell lines, conferences that often featured a presentation by Nelson-Rees.

For two friends, both interested in stirring things up, it was funny how different their styles were. Stevenson was easy-going, informal, naturally likeable. He was fond of telling stories, such as the one about the New York medical examiner who performed rectal autopsies so that relatives of the deceased wouldn't notice the incisions. Nelson-Rees on the other hand was stiff, painfully honest, and sometimes holier-than-thou; he just automatically got certain people's dander up. If tact consists in knowing how far to go in going too far, as Cocteau once wrote, then Nelson-Rees was tactless.

Stevenson had tried to explain the benefits of a little diplomacy to Nelson-Rees. "You're very valuable, Walter," he once said to him. "You have a real talent for sniffing out trouble. But it might be advantageous to talk more softly and still carry that big stick."

"You cannot ask a leopard to lose his spots, Bob," Nelson-Rees would respond, "I am what I am."

***

And what he was looked very much like obnoxious to some people. He was so high and mighty about HeLa cells and so full of himself. In his talks he had started to make comments like, "This slide shows the now well-known marker chromosomes of HeLa. . . ." or, "In the case of the now well-known Russian HeLa calls .... " as if to say, "You've all heard of me, no doubt, and my very important and well publicized work."

He described his 1974 trip to Russia like a missionary returning from the jungle with word that the heathen had seen the light. Not only that, he said, they didn't have any of this trouble with ego or funding worries.

"Contrary to being an insult to our Russian colleagues, our data resulted in a most generous invitation to Moscow last November," he told the crowd at one meeting. "There I visited five major institutes and at least fifteen individual laboratories, gave three seminars (translated simultaneously), and witnessed the initiation of at least four new cytogenetic control sections for cell line monitoring at different institutes. As you can see, I was also allowed to return after a very warm and friendly reception. "

All right, Walter, you're terrific! Could we move on to something interesting now? Some thought all this talk about sterile conditions and strict monitoring sounded more like a fifth-grade personal hygiene class than serious scientific discussion: never swap cells, boys and girls, and remember to floss every day. Whenever he rose at a conference to ask a question or make another one of his public service announcements for care and quality, these people would glance at each other and roll their eyes.

There were many, of course, who took his talks seriously. Even Duff and the other bureaucrats at the institute were convinced his work was important and his motives were unselfish, though he did make them nervous. But there were also a few who saw Nelson-Rees as a dangerous publicity hound, a Joseph McCarthy of the cell culture circuit who wanted to further his own career by ruining the careers of others. They bristled every time he challenged colleagues about the identity of cells used in experiments or asked how that identity was determined. And he was doing that a lot these days.

***

One afternoon in June of 1975, Walter Nelson-Rees and Relda Cailleau were standing in the lobby of Montreal's Queen Elizabeth Hotel screaming at each other. The occasion was the annual meeting of the Tissue Culture Association; it had been about a year since the publication of Nelson-Rees's report in Science.

Nelson-Rees had begun the conversation by asking about a cell line Cailleau had initiated sixteen years earlier, a lung culture called MAC-21. Cailleau exploded.

"You're an assassin. You're vindictive. You're just out to wreck people's reputations!"

"Relda, there's no need to scream at me," Nelson- Rees screamed back at her.

"We know all about cell culture. You have no right to attack us."

"Relda, the very way you're telling me to mind my own business is what makes it my business!" Nelson- Rees's Dudley Doo Right tenor rose and fell in pitch, sounding over the din in the lobby like an air raid siren.

They were quite a sight. Nelson-Rees, six-foot-three and gangly, towering over Cailleau, all of five feet, a round, tough-talking bundle of volatility. She had a large nose, darting eyes, and thick, dark eyebrows. As she argued in her high scratchy voice, she tossed her head from side to side and waved her arms around.

"Relda" was her mother's maiden name spelled backwards. The former Rose Adler, her mother adopted "Rose Relda" as a stage name while singing at the Opera Comique in Paris. "I can curse real well in French," Relda sometimes told new acquaintances to whom she was describing her background.

Walter and Relda went way back, though Relda went back even farther on her own. In the early 1960s, when the Oakland lab and Nelson-Rees were both just starting out, she was already an accomplished cell culturist working at the D.C. San Francisco Medical School. They would bump into each other all the time at local and national scientific gatherings, always as courteous colleagues, if not close friends. In 1966, however, they had a falling out over the question of an evening's entertainment -- an incident that Nelson-Rees believed to be the start of a cold war between them.

The way Nelson-Rees told the story, he was chairman of the organizing committee and she was in charge of rooms and facilities for a San Francisco meeting of the Tissue Culture Association. Nelson-Rees had proposed a cruise on the bay with a buffet dinner and California wine tasting. Cailleau, sure that the summer fog would spoil the festivities, fought the idea but was overruled. The cruise, complete with a watercolor sunset and radiant full moon, was a rousing success. Although they lived less than a mile from each other in San Francisco, Nelson-Rees and Cailleau rarely spoke again. In 1970 she moved to Houston to work at the M.D. Anderson Hospital and Tumor Institute.

But of course it wasn't the cruise that was bothering Cailleau at the moment. In the past year she had watched Nelson-Rees crusading, making speeches, basking in the limelight, and she didn't like what she saw. His methods were antagonistic, and his goals appeared more destructive than constructive. On top of that she couldn't stand his attitude.

"He's so busy showing off what a great guy he is, how he's always right," she once complained. "He's so supercilious it riles the hell out of me."

And now he was trying to pull that stuff on her, trying to attack her cell line, MAC-21.

Nelson-Rees always insisted he wasn't attacking anyone, really. He said it was just part of his normal information-gathering routine. He scrutinized every cell line sent into Oakland for analysis, he scanned the journals for signs of mix-ups, and he walked into every conference with his eyes and ears wide open, constantly searching for clues, continually trying to make connections.

In the case of MAC-21, he had stumbled across its suspicious nature about a month earlier while checking the validity of two other cell lines developed by a colleague of Cailleau's at the Anderson Hospital. It happened at a meeting of cancer researchers in San Diego. Nelson-Rees noticed in the program an announcement of two new breast tumor lines, SH2 and SH3. Even in 1975 breast cells were very hard to cultivate. Researchers had put hundreds, perhaps thousands of tumor fragments into culture, hoping for a long-lived cell line to take root, but there were only one or two established cultures. Bassin's HBT3 and Plata's HBT39B had been celebrated additions to the short supply until Nelson-Rees came along. The report of two brand new breast cancer lines was therefore of great interest.

What made Nelson-Rees suspicious of SH2 and SH3 were a couple of characteristics described in the program. To begin with, both cultures were said to have come from Caucasian patients, yet both carried G6PD type A, the enzyme variant that is virtually nonexistent among Caucasians. Their early histories were strange too. SH3 had sat dormant, barely growing in the culture dish for thirteen months before it bloomed into a fast-expanding cell line; SH2 had taken three years to do the same. Nevertheless, the announcement took pains to point out that the chromosomes and several other characteristics of the SH lines were clearly different from those of HeLa cells.

Nelson-Rees went to a talk given by a member of the Anderson group, patiently sat through it, and rose during the question-and-answer period. "Have you any information," he asked, "on whether chromosome banding techniques have been applied to these cells to exclude the possibility that SH2 and SH3 are, first of all, not one and the same cell line, and secondly that they do not have a whole complex of marker chromosomes in common with many now well-known HeLa strains?"

Gabriel Seman, the French-born virologist who had initiated the SH lines, answered that in fact banding had been done by collaborators at the Institute for Medical Research in New Jersey, who assured him that they were not HeLa cells.

One of the advantages of Nelson-Rees's public displays was that it turned up extra clues, often volunteered by colleagues who smelled something rotten but were not as ready as he to stand up and make a spectacle. The SH cells might have wriggled off Nelson-Rees's hook except that as the audience was filing out, Lewis Coriell, the director of the New Jersey institute, pulled Nelson- Rees aside to say that he too was bothered by the unusual characteristics of these cell lines. He said he would discuss them with his karyologist, Bob Miller, who had done the analysis for Seman.

Back at Oakland a couple of days later, Nelson-Rees received a letter from Coriell explaining that months earlier Miller had indeed concluded from his analysis of banded chromosomes that SH2 and SH3 were identical cell lines. Why Seman had reported them to be two independent lines was a mystery. In addition Miller had seen some similarities between the SH chromosomes and those of a HeLa culture he had in his lab, but he had decided the SHs were not HeLa because there were also many dissimilarities. "You have looked at more HeLa cell lines," wrote Coriell, "and he [Miller1 would like to have your opinion of his interpretation." Mug shots of the cells were enclosed.

Three chromosomes in each SH line were so obviously HeLa markers that Flandermeyer "the Slow and Deliberate" made his pronouncement the very same day the letter arrived. The markers, coupled with the observation that these cultures from Caucasian women were both carrying the black type A enzyme, convinced Nelson-Rees that both cell lines had been taken over by HeLa. He notified Coriell and Miller.

"In view of this additional information it would seem probable that the lines are contaminants," Miller then wrote to Seman. "I realize this differs somewhat from my original interpretation but hope that the situation is now clarified."

Yes, there was somewhat of a difference between saying SH2 and SH3 are cultures of breast cancer and saying, "Oops, they're HeLa."

Seman was incensed, though it is hard to say what upset him more: Miller's new diagnosis that the precious breast cultures were Henrietta Lacks's well-traveled cervical tumor cells, or the fact that Miller dared send the SH photographs to another investigator. "Wizzout my permission!" as he put it. Seman was even angrier with Nelson-Rees for using information that Seman decided had been obtained unscientifically and illegally. "Eeet's a case for my attorney," he declared.

Seman's colleagues at Anderson, Relda Cailleau among them, were equally outraged. Nelson-Rees had acted irresponsibly and unethically, they all agreed. It was the same trick he had played on other unsuspecting researchers. To them the question of HeLa contamination was somehow a secondary concern.

Not to Nelson-Rees, though, who for the moment was oblivious to the reputation he was building at the Anderson Hospital. Having nailed SH2 and SH3, he turned his attention to MAC-21. He remembered seeing a reference to MAC-21 in the paper announcing the SH lines. Specifically, the report had said that the SH cells were like MAC-21 cells in that they had the type A form of G6PD.

Now MAC-21 had been around a long time, used extensively in lung cancer research, yet Nelson-Rees didn't recall ever hearing that it had this unusual trait. So it was that a month later when he saw Relda Cailleau descending the stairs at the Queen Elizabeth Hotel in Montreal he walked up and said, "What's this I read about MAC-21 being type A? Have you ever checked it?" That's when Cailleau erupted.

"You're doing this to everyone, are you? You're assassinating people!"

"Look, we know there's a problem with SH2 and SH3. They were both type A G6PD and we found out they have HeLa chromosomes."

"MAC-21 is not HeLa. I'm absolutely positive."

They went on for ten minutes like that. Finally Nelson- Rees went into one of his commercials. Cailleau might be interested to know that he and Bob Stevenson were firing up the cell standards committee of the association to advertise the need for careful controls and regular monitoring of cell identities, he said. "But I don't have to tell you the importance of keeping things straight. As you know, Relda, I'd be glad to check any of your cultures for you."

At that, Cailleau stomped out of the lobby.

***

It was during this early period of the crusade, within a year of the Science article, that an amusing telegram arrived at the Oakland lab. The unsigned message offered Nelson-Rees a high-paying and prestigious new job in research. It also offered a one-way ticket to get him to this very attractive position -- in Uganda.
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Re: The Rapeutation of Walter Nelson-Rees, An Excerpt From "

Postby admin » Wed Mar 09, 2016 7:55 am

9: Damage Report

It was too bad that Nelson-Rees didn't have an accountant traveling along on the crusade, someone to assess the piles of spoiled research he had dug up and left rotting in his wake, a bookkeeper of the bereaved to record the number of hours squandered and research dollars frittered away. The newspapers said millions had been wasted by HeLa's unexpected spread, but there were no official damage reports. The bureaucrats at the National Cancer Institute never tried to survey the wreckage, and they probably would have failed had they attempted it. Very few of HeLa's victims, even those asked point-blank, would ever detail how much time, effort, and money they had wasted, or how many colleagues they had led astray.

Take Ernest Plata, for instance, the cultivator of HBT39B, one of the two HeLa-contaminated breast cultures that came out of the institute's building 41. Plata established HBT39B in the summer of 1971 and studied it on and off over the next two and a half years, hoping to find viral clues to the cause of breast cancer. After Nelson- Rees convinced him that his malignant breast cells were in fact malignant cells of the cervix, Plata put them permanently on ice and notified the six or seven people with whom he had shared them. No harm done, though, according to Plata.

"I wouldn't necessarily count it as a loss," he said many years later. "In some ways, it served to accelerate awareness and prevent many more losses."

Not every researcher who had a run-in with Henrietta Lacks considered it such a lucky break. Some simply refused to believe it. They continued working with their cultures, many of them honestly convinced the cells were bona fide in spite of Nelson-Rees's test results. Needless to say, they offered no damage estimates.

Nelson-Rees himself never cared much for figures. Besides, he was too busy tracking HeLa down, convincing people he was right, and lobbying for reform. "The enormity of the problem is obvious," he would say, pointing to his growing list of HeLa-contaminated cultures. "There's many a sad tale here."

One of the few sad tales that did have a price tag was the story of HBT3, the other breast cancer culture from building 41. The mix-up of HBT3 was no more scandalous than the mix-up of any other cell line. It's just that Bob Bassin and his group reacted pretty reasonably to the news about their culture, and they candidly discussed details of the mishap.

Like Plata and many others, Bassin and co-worker Brenda Gerwin had been looking for signs that a virus was the cause of breast cancer. First, they needed a living piece of breast cancer. After trying nearly 100 tumor fragments, Bassin finally got one to spawn a cell culture, which he christened HBT3. Although there were no viruses to be found in HBT3, Bassin and Gerwin hoped to turn up a clue that viruses had been in the tumor at an earlier time and had left their mark on the cells. What they hoped for was a telltale enzyme produced by the virus, an enzyme called reverse transcriptase.

It took Gerwin and a technician a year, devoting half their days to the effort, to purify and isolate an enzyme from the cells and to identify it as reverse transcriptase. In August 1973, Gerwin, Bassin, and their associates published a report in the Proceedings of the National Academy of Sciences announcing the find.

Gerwin and her assistant then spent another three or four months combining the enzyme with blood serum drawn from patients with breast cancer. The theory was that if this enzyme were really manufactured by viruses that caused breast cancer, then there would be antibodies in the blood of breast cancer patients that would attack the enzyme. But they found no evidence of an attack. It was about that time, toward the start of 1974, that Nelson- Rees's letter arrived, making it clear why the blood of the breast cancer patients had not reacted to the enzyme from their "breast cancer" cell line.

It took another month or two, but Gerwin and Bassin became convinced that HBT3 was actually HeLa, and that they had been wasting their time. Had it been another kind of cancer cell that slipped in, they might have salvaged a few observations about that cancer's relationship to viral enzymes. But not in the case of the HeLa line. HeLa had been in so many labs and so many different microbiological environments, they knew, that it could have picked up all kinds of things that had nothing to do with viral cancer. They abandoned their work on HBT3.

Nearly a decade later, Gerwin and Bassin calculated the damage. Gerwin figured the half year of purifying and identifying the enzyme was not a total waste since some of the techniques developed she applied to other experiments. Perhaps three of those six months were for naught. She and her technician wasted another three months testing the patients' serum against the enzyme. The combined six-month salary of a Ph.D. and a technician in the mid-1970s was about $20,000. To that Gerwin added support costs, anything from the plumbing and heating to cell culture materials, which she guessed came to another $20,000. The overall loss for Gerwin's end of the project, then, was about $40,000.

Before Gerwin even started her work, however, Bassin had spent three months nursing the cells into a sustainable culture and performing various tests. He thought it cost at least $25,000 in wages and laboratory expenses. In other words, the HeLa cells that overran HBT3 inflicted $65,000 worth of damage on the group that initiated the culture.

The damage didn't end there, of course. Bassin had sent twenty-seven shipments of what he thought was HBT3 to twenty research teams around the world, from the University of Rhode Island to the Karolinska Institute in Stockholm. The shipments went out between August 1972 and August 1973, meaning that scientists could have been working with the cells for anywhere from eight months to a year and eight months before they received Bassin's notice about the HeLa contamination. If only four of the twenty groups invested as much effort in this culture as did Bassin's team, the total damage would be $260,000.

There were only two other sad tales for which anyone would try to figure a tab. One was the Russian caper, Nelson-Rees's first brush with HeLa. According to Wade Parks, leader of the team of American virologists who analyzed the viruses in the six Russian cultures -- the viruses some hoped would include the human tumor virus, the ones that turned out to be monkey viruses -- the laboratory work alone cost $50,000 to $60,000. As for the total cost -- including such incidentals as investigators' time, the price of hiring a commercial lab to grow large quantities of the cells, and the expense of elaborate meetings to discuss the findings -- Parks estimated it many years later at a quarter-million dollars.

And then there was MA160, a bogus culture of human prostate cells that Nelson-Rees featured in his 1974 Science article. Seeing that article, a cell biologist at the University of Colorado named Mukta Webber scanned the scientific literature and found that six recently published studies had been based on the erroneous assumption that MA160 was a prostate cell. At the time she gave no quantitative assessment of the harm done, though researchers familiar with the field would eventually estimate the price of those six misguided studies at $30,000 to $40,000 apiece -- a total of $210,000 to $280,000. As it turned out, Webber had found only a portion of MA160's misadventures.

But taking even the minimum damage estimate of $210,000 for MA160 and adding a quarter-million each for HBT3 and the Russian cells, the cost of just three instances of HeLa contamination approached $1 million. To say nothing of the two other cultures on Nelson- Rees's 1974 list or the monetary equivalent of fifteen years of work lost in the early era of cancer research, long before Nelson-Rees joined the chase. Or the waste due to holdover cultures from those days, lines such as the Chang liver and the WISH amnion, which, despite being branded as HeLa, were still surfacing periodically in published research reports as bona fide liver and amnion cells. Or, finally, the many new HeLa-contaminated cell lines that Nelson-Rees and Flandermeyer were still uncovering.

Bob Stevenson, at one time the man in charge of cell cultures for the institute, was one of the few officials ever to guess at the total damage HeLa had done. Some ten years after the newspapers first reported the story, Stevenson would look back and figure they were right. By the time Nelson-Rees had published his report in Science, the losses added up to millions of dollars.

"Millions is probably the right ballpark," Stevenson would say. "But you're not going to get anyone to admit that. It's like something uttered in a confessional."
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Re: The Rapeutation of Walter Nelson-Rees, An Excerpt From "

Postby admin » Wed Mar 09, 2016 7:56 am

10: Provenance

Walter Nelson-Rees is driving away from Oakland, toward the East Bay hills and Orinda. Although the passenger has requested no tour, he is from out of town and Nelson-Rees cannot resist. "This street we're on was the old Broadway of Oakland. That's old Tunnel Road over there. The freeway on our left was built with all the fill they dug out of the hill up ahead to make the Caldecott Tunnel. On the right here is a new sports complex, a recreation area I guess you'd call it, that is in the process of being constructed .... "

He turns the car back in the direction of Oakland and heads downtown.

"This church over here on Castro Street used to sit one block that way on Brush Street. They actually lifted the entire thing and moved it for the freeway."

The visitor begins to speak, but -- "Originally, I think it was Greek Orthodox. Then some kind of congregational church, and then a synagogue. I believe it's Methodist at the present time. Though I'm not sure of that .... "

The man's incredible, thinks Nelson-Rees's prisoner, he even spells out what he doesn't know.

"These are all original little Victorian houses. Bret Harte lived and worked in this area. He had a place with his stepfather here on Fifth and Clay, his stepfather who was the model for the character of Colonel Starbottle in his short story "The Romance of Madrono Hollow." Do you know Bret Harte, the writer?"

The passenger hesitates, then nods, wondering whether a nod will bring momentary relief or only encourage his captor.

But it makes no difference. Nothing can stop Nelson- Rees now. He is free associating in high gear, interrupting himself at every turn, improvising around the one tune his brain is always humming: Origins. Histories. Where things came from and what they used to be.

It wasn't just buildings and roadways. He read postmarks, for instance, as if they were clues to buried treasure. If a letter arrived in his office without one, he would take out his red felt tip pen, circle the stamp boldly, and write "NO CANCELLATION!" across the envelope before filing it away. It really bothered him not to know where and when the thing had been mailed.

On napkin strips and pages tom from scientific journals, he often wrote up little summaries of conversations he'd had. Some of his notes of telephone calls even specified that he'd been called collect. These along with letters, telegrams, memos, conference agendas, lab notes, articles, and scrawled copies of questions he had asked speakers at certain meetings -- as well as the speakers' replies -- all of these were stuffed into thick, black notebooks that filled the bookshelves in his office. As one of Nelson-Rees's close friends once explained, "When Walter says something to anybody, he's got a piece of paper with his name on it, with the date, and what was said. It's there. You can criticize it. You can impugn it. You can say it's a lie. But it's there."

It was the same with his interest in art. Provenance is the term for a painting's history, and until he knew the provenance in full detail, Nelson-Rees felt he couldn't appreciate the work. When was it painted, where was the artist working at the time, who were his teachers, was it ever exhibited? It was probably under the force of this compulsion in 1953 that he had changed his very name, from Rees to Nelson-Rees, thereby specifying both parents, his own provenance. And so it was with cell cultures. If you didn't know everything about the cell line you were experimenting on, especially its true identity, then you didn't know anything.

So it was probably inevitable that Nelson-Rees would publish a second list of HeLa casualties. As it turned out, though, there were other good reasons.

Since the first article, the one that had "indicted" five cell lines as HeLa, he and Flandermeyer had discovered eleven more HeLa-contaminated cultures. When Nelson-Rees told the people who established these cultures that the cells were spoiled, many simply refused to believe him. It was not enough to deliver the bad news to these researchers and hope that, like Bob Bassin, they would do the honorable thing. As each of the eleven turned up, it became increasingly apparent to Nelson- Rees that he would have to warn the world himself through another published list.

***

SH3, the purported breast cancer culture established by Gabriel Seman at the M.D. Anderson Hospital, was one such case. Seman consulted a karyologist at Anderson who disagreed with Flandermeyer and Nelson-Rees's judgment that SH3 had HeLa chromosomes, the conclusion with which Miller had concurred. Seman also changed his story about the origin of the culture. After examining a photograph in his files, he concluded that the woman who donated the cells was not Caucasian as he had originally reported, but Mexican. Since the rare type A enzyme occasionally appears among interracial Mexican-American populations, according to Seman, this meant SH3 was not necessarily a HeLa culture.

Having satisfied himself that SH3 was a bona fide breast culture, Seman saw no reason to curb its use or warn colleagues. While Nelson-Rees was preparing his new list, which would blow the whistle on SH3, the cell line was already being used by breast cancer researchers at the Memorial Sloan-Kettering Cancer Center in New York and by a group in the Soviet Union, as well as by an investigator who worked separately from Seman at Anderson. Indeed Seman would soon publish his own report in the journal Cancer, officially announcing the availability of this new breast cancer culture to the scientific community at large. In the article, he would rule out the possibility of HeLa contamination.

***

ElCo was a cell line that Nelson-Rees chose to include on his second list for similar reasons. According to oncologist Roland Pattillo, the man who established it,  ElCo was yet another breast cancer culture. In his laboratories at the Medical College of Wisconsin, Pattillo had used the cells to test the effects of certain chemotherapy drugs before administering them to the woman from whom the cells had supposedly been taken. He also studied the reaction of other breast cancer patients to this "breast culture" using a skin test something like the ones used to check for allergies. These patients showed an immune reaction to ElCo as if their bodies had already been primed against it, suggesting to Pattillo that different breast tumors might have certain characteristics or antigens in common.

The only problem, Nelson-Rees told Pattillo after checking the culture, was that ElCo was neither a breast cancer culture nor representative of Pattillo's patient. It was a HeLa culture.

Pattillo protested that breast cancer patients would never have had an immune reaction to the cells unless they were genuine breast cancer cells. He would later concede, however, that he never tested that assumption by checking patients' reactions to HeLa cells. Pattillo also explained that he knew all about the infamous tumor of Henrietta Lacks, having been an associate in the laboratory of George and Margaret Gey soon after Mary Kubicek placed that fateful bit of tissue into the roller tube. But he said he had no HeLa cells in his own lab, so contamination was impossible.

Nothing in his lab was labelled HeLa, that was true. But he had been experimenting with another breast culture, Bassin's HBT3, and apparently HBT3 had dropped in unexpectedly on some of the other cell lines in the neighborhood. Nelson-Rees and Flandermeyer's analysis showed that ElCo contained the very same banded chromosomes as did HBT3 and its HeLa-contaminated cousins: the four standard HeLa markers and the newer pair, the Mickey Mouse and Zebra.

Pattillo just didn't believe it. He kept working with  ElCo as if it were a breast tumor culture and kept publishing results. "That's what really got the wheels rolling around here," Flandermeyer recalled. "That's the kind of thing that got Walter all fired up and publishing new lists and writing letters to editors."

But what worried Nelson-Rees more than Pattillo's reaction was the fact that HeLa cells were still circulating in their old HBT3 disguise. HBT3 had been on the first list in 1974. For the last two years he had been making as much noise as he could about HBT3 and the four other HeLa-contaminated cultures he had originally stumbled upon. And yet here was Pattillo claiming he had no HeLa in his lab -- although, yes, he did use a little HBT3 now and then.

***

Unfortunately, HBT3 was not the only active alumnus from the first list. Nearly everyone, it seemed, was still using MA160, the bogus prostate cell that by the mid-1970s had helped researchers waste an estimated $200,000.

MA160 was developed in 1966 through the collaboration of several scientists and a man named Monroe Vincent, a partner in the biomedical supply firm of Microbiological Associates, Inc. The scientists provided some of the technical expertise; Monty Vincent personally contributed the cells, a lump of tissue that had been taken from his own prostate. The lump was biopsied by his doctor who was concerned that it might be malignant. Fortunately, it turned out to be benign.

It also turned out to be a good source of culturable cells, or so it seemed. The cells from Vincent's prostate languished in the laboratory for a few months and then, touched by the miracle of "spontaneous transformation," were reborn as a bunch of frantic cancer cells. The first long-lived line of prostate cells, MA160 became a bestseller.

As it happened, though, the cells were exactly the opposite of what they were supposed to be. Not male, but female. Not prostate, but cervix. Not from a white donor, but a black one. MA160 was arguably the most tasteless of HeLa's practical jokes.

Stan Gartler was the first to nail MA160 as HeLa in 1968. He added it to his list of the original 18 HeLa-contaminated cultures based on its having the A type of G6PD enzyme. Vincent and his co-cultivators, however, dismissed the finding in their first official description of the cell line in Science in 1970. They theorized that Vincent might have had Negro ancestors from whom he inherited the rare, black enzyme. They added that the cells contained Y chromosomes, proof they had come from a male donor.

In 1973, however, Ward Peterson, Nelson-Rees's regular partner in Detroit, reported that he could find no Y chromosomes even in the earliest samples of MA160 available. Furthermore, having tested Vincent's own blood, he could say without a doubt that whatever exotic ancestry Vincent claimed, he had ended up with type B G6PD, the type expected for whites. "His" cell line was certainly not his anymore, but that of a black woman. In 1974, Nelson-Rees and Flandermeyer found that MA160 not only had the type A enzyme and was missing its Y chromosome, it also displayed banded marker chromosomes identical to HeLa's. Another group that had been working with the culture, a team at the Anderson Hospital, no less, soon came to the same conclusions.

It wasn't long before Mukta Webber, the Colorado cell biologist, reported that MA160 failed to produce prostatic acid phosphatase, a chemical normally manufactured by prostate cells. Webber also reported that despite its unprostatelike behavior, MA160 was still a popular research culture, describing and giving references to the six recently published "prostate" studies based on the cell line.

But that was not the end of MA160. Early in 1975 a West German scientist named Frederick Schroeder announced the establishment of a new prostate culture at a conference in Italy. Nelson-Rees, who happened to be attending, heard Schroeder describe the new cell line, EB33, as quite similar to the prostate culture MA160, which Schroeder also had been studying in his lab. In certain respects, EB33, like MA160, behaved strangely, according to Schroeder. Nelson-Rees began asking questions. A few months later, having analyzed several samples of EB33, he wrote Schroeder that his new prostate culture was definitely MA160, which was definitely HeLa.

And still MA160 and its alter ego endured. Many researchers went ahead and worked with EB33 just as others had continued to use MA160. Some, including Schroeder, used both cultures in their experiments, presumably to increase the validity of the observations. Here it was again, Nelson-Rees's nightmare come true. Researchers were not only using MA160 as if it were a bona fide prostate culture, they were exposing all the other cell lines in their labs to a HeLa culture in disguise. Which was why in addition to warning people away from the eleven newly spoiled cultures like EB33, ElCo, and SH3, Nelson-Rees decided his second list would have to remind them of the contaminated cultures he had publicized earlier.

In fact he decided not only to repeat the lines he indicted in 1974, but also to quote every report he could lay his hands on that branded any cell culture HeLa. Among others, he cited Gartler's original findings, recent studies by Ward Peterson in Detroit, and a paper soon to be published by researchers at the American Type Culture Collection. In the manuscript he submitted to Science accompanying this second list, he explained how widespread "secondary contamination" had become and admonished his colleagues, "HeLa by many other names can spell trouble."

The list, published in January 1976, was an impressive and distressing inventory. More than forty different research cultures had been commandeered by HeLa. If each culture had cost science a quarter-million dollars, as had the few for which damage estimates could be made, this list represented $10 million in losses. The evidence against each culture, organized in a detailed chart, was overwhelming too. Not only were these cells identical to HeLa on the basis of their C6PD enzymes and marker chomosomes, but many had also been tested for the enzyme PCM and for a few more biochemical traits called HLA antigens. These all matched HeLa's characteristics.

And, oh yes, there was one other piece of information under each cell line in Nelson-Rees's second list: a researcher's name. Any scientist who had supplied him with a culture that turned out to be HeLa was immortalized in what came to be known as Nelson-Rees's hit list. No one considered this an honor. A few felt deeply betrayed and never spoke with him again. And the ranks of those who thought Nelson-Rees was just out to make a name for himself grew considerably.

Jim Duff and some of the other bureaucrats at the institute wanted to know why he had to name names. "You keep pushing people," Duff complained. Why, they wondered, couldn't Nelson-Rees simply give the sample's "passage level" -- the number of times it had been grown out, cut up, and transferred to new flasks, which would give a relative indication of how close it was to the original culture-or identify it in some other way, and leave off the scientist's name?

Leave off the name? That, to Nelson-Rees, was like -- well, like asking the Post Office to forget about the postmark. Why not take down all the road signs and let drivers fend for themselves? Why not ask the telephone company to publish just the phone numbers and have people guess to whom they belong? To leave off the names would defeat the whole point of his list. He was trying to be as precise and complete as possible, to specify where the HeLa-contaminated cultures had come from and, by implication, where they had not come from. After all, he hadn't tested every sample of these cell lines. Not yet.

"After the 1974 paper," Nelson-Rees explained to Science News magazine, "some researchers analyzed cultures they had been using of the same type we 'fingered' and found them to be bona fide bladder carcinoma cells, or whatever. Therefore, the source of these cultures becomes an important piece of data. I felt obligated to state from whence these cultures came and let the other shoe drop where it may."

Of course, Nelson-Rees must have known that naming names was, as the magazine put it, "an action sure to be interpreted by some as unfriendly." But maybe he figured that in order to make any progress, the second list had to be more combative than the first.

"At this point," he told Science News, "I'm going to go hide."

Naturally, he did nothing of the kind.
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Re: The Rapeutation of Walter Nelson-Rees, An Excerpt From "

Postby admin » Wed Mar 09, 2016 7:56 am

11: Another Run-in With Relda

Nelson -Rees was browsing through the April 1977 issue of the Journal of the National Cancer Institute when he caught the scent of something mildly suspicious on page 863.

Now some people would have said he went looking for bad news, especially after that hit list with the names on it. But the way he saw it, he just wanted to make sure the reports of good news were genuine. And starting on page 863 of the institute's journal there appeared to be very good news indeed, a report of some promising findings about lung cancer.

Richard Akeson of UCLA medical school had found several specific antigens associated with the cells in a lung tumor culture. Antigens are like biochemical dog tags, tiny nameplates on a cell's surface that can be recognized by the body's immune system, though in the case of cancer the system seems unable to successfully attack even when it recognizes the enemy. While the discovery was preliminary, Akeson wrote, such tags might eventually help doctors diagnose this particular kind of lung cancer more effectively, perhaps earlier, and help them monitor its progress. Furthermore, though Akeson didn't actually mention it, there was always the hope that if tumor-specific antigens could be found for certain cancers, they might serve as signposts to be followed by cancer- killing drugs or other selective assaults to be developed in the future. Such was the promise of an antigen found to be unique to a particular kind of tumor cell.

Nelson-Rees knew from experience, however, that antigens specific to a certain type of cell -- and only that type -- were practically impossible to find. And, of course, finding a unique antigen wasn't enough. You also had to be certain of the tumor cell's identity in the first place, so that later when you saw that antigen on the cells of an unidentified culture you could say, for example, aha, we must have a dishful of Russian bladder cancer here. Given the proclivities of biologists to mix things up, Nelson-Rees figured, any tissue-specific claim was worth a closer look.

According to the "Materials and Methods" section of Akeson's report, the lung tumor cell line was called 2563 and had come from Litton Bionetics, Inc., in Kensington, Maryland. Following that bit of information was a footnote that directed Nelson-Rees to the bottom of the page. There amid the fine print, quite unexpectedly, he bumped into none other than Relda Cailleau. It seemed that 2563 was another name for MAC-21, the lung culture that had triggered the hotel lobby screaming match in Montreal two years earlier. Despite Cailleau's assurances at the time that MAC-21 was not HeLa, the matter was left very much unresolved in Nelson-Rees's mind. Now that someone was basing important new claims about lung cancer on this twenty-year-old culture, he decided the question had to be settled.

He ordered samples of 2563 from Litton Bionetics that summer, and by September the verdict was in. According to Ward Peterson's analysis, the culture's G6PD enzyme was type A, the same as HeLa's. Although Cailleau had taken the original MAC-21 cells from a fifty-three- year-old man, Nelson-Rees and Flandermeyer saw no male Y chromosomes. And in nearly every cell, they found abnormal chromosomes whose shapes and banding patterns matched three of HeLa's best-known markers. Nelson-Rees wrote a detailed letter to John C. Bailar III, editor of the Journal of the National Cancer Institute, asserting that 2563 had been contaminated by HeLa and was not at all characteristic of lung tumor cells. He asked that the letter be published as soon as possible, saying his results were "an important addition to the increasing knowledge of what does and does not constitute organ-or tissue-specific antigens."

Three months passed with no word. Nelson-Rees called the journal, and someone there explained they had sent copies of his letter to Akeson and Cailleau so that they could write rebuttals to be published along with his letter. Akeson's reply had come in, a cautious concession that the "lung cancer cells" were apparently HeLa, but they were still waiting for Cailleau's.

Convinced as ever that the cells were pure, Cailleau had pulled out the one remaining ampule of MAC-21 in her possession and had it analyzed by Sen Pathak, a chromosome banding expert at the M.D. Anderson Hospital, and Michael Siciliano, the local enzyme man. She was "disappointed" as she later described it, when Pathak reported seeing HeLa's marker chromosomes in MAC-21 and Siciliano said not only was the G6PD HeLa's type, but thirteen other enzymes he tested matched as well.

Disappointed, but undaunted, Cailleau still insisted that MAC-21 had been a genuine lung cancer cell for many years, and if it was HeLa now, well, that was somebody else's doing. She explained to her colleagues, as she would afterwards to anyone who asked about MAC-21, that she had shared the cells with other researchers in San Francisco before coming to Houston. When she moved she took with her two different batches: one of live samples from those researchers and another of frozen samples that only she had used. Then in the early 1970s some fool technician had failed to keep the freezer stocked with dry ice, and she lost seventeen years' worth of cell cultures, including the bona fide MAC-21 cells. Her only remaining samples were those that she had had no direct quality control over.

"The problem," Cailleau said, "is that often contamination of cells is a secondary or a tertiary thing done by associates or technicians, and the originator knows damn well he or she didn't contaminate them. In my hands, there never has been a HeLa contamination." In other words, yes, the MAC-21 cells she had now were indeed HeLa, but it wasn't her fault. The same was true for the samples she had sent a few years earlier to a variety of colleagues, including one who must have passed them on to Litton Bionetics under the name 2563, some of which had ended up in Richard Akeson's lab.

All this Cailleau explained in a letter to Bailar at the Journal of the National Cancer Institute in January 1978. Painful as it must have been, she also telephoned Nelson- Rees -- collect -- and laid out the whole mess for him. She sounded uncharacteristically subdued, Nelson-Rees thought, even a bit sad. As pleased as he was to hear his suspicions confirmed, he kept his I-told-you-so's to himself.

Still, it would be another six months before anyone but Nelson-Rees, Akeson, Cailleau, and Bailar had any inkling that MAC-21 and 2563 were not what they were advertised to be. Scientists who had read Akeson's report on lung tumor-specific antigens would have had an entire year to follow his lead without any indication it might be a dead end. Finally Nelson-Rees's letter appeared in the journal's June 1978 issue. It was followed by a letter from Akeson who said that based on Nelson-Rees's work "it seems reasonable to presume" the cells he had thought were from a lung tumor were actually HeLa cells. He added, however, that some of the antigens in these cells did seem to resemble those seen in genuine lung tissue, concluding that further studies were needed to sort through the confusion. Following Akeson's letter was a one-paragraph statement from Cailleau conceding that the single culture of MAC-21 she had checked was HeLa. "I am certain that the original MAC-21 was obtained from a mucoid adenocarcinoma of the lung and remained uncontaminated for several years," she wrote with a trace of controlled testiness. "The addition of HeLa cells to our cultures occurred later and the transfers were carried out by several people."

Around the time of this embarrassingly public cell mix-up, Nelson-Rees received a draft report from a microbiologist at the National Cancer Institute. It was an extensive review of breast cancer cultures available to experimenters, and, as Nelson-Rees was so experienced in telling good cells from bad, the microbiologist asked him to critique the report before it was submitted for publication. Oddly enough, there were several cultures listed whose technical descriptions were missing. The cover letter explained these were Cailleau's cultures, and Cailleau had asked that all information on them be deleted from any copies of the draft that Nelson-Rees might see. Nelson-Rees was amazed-first that Cailleau would have the gall to dictate the handling of someone else's manuscript and, second, that the author would knuckle under. When Nelson-Rees called to find out more, the microbiologist said that Cailleau had been "adamant on this point; abusive, in fact."

Obviously Cailleau had been more than just miffed by her latest dealings with Nelson-Rees. She was determined to make the MAC-21 incident the last time he would ever stick his nose into one of her cell cultures.

***

A few weeks later, the Tissue Culture Association asked Nelson-Rees to evaluate a stack of abstracts, or summaries, of research reports submitted for the association's upcoming annual meeting. He found one of the abstracts particularly interesting. It described an unusual sample of breast tumor cells that exhibited a surprising number of characteristics similar to HeLa's. In fact, many of its chromosomes looked like HeLa's banded markers, the authors said. Yet they knew for certain the cells were not HeLa because they had come directly from the patient -- out of a pleural effusion, a sample of liquid taken from the patient's chest cavity, to which the tumor cells had spread. The cells in the effusion were analyzed immediately; they had not sat around the lab and had a chance to become contaminated. The implication was that in this case the standard techniques of identifying a HeLa cell might well have led to a false diagnosis. These bona fide breast cancer cells might have been wrongly branded as having been overtaken by HeLa.

The abstract had been submitted by three scientists at the Anderson Hospital: Sen Pathak, Michael Siciliano, and Relda Cailleau.

Nelson-Rees almost laughed. He doubted the claim, of course, but was intrigued at the same time. In part because he wanted a chance to examine the supporting data, he recommended that the report be accepted and presented at the meeting. He then wrote to Pathak, the karyologist, asking to see photographs of these curious chromosomes.

When the photos arrived two months later, eagle-eye Flandermeyer looked and looked but he couldn't see any known HeLa markers among the banded chromosomes. Nor could Nelson-Rees. They sent copies to K.S. Lavappa, the chromosome expert at the American Type Culture Collection, who wrote back that there certainly were some strange-looking chromosomes in these cells, but none of them were HeLa markers.

Two weeks before the annual meeting, at which Pathak was scheduled to discuss this cell line, Nelson- Rees received yet another manuscript, this one from Bailar, who asked him to review it for the Journal of the National Cancer Institute. It was a much more detailed report on the very same cell line. In addition to Pathak, Siciliano, and Cailleau, T. C. Hsu, head of the Tissue Culture Department at Anderson and described by some as the guru of cytogenetics, had put his name on the manuscript, adding considerable credibility to it.

What the earlier abstract had only implied, this twenty-five-page paper announced loud and clear: that the standard tools of HeLa hunting -- enzyme tests and chromosomal analysis -- were of questionable value. What's more, it suggested that every HeLa case Nelson-Rees had ever published was now in doubt. In short, it looked like the gang from Anderson was indicting the indicter himself.

"The HeLa markers have been extensively used by investigators to identify ... cell line contamination," the authors wrote, citing references to three of Nelson-Rees's published reports, including the latest hit list. Because they found chromosomes similar to HeLa markers in this tissue sample that was clearly not HeLa, they concluded that "marker chromosomes alone are not unequivocal evidence for identifying cellular contamination."

Wait a minute, thought Nelson-Rees. From what he and Flandermeyer and Lavappa could see, there were no HeLa markers in these cells. Sure, maybe there were a few that looked vaguely similar. Maybe someone with an untrained eye might be confused. But there was not a single marker chromosome in the photographs that a qualified karyologist should mistake for HeLa's. And even if there were, Walter Nelson-Rees would never label a cell culture HeLa based solely on marker chromosomes. There were also the enzymes to consider.

But the folks from Anderson observed that twelve out of fourteen isoenzymes tested in their breast cancer cell were identical to HeLa. "The suggestion, then, that cell lines showing only three enzyme characteristics in common with HeLa be considered de facto strains of HeLa may be overly simplistic," they wrote, offering footnotes to two more of Nelson-Rees's reports.

Overly simplistic{ Nelson-Rees couldn't believe this stuff. He'd never claimed that enzymes alone are enough. But if the marker chromosomes match and you pick the proper enzymes to check, then three can be plenty. The single finding of type A G6PD, for instance, in a culture thought to be from a white donor is more than enough to tell you something is awry.

Ah, but the "HeLalike" cells reported in this manuscript had come from a black woman who carried the A form of G6PD. How convenient, thought Nelson-Rees.

As he scanned the rest of the report, he noticed the authors neglected to point out that many of the other enzyme variants they tested were common to 90 percent of the human population, including the late Henrietta Lacks, and therefore of relatively little value in distinguishing their breast cancer cell from HeLa. And by a strange coincidence the two enzymes that turned out different from HeLa's happened to be the very last two they tested, even though one of them, called PGM3, had become a standard test for HeLa, and therefore likely to be among the first checked by anyone really interested in finding out if he had a HeLa cell or not.

The handling of the enzyme data was not the only thing Nelson-Rees found suspicious. He called Flandermeyer in and they pored over the chromosomal mug shots Pathak, Cailleau, and colleagues had used to prove their case. In an hour or so they were both shaking their heads, astonished.

The mug shots were photographic composites, constructed from bits and pieces of various photos of various cells in this breast cancer culture.

When you're comparing banded chromosomes from two different tissue samples -- a known HeLa culture, say, and one you suspect may be contaminated with HeLa -- there's only one correct way to do it, according to Nelson-Rees and most other karyologists. After searching hundreds of photos you begin to recognize the same complex of markers, the same group of defective chromosomes, appearing in every one of your suspect cells. You then pick a single cell in which this group displays its banding patterns clearly, cut out the defective chromosomes, and put them next to markers from a HeLa cell you've analyzed and photographed. In a given side-by-side comparison, the mug shots must come from only two individual cells. If you have to pick one chromosome from this suspect cell and another from that in order to find ones that match those from the HeLa cell, then you're reaching.

Apparently that's just what the Anderson workers had done. After a close look at all the photographs, Nelson- Rees and Flandermeyer were convinced that the comparison photo didn't display mug shots of chromosomes from only two cells, one from the breast culture and one from a HeLa line, but from at least seven, four different breast cells and three HeLas. What's more, the HeLa mug shots appeared to have been taken from another researcher's published report, not a culture the Anderson workers had studied themselves. In fact, they had cut out and reassembled so many mug shots in order to make their comparison look convincing they had even pasted in the image for one of the breast chromosomes upside down and reversed.

So this was what Pathak was going to unveil at the annual meeting. And as if that wasn't enough, the Anderson gang wanted to publish the entire argument in the Journal of the National Cancer Institute as well. Nelson- Rees was astonished that anyone would go to such lengths to discredit him.

For the next few days he drove Flandermeyer and the others hard. He demanded a detailed analysis of the mug shots showing that they weren't HeLa markers despite the photographic sleight-of-hand and offering alternative descriptions for each chromosome. When that was done, Nelson-Rees sat down, poison pen in hand, to draft a statement he planned to deliver at the annual meeting following Pathak's presentation. When it was done, he took it to Flandermeyer, who recommended toning it down. They removed a few of the more offensive passages, including a final statement Flandermeyer felt was particularly undiplomatic. Nelson-Rees then returned to his office and wrote the venom back in.
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Re: The Rapeutation of Walter Nelson-Rees, An Excerpt From "

Postby admin » Wed Mar 09, 2016 7:56 am

12: Showdown

As soon as Nelson-Rees arrived at the Denver Hilton Hotel, where members of the Tissue Culture Association were gathering for their twenty-ninth annual meeting, a friend came up and said, "The Anderson group is out to get you."

The following morning, the inflammable Relda Cailleau ignited at the mere mention of his name. She was at a meeting of the association's executive council, where she learned that Nelson-Rees had been elected vice-president of the association and was to be officially installed the next day -- and she couldn't contain the fire and fury.

"My god," she blurted out. "How could he have been elected vice-president?"

She told her startled fellow council members how Nelson-Rees had attacked her work and that of many well-respected scientists, how his methods were antagonistic and his motives destructive. "His approach is always, 'I've got to prove I'm right and you're wrong,'" she railed. "My God, the man is paranoid!"

Cailleau urged the council to demand his resignation, a difficult task as Nelson-Rees didn't yet hold the office. But she had made her point, several times over, and set the stage for the main event later in the day.

Nelson-Rees was set himself. Earlier he had told Bob Stevenson, "I'm really going to throw the fat on the fire." "Think about it," Stevenson answered.

Some 200 people packed into the Hilton's Vail Room that afternoon, considerably more than were originally expected for the session on mammary tissue. They had heard there was quite a show in store. Some said it might really get ugly. They came not necessarily to root for one side or the other, but -- like decent townsfolk gathering on rooftops and peering out of windows at a gunfight in the street -- just to watch.

The paper by Pathak, Cailleau, and Siciliano, entitled "Fresh Pleural Effusion from a Patient with Breast Cancer Showing Characteristic HeLa markers" was scheduled third. The audience sat restlessly through the first and second talks. Then, about thirty minutes into the program, the chairman read the title of the paper, and Pathak, the young Indian-born chromosome expert, came forward to deliver the talk. Cailleau was sitting in the audience about halfway to the back of the room. Nelson- Rees was about ten rows behind her. Siciliano had stayed home in Houston.

Pathak seemed apprehensive. He spoke a bit shakily in that British-educated, Indian accent, painfully formal and polite. Whenever he mentioned a colleague, it was always by title and full name, pronounced in one breath as if it were all a single word: "We examined chromosomes in cells obtained by DoctorReldaCailleau .... Isoenzymes from DoctorReldaCailleau's cells were analyzed by DoctorMichaelSiciliano."

Pathak described the findings in a straightforward way: These cells taken directly from a breast cancer patient showed chromosomes that were quite similar to HeLa's characteristic marker chromosomes, he said. And thirteen of fifteen isoenzymes checked were identical to HeLa's. (They had tested one additional enzyme since writing the draft report Nelson-Rees had seen.) There were no direct references to Nelson-Rees, no outright challenges to the usefulness of his techniques as there were in the written version of this talk that had been sent to the Journal of the National Cancer Institute. Still, to some members of the audience, especially to Nelson- Rees, the insinuation was clear.

When Pathak had finished, Nelson-Rees raised his hand. The session chairman, having heard that trouble was afoot, quickly called on someone else. Nelson-Rees decided he would not be denied again. When that question was answered, he simply stood up and without the aid of a microphone began to read his statement.

"As a referee of this abstract for the program committee, I questioned the use of the adjective 'characteristic' in the title, followed by 'similar' in the text. Dr. Pathak subsequently sent me a photograph of the 'marker' chromosomes stating he 'was puzzled regarding this observation.' I am puzzled as well, not with the observations but with the way they are being presented."

The chairman tried to cut him off, explaining that the presentation had gone over its allotted time.

"John," said Nelson-Rees as he moved from his seat up towards the front of the room, "I'm going to finish this."

He went on to explain that as a reviewer of the manuscript submitted to the cancer journal, he had looked over all the photographic evidence the authors used to prove their case. He said he discovered they had "fished about" in several different cells from this breast tumor to find their so-called HeLa chromosomes, a highly irregular procedure.

"We have carefully studied many different cell lines and observed a good number of marker chromosomes, but we've never had to create a collage such as has been done here in order to sensationalize a point."

The audience was aghast at such a flagrant breach of etiquette. One simply didn't use such language. Perhaps a point would need to be confirmed; maybe there was a chance for an alternative interpretation. But publicly accusing learned colleagues of fishing about for data and sensationalizing their results -- Nelson-Rees might just as well have been spoon-launching mashed potatoes at a formal dinner.

"As a matter of fact," Nelson-Rees proclaimed, "without stating so and without giving credit to the published work, the authors lifted HeLa marker chromosomes from the work of Heneen which best matched their altered chromosomes and presented them in the manuscript apparently as their own observations." He held up the Swedish journal Hereditas that contained the report he claimed they had "lifted" from and added that despite such pilferage, the chromosomes they presented are not convincing matches.

There was a sudden disturbance in the audience. Relda Cailleau was becoming agitated, muttering loudly to the people around her, but Nelson-Rees pressed on.

"Aside from the above-mentioned irregularity in scientific procedure, it appears that some normal chromosomes were erroneously classified as markers and vice versa, and at least in one instance a chromosome's image was used twice, albeit after reversal of the negative."

Nelson-Rees explained that he had sent photographs of the "so-called characteristic" HeLa markers to Lavappa for a second opinion and quoted Lavappa's written reply: "'There is no trace of the four distinct HeLa markers, which are well characterized and have appeared in many publications.'" And then it was time for the finale, the part that he had typed out in capital letters.

"I suggest you stick to pleural effusions," he said to a dumbstruck Pathak at the podium, "and leave the HeLas to us."

A momentary silence.

Pathak broke the trance and added a touch of absurdity to the scene by thanking Nelson-Rees profusely. Nelson-Rees dropped 150 copies of his statement in a pile at the back of the room and left. Suddenly pandemonium broke out over the Vail Room. There were five more speakers scheduled for the afternoon, but half the audience was rushing for the exits.

Relda Cailleau made her way over to the stack of statements, grabbed about thirty of them, and headed out. In the hallway she worked the crowd, offering copies of Nelson-Rees's attack as proof the man had no business reviewing manuscripts for technical journals (Just look at how he violated the reviewers' oath of confidentiality!) or serving as vice-president of the Tissue Culture Association (Was such rude, unethical behavior fitting for an officer of our group?).

Pathak returned to his hotel room, where he remained for most of the last two days of the conference. He would later explain his absence by saying he was deeply shaken by the wrongful accusation and public humiliation he had suffered. In fact, he was so distraught that he refused to do any research on the chromosomes of tumor cells for an entire year. Long after the incident, Nelson-Rees offered an alternative explanation: "A little dog -- or even a big one -- who has peed on the living room rug and is found out always hides underneath the couch for a couple of hours."

At the moment, however, Nelson-Rees was fresh out of quips. His feistiness was suddenly spent. He was exhausted and, he noticed, quite alone. As he moved through meetings the rest of the day, people avoided him, looked away disapprovingly when he approached, and whispered about what a disgraceful performance he had given.

One of those who attended his performance and had been especially shocked was Colonel Albert Leibovitz. A tissue culturist formerly with the U.S. Army and now at the Scott and White Clinic in Temple, Texas, Leibovitz had at one time respected Nelson-Rees's crusade to clean up the field, or so he said. Recently, though, it looked to Leibovitz like Nelson-Rees was on a maniacal rampage. His means were extreme. And what he had done this afternoon could only be described as scandalous. "It is so unethical to be a reviewer of a paper and use confidential information to tear its authors down," Leibovitz declared.

Leibovitz, of course, may have felt some special sympathy for the Anderson crew, having had a recent tussle with Nelson-Rees himself. The matter involved eight cell lines Leibovitz had established: colon, bladder, and six other kinds of cancer, all of which had apparently been taken over by a single line of colon cancer cells. Although it was not a case of HeLa contamination it was still a large mix-up, and Nelson-Rees felt it ought to be publicized immediately. When Leibovitz balked, preferring to double-check the findings and investigate the source of the contamination further, Nelson-Rees tried to publish a letter in the Journal of the National Cancer Institute, warning other scientists away from these cells. Leibovitz was furious. Certainly Nelson-Rees had been right to want to get the word out, but he was jumping the gun for no reason, Leibovitz believed, and worse, he had broken an implicit confidence -- just as he had in attacking the report by the Anderson group.

Later that afternoon at the general membership meeting of the Tissue Culture Association, it was Colonel Leibovitz who stood up and with great indignation urged that the association create an ethics committee to deal with one of its members who, at a presentation earlier in the day, "committed unscientific acts without presenting proof to support his statements." This person's verbal assault was all the more inappropriate, the colonel explained, because he had been the confidential reviewer of a manuscript covering this particular presentation.

Seated out in the audience, Nelson-Rees kept a faint, nervous smile on his face as the members discussed the need to discipline the unnamed outlaw and eventually voted to have Leibovitz draft a resolution for consideration at the next meeting of the executive board.

That night Bob Stevenson took Nelson-Rees out for a drink and a little lecture. It was another version of the friendly advice Stevenson had regularly offered: Diplomacy gets you farther. When you do something, you do it properly, or you risk losing credibility.

On the other hand, Stevenson added, he was only too glad somebody was stirring things up. He grinned his mischievous grin and said, "It's always nice to drop a turd in the punchbowl once in a while."

***

Then came the letters. In a few weeks Keith Porter, the president of the association, was hip deep in testimonials of outrage, shock, and dismay:

I am writing to express my concern about the incident that took place in Denver. Nelson-Rees has abused his privilege as a reviewer and impugned the integrity of Ors. Pathak et a1. I hope the Executive Board will treat this matter with the utmost seriousness, because I believe the credibility of the Board, as well as the good reputations of Ors. Pathak, Siciliano, and Cailleau are at stake.

I am but one of many who was shocked and offended by the serious and unethical activities of Walter Nelson-Rees.

Dr. Nelson-Rees behaved ungentlemanly and indiscreetly. He appeared to lack the objectivity and open-mindedness of character important for a good scientist.

Leibovitz's resolution also arrived, four pages documenting Nelson-Rees's crimes. "It is immaterial whether his findings are right or wrong," the statement read and urged that he be censured, stripped of his vice-presidency, and banned from reviewing manuscripts for the association.

It was all very well, all this righteous wrath, but as far as Porter could see, there was no way to satisfy it. Nelson-Rees was guilty of no malfeasance, and even if he were, he was not in office when he so distressed everyone; that was the day before he was officially installed. When the officers, including Nelson-Rees, met a few months later, Nelson-Rees agreed to apologize to Pathak in writing, but only for the manner in which he "disagreed" with Pathak's report. The president hoped that would satisfy the offended.

It didn't come close to satisfying Relda Cailleau, who wrote the president four months after the incident:

Dear Dr. Porter,

If he [Nelson-Rees] and other members of the Council consider that his apology for having "offended" Dr. Pathak is enough to close the subject, they are mistaken. His unethical conduct is not mentioned and it is the only subject of importance for the T.C.A. and for any reputable scientist .... If he cannot be removed from office or forced to resign because the T.C.A. constitution has no provision for such a contingency, he must at least be made to acknowledge his unethical conduct (which incidently is not limited to this occurrence) .... He should no longer be allowed to review articles without some safeguard to the people whose reputations he tries to destroy. The only way this can be done is to inform the membership as well as the editors of the various journals in which his comments have appeared.


Porter answered:

Dear Relda:

... Unofficially, I would urge you to forget the incident. I have heard some very abusive attacks of one scientist on another at scientific meetings and they did not become the business of the society or institution sponsoring the meeting to settle. The TCA cannot undertake to censure, punish or even reprimand its members for "unethical" behavior. As I have said before, Walter was duly elected and cannot be removed from office without going back to the membership of the Association. I do not think that you could influence the required plurality.


What amazed Nelson-Rees was that nowhere amid the outcry over his Denver recital was there any discussion of the merits of his argument, no effort made to determine whether the results reported by Pathak and company were trumped up, as Nelson-Rees had charged. They had claimed that his methods for fingering a cell line as HeLa were in doubt and, by extension, his hit lists and other reports were wrong. Well, Nelson-Rees couldn't stand being told he was wrong. And it incensed him that no one cared to consider the substance of his rebuttal.

After the Denver meeting, he sent off a stinging critique to Bailar at the Journal of the National Cancer Institute, restating and amplifying his argument. The Pathak report, he wrote, should not be published unless its" entire concept is altered." He questioned its" sincerity and merit" and accused the authors of resorting to "photographic trickery" in their presentation of the chromosome evidence. As for the enzyme data, he wrote, it may well be true that thirteen out of fifteen enzymes tested were the same as HeLa's; but two of them, the two most useful for identifying HeLa cells, were different. Hence the system holds up. No one would mistake these cells for HeLa, as Pathak and Cailleau's gang had claimed.

Like his speech in Denver, however, these criticisms were ignored. The paper, barely revised from the original version, was published in February 1979.

***

When they were asked about it years later, the Anderson researchers insisted they had never set out to discredit Nelson-Rees's years of crusading, as he had somehow imagined. It had all begun as a simple study of chromosomes in fresh tissue samples. Pathak wanted to find out if the defects observed in chromosomes of cultured cells resulted somehow from extended growth in a laboratory dish. Or did cancer cells taken fresh from a patient show abnormal chromosomes as well? Cailleau had been supplying him with fresh pleural effusions. And one day they simply stumbled across this unusual bunch of cells containing chromosomes similar to HeLa markers.

T.C. Hsu, the senior cytogeneticist in the lab, thought the finding raised an important point in the current climate of HeLaphobia. In most cases, said Hsu, perhaps 98 percent of them, a cell that has HeLa markers is probably HeLa. But these markers may not be unique to HeLaj they may be created by some general mishap of cancer. So there may be a few cases in which a cell carries HeLalike chromosomes and yet is something quite different.

"The findings so shocked Nelson-Rees, he thought it rocked the foundations of all his work," explained Hsu. "But it didn't really rock his foundations."

"We were simply urging caution when diagnosing cells as HeLa," added Pathak, "not disagreeing with all his results."

Still, in their published report, and in follow-up articles and letters to certain colleagues, Hsu, Pathak, and Cailleau repeatedly warned against relying on chromosomes alone, referring to Nelson-Rees by name and suggesting that he had foolishly done just that -- when he never had.

As for their need to create a collage, breaking with the traditional means of comparing chromosomes in cell lines, they explained that fresh tissue samples are always a mess, carrying all kinds of bodily by-products that are eventually filtered out of a long-term cell culture. So in a fresh pleural effusion, it is difficult to take clear photographs of all the marker chromosomes within a single cell.

"The composite was prepared in order to show an example that was clear-cut to the untrained eye," said Cailleau.

As Flandermeyer often pointed out, though, the identification of chromosomes entails a great deal of judgment. "There's an art to it, and if you're not careful you can incorporate your bias," he once said, thinking of the Denver affair. "You can make things conform to what you expect, especially if you pick around, choosing one chromosome from one cell and another from the next."

The presence of specific enzymes inside a cell is much less ambiguous, and therefore far more resistant, to an investigator's bias. On the other hand, the choice of which enzymes to look for and the order in which they are checked can create certain impressions.

Stephen O'Brien, an enzymologist at the National Cancer Institute and an ally of Nelson-Rees, was puzzled by the Anderson group's need to check fifteen enzymes in their breast cells before finding two that differed from HeLa's. Had he been doing the experiment, O'Brien said after reading the report, he would have analyzed perhaps six, and the two mismatches would have been among the first.

But according to Siciliano, the enzyme expert who collaborated with Hsu, Pathak, and Cailleau, the fifteen enzymes they checked were the standards, the ones they checked routinely. It just so happened the two that turned out to be different from HeLa's were tested last.

Like his colleagues, Siciliano insisted the report was not intended to discredit Nelson-Rees or to knock the foundation out from under his published findings. "I don't think there was any stuff in his published papers that's refutable," he said.

Why, then, the comment that Nelson-Rees's use of only three enzymes is overly simplistic?

"Most of the cells on his lists came from white donors," explained Siciliano, "so testing for only one enzyme, G6PD, was enough. But I was concerned for the future as more blacks donate tissue cultures, as they're doing in institutions like ours."

***

Motives, alas, are slippery fish. It is difficult for anyone to know what the Denver blowup was really all about. Anyone but Nelson-Rees, that is, to whom the truth was obvious.

"This group of people was trying to destroy years of careful work and promotion of the use of good cell lines, " he said after the incident. "It was just a case of this being put together to cause embarrassment to me.

"You know, I didn't 'impugn their reputations,' as some said of my rebuttal at Denver. I called them absolute shits. They were the ones who were unscientific and unethical, not me."
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