The Cytopathic Effect

in cytopathiceffect •  2 years ago 

COVID-19 and Ireland – Part 24

~Part 1~

As we have seen, the standard way of “isolating” viruses is to grow them in tissue cultures and observe the phenomenon known as the cytopathic effect, or CPE—biochemical and morphological changes that occur in host cells when they are infected by pathogenic viruses. The commonest cytopathic effect is lysis, the breaking down of the cell membrane. An infected cell may also die without experiencing lysis due to its inability to reproduce by cell division. Two common manifestations of the CPE are:

  • The appearance of large spherical syncytia
  • The formation of plaques

A syncytium is a mass of cytoplasm containing many nuclei, or a multinucleate cell resulting from the fusion of several mononuclear cells.

A viral plaque is a patch of abnormal tissue distinguishable from surrounding tissue, which is formed when a cell culture grown on some nutrient medium is infected with a sample of a virus. The virus replicates and spreads, generating regions of cell destruction known as plaques.

The Cytopathic Effect

In order to eliminate the possibility that the results of an experiment are due to confounding variables rather than the independent variable under study, scientific controls are used. Controls are a necessary part of the scientific method. Many peer-reviewed papers that describe the “isolation” of a virus by observing the CPE mention mock-infected controls: a tissue culture that does not contain the alleged virus and in which the CPE is not observed.

Opponents of the conventional viral theory argue that these mock-infected controls are not proper controls, and that most peer-reviewed papers that mention them at all never provide enough detail to allow other researchers to replicate the alleged findings. There is, furthermore, a suspicion that the CPE is being caused by the way in which the tissue culture is treated and not by the presence of any virus:

The only way it would possibly be a proper control is if the virologists started with only the purified/isolated particles assumed to be the “virus” as the independent variable used for experimentation. As they do not do so, the unpurified human sample already has many host and foreign materials present along with many others added to it which need to be factored into the equation as potential causes of any observed effect. Thus, using only a cell without a human sample is not a proper control when an unpurified sample without isolated particles is utilized to try and prove cause and effect. In order to have a proper control, the virologists would need to use samples from both healthy humans and those who are sick with similar symptoms but said not to have the assumed “virus.” (Mike Stone, ViroLIEgy)

Stefan Lanka

Stefan Lanka

German virologist—or ex-virologist, as he now refers to himself—Stefan Lanka claims to have carried out the correct control experiments:

Virologists primarily believe in the existence of viruses, because they add allegedly “infected” blood, saliva or other body fluids to the tissue and cell culture, and this, it must be stressed, after having withdrawn the nutrients from the respective cell culture and after having started poisoning it with toxic antibiotics. They believe that the cell culture is then killed by viruses. The key insight, however, is that the death of the tissue and cells takes place in the exact same manner when no “infected” genetic material is added at all. The virologists have apparently not noticed this fact! According to the most basic scientific logic and the rules of scientific conduct, control experiments should have been carried out. In order to confirm the newly discovered method of so-called “virus propagation”, in order to see whether it was not the method itself causing or falsifying the result, the scientists would have had to perform additional experiments, called negative control experiments, in which they would add sterile substances or substances from healthy people and animals to the cell culture. This, of course, to check whether it is not the method itself that yields or falsifies the results.

These control experiments have never been carried out by the official “science” to this day. During the measles virus trial, I commissioned an independent laboratory to perform these control experiments and the result was that the tissues and cells die, due to the laboratory conditions, in the exact same way as when they come into contact with allegedly “infected” material.

The entire purpose of control experiments is to exclude the possibility that it is the applied method or technique which may cause the result. Control experiments, then, are the highest duty in science and also the exclusive basis of claiming that one’s conclusion is scientific. During the measles virus trial it was the legally appointed expert—Dr. Podbielski, see further in this article—who stated that the papers which are crucial for the entire science of virology contain no control experiments. From this we can conclude that the respective scientists have been working extremely unscientifically, and this without even noticing it. (Lanka 1:3)

Andreas Podbielski

Lanka believes that the antibiotics added to the tissue cultures are partly responsible for the cytopathic effect:

Virology claims to isolate viruses in the laboratory and from claimed isolated particles, claims to find the genetic material to determine their structure. In no publication claiming an isolation of a virus is there a description of an actual structure that has been isolated. on the contrary, experimentally produced death of tissues in the laboratory is misinterpreted as the effect of viruses because it is assumed that the tissues would die because supposedly infected body fluids are added. In reality, the tissues die because they are no longer nourished and are killed by toxic antibiotics. (Lanka 3:4)

Pen-Strep, a mixture of two common antibiotics, penicillin G and streptomycin, is widely used in mammalian cell culture media to prevent bacterial contamination.

I don’t understand why Lanka also implicates malnourishment as a factor in the CPE. Tissue cultures are grown in a medium that is supposed to provide the cells with all the nourishment they need. In an earlier article we saw that popular choices are Eagle’s Minimum Essential Medium and Dulbecco’s Modified Eagle Medium:

Minimal Essential Medium (MEM) is a synthetic cell culture medium developed by Harry Eagle first published in 1959 in Science that can be used to maintain cells in tissue culture. It is based on 6 salts and glucose described in Earle’s salts in 1934 (calcium chloride, potassium chloride, magnesium sulfate, sodium chloride, sodium phosphate and sodium bicarbonate), supplemented with 13 essential amino acids, and 8 vitamins: thiamine (vitamin B1), riboflavin (vitamin B2), nicotinamide (vitamin B3), pantothenic acid (vitamin B5), pyrodoxine (vitamin B6), folic acid (vitamin B9), choline, and myoinositol (originally known as vitamin B8). (Eagle’s Minimal Essential Medium)

Harry Eagle

In 2020, during the Covid-19 Pandemic, Stefan Lanka decided to carry out for “SARS-CoV-2” the control experiments which he claims have never been properly conducted by virologists. These experiments were conducted in three phases. In Phase 1, Lanka claims to have shown that the cytopathic effect occurs even in the absence of any virus-infected material. Lanka’s paper describing these experiments has not yet been published, so far as I can tell:

  • Stefan Lanka, Preliminary Results: Response of Primary Human Epithelial Cells to Stringent Virus Amplification Protocols, April 2021 (unpublished)

Lanka’s colleague Tom Cowan, however, describe the results of these experiments in his book Breaking the Spell:

Here is the essence of Lanka’s experiment, done by an independent professional laboratory that specializes in cell culturing. As seen in this series of photographs, each of the four vertical columns is a separate experiment. The top photo in each column was taken on day one, and the bottom photo was taken on day five.

In vertical column one, normal cells were cultured with normal nutrient medium and only a small amount of antibiotics. As you can see, on neither day one nor day five was any CPE found; the cells continued their normal, healthy growth.

Lanka’s Control Experiments and the Cytopathic Effect

In vertical column two, normal cells were again grown on normal nutrient medium and a small amount of antibiotics, but this time, 10% fetal calf serum was added to enrich the medium. Still, the cells in the culture grew normally, both on day one and day five.

The third vertical column shows what happened when Dr. Lanka’s group used the same procedures that have been used in every modern isolation experiment of every pathogenic virus that I have seen. This included changing the nutrient medium to “minimal nutrient medium”—meaning lowering the percentage of fetal calf serum from the usual 10% to 1%, which lowers the nutrients available for the cells to grow, thereby stressing them—and tripling the antibiotic concentration. As you can see, on day five of the experiment, the characteristic CPE occurred, “proving” the existence and pathogenicity of the virus— except, at no point was a pathogenic virus added to the culture. This outcome can only mean that the CPE was a result of the way the culture experiment was done and not from any virus.

The fourth and final vertical column is the same as vertical column three, except that to this culture, a solution of pure RNA from yeast was added. This produced the same result as column three, again proving that it is the culture technique—and not a virus—that is causing the CPE.
(Cowan 13-4)

The fourth column is not particularly relevant to the question under discussion. It was included as a control for the method used by virologists to derive the genome of a virus—a subject for another article.

In the following video—with English dubbing and Italian subtitles—Stefan Lanka briefly describes the results of these control experiments as far as the CPE is concerned (the entire interview in German may be watched on Odysee:

The Empire Strikes Back

For the most part, mainstream academia does not attempt to refute counterarguments to any of its received orthodox teachings. Researchers like Stefan Lanka or Andrew Kaufman are simply ignored and never mentioned. But one scholar at least has taken up the challenge and offered a rebuttal to Lanka’s control experiments. Frank Visser is a psychologist of culture and religion, who founded IntegralWorld in 1997. He has worked as production manager for various publishing houses and as service manager for various Internet companies. He lives in Amsterdam. On his twitter account, he is described as a sailing instructor, a skeptic, and a blogger.

The title of his article, Stefan Lanka’s Counterfeit “Control Experiments, and his description of Lanka as a virus denialist tells one clearly where he stands on the question. Denialist is a nonsensical term used to imply that the person so described is a crank, a “conspiracy theorist”. Elsewhere, Visser refers to other skeptics of mainstream virology as The Silly Sixteen. No self-respecting scientist should need to indulge in ad hominem attacks of this sort if he truly believes that he has won the argument.

Visser begins his rebuttal by misrepresenting the situation:

Frank Visser

Stefan Lanka, the Godfather of Virus Denialism ... has announced two experiments that, in his opinion, are “going to be the definitively experimental disproval of virology.” For decades he has claimed that viruses don’t exist, and virology is one huge mistake ... But he did not provide any empirical proof for his extravagant opinions. Claiming something is not the same as empirically demonstrating it. This time, he seems to have understood this is paramount to make any impression on the scientific community.

Is this true? In 2015-2016, according to at least one online source, during the celebrated Barden-versus-Lanka measles trial, Lanka commissioned an independent German laboratory to carry out control experiments for the alleged isolation of the measles virus. But I have not been able to confirm this. Nevertheless, Visser has failed to realise that the burden of proof in such a case lies with those who allege the existence of the thing, not with those who dispute its existence. One cannot claim that the Flying Spaghetti Monster exists unless someone proves that it does not exist. In science, Occam’s Razor tells us not to posit the existence of something until its existence has been proved with scientific rigor.

After much irrelevant criticism of Lanka and his Projekt Immanuel, Visser finally gets to the control experiments. To be fair to Visser, Lanka’s paper on these experiments has not yet been published, which makes it all but impossible for anyone—for or against Lanka—to properly assess them. Nevertheless, Visser believes that he can show that Lanka’s claims are spurious. His first argument is a sort of tu quoque. He accuses Lanka of failing to include proper controls:

Tu Quoque

His suggestion is that the so-called cytopathic effect (CPE) or cell damage usually said to prove the existence and impact of viruses is actually an artifact caused by the chemicals that are added to the viral cell cultures. Cell cultures without virus show the same damage—or so he claims.

The picture he shows in this short video displays damage done to healthy cells over 1 and 5 days, with different chemicals added to the mix (antibiotics, nutrients and yeast RNA). The same that are added to virus-infected cells. Ergo: there is no evidence for a virus.

This presentation is misleading for he should have given also pictures of SARS-CoV-2 virus infected cells as a positive control (but most probably he does not have access to a lab that has the proper safety level to work and experiment with this virus).

This is a fair comment. As far as I can tell, Lanka’s experiments lack the required controls. But is this enough to refute Lanka’s conclusions? I think not. There are plenty of peer-reviewed papers that do contain such images. Still, it is a fundamental mistake on Lanka’s part.

Visser proceeds to his second argument:

Secondly, he erroneously suggests that in regular control experiments these added chemicals are not present. This is most definitely not the case: mock-infected cells get the same treatment as virus-infected cells, EXCEPT for the virus. You won’t have to take my word for it.

MOCK-INFECTED: “Two specimens are used one that is infected with the virus/vector of interest and the other is treated the same way except without the virus. Sometimes a non-virulent strain is used in the mock-infected specimen.” [Molecular Biology Glossary, www.genscript.com]

The key sentence here is: mock-infected controls are “treated the same way” as those infected with viruses. And yet they show clear differences in all the papers I have consulted presenting CPE effects caused by SARS-CoV-2.

Positive and Negative Controls

This is, again, a fair point—if it is true. It is very difficult to confirm this, however, as most peer-reviewed papers that mention mock-infected controls don’t give any details of how they were treated—meaning that it is impossible for independent researchers to replicate their findings. Note, in passing, how the definition quoted by Visser assumes that the first specimen is “infected with the virus/vector of interest”. But aren’t we trying to prove that very point?

In response to Visser’s criticism, Lanka might argue: I treated my mock-infected controls exactly the same way that infected samples are treated when viruses are being isolated, but I still observed the CPE. To this, Visser’s reply shocked even me:

But here’s the catch: these mock-infected cells also (often, but not always) show cell damage over time, like the virus-infected cells do, but different both in quality and quantity. So it requires a trained eye to spot the difference. Just showing a couple of cells with some damage won’t do here. [Sébastien Eymieux et al]

The acknowledgment that mock-infected cells can experience the CPE should give us pause. The CPE, after all, is the very thing which, it is claimed, proves the presence of a pathogenic virus. Now we require trained eyes to tell the difference between the CPE caused by a virus and the CPE caused by something else? How were these trained eyes trained in the first case? How did John Franklin Enders, the biochemist who established this method of isolating viruses in 1954, know that the CPE he observed was caused by the measles virus?

John Franklin Enders

The historical sequence of events seems to be:

  • Enders observes the CPE in tissue cultures infected with measles. He concludes that a pathogenic virus is causing the CPE, thereby “isolating” the measles virus. He does not perform any negative controls.

  • Other researchers apply Enders’ technique to “isolate” other viruses. Sometimes they carry out mock-infected controls, observing no CPE, thereby confirming the CPE they observed in the infected samples was caused by the pathogenic virus.

  • Other researchers apply Enders’ technique to “isolate” other viruses. They carry out mock-infected controls, but observe the CPE. They conclude that the CPE can also be caused by something other than a pathogenic virus. But, don’t worry, they can tell the difference between the two.

In the last article we saw how the peer-reviewed literature on the isolation of viruses is replete with papers in which the authors have misidentified nonviral subcellular bodies with viruses. Are these the same experts whose eyes are trained to tell the difference between the two types of CPE?

Mike Stone

Visser’s third criticism is one that I have already raised myself:

And what about the claim, made by both Lanka and Cowan (see Part 27), that both mock- and virus-infected cells are starved to the point of dying? Says a random page on viral culture (Simulab):

“Cell culture media often includes a range of salts, vitamins, hormones and other growth factors, amino acids or proteins, glucose, antimicrobial agents, a buffering system, a pH indicator, and non-specific sources of nutrients such as foetal bovine serum.”

Doesn’t really look much like a starvation diet to me. These are all nutrients.

This is fair comment. I have little so say in support of Lanka and Cowan on this point. But I do wonder why the bovine foetal serum is added in the first place. Eagle’s Minimum Essential Medium was designed as sufficient nourishment for cells. That’s what the words Minimum Essential mean. So why the bovine foetal serum?

Mike Stone of ViroLIEgy has this to say on the matter of cell nourishment:

It is clear that the media consists of many different compounds said to keep the cell alive and aid in the growth of any “viruses” present. However, there are many issues related to the cell culture media that can influence the outcome of the culture and the results obtained. These include: the unknown and variable composition of the vitamins/micronutrients within the media, the unknown interactions between the various components in the media and the stability of the cell, the inclusion of antibiotics and fetal bovine serum in many media, etc. (ViroLIEgy)

Pen-Strep Chemical Formulas

Visser also takes issue with Lanka’s claim that the antibiotics added to the tissue culture are causing the CPE:

And antibiotics? These are not added to kill cells, but bacteria that might proliferate as well in the cell cultures.

Well, that may be why they are added, but once again the onus is on the mainstream virologists to prove that these antibiotics themselves are not causing the CPE. This they have never done, let alone investigated, so far as I know.

In Conclusion

Those of us who just want to know whether pathogenic viruses exist or not are left in limbo:

  • Stefan Lanka tells us that tissue cultures uninfected with any viral matter experience the cytopathic effect.

  • Some virologists tell us that their mock-infected tissue cultures experienced no cytopathic effect even though they were treated the same as the infected cultures, which did experience the CPE.

  • Other virologists tell us that their mock-infected tissue cultures did experience the cytopathic effect, but they have trained eyes that allow them to distinguish this nonviral-induced CPE from viral CPE.

This is a wholly unsatisfactory situation. It actually makes no sense.

Frank Visser’s Parting Shot

Finally, I was pleasantly surprised by the comments section of Visser’s rebuttal. I expected a litany of ad hominem attacks on Lanka by Visser’s subscribers, but most of the comments are sympathetic to Lanka and what he is trying to do.

And that’s a good place to stop.


References

  • Stefan Lanka, The Virus Misconception: Part 1: Measles as an Example, WissenschafftPlus Magazin, Issue 01/2020, LK-Verlags UG, Werder (Havel) (2020)
  • Stefan Lanka, The Virus Misconception: Part 2: The Beginning and the End of the Corona Crisis, WissenschafftPlus Magazin, Issue 02/2020, LK-Verlags UG, Werder (Havel) (2020)
  • Stefan Lanka, _The Virus Misconception: Part 3: _, WissenschafftPlus Magazin, Issue 03/2020, LK-Verlags UG, Werder (Havel) (2020)

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