PURITY: Relative freedom from extraneous matter in the finished product, whether or not harmful to the recipient or deleterious to the product. (21 CFR 600.3(r))FDA Guidance Definition for vaccine purity
The above definition for purity is taken from the FDA’s Guidance for Industry. According to the FDA, purity is considered relative freedom from extraneous matter. What is “relative freedom?” Relative implies that something is true to a certain degree. In other words, there is freedom from extraneous matter to a certain degree. However, in relation to purity, there can be no degrees of freedom from extraneous matter. A substance is either pure, meaning that it is free of all contaminants and foreign matter, or it is impure, meaning it is mixed with contamination and foreign matter. There is no such thing as purity to a certain extent yet you will commonly find discussions of purity within the “scientific” papers as if this can be the case. Terminology such as low purity, partially purified, somewhat purified, relatively purified, highly purified, etc. are thrown about as vague descriptors used to wiggle around a concept that is so simple and straight forward that most children can understand it. Something is either pure or impure. There is no in-between.
The issue of purification is absolutely vital to the foundational problems regarding virology. In order to prove the very existence of the fictional creations we are supposed to fear, it must be shown that “viruses” exist in a purified and isolated state. However, purification is mostly ignored in any original “virus” study when it should be a prominent focus. If we are to believe that these invisible intracellular parasitic entities exist, we need to be able to observe them alone. As we can not observe “viruses” in nature nor with our own eyes, this means that it is essential that the particles assumed to be “viruses” are taken directly from the fluids of a sick human and then purified free of any contaminants, pollutants, foreign materials, etc. contained within so that nothing else remains other than the assumed “viral” particles. Only then could pathogenicity be proven by having a valid independent variable separated from everything else in order to establish a cause-and-effect relationship for the particles claimed as the causative agent. Only then would electron microscope images of just those particles with nothing else contained within the sample be valid as evidence.
However, as you will see, virologists have admitted numerous times that “viruses” can not be purified and isolated directly from the fluids of a sick human. They state that “viruses” must be cultured with a host cell first before purification and isolation can occur. This creates many problems as:
- The methods used to purify and isolate “viruses” are ineffective
- “Viruses” can not be separated from exosomes and other MVB’s within the sample
- The cell culture method is impure and the media and host cell DNA can not be purified away from the invisible “viruses”
In this article, I will break these various problems down and show that, from their own sources, virology admits that complete purification and isolation of the assumed “viral” particles is an impossibilty.
A big hindrance to obtaining evidence of purification of the particles assumed to be “viruses” is that there is no standardized universal purification method. The techniques used vary by “virus” and the researchers performing the study. What “works” for one “virus” may not work for another as noted in this source on purifying plant “viruses:”
“Purification refers to the separation of virus particles from host components in a biologically active state. Purified virus is required for the production of antibodies, physical, biochemical and molecular characterization of virus isolates. Purification of virus involves several steps such as propagation of the virus in the host, extraction of sap, clarification, concentration and further purification. Purity of purified preparation can be checked through UV absorption spectra and its infectivity by inoculating to a susceptible host under optimal environmental conditions in an insect-proof glasshouse. Purification methods vary with different viruses, and there are no universal methods of virus purification. Procedures that are effective for one virus may not work with the other.”
Complicating the issues related to the lack of any standardized methods or procedure to achieve purification is the existence of various methods virologists can choose from in order to attempt to obtain purification. These include but are not limited to:
However, you will be hard pressed to find much mention of any of these methods being used in “virus” studies at all. If you do happen to find purification methods being performed, it is typically just a spin on the ol’ centrifugation for a bit. While this may separate some of the larger substances from the smaller ones, it does not offer complete purification of all particles.
“Viruses”/Exosomes: Inseparable “Cousins”
How can we be sure that we are isolating and quantifying extracellular vesicles rather than enveloped viruses present in the sample? Equally, how can viral researchers know that they are not detecting similarly sized non-viral vesicles or empty vectors during vaccine production?”https://www.google.com/amp/s/www.nanoviewbio.com/exosome-blog/2020/5/5/extracellular-vesicles-and-viruses-two-sides-of-the-same-coin%3fformat=amp
We can easily find this lack of complete purification to be the case for all of the methods used for “viruses” if we look to exosome research as it shares the exact same procedures as those used in virology. For those who may not know, exosomes are “viruses” in every sense of the word. They share the same biochemical and morphological characteristics. The main difference (beyond the name) is that exosomes are considered “non-infectious” and they are assumed to play a role in intracellular communication. In other words, they are the exact same particles as “viruses” but they were given different names and different functions than their “viral” cousins. Fortunately, as I noted earlier, the same methods of purification are used for these identical particles. If the purification methods are unable to completely purify exosomes, the same can be said of “viruses.” For instance, in this 2020 study on the attempts to separate “viruses” from exosomes, we can see that ultracentrifugation, the “gold standard” method of purification for both entities, is considered inadequate:
“The method is not ideally suited for the separation of EV and viruses, particularly from infected cultures and/or body fluids and it is difficult to adopt ultracentrifugation to large-scale production environments or to environments that present biosafety concerns.”
This inability to purify exosomes from “viruses” does not stop with ultracentrifugation. No matter what method is utilized, each one has its own drawbacks and limitations which lead to a lack of complete purification and the inability to separate “viruses” from exosomes. These next two sources show that the same methods shared between “viruses” and exosomes are all equally unable to successfully purify and isolate these particles from each other:
Exosomes and viruses
“Because of their similar physical properties, the techniques used by researchers for purification of exosomes and virions are also similar.
Ultracentrifugation at greater than 100,000 x g is used to concentrate both exosomes and viruses. This technique retains intact, functional viruses and exosomes. Because exosomes and viruses are similar in size and density, separation of the two is not possible using this technique.
Polyethylene glycol (PEG) can be used to precipitate both exosomes and viral particles. PEG precipitation is commonly used in commercially available exosome isolation kits and has been harnessed for isolation of many different viruses for many years. Again, you cannot separate viruses and exosomes with this protocol.
Exosomes and viruses can also be purified using chromatography techniques based on size, affinity, hydrophobicity, or other characteristics. Size exclusion chromatography (SEC) purification such as Exo-spin can be used to isolate both exosomes and viral particles. Again, exosomes and virions cannot be distinguished from one another using this method.”
Review on Strategies and Technologies for Exosome Isolation and Purification
“A method that can efficiently provide intact and pure exosomes samples is the first step to both exosome-based liquid biopsies and therapeutics. Unfortunately, common exosomal separation techniques suffer from operation complexity, time consumption, large sample volumes and low purity, posing significant challenges for exosomal downstream analysis.”
“At present, common exosome isolation technologies, such as ultrafiltration, immunoaffinity, ultracentrifugation (“gold standard” for the isolation of exosomes) are expensive instruments, large volumes of sample, possible protein contamination, complete isolation steps, but they result in low isolation efficiency, sample loss, low exosome recovery and purity (LeBleu and Kalluri, 2020).”
“Although exosomes play an irreplaceable role in early detection and treatment, they are small in size (30–150 nm), low in density (1.13–1.19 g/ml), and mixed with similar components (e.g., cell fragments, proteins) in the body fluids, which pose tremendous challenges for their separation (Cui et al., 2018; Lin et al., 2020). In addition, the biological activity of exosomes will be affected by different separation techniques (Paolini et al., 2016).”
“Although the above-mentioned traditional separation methods are the most widely used, there are also many disadvantages, such as large sample consumption, risk of damage to exosomes, low purity, and long time consuming, which are hard to meet the current increasing scientific research needs.”
As can be seen, the methods listed above do not offer the ability to actually separate the particles believed to be exosomes from those believed to be “viruses.” This is a major problem as purification is absolutely necessary not only in order to prove cause and effect but also to characterize the “virus” physically, molecularly, and biochemically. Without having just the assumed “viral” particles alone, the “virus” can not be studied independently nor differentiated from the millions of similar and/or identical particles within the sample. There can be no direct proof for the existence of any “virus” as there are numerous other microorganisms, substances, and contaminants within a sample that could also be the potential cause of the disease rather than the assumed-yet-never-seen “virus.” Exosome literature is rife with papers explaining the impossibility of separating these particles from their “viral” cousins. As exosomes and “viruses” are the same size and shape, this means that complete purification and isolation of any “virus” is impossible. There will always be exosomes and/or any other particle that is either the same size or smaller than the assumed “virus” particles left remaining. Highlights from the below article from 2019 presents further evidence to this effect:
How are Exosomes Isolated?
“Exosomes are extracellular vesicles that are believed to play a role in communication between cells by transporting materials inside the vesicle.
Exosomes can be isolated from cells using typical methods for purifying cell fractions. The challenge in isolating vesicles is differentiating them from other types of membrane material in the cell culture supernatant.
This is done through successive steps of centrifugation at increasing speeds. The final supernatant is ultracentrifuged at 100,000g to pellet the exosomes. This is then washed to remove contaminating proteins and centrifuged at high speed one more time.
However, that type of preparation is more an enrichment of the sample than a purification. Further analysis of the sample through biochemistry or microscopy is still required to characterize the vesicles.
In addition to exosomes, the extracellular milieu contains extracellular RNA, other types of vesicles, protein complexes, and lipoproteins. These are not fully separated from exosomes through the centrifugation protocol. There are also commercial kits available to isolate exosomes that make use of polymers, but these kits tend to co-isolate other molecules, particularly RNA-protein complexes.
The Executive Committee of the International Society for Extracellular Vesicles (ISEV) proposed criteria for characterizing exosomes to aid in consisting reporting of experimental results.
The criteria are that all exosome samples must adhere to are given below:
- Be isolated from extracellular fluids like cell culture medium or body fluids. The collection should be gentle to minimize cell wall disruption that could contaminate the sample with intracellular compartments.
- Have an overview of protein composition included in its description. The amount of 3 or more relevant proteins should be reported in a semi-quantitative manner in any exosome preparation.
- Have levels of proteins not expected to be enriched to determine the extent of co-isolation of intracellular vesicles.
- Have single vesicles characterized as an indication of heterogeneity.
- Have a quantitative analysis of dose-function relationships in the case that in vitro functional studies are carried out.
- Have demonstration of association of molecules with the exosome, in the case that functional changes are ascribed to single molecules or clusters of molecules.
Study of exosomes is a rich area of research due to the large number of unanswered questions, including exactly how they are formed, what their functions are, how they communicate with acceptor cells, and their role in various diseases.
Consistent methods for isolating and characterizing exosomes and distinguishing them from other types of extracellular and intracellular vesicles are needed to enable these advances.”
There are no consistent methods used for purifying (i.e. “isolating”) exosomes nor ways for distinguishing them from other extracellular and intracellular organisms. In other words, exosomes (the “non-infectious virus”) can not be completely separated from everything else nor can they be differentiated from everything else within the cell culture supernatant. This should tell you everything you need to know regarding the impossibility of complete purification and isolation of these entities.
Purifying “Viruses” from Culture for Vaccines?
Beyond being unable to completely purify and isolate exosomes (cough “viruses” cough) from everything contained within a sample, it is also admitted that “viruses” can not be completely purified from the cell culture materials that they are supposedly grown in. Take, for instance, the Jynneos vaccine for monkeypox. It is claimed that the vaccine, which utilized chicken embryo fibroblast cells for culturing the assumed “virus,” was purified and that it contained no animal material. However, it is later revealed within the same paragraph that not only does the vaccine contain host-DNA (i.e. chicken DNA from the CEF cell), it contains other proteins (source unknown) as well as antibiotics used for culturing and an enzyme used for purification:
- Benzonase – a genetically engineered endonuclease from Serratia marcescens produced and “purified” from E. coli strain
- Gentamicin – an antibiotic that can cause severe damage to the kidneys
- Ciprofloxacin – another antibiotic used to treat urinary bacterial infections
“JYNNEOS is a live vaccine produced from the strain Modified Vaccinia Ankara-Bavarian Nordic (MVA-BN), an attenuated, non-replicating orthopoxvirus. MVA-BN is grown in primary Chicken Embryo Fibroblast (CEF) cells suspended in a serum-free medium containing no material of direct animal origin, harvested from the CEF cells, purified and concentrated by several Tangential Flow Filtration (TFF) steps including benzonase digestion. Each 0.5 mL dose is formulated to contain 0.5 x 108 to 3.95 x 108 infectious units of MVA-BN live virus in 10 mM Tris (tromethamine), 140 mM sodium chloride at pH 7.7. Each 0.5 mL dose may contain residual amounts of host-cell DNA (≤ 20 mcg), protein (≤ 500 mcg), benzonase (≤ 0.0025 mcg), gentamicin (≤ 0.163 mcg) and ciprofloxacin (≤ 0.005 mcg).”
This is also seen in the Johnson & Johnson “Covid” vaccine which contains trace amounts of host DNA belonging to the PER.C6 TetR cell line derived from human embryonic retinal cells from an aborted fetus supposedly genetically modified with “adenovirus.” This cell line is known to cause tumors in mice:
“The Ad26 vector expressing the SARS-CoV-2 S protein is grown in PER.C6 TetR cells, in media
containing amino acids and no animal-derived proteins. After propagation, the vaccine is
processed through several purification steps, formulated with inactive ingredients and filled into
Each 0.5 mL dose of Janssen COVID-19 Vaccine is formulated to contain 5×10^10 virus particles (VP) and the following inactive ingredients: citric acid monohydrate (0.14 mg), trisodium citrate dihydrate (2.02 mg), ethanol (2.04 mg), 2-hydroxypropyl-β-cyclodextrin (HBCD) (25.50 mg), polysorbate-80 (0.16 mg), sodium chloride (2.19 mg). Each dose may also contain residual amounts of host cell proteins (≤0.15 mcg) and/or host cell DNA (≤3 ng).”
Click to access Janssen+COVID-19+Vaccine-HCP-fact-sheet.pdf
Remember, purification means to free from contaminants, pollutants, foreign materials, etc. Do these vaccines sound like purified concoctions to you? If they can not separate out the cell culture additives as well as the benzonase substance used for “purification” as in the Jynneos vaccine, how can they purify a “virus” to study for cause and effect, let alone for use in a vaccine? If they can not separate the host cell DNA as noted in both vaccines, whatever cell line they use, from cancer cells to aborted fetuses to monkey kidneys, will remain within the vaccine and injected into the body. This is why it is imperative to be able to show that these assumed “virus” particles exist directly in the fluids of a sample from a sick human first before it is subjected to the cell culture process and mixed with all sorts of outside contaminants and foreign materials that are subsequently unable to be purified away from the assumed “viral” particles.
We can gain further insight into this inability to separate the “virus” from the culture materials in these highlights from a 270-page “virus” purification manual:
VIRUS PURIFICATION, DETECTION AND REMOVAL
“In vaccine manufacturing, the cell culture is commonly accompanied with cellular debris, unwanted media proteins, adventitious agents, residual DNA, nucleic acid and many process related leachable contaminants. As per the FDA vaccine approval requires the freedom from extraneous material whether or not harmful to the recipient . Viruses have a unique size, shape and surface chemistry, i.e. hydrophobicity and charge. Most often a series of unit operations are required to increase the yield of virus particles. The currently used operations involve a combination of precipitation, centrifugation, filtration and chromatography [13, 14].”
“Vaccine production in therapeutic industry is currently performed on a case-by-case basis for each virus and a single platform system to purify viruses is still lacking. A traditional ATPS system was used for virus precipitation but lack of selectivity for virus against protein contaminants and difficulty in separation from polymer phase inhibited ATPS progress towards vaccine development systems.”
“Traditionally, virus is purified by ultracentrifugation on cesium chloride (CsCl) or sucrose gradients [10, 11]. However, the shear force has been known to reduce virus infectivity and moreover, it is time consuming, labor intensive and difficult to scale up . Virus precipitation with modest results have been obtained using salts ammonium sulfate  or calcium phosphate . Polyethylene glycol (PEG)/aqueous two-phase system has also been studied for virus purification , but the technique lacks the ability to purify virus from many cell culture contaminants.
Chromatography a popular technique for biomolecular purification has been seldom used for virus recovery due to diffusional limitations and large virus size, restricting access to internal surface area of beads. The technique has not provided high virus yield but it is known to work well for protein biomolecules < 5 nm in size . Chromatography has gained wide spread attention due to its ability to create specificity for a biomolecule based on several variables as charge, size and hydrophobicity [17-19]. The virus produced in the lab is heavily contaminated with cell media proteins and our goal is to consider each variable individually (charge, size and hydrophobic) for maximum protein removal with best possible recoverable virus. The purified virus can be very useful to analyze cell protein and contaminant interference during lab scale experimentation.”
“Purification of virus is usually performed from density gradient centrifugation or salt or polymer precipitation techniques. The methods are difficult to scale up or lack specificity from co-precipitating impurities. Filtration is another technique for virus purification but this may cause virus degradation from shear stress, especially in tangential flow filtration conditions. An efficient and fast paced system to achieve high purity virus is required.”
It should be clear from the above passages that it is an impossibility to purify “viruses” from the cell culture supernatant and the material used for propagation. This should be the final nail in the coffin for both cell culture and virology as it has been admitted numerous times that the only way virologists can “isolate” a “virus” is by means of cell culture.
From University of Auckland associate professor and microbiologist Siouxsie Wiles in an AAP “Fact” check:
“Viruses are basically inanimate objects which need a culture to activate in. But the way they are phrasing the requests is that the sample must be completely unadulterated and not be grown in any culture – and you can’t do that,” she told AAP FactCheck in a phone interview.
“You can’t isolate a virus without using a cell culture, so by using their definition it hasn’t been isolated. But it has been isolated and cultivated using a cell culture multiple times all around the world.”
From the CDC:
For more on this impossibility of purifying the “virus” from the cell culture supernatant, let’s look at the FDA’s Guidance for Industry for vaccines which you will find highlights for below. The FDA states that live attenuated “viruses” can not be purified as rigorously as subunit vaccines (a vaccine that contains “purified” parts of the pathogen that are used to elicit an immune response) which leads to greater contamination. Due to this, it is not possible to validate the clearance of adventitious agents in the live “virus” vaccines which means that purification is impossible. For inactivated “virus” vaccines, there is the possibility that not all of the adventitious agents are inactivated as was seen in the case of the polio vaccine.
The FDA admits to numerous sources of contamination just from the cell culturing process itself such as:
- Exposure to the person or animal from which the “virus” was “isolated”
- The cells and raw materials (e.g., serum or trypsin) used
- Materials used in banking and propagation of cells for “viral” seed growth
- Other materials used during production and filling of the seed
- Any infectious “viruses” (including those that infect nonhuman species)
- Retroviruses that may either be endogeneous (coming from within the host) or exogenously acquired
The FDA’s laundry list of sources of contamination establishes that culturing the sample immediately invalidates any claims of purity. Even when checking for contamination, the FDA allows for the use of “non-infected control” cultures to establish whether or not the culture is free of contamination rather than checking the actual “infected” culture to be used for the vaccine. While it should be obvious that a stand-in culture should not be used to judge whether or not a separate culture is free of contamination, the conditions used for the control culture only need to be similar to the “infected” culture rather than identical which immediately invalidates this process as a control.
It is also admitted by the FDA that cell lines such as the Vero cells used in culturing many “viruses” (including “SARS-COV-2”) have biochemical, biological, and genetic characteristics that are different from primary cell lines which may be a result of transformation caused by an adventitious agent. There also may be potential risks from residual DNA within the vaccine as well. As the mechanism by which most cell lines become immortal is unknown, and due to the fact some cell lines cause tumors in inoculated mice, there are many concerns with using these cell lines in vaccines. This is yet another example which shows that complete purification of “viruses” coming from these cell lines is an impossibilty.
Finally, it is admitted by the FDA that the cytopathogenic effect, which is used to determine whether or not a “virus” is present in the culture, can in fact be caused by substances other than a “virus:” Obviously, if other factors can cause the same effect ascribed to a “virus,” there is no need for the virus” to enter into the equation as a possible explanation as the cause of the effect, especially when the particles assumed to be “viruses” are unable to be seen in a completely purified and isolated state without culturing first:
Characterization and Qualification of Cell Substrates and Other Biological Materials Used in the Production of Viral Vaccines for Infectious DiseaseIndications
“Live attenuated viruses, whole inactivated virions, or virus-like particles often cannot be purified as rigorously as viral subunit vaccines; as a consequence, the potential for contamination is greater than that of subunit vaccines. Generation of
live viral vaccines often involves cell disruption, which may add cellular components to the vaccine bulk. In addition, such vaccines often are minimally purified and are not subjected to inactivation steps. Comprehensive testing and qualification of the biological starting materials and lot-by-lot testing for adventitious agents are particularly important because it is not possible for a manufacturer of live viral vaccines to validate the process for clearance of adventitious agents.
In contrast, for more highly purified products where viral clearance can be demonstrated during downstream processing, you can place more reliance on process validation (Ref. 2). For inactivated vaccines, the concern is that the process used to inactivate the vaccine virus may not inactivate all adventitious agents potentially present (as occurred with early inactivated poliovirus vaccines [Ref. 4]). Therefore, you should validate your process for inactivation of adventitious agents using different model viruses (Ref. 2). The choice of tests and the stages at which the tests are applied will depend on your inactivation process. The degree of viral clearance that is feasible may influence the sensitivity of the testing you should perform to demonstrate the absence of contaminants in your product.”
2. Potential Sources of Contamination
“It is important that you identify and examine all potential sources of contamination of your product. For example, a viral seed could be exposed to the following potential sources of contamination: the person or animal from which it was isolated; the cells and raw materials (e.g., serum or trypsin) used in its isolation and attenuation; materials used in banking and propagation of cells for viral seed growth; and other materials used during production and filling of the seed. You should also consider the species of origin of your cell substrates, viral seeds, and other biological starting materials in selecting your tests to ensure the absence of contaminants. Furthermore, you should consider any infectious viruses (including those that infect nonhuman species) as potential contaminants if there is the possibility of contact with your product or cell substrate at any time during development or production. Retroviruses may be either endogeneous (i.e., encoded by the cell substrate genome) or exogenously acquired. Retrovirus testing should address the possibility that either type of retrovirus could contaminate a product. Finally, you should consider the possibility of contamination from unusual sources, as exemplified by the reported presence of minute virus of mice (MVM) in some lots of recombinant proteins (Ref. 5). The susceptibility of the cell substrate to infection by agents of potential concern can influence the tests needed to assure absence of contamination.”
4. Use of Control-Cell Cultures
“We recommend the use of control cells when it is not feasible to directly test cells or product at various stages of manufacture. For example, if you are using primary cell cultures to propagate your vaccine virus, complete testing of the primary culture might not be feasible prior to inoculation of virus. In this situation, when possible, you should produce and test uninfected control-cell cultures that are derived in parallel with, and handled in the same manner as, the production culture.”
“Control cell cultures should be propagated under conditions similar to conditions of manufacture for a time appropriate to the reason for performing the cultures. This time period may be 14 days or more, when needed to allow for detection of potential adventitious agents that may be latent, endogenous, or poorly replicating. Control-cell cultures should be processed simultaneously with your production culture, but left uninfected. Control cells should be evaluated for the presence of adventitious agents using the same tests that would have been performed on the production cultures, if it were feasible. Tests for adventitious agents should be performed on control cells at times at which you would perform similar tests on your manufactured product.”
5. Special Considerations for Continuous Cell Lines
“Some continuous cell lines, including Vero cells and CHO cells, have been used as substrates for licensed biologicals. Cell lines might have biochemical, biological, and genetic characteristics that differ from primary or diploid cells (e.g., they are typically aneuploid and have accumulated genetic changes). Because the mechanism by which
most cell lines become immortal is generally unknown, and because some cell lines form tumors when inoculated into immunodeficient rodents, there are additional concerns for continuous cell lines compared with diploid cells, including the potential that transformation was caused by an adventitious agent and potential risks from residual DNA.
Products prepared in cell lines (including viral vaccines) should be purified to be free (see Section V. Glossary) of adventitious agents and residual cells and should have low levels of cell-substrate DNA. When potential biological activity of residual cell substrate DNA is a concern, you should introduce procedures that
extensively remove or degrade DNA. If you are considering the use of cell lines, we encourage you to develop and evaluate efficient methods for the purification of your product as an essential element of any product development program.”
“Your biological starting materials should be characterized sufficiently to ensure that they do not contaminate the final product with extraneous infectious organisms, such as bacteria, fungi, cultivatable and non-cultivatable mycoplasmas and spiroplasma, mycobacteria, viruses, and the agent(s) responsible for
transmissible spongiform encephalopathies (TSEs). For a substance to be considered free of a contaminant, your assay should demonstrate, at a predefined level of sensitivity, that a certain quantity of the substance is free of that contaminant. Alternatively, a validated process that is known to remove a contaminant to a defined level may be used to demonstrate the absence of that contaminant.”
“An appropriate volume should be inoculated onto monolayer cultures of at least 3 cell types. The sample to be tested should be diluted as little as possible. The cell cultures should be observed for at least 2 weeks. After 2 weeks of observation, the original cultures of supernatants or lysates from cell banks are subcultured onto fresh cells and observed for at least an additional 2 weeks. This subculture onto fresh cells might help to distinguish between non-specific CPE and virus-induced CPE, as toxic effects of the initial specimen or length in culture will be diluted, whereas virus-induced CPE should be amplified.”
- Purification refers to the separation of “virus” particles from host components in a biologically active state
- Purified “virus” is required for the production of antibodies, physical, biochemical and molecular characterization of “virus” isolates
- Purification methods vary with different “viruses,” and there are no universal methods of “virus” purification
- Procedures that are effective for one “virus” may not work with the other
- As the most common purification method, ultracentrifugation is not ideally suited for the separation of exosomes (i.e. non-infectious “viruses”) and “viruses,” particularly from infected cultures and/or body fluids
- The techniques used by researchers for purification of exosomes and “virions” are similar
- None of the methods, including ultracentrifugation, precipitation, and chromatography, can separate exosomes from “viruses”
- Common exosomal separation techniques suffer from operation complexity, time consumption, large sample volumes and low purity
- At present, common exosome isolation technologies, such as ultrafiltration, immunoaffinity, ultracentrifugation (“gold standard” for the isolation of exosomes) are expensive instruments, large volumes of sample, possible protein contamination, complete isolation steps, but they result in low isolation efficiency, sample loss, low exosome recovery and purity
- Like “viruses,” exosomes are small in size (30–150 nm), low in density (1.13–1.19 g/ml), and mixed with similar components (e.g., cell fragments, proteins) in the body fluids, which pose tremendous challenges for their separation
- Exosomes are extracellular vesicles that are believed to play a role in communication between cells by transporting materials inside the vesicle
- The challenge in isolating vesicles is differentiating them from other types of membrane material in the cell culture supernatant
- This is done through successive steps of centrifugation at increasing speeds
- However, that type of preparation is more an enrichment of the sample than a purification
- In addition to exosomes, the extracellular milieu contains extracellular RNA, other types of vesicles, protein complexes, and lipoproteins
- These are not fully separated from exosomes through the centrifugation protocol
- The Executive Committee of the International Society for Extracellular Vesicles (ISEV) proposed criteria for characterizing exosomes to aid in consisting reporting of experimental results and within these criteria, they state:
- Be isolated from extracellular fluids like cell culture medium or body fluids. The collection should be gentle to minimize cell wall disruption that could contaminate the sample with intracellular compartments
- Have levels of proteins not expected to be enriched to determine the extent of co-isolation of intracellular vesicles
- In other words, it is well-known that other intracellular components will be “isolated” along with the exosomes
- Study of exosomes is a rich area of research due to the large number of unanswered questions, including:
- Exactly how they are formed
- What their functions are
- How they communicate with acceptor cells
- Their role in various diseases
- Consistent methods for isolating and characterizing exosomes and distinguishing them from other types of extracellular and intracellular vesicles are needed to enable these advances
- In vaccine manufacturing, the cell culture is commonly accompanied with:
- Cellular debris
- Unwanted media proteins
- Adventitious agents
- Residual DNA
- Nucleic acid
- Many process related leachable contaminants
- Vaccine production in therapeutic industry is currently performed on a case-by-case basis for each “virus” and a single platform system to purify “viruses” is still lacking
- A traditional ATPS system was used for “virus” precipitation but lack of selectivity for “virus” against protein contaminants and difficulty in separation from polymer phase inhibited ATPS progress towards vaccine development systems
- Polyethylene glycol (PEG)/aqueous two-phase system has also been studied for “virus” purification, but the technique lacks the ability to purify “virus” from many cell culture contaminants
- Chromatography a popular technique for biomolecular purification has been seldom used for “virus” recovery due to diffusional limitations and large “virus” size, restricting access to internal surface area of beads
- The “virus” produced in the lab is heavily contaminated with cell media proteins
- Purification of “virus” is usually performed from density gradient centrifugation or salt or polymer precipitation techniques
- The methods are difficult to scale up or lack specificity from co-precipitating impurities
- An efficient and fast paced system to achieve high purity “virus” is required
- According to University of Auckland associate professor and microbiologist Siouxsie Wiles, “Viruses are basically inanimate objects which need a culture to activate in. But the way they are phrasing the requests is that the sample must be completely unadulterated and not be grown in any culture – and you can’t do that. You can’t isolate a virus without using a cell culture, so by using their definition it hasn’t been isolated. But it has been isolated and cultivated using a cell culture multiple times all around the world.”
- According to the CDC, purifying “viruses” without cell culturing is outside of what is possible in virology
- In other words, it is impossible to purify “viruses” without culturing and it is also impossible to purify and isolate them with culturing
- From the FDA’s Guidance for Industry, we learn that live attenuated “viruses,” whole inactivated “virions,” or “virus-like” particles often cannot be purified as rigorously as “viral” subunit vaccines; as a consequence, the potential for contamination is greater than that of subunit vaccines
- Generation of live “viral” vaccines often involves cell disruption, which may add cellular components to the vaccine bulk
- In addition, such vaccines often are minimally purified and are not subjected to inactivation steps
- Comprehensive testing and qualification of the biological starting materials and lot-by-lot testing for adventitious agents are particularly important because it is not possible for a manufacturer of live “viral” vaccines to validate the process for clearance of adventitious agents
- For inactivated vaccines, the concern is that the process used to inactivate the vaccine “virus” may not inactivate all adventitious agents potentially present (as occurred with early inactivated poliovirus vaccines)
- The degree of “viral” clearance that is feasible may influence the sensitivity of the testing that should be performed to demonstrate the absence of contaminants in the product
- It is important to identify and examine all potential sources of contamination of the product such as:
- The person or animal from which it was isolated
- Tthe cells and raw materials (e.g., serum or trypsin) used in its isolation and attenuation
- Materials used in banking and propagation of cells for “viral” seed growth
- Other materials used during production and filling of the seed
- Furthermore, any infectious “viruses” (including those that infect nonhuman species) should be considered as potential contaminants if there is the possibility of contact with the product or cell substrate at any time during development or production
- “Retroviruses” may be either endogeneous (from the host genome) or exogenously acquired
- The susceptibility of the cell substrate to infection by agents of potential concern can influence the tests needed to assure absence of contamination
- If using primary cell cultures to propagate vaccine “virus,” complete testing of the primary culture might not be feasible prior to inoculation of “virus”
- In this situation, when possible, they are to produce and test uninfected control-cell cultures that are derived in parallel with, and handled in the same manner as, the production culture
- In other words, if they decide that they can not check the uninfected culture used for the vaccine for contamination, using another uninfected culture as a stand-in suffices
- Control cell cultures should be propagated under conditions similar (i.e. not the same) to conditions of manufacture for a time appropriate to the reason for performing the cultures
- Control-cell cultures should be processed simultaneously with the production culture, but left uninfected
- Cell lines, such as Vero and CHO, might have biochemical, biological, and genetic characteristics that differ from primary or diploid cells (e.g., they are typically aneuploid and have accumulated genetic changes)
- Because the mechanism by which most cell lines become immortal is generally unknown, and because some cell lines form tumors when inoculated into immunodeficient rodents, there are additional concerns for continuous cell lines compared with diploid cells, including the potential that transformation was caused by an adventitious agent and potential risks from residual DNA
- Products prepared in cell lines (including “viral” vaccines) should be purified to be free of adventitious agents and residual cells and should have low levels of cell-substrate DNA
- When potential biological activity of residual cell substrate DNA is a concern, procedures that extensively remove or degrade DNA should be introduced
- If considering the use of cell lines, it is encouraged (not required) to develop and evaluate efficient methods for the purification of the product as an essential element of any product development program
- Biological starting materials should be characterized sufficiently to ensure that they do not contaminate the final product with extraneous infectious organisms such as:
- Cultivatable and non-cultivatable mycoplasmas and spiroplasma
- Agent(s) responsible for transmissible spongiform encephalopathies (TSEs)
- For a substance to be considered free of a contaminant, the assay should demonstrate, at a predefined level of sensitivity, that a certain quantity of the substance is free of that contaminant (i.e. not all of it has to be shown free of contamination)
- Alternatively, a validated process that is known to remove a contaminant to a defined level may be used to demonstrate (i.e. to infer) the absence of that contaminant
- The cell cultures should be observed for at least 2 weeks and after 2 weeks of observation, the original cultures of supernatants or lysates from cell banks are subcultured onto fresh cells and observed for at least an additional 2 weeks
- This subculture onto fresh cells might help to distinguish between non-specific CPE and “virus-induced” CPE, as toxic effects of the initial specimen or length in culture will be diluted, whereas “virus-induced” CPE should be amplified
- In other words, it is admitted that the CPE used to identify “viruses” can be caused by factors other than “viruses” thus destroying this effect as an indirect indicator of the presence of any “virus”
Obviously the main objective in the purification and isolation of viruses is the separation of the virus from the host tissues and cell organelles.”DOI: 10.1007/978-94-009-5876-0_3
It should be clear by now that virology can not completely purify and therefore isolate the particles assumed to be “viruses” from everything else that is sure to be within the sample. Virologists openly admit that they:
- Can not purify directly from the fluids of humans
- Can not purify the particles from cell culture supernatant
- Can not separate “viruses” from exosomes and other similar MVB’s
- Can not separate the “viruses” from the cell culture host material and DNA
- Can not separate the cell culture media from the sample
As purification can not be achieved, virology lacks the physical particles necessary for use as an independent variable to vary and manipulate in order to determine cause and effect. There is no direct evidence as to the existence of the particles assumed to be “virus” ever being found within humans. The only evidence is from indirect cell culture experimentation citing patterns of cell death known as the cytopathogenic effect (CPE) seen in the cells after being poisoned and starved for days which is then blamed on an invisible “virus.” As other contaminants, chemicals, and impurities remain within the cultured sample, there can be no certainty that the other substances within may be the potential cause of the CPE seen within the culture over the assumed “virus.” It is a fact that other substances will be there within the fluids, especially in any sample processed through cell culture, as can be seen from the exosome studies as well as from attempts to purify “viruses” from cell culture supernatant for vaccine use. The FDA admitted that there are other factors besides a “virus” which can cause the CPE seen which is instead blamed on the invisible “virus.” The cell culture method is by definition an impure process as many foreign elements, chemicals, and contaminants are added to the culture which can not be separated out. As virology admits that “viruses” can not be purified and isolated directly from the fluids of a sick human and must be cultured, there is no purification that can ever be achieved by subjection to an impure process.
Without purification, virology can not adhere to the scientific method and is thus a pseudoscience. We know this and virologists know this which is why purification methods are rarely performed and why this criteria is rarely discussed in virology papers. Purification is the black sheep of the virology family and it will remain as such until virology can find a way to do what it has been unable to do for well over 100 years.
Perhaps a psychologist could explain why virologists feel free to abuse the word “isolation” so freely, but are scared to death of even writing the word “purification”
wow ! Excellent Mike .
Simple and clear, let me repeat it :
“Purification is the black sheep of the virology family and it will remain as such until virology can find a way to do what it has been unable to do for well over 100 years.”
Let me add a phrase from David Crowe : “Perhaps a psychologist could explain why virologists feel free to abuse the word “isolation” so freely, but are scared to death of even writing the word “purification”. ( “Isolation versus Purification” , David Crowe May 21, 2020 )
* From the FOIA answer “According to the CDC, purifying “viruses” without cell culturing is outside of what is possible in virology” …… So I wonder isnt the human being the better place for “viruses” to grow ? Take a direct sample, purify and show them !!!! … What a joke
* Interestingly , when Michael from New Zeland sent FOI requests for viruses ( taken directly form human sample……) all the answers NO RECORDS, when he sent the same FOI request but for exosomes instead , he recieved from The University of Otago 331 documents relating to the isolation of human exosomes (supposedly from unadulterated samples, need to check that ,but at least he received records… )
So , they can attempt ( without success I guess ) the purification for exosomes, but not for “exosomes” 🙂
In their pseudoscience sounds correct : because there will be no CPE , so no “virus isolation” for them….I know …I hardly understand myself… but´s thats ViroLIEgy !
And here and interesting flyer :https://media.beckman.com/-/media/pdf-assets/technical-application/centrifuge-technical-application-exosomes-interview-shiba-isolation-ultra.pdf
Misha from gamzuletova.org
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Thanks for the kind words and the links Misha! I love David Crowe’s work as he was a huge influence on me. One thing he said was that virology could try to wiggle around isolation but it had no answer for purification (my paraphrasing). I miss his insight.
The mind set in fear seeking ‘protection’ buys magical solutions that are only a repackaging of conflict in further complexity and diversion.
This masking of control, protects the means by which to gain function as the arbiter of what cannot be addressed as a matter of national or biological ‘security’.
This pattern runs what can be called a split mind.
brilliant – thank you very comprehensive x
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Thanks for the kind words Jo! 🙂
Welcome back. Mike, feels like it’s been a while. And brilliant indeed. I’m doing a presentation to my group this coming week regarding the lack of scientific proof for the existence of SARS-COV-2, getting a lot of resistance. Including this absurd hit piece by Steve Kirsch. him interviewing Toby Rogers, getting into the “no virus people” at around 1:19:00.
I’m using your response to Kirsch from January as an item in my presentation, by the way.
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Great clip from 1:19 onwards. I really don’t see why you called it a hit piece. And unless these guys have been untrustworthy opponents in the past I agree with them that Massey saying she needs to talk to her people first does kinda come across as shady or insecure at the very least. It’s certainly not open. I mean she should be up for it herself even if she can’t get anybody else in the gang to join in, right? It’s not like any of the nuances of this stuff are beyond anybody’s pay grade.
It’s an easy debate to win. Not that they will necessarily acknowledge that they lost.
It goes back to the first and second inches. They are both self-evident. Anaerobes cannot be around healthy tissues and viruses can’t do anything.
Toby’s tuba analogy that’s his refutation of the “viruses can’t do anything” argument is self-defeating. The person choosing to play the tuba in the orchestra — who he conveniently ignores — can only be analogous to the body choosing to play (acknowledge by reproducing) the virus. His analogy unwittingly acknowledges that the body has all the agency. The problem for you guys here is that you refuse to even talk about the false internal logic of viral theory because you think it means you’re defending exosomal theory. Using exosomal theory to discredit viral theory is a no-brainer. All you have to do is add the disclaimer that you don’t actually believe in exosomes but that the internal logic of exosomal theory is true and consistent whereas with viruses it is obviously not because RNA/DNA can’t do anything ever, no matter how sick the body is. Nucleic acids are an informational format, period.
Given their tacit acknowledgement that the terrain plays the leading role in disease by making the body susceptible, their thinking reinforces my contention that it’s a Thin Blue Line between GT and TT.
You guys really need to start using the anaerobe/saprophytes argument for bacteria and fungi. It’s a giant killer. It exists on the creative commons. Savor the looks on their faces when they hear it for the first time lol. Basic biology apparently long forgotten.
If Steve Kirsch didn’t set a record for BS and untruths per 5 minutes, he sure gave that record a close chase. He completely misstates the case put forth by the people he names as “the virus does not exist,” when he has been corrected hundreds of times about that, the case being “there is no scientific proof for the existence of the virus.” He claims that the “no virus” people make all sorts of claims about what causes the alleged “Covid” disease, when in fact they state that the very idea of one unique cause is part of the problem. He completely lies about the record of his debate challenges with Christine Massey. She saved copies of the communications, see her update towards the bottom of this page, https://www.fluoridefreepeel.ca/open-letter-to-steve-kirsch-january-10-2022/
He claims to not be the expert, yet he posted a blog entry stating “Has the virus been isolated? Yes!” And he was presented by Dr Joseph Marcela as his source for information that the virus has been proven to exist via isolation. Mike responded to Steve with “Has the virus been isolated? No” in January as well, pretty much shredded Steve’s case.
Toby likewise claims the virus exists, saying “of course.” When did he prove it? And he makes it about “terrain theory vs germ theory,” and claims to be combining them, when he is trying to co-opt terrain theory and incorporate it into germ theory, as if “terrain” is just about “environmental factors.” the same old game, as if these could be blended like coffee and cream, rather than being two mutually contradictory theories. Have viruses in fact been proven to exist? What if we were to assert that people get sick because their environment makes them susceptible to unicorns?
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Thanks for the additional context. Yeah Steve’s not a straight shooter. No point dealing with people like that.
As for Toby, sure you can see that he his boyish lack of a center of gravity when he says “of course” viruses exist.
The terrain *is* just about environmental factors, though, Jeffrey. That’s the terrain in Terra Theory. I mean, what else is there?
And that part about you guys going on about how you never said viruses don’t exist. I just don’t get that man. For starters it contradicts the argument that they can’t *do* anything, because if they can’t do anything and they’re just nucleic acids then they damn sure ain’t viruses as they define them to be. Why you guys can’t own that I will never know unless you explain it to me.
People will probably object when it comes to facultative anaerobes because they do represent a grayish area insofar as life and death are two sides of the same coin and the coin’s edge is the thin line between them (though not a blue line).
While atmospheric oxygen levels average around 21pc and dropping, and lower in cities, they used to be several pc higher. Core cell tissue oxygenation is higher than peripheral tissue oxygenation which can get down to 5pc oxygen. At this lower level of oxygen facultative anaerobes can perform aerobic respiration in the interstitial water spaces between the cells that have the same oxygen levels but the facultative anaerobes are still saprophytes which means they are eaters of dead organic matter. You can find statements out there saying most families of this facultative anaerobe or that one are saprophytes but a few are not but there are never any accompanying details as to which ones and what healthy tissues they are eating. And this unsubstantiated claim is the microbial millimeter we can’t give them; the facultative yogurt fermenters are eating dead tissues, even in raw milk, and in raw milk they won’t be eating the macrophages until they die.
Bacteria are about a tenth the size of a cell, they’re not about to take on a cell that can defend itself. Facultative fungi can grow long lengths but they don’t squeeze cells to death like pythons; they’re also saprophytic. What the mainstream doesn’t seem to understand is that we commonly have localized death occuring in our bodies and therefore we have localized saprophytic activity between states of dormancy.
A useful analogy for saprophytes not eating healthy cells is that of carrion birds nut hunting prey. They don’t do it because they’re not built for it. When we have new lambs or kids the livestock guardian dogs will make like Hillary Clinton and want to impose a no fly zone on all big birds including the turkey vultures because presumably they can’t tell the difference between the vultures and the hawks and eagles. But we humans have figured out which ones are predators.
Humans have no natural predators. That’s a big part of what defines us. A lone, small child may be vulnerable to predation out in the wild but that does not qualify humans as natural prey because that itself is an unnatural situation.
reante, prove to us that unicorns don’t exist.
science becomes the false idol known as Big Science the minute we let its provincial diktats (such as the laboratory scientific method) supercede Reason, however reason-based are the diktats themselves. Reality has sufficiently proven to humans that unicorns don’t exist. Same goes for submicroscopic terrorists without boxcutters.
I want to say live a little, brotherman. Spending months on end arguing that there’s zero evidence of viruses existing yet refusing to realize that they don’t exist is just no way to live. That looks like purgatory to freewheeling ole reante. Still, different strokes for different folks!
Thanks Jeffrey! I’m on vacation until the 23rd so I’ve been a little slower than usual as it has been a busy two weeks. I hope to get back to a more regular pace once I return. I will have to watch the Kirsch interview but it does not surprise me that he is misrepresenting information. Par for the course with him.
Thanks, Mike, and please, do basically stay on vacation, you have more than earned a break from this! Enjoy.
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Thank you so much Jeffrey! 😁
Exosomes are indeed isolated and purified. There are many scientific papers on this subject matter. Here is a good review paper – https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5327650
The claims that viruses containment exosome purification are not provable (as viruses have never been proven to exist). If that was the case, exosome research would be impossible (as how would they know if they were dealing with viruses or exosomes).
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The article you shared actually came out before the papers I shared stating that purification of exosomes from “viruses” is impossible and that better purification methods are necessary. Exosomes are the same exact particles seen in EM as “viruses” but have been given different names/functions.
The problem with the claim in the papers that viruses cannot be separated from exosomes is that viruses have never been proven to exist. Yet, since 1983, exosomes have been proven to exist. So the claims made in the papers you shared are speculative. How do they know that viruses cannot be separated from exosomes? Of all of the purified samples of exosomes, have they ever found viruses in their study? I haven’t read any papers that ‘found’ viruses during their isolation and purification methods for exosomes. Surely if viruses and exosomes were inseparable, those researching exosomes would have discovered viruses.
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How were exosomes proven to exist?
A common theme to the idea of both virus and exosome is of communication or transmission and reception to information energetically encoded or carried by cell debris. Either to and from ‘cells’ or within a Biome that is greater than the sum of its parts (Terrain as total environment always embraces its seeming parts)
Another common theme to both is of communication occurring outside the mind – as the posit of separate minds in, as or through separate bodies, though our own mind is no more ‘isolatable’ than the postulated virus.
My current assumption is that some extracellular communication or process has been observed that involves or is associated with exosomes. I haven’t read up on that. But is there anything to support any of the dramatic functions assigned to virus as exogenous hijackers of cells, by which to propagate a contagion of cell functioning as some kind of attack on the body? The cytopathic effect is a cytopath-etic prop for a dying cause, given solemn worship within the church founded on a War against Disease. Only a trained monkey could believe it demonstrates the existence of its god.
BUT the virus theory carries all the signature characteristics of the mind of its maker. It is this ‘jump’ from an externalise casting out of our mind to an awakened responsibility for thought that I give most focus. When fear is denied and masked over, a state of dis-integrity runs as a masking over, supported by a diversionary defence, that is itself masked over, such as to have not just turned a blind eye, but compounded a mindset by which blindness dictates what can be seen or at least interprets and accepted.
The primary issue to me in all this is not that people for their own reasons believe or act from beliefs that may be self-destructive (or educationally instructive) to their own living, but that such beliefs are weaponised as a cultic demand for sacrifice or tyranny over those who freely choose not to share those beliefs and behaviours.
As I see it the invested and defended lie has run out of time and space to maintain an ability to pass off as true, but many will die with, rather than release, the identity it gives them.
This appears to be a good development for reante here. Thank you. Can you confirm to me that exosomes are most easily and legitimately batch-isolated (as classified by particle size and subsequent chemical analysis) by filtering and centrifuging plasma, and without any processing that would physically FUBAR the proceedings?
If so, dang, why didn’t I think of that? The plasma, of course.
Mike, to answer your question “how were exosomes proven to exist,” that would be by separating the hypothesized particles by size, chemically analyzing them in order to cross-reference their constituent parts with all existing known human chemical compounds and molecules, and patterning the results in accordance with existing patterns. Which is to say, they ‘proved’ exosomes the same way they ‘proved’ the human species, or anything else. It’s just that some things are more difficult to prove than others. But as long as Reason be our guide we have nothing to fear.
Yet, as with “viruses,” they have not done this for exosomes. Reason can be a good guide but we need to not allow it to take us down false paths based on what we think makes sense.
Thanks Mike. What did they not do that they would’ve needed to do? Or what did they do that they shouldn’t have done?
Mike Stone Wrote: ‘How were exosomes proven to exist?’
The same way bacterio phages were proven to exist. Exosomes have been isolated and purified using standard scientific methods.
Here is one such paper that purifies exosomes from human urine – https://www.spandidos-publications.com/10.3892/ijmm.2018.3944
If you dispute this paper didn’t purify exosomes, than please point out where in the methods section the scientist failed to use the scientific method.
This what Cowan and others point out is NOT done regarding postulated viruses.
I’m more than open to the unfolding our our closed system of objects in space to an intercommunicating whole of which the parts are never truly apart from, but can operate as distinct facets within a greater whole.
I skimmed the study and not pathological linguistics as in ‘target cells’, or disease information instead of adaptive communication to resolving or rebalancing within environmental stresses.
The finding of the ‘packets’ may be from bacterial exosomes?
I haven’t followed the reference links but finding a packet of proteins is part way. Demonstrating the functions assigned to it is the other part.
Invested belief in hypothesis can fund the results that attract more investment, particular where patents and profits can accrue under ‘serving humanity’. I don’t mean to be cynical, but such a machine operates from aligned self-interest.
I feel the exosomal theory holds some promise for the release of a pathological modelling of an organism as a death trap, a killing and mating machine or a captured asset to be remade and reconfigured according to … attempts to delay or escape death, align with the predators not the prey, and maintain mating rights and other similar privileges.
The specialism for the use and interpretations of some of the ‘extended sensing’ used as measuring observed event and reaction, could as well be a black box under a high priest.
But I found the paper interesting. Thankyou.
Where did the researchers fail to follow the scientific method? For starters, the researchers never observed a natural phenomena which is the very first step of the scientific method:
1. Observe a phenomenon
2. Alternate hypothesis
*Independent variable (the presumed cause)
*Dependent variable (the observed effect)
3. Null hypothesis
5. Analyze the observation/data
6. Validate/invalidate hypothesis
Secondly, this study is from 2018 and presumes such a thing as an exosome exists based on a concept that was developed first in 1981 before any exosomes were ever observed as stated here:
“The CONCEPT of exosomes was first proposed by Trams et al (1) in 1981, while soon after, exosomes were identified in a study of reticulocyte differentiation as a consequence of multivesicular endosome fusion with the plasma membrane.”
This 2018 study was comparing purification and isolation procedures by using indirect methods to suggest the presence of exosomes:
“The EXISTENCE, PURITY and production of the exosomes isolated by OUF and conventional ultracentrifugation (UC) were systematically COMPARED by TRANSMISSION ELECTRON MICROSCOPY, WESTERN BLOTTING and NANOPARTICLE TRACKING ANALYSIS.”
The existing methods for exosome “isolation” were admitted to produce low purity:
“A large number of strategies have been applied for the isolation of urinary exosomes. However, the high cost of these methods and the LOW PURITY OF THE OBTAINED EXOSOMES remain important challenges, limiting further development of exosomes (22).”
The authors admit that there is a need for a method that produces high purity. What is “high purity?” That does not seem to mean complete purification, only a higher level of purifucation by comparison:
“Thus, there is an URGENT NEED to establish a simple and rapid method of isolating urinary exosomes WITH HIGH PURITY, production and biological activity for further applications in research and clinical practice (24,25).”
“The present study reports an optimized ultrafiltration (OUF) method that can effectively isolate urinary exosomes with HIGH PURITY and quality in a much more simplified way compared with UC.”
The OUF method which is claimed to offer “higher purity” is mixed with ExoQuick-TC:
“Next, the concentrated supernatant (CSN) was INCUBATED WITH ExoQuick-TC™ EXOSOME PRECIPITATION SOLUTION (cat. no. EXOTC50A-1; System Biosciences, Palo Alto, CA, USA) for 30 min at 4°C.”
From System Biosciences, we can see that this is a proprietary polymer, which can mean it is either natural or synthetic substances which were added to the sample and for which the ingredients remain unknown:
“ExoQuick-TC is a proprietary polymer that gently precipitates exosomes. First, pre-clear your samples of cells and cellular debris, and then simply add the appropriate amount of ExoQuick-TC to your cleared biofluid, refrigerate, and centrifuge (see the product manual for protocol details). Your exosomes will be in the pellet, ready for resuspension in an appropriate solution.”
This addition and incubation in ExoQuick-TC destroys any claim of purification/isolation in the OUF group.
Next, we can look at the indirect methods used to suggest the presence of the presumed exosomes. For NTA, the samples were diluted in PBS which once again destroys purity and isolation:
Nanoparticle tracking analysis (NTA)
“A NanoSight LM10 (Malvern Panalytical Ltd., Malvern, UK) was used to detect the size of the exosomes. Briefly, 10 µl exosomes samples WERE DILUTED TO 1:100 IN 1X PBS.”
For TEM, the usual fixing, dehydration, and staining methods were employed. The sample can only be viewed dead and deformed which means no information can be obtained about what the particles are nor how they function. The particles may only be artefacts created by the TEM prep as suggested by Harold Hillman:
“Exosome samples were FIXED WITH 1% GLUTARALDEHYDE in PBS at an optimal concentration. The mixture was then spotted onto a 300 mesh carbon/formvar-coated grids and DRIED AT ROOM TEMPERATURE. Next, the grids were WASHED WITH PBS and STAINED FOR CONTRAST USING URANYL ACETATE in water for 10 min.”
The next test was western blotting which is an antibody test. It should be obvious that one fictional entity can not be used to identify another, especially when neither had been properly purified nor isolated before this study in order to ensure they both exist and that the reaction is specific. You can also note the various chemicals and substances used:
“For western blotting, the protein blot was blocked with 5% SKIMMED MILK for 2 h at room temperature and incubated overnight at 4°C with the following primary antibodies, according to manufacturer’s protocol: Anti-CD63 (cat. no. Ab134045; 1:500; Abcam, Cambridge, MA, USA) and anti-heat shock protein 70 (anti-Hsp70; cat. no. Ab2787; 1:500; Abcam). Following WASHING IN 0.1% PBS-Tween 20, the protein blot was INCUBATED WITH HORSERADISH PEROXIDASE-CONJUGATED GOAT ANTI-MOUSE IgG1 SECONDARY ANTIBODY (cat. no. SA00001-1; 1:5,000; ProteinTech, Chicago, IL, USA). Subsequently, the protein blot was WASHED FOUR TIMES IN 0.1% PBS-Tween 20 on an orbital shaker. ENHANCED CHEMILUMINESCENCE MIXTURE (Sigma-Aldrich; Merck KGaA) was then added to the blot, and images were captured using a gel imaging system (Bio-Rad Laboratories, Inc., Hercules, CA, USA).”
The pellets obtained by both the UC and OUF methods were not identical with it said the OUF were larger than the UC:
“UC and OUF were used to isolate exosomes from the same volume of urine. The results demonstrated that pellets (denoted by the black arrow in Fig. 2A) isolated from the OUF group APPEARED TO BE LARGER IN COMPARISON with those from the UC group.”
The OUF particles were “cup-shaped” with various sizes while the UC group had fewer particles that were then said to be larger than the OUF group (contradicting the earlier statement) and did not resemble exosomes. The time of collection (i.e. morning) also determined if particles were found in both groups.
“TEM analysis was used to observe the morphology and size of exosomes. Notably, the TEM images of the OUF group revealed that exosomes DISPLAYED A CUP-SHAPED MORPHOLOGY, with relative sizes ranging BETWEEN 30 AND 100 nm (Fig. 3A and B). However, FEWER EXOSOMES WERE DETECTED IN THE FIELD OF VISION IN THE UC GROUP when compared with the OUF group. The majority of the vesicles isolated from the UC group PRESENTED AS LARGER PARTICLES that were white, WITHOUT THE CLASSICAL EXOSOME MORPHOLOGY and with a diameter of 50-200 nm (Fig. 3A). In addition, the TEM images of exosomes isolated in the three different collection periods were compared. The results indicated that the EXOSOMES IN THE MORNING GROUP EXHIBITED BETTER MORPHOLOGY in both UC and OUF, and it is thus proposed that morning would be the best time for collecting urine samples for exosome isolation.”
The size range of presumed exosomes was used to claim the presence of exosomes, which also happens to be the same size range as “viruses” yet they assumed these particles to be exosomes:
“Next, NTA was used to measure the size distribution of particles CONSISTENT WITH THE SIZE RANGE OF EXOSOMES.”
“These results clearly SUGGESTED THE EXISTENCE of exosomes in urine and the superiority of OUF over UC for isolating these exosomes.”
The authors admit that research can only suggest what exosomes do (as they are never observed alive and functioning within a living body). The nonstandardized “isolation” methods remain unresolved which limits research:
“At present, increasing research SUGGESTS that exosomes serve a significant role in cell-to-cell interaction and have great potential as biomarkers for a number of diseases. However, severe issues associated with the INEFFICIENCY OF NONSTANDARDIZED EXOSOME ISOLATION PROTOCOLS REMAIN UNRESOLVED, limiting associated research and applications.”
Again, this next section shows that the filtration step included incubation in ExoQuick-TC resulting in “higher purity” based on indirect evidence:
“Next, a dialysis membrane with MWCO of 10,000 kDa was introduced to concentrate the filtered fluids up to 1/50 of the initial volume. This concentration step was aimed to REDUCE THE CONSUMPTION OF ExoQuick-TC AND THE INCUBATION TIME OF CSN AND ExoQuick-TC MIXTURE. Subsequently, ExoQuick-TC was incubated with CSN to gently precipitate the exosomes from the mixture (Fig. 1). The aforementioned improvements included in the OUF method resulted in the production of more exosomes, which were of HIGHER purity and biological stability, as demonstrated through various characterization procedures.”
In this next section, another indirect marker is used to suggest exosomes were detected. It is also stated that the OUF pellets were larger than the UC pellets which once again contradicts and flip-flops back to their previous statement:
“The exosome marker proteins CD63 and Hsp70 were detectable in the exosome pellets, indicating that UC and OUF were both able to isolate a considerable amount of exosomes (Fig. 2B). However, according to the results shown in Fig. 2, the exosome pellets harvested from the OUF group WERE CLEARLY LARGER IN COMPARISON with those from the UC group (Fig. 2B).”
Lastly, few “exosome-like” vesicles were identified in the UC group and they were not similar. The OUF were “relatively” homogeneous. This indicated a “higher purity” in the OUF group by comparison:
“FEW TYPICAL EXOSOME-LIKE VESICLES WERE IDENTIFIED IN THE UC GROUP, and the majority of these particles were HETEROGENEOUS (50-200 nm), WITHOUT A CLASSICAL EXOSOME STRUCTURE (Fig. 3A). By contrast, TEM images of the OUF group demonstrated a high concentration of exosomes, which were RELATIVELY homogeneous with a size of 100-130 nm (Fig. 3B). These results indicated that the purity of the exosomes isolated by OUF WAS HIGHER IN COMPARISON with that of UC-isolated exosomes. Subsequently, NTA was applied to measure the proportion of vesicles CONSISTENT WITH THE SIZE RANGE OF EXOSOMES. This revealed that the mean proportion of exosomes in the OUF group (95.2%) was higher compared with that in the UC group (33.33%; Fig. 4). This SUGGESTED that the abundance of exosomes isolated by OUF was higher as compared with that obtained by UC.”
In summary, nowhere in this study was a natural phenomena observed nor was cause and effect ever determined utilizing the independent and dependent variable. No controls were discussed. The evidence was indirect (NTA, TEM, Western Blotting) and could only suggest the presence of exosomes (which were already presumed to exist) based on size and morphology. All the researchers did was prove their OUF method created different particles in TEM than the UC method. This study did not follow the scientific method and does not show proof that exosomes exist.
I recommend that comment being its own page!
One day I hop to gain more perspective on natural genetic modifications – (I understand bacteria such as e-coli is one such means) and the ‘spanner in the works’ of techniques such as CRISPR. Hugely invested narratives can run on hype…
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This is a response to Mike Stone’s comment regarding his analysis on the Exosome Purification Paper that I posted. I cannot reply directly to his comment (I guess due to the threading of this forum).
How does Mike Stone define ‘scientific method’?
The assumption from the paper is that Ultra Centrifugation is the gold standard for purifying very small things. This is widely accepted in the scientific community. It appears that Mike Stone rejects Ultra Centfiguation as a purification method, is that correct?
1. The scientific method requires observations but why do you use the word ‘phenomenon’?
The authors have observed exosomes via ultra centrifugation (gold standard) as a collective.
2. The second step is to propose a hypothesis. Why do you use ‘alternate’ hypothesis’. That assumes there is already an established hypothesis.
Clearly they proposed a hypothesis that one technique of purification has better results of an another technique.
3. What is the point of a ‘null hypothesis’? Null means nothing or void, so what is the point of a non-existent hypothesis?
They tested 1 purification method. The other method is the control and is the gold standard ultra centrifugation.
Yep, they did that (see results).
6. I think you meant to write, repeatability as that is a requirement for the scientific method
They ran the experiment multiple times. But it is required for others to repeat their experiment in order to validate their results (as any scientific work requires).
My challenge to Mike Stone is to find any scientific paper that has achieved 100% purification (complete purification) ie particles in a vacuum with no other different particles in the presence of the studied particle. If you cannot find 100% purification of anything, than please rectify your claims as what you propose to me seems impossible.
This notion of 100% purification is often brought up by the pro-virus people when we inquire about isolation and purification of SARS-COV-2. When we compose our FOIA requests, we specifically mention we are NOT looking for particles in a vacuum (which is what Mike Stone is suggesting with complete purification). However, if Mike Stone can produce ‘particles in a vacuum’, IE complete purification, than I would happy to stand corrected.
I also question why it is necessary to have ‘complete purification’ in order to prove the existence of something. What level of purification is sufficient to observe cause and effect?
First of all, the scientific method I listed was not “my definition” of the scientific method. It is THE scientific method.
1. Ultracentrifugation, as I have shown in many articles, can not properly purify and isolate anything. This is admitted numerous times in various sources including this article I wrote which you commented on:
“The method is NOT IDEALLY SUITED for the separation of EV and viruses, particularly from infected cultures and/or body fluids and it is difficult to adopt ultracentrifugation to large-scale production environments or to environments that present biosafety concerns.”
“Ultracentrifugation at greater than 100,000 x g is used to concentrate both exosomes and viruses. This technique retains intact, functional viruses and exosomes. Because exosomes and viruses are similar in size and density, SEPARATION OF THE TWO IS NOT POSSIBLE USING THIS TECHNIQUE.”
As well as in the paper you shared:
“Among the available strategies, ultra-centrifugation (UC) is the most widely used method, which exhibits a low throughput and IMPURE ISOLATED EXOSOMES due to high-molecular-mass proteins, sample heterogeneity and low stability, given the intensiveness of sequential UC steps (23).”
Do you dispute this?
Exosomes were never observed in nature which is necessary to form a hypothesis. As your paper stated, exosomes were a concept first (never observed) and then researchers searched for evidence to fit their fictional concept:
“The CONCEPT OF EXOSOMES WAS FIRST PROPOSED by Trams et al (1) in 1981, while soon after, exosomes were identified in a study of reticulocyte differentiation as a consequence of multivesicular endosome fusion with the plasma membrane.”
2 & 3. I use alternate/null hypothesis as these are a part of the scientific method:
“An alternative hypothesis is one in which a difference (or an effect) between two or more variables is anticipated by the researchers; that is, the observed pattern of the data is not due to a chance occurrence. This follows from the tenets of science, in which empirical evidence must be found to refute the null hypothesis before one can claim support for an alternative hypothesis (i.e. there is in fact a reliable difference or effect in whatever is being studied). The concept of the alternative hypothesis is a central part of formal hypothesis testing.”
4. You are correct in that the study you presented was a comparison paper between two purification methods and both produced different results. This paper does not prove the existence of exosomes. At best, it showed differences in two methods and then determined the differences based on indirect evidence suggesting the presence of exosomes.
5. Yes, the results were comparing different methods. They do not show direct proof that the particles in TEM images are exosomes. There is no determination as to how these particles function as this can not be observed in a dead state in TEM.
6. Repeatability is definitely important but step 6 is testing the hypothesis by the original researchers through experimentation as necessary in order to confirm whether or not the proposed hypothesis holds up.
“My challenge to Mike Stone is to find any scientific paper that has achieved 100% purification (complete purification) ie particles in a vacuum with no other different particles in the presence of the studied particle.”
This is a burden of proof reversal fallacy. I am not claiming such a paper with complete 100% purification exists. I state the opposite. You are seemingly admitting that 100% purification can not be achieved. It appears that we agree this evidence of 100% purification does not exist. Why should I produce proof of something we both agree is impossible for exosomes?
You also state this:
“I also question why it is necessary to have ‘complete purification’ in order to prove the existence of something. What level of purification is sufficient to observe cause and effect?”
Why would complete purification not be necessary? How would one know which particles are the ones being searched for if there are other substances contained within the sample?
A reminder on what purification means:
“Purification: the act or process of making something pure and free of any contaminating, debasing, or foreign elements”
In regards to chemistry:
“Chemical purity refers to an element contains a SINGLE SUBSTANCE, without any other element tarnishing its STANDALONE EXISTENCE.”
In order to know the thing you are looking for exists, it must be seen in a purified/isolated state free of all contaminants, foreign elements, pollutants, etc. This is the only way to have a valid independent variable in order to prove cause and effect. This is the only way to biochemically and molecularly characterize the particles alone. This is the only way any TEM images would have any meaning.
Without purification and isolation, problems arise in identifying the identical particles as stated in the exosome article I shared:
“How can we be sure that we are isolating and quantifying extracellular vesicles rather than enveloped viruses present in the sample? Equally, how can viral researchers know that they are not detecting similarly sized non-viral vesicles or empty vectors during vaccine production?”
You can take out “virus” and add any MVB’s or extracellular and intracellular particles and the meaning remains the same. They can not idistinguish between the particles as they are all of similar size/shape.
The problem with your paper is that it assumes exosomes exist and that they have a certain size and shape. Where was this determined originally? Obviously not in 1981 when the exosome concept was first ntroduced. Just as with “viruses,” the original DIRECT proof of purified/isolated exosomes must be shown first before any paper from 2018, which starts off by assuming their existence, would have any merit.
I again cannot reply to Mike’s comment directly. So here is another response.
My question for him was the following:
“My challenge to Mike Stone is to find any scientific paper that has achieved 100% purification (complete purification) ie particles in a vacuum with no other different particles in the presence of the studied particle.”
Mike responded with the following:
“This is a burden of proof reversal fallacy. I am not claiming such a paper with complete 100% purification exists. I state the opposite. You are seemingly admitting that 100% purification can not be achieved. It appears that we agree this evidence of 100% purification does not exist. Why should I produce proof of something we both agree is impossible for exosomes?”
In my question to him, I am not asking for a scientific paper that purifies exosomes 100%. I am asking for any scientific paper that purifies ANYTHING 100%. Could be gold, could be water, could be anything. I don’t think it is possible to have 100% purity of anything. This would of course require a vacuum to be used as you need to ensure there is zero possibility of contamination including the container that is being used.
Basically, I think Mike is asking for something that is impossible but I am open to being challenged to this and will accept even 1 scientific paper that demonstrates 100% purity of a thing.
I think the difference Mike and I have is that he requires 100% purification of a thing to prove it’s existence. I do not agree with that notation and believe it is impossible to have 100% purification.
Also citing papers that claim that viruses and exosmomes are inseparable is no different than stating unicorns and exomsomes are inseparable. I accept the concept of different degrees of purity. If I have 99.99999% gold, I don’t dispute the fact that gold exists, has properties that can be observed, and so forth.
It also seems that Mike is disputing that Ultra Centrifugation is the gold standard for purification of small things. I have read this in many papers and even Andrew Kaufman has said it numerous times. Does Mike have any scientific papers disputing this?
In order to prove the particles claimed as exosomes (or “virises”) are what they are claimed to be, they must be purified and isolated away from everything else within the sample. This must be done in order to determine their biochemical, morphological, and molecular composition. This can only be done using completely purified and isolated particles. Regardless of whether they are called exosomes and/or “viruses,” the same principles apply to both. If not purified and isolated, any of the other substances contained within the sample could be the thing that is being looked for and potentially the cause of any observed effect (if any).
As for purification, I already provided the definitions showing that the substance must be alone. If this can not be done, then that is a problem for exosome researchers and virologists.
In any case, this first source offers many methods commonly used to obtain pure chemical compounds, which they define as:
“A pure substance contains ONLY ONE KIND of molecules while an impure substance is a mixture of different kinds of molecules. Thus, to purify an impure substance is to SEPARATE THE DESIRED MOLECULES from the mixture and therefore, the purification is the SEPARATION.”
There is also a section relating to distillation in a vacuum such as used when obtaining salt from salt water.
According to this next source, purity is required for identification. The melting and boiling points of a substance is used to determine purity:
“FOR THE IDENTIFICATION OF A COMPOUND, qualitative analysis of pure substance is required. Therefore, first we have to purify the substance and then check
its purity. There are many techniques namely, crystallisation, distillation
sublimation, chromatography etc. available for purification of a compound. In this unit you will learn about crystallisation as a technique for purification of a
compound. The purity of a compound may be checked by determining its melting or boiling point. The technique for determination of melting and boiling points will also be described in this unit. Pure solid and liquid compounds possess
sharp melting and boiling points. Therefore, melting and boiling points of a compound can be used as a criteria of purity.”
Regardless, even if they were somehow able to obtain 100% purification, this still would not prove the particles in the images do what is claimed of them. As the particles are only seen in a dead and heavily altered state in TEM, there is no ability to observe how they function within a living organism nor whether they exist as imaged within a living organism.
So if you are going to claim exosomes exist, you must show them in a purified/isolated state and show how their functioning inside a living organism was determined. It’s funny how you can clearly see that “viruses” are unicorns yet then claim the same exact unicorns are actually called exosomes.
What do you possibly hope to gain from such a position?
Only the perception of an undermining of another’s claims?
So you cant isolate rats from mice or frogs?
Sugar from salt?
So as to characterise unique identifiable characteristics?
At far from human scales, the means to observe become artefacts to their instrumentation and interpretation of data finds according to what is sought..
As there are always more factors than can be 100% guaranteed to be currently known, you can assert knowledge impossible whenever it suits you not to accept the practical import of a situation.
The theories that are being questioned and found lacking are foundational to a global coup, set over global mass poisoning and live human experimentations whose results may be far worse than death.
If a particle or packet cannot be definitively demonstrated, its supposed function returns to the mind that made it. You can have your own belief, but you have no power to make it true.
That you (and many others) WANT to believe it, is to my view an interesting symptom of a narrative identity of personal & social masking that can be seen as a conditioned mindset operating as autonomous habit instead of relational awareness. Humanity sets conflicts as the basis of an identity of protected conflict. It protects by hiding or masking over a lack of true foundation. That’s it’s purpose, to run on its ‘own’ premises rather than share in reality. Of course no one can achieve this 100%. Or in truth at all, but a distortion of function can seek to gain a self from a sense of broken function, as in All the king’s horses and all the king’s men, and that is the nature of ‘seek and don’t find’ running the form of a reality while actually running an obfuscation. Good Day!
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“It’s funny how you can clearly see that “viruses” are unicorns yet then claim the same exact unicorns are actually called exosomes.”
Mike, that’s a low blow. Viruses as submicroscopic terrorists without boxcutters are unicorns.
Michael S, Mike doesn’t believe in genes or genomics, so you’re not going to be able to reason with him regarding RNA/DNA messaging. He’s not interested. That’s just where he’s at.
He also has an extremely narrow view of the scientific method that doesn’t admit simple deduction as part of the method. If you tell him that the ultra centrifugation of exosomes clearly yields exosomes and other things that according to the chemical and morphological analyses are obviously not exosomes, then it’s all null and void to him. That’s where he’s at, and I’m sure he’s not the only one.
It’s a good thing that you mentioned Kaufman is down with ultra centrifugation as an isolation method.
And it’s great you brought this information to the conversation. I’ve been talking up proteomics for a bit here but I have no background in biochemistry and haven’t really even researched exosomes. I just operate on feel.
When we urinate exosomes onto the pasture we are participating in the homeostatic regulation of the cellular pool. In springtime those exosomes have a great chance of ending up in microbes with their informational formats intact, for whatever they’re worth.
Can you please show where exosomes were originally purified and isolated from the fluids and then characterized biochemically/molecularly?
Can you please show where the functions ascribed to exosomes was proven?
Can you please share how the particles known as exosomes are different from those claimed to be “viruses” without using theoretical functions to separate them?
Ultracentrifugation is considered the “gold standard.” I never disputed this. However, I have shown in numerous sources that Ultracentrifugation can not properly purify nor isolate the particles as claimed.
Mike, can you please provide a single scientific paper that achieves 100% purity of a substance?
Regarding chromatography (also known as immunoaffinity), that is a common purification method used for studying exosomes.
This paper is on topic to your concerns: “Isolation of Exosomes from Blood Plasma: Qualitative and Quantitative Comparison of Ultracentrifugation and Size Exclusion Chromatography Methods”
Did you read this paper before sharing? It pretty much destroys your contention that ultracentrifugation is valid for purification/isolation and the researchers limitations stating other proteins and impurities may have been in their SEC preparations show that the findings are invalid:
“Reproducible isolation of biologically active, intact exosomes with high efficiency and WITHOUT IMPURITIES from as complex matrices as plasma IS TECHNICALLY CHALLENGING AND HAS NOT BEEN STANDARDIZED. Exosome isolation from plasma, as from cell cultures, is often performed with UC-based methods, ALTHOUGH CONCERNS ABOUT EFFICIENCY AND PURITY HAVE BEEN RAISED . Recently, it has been proposed that with size exclusion chromatography (SEC) exosome isolation from blood plasma can be performed without significant impurities [16–19]. HOWEVER, THE EFFICIENCY OF EXOSOME ISOLATION BY SEC IS STILL DEBATED.”
“To assess the amount of impurities in isolates, albumin was chosen as a characteristic marker, since it is the most abundant plasma protein. In addition, liquid chromatography–mass spectrometry (LC-MS)-based proteomics experiments revealed that ALBUMIN AS WELL AS A NUMBER OF OTHER PLASMA PROTEINS ARE OVERREPRESENTED IN THE UC-DERIVED PREPARATIONS (Table A in S1 File). Accordingly, UC- derived isolates CONTAINED A SIGNIFICANT AMOUNT OF ALBUMIN as assessed by Western blots (Fig 1B).”
“Furthermore, ALBUMIN WAS PRESENT IN ALL SUBSEQUENT ISOLATES. Then we analyzed the efficiency of UC while modulating various parameters (such as sample viscosity and sedimentation distance) which might potentially influence the efficiency of the exosome isolation. UC of 10-fold diluted blood plasma (experiment (f)) or UC with smaller sedimentation distance (1 mL sample; experiment (g)) resulted in approximately the same yield as UC of 7 mL 2-fold diluted plasma. ALBUMIN WAS ALSO DETECTABLE AFTER EACH UC CYCLE IN THESE EXPERIMENTS (Fig 2C). The efficiency of UC of approximately 7 mL plasma at 37°C (experiment (e)), however, proved to be higher than that of the UC performed at 4°C since after each UC cycle less CD63 was present in the consecutive pelleted exosome isolates. HOWEVER, ALBUMIN CONTENT OF THE ISOLATES WAS STILL SIGNIFICANT (Fig 2C).”
“Some vesicular structures could also be identified after 6h and 14h UC, although a HIGH AMOUNT OF AMORPHOUS MATERIAL OBSCURED THEM. DLS analysis established that 1h UC samples contained predominantly exosomes with expected size range, however, after 3h, 6h and 14h of UC a smaller proportion of particles fell into the exosomal range (Fig 3C).”
“To assess efficiency of the isolation, equal volumes of SEC fractions were analyzed by Western blots, while purity is tested by loading equal amount of protein. In case of Sepharose 2B, CD63 and TSG101 were detected predominantly in fractions 6–7, BUT ALBUMIN WAS ALSO PRESENT IN THESE FRACTIONS (Fig 4A). However, fraction 6 from Sepharose CL-4B and Sephacryl S-400 columns presented exclusively CD63 and TSG101 signals without significant albumin impurity (Fig 4A). By loading equal volumes of SEC fractions, the vast majority of CD63 and TSG101 WAS CO-ELUTING WITH ALBUMIN IN CASE OF EVERY MEDIUM, WHICH INDICATES THAT SEPARATION EFFICIENCY WAS LOW, approximately below 1% (Fig 4A).”
“Here we evidenced that exosomes can be isolated by SEC from blood plasma without significant amount of albumin BUT NOT BY UC, and that ONLY A MINORITY OF EXOSOMES CAN BE ISOLATED BY SEC OR UC FROM THE BLOOD.”
“Efficiency of exosome isolation methods and the purity of isolates HAVE NOT BEEN THOROUGHLY PREVIOUSLY CHARACTERIZED FOR BLOOD PLASMA. Although UC has been the most commonly used method to isolate exosomes, here we found that only a minority of exosomes could be isolated from the blood plasma. This is in agreement with previous reports, WHERE THE EFFICIENCY OF EXOSOME ISOLATION FROM HIGHLY VISCOUS CELL CULTURE MEDIA WITH UC WAS LESS THAN 5% .”
“Nevertheless, it should be mentioned that the determination of efficiency in our and abovementioned studies is ONLY AN APPROXIMATION, since, to date, there is NO METHODOLOGY PROVIDING EXACT QUANTITATIVE ANALYSIS OF EVs THAT MEETS MOST CRITERIA, e.g., SELECTIVITY, REPRODUCIBILITY AND ROBUSTNESS. To assess the presence of impurities, here we demonstrated that ALBUMIN CONTENT OF UC-DERIVED ISOLATES WAS SIGNIFICANT. This is in agreement with previous findings where the majority of proteins, including albumin, detected in UC-isolated exosomal preparations, was unrelated to the conventional exosomal proteome . Since albumin is the major soluble plasma protein and a predominant component of the isolates, WE CONCLUDE THAT UC IS NOT AN OPTIMAL METHOD TO OBTAIN A PURE EXOSOMAL PREPARATION i.e. as needed for in vivo experimentation or for analytical assays (e.g., proteomics or RNA analysis), however, exosomes isolated by UC might be appropriate for analyses WHERE CONTAMINATING MATERIALS DO NOT INTERFERE WITH MEASUREMENTS.”
“In contrast, DLS analysis revealed that the mode diameter of the predominant particle population in 3h UC isolates was 18 nm indicating that the MAJORITY OF THE PARTICLES MIGHT BE PROTEIN AGGREGATES RATHER THAN VESICLES. This was further confirmed by TEM, WHERE SIGNIFICANT AMOUNT OF AMORPHOUS SUBSTANCE OBSCURED THE VESICULAR STRUCTURES. Nevertheless, these results highlight that experiments with exosomal isolates obtained with UC require careful designing and well-defined controls, such as applying detergent treatment .”
“Interestingly, in our experiments, the purity of exosomal isolates obtained by the most commonly used media Sepharose 2B WAS INFERIOR TO OTHER INVESTIGATED MEDIA. This observation highlights that systematic analysis and optimization of SEC-based assays is still warranted. Here, we also found that SEC performed on 10 mL columns, is NOT AN EFFECTIVE METHOD FOR THE ISOLATION OF EXOSOMES, its efficiency is comparable to those of the UC-based methods. This finding CONTRADICTS RECENT RESULTS which show that commercially available 10 mL ExoSpin or qEV columns separate exosomes from soluble contaminants with high efficiency [19, 24].”
“In this regard, here we show that SEC performed on a 120 mL Sephacryl S-400 column does not result in a significantly improved yield, suggesting that SEC MAY HAVE ITS INHERENT LIMITATIONS FOR EXOSOME ISOLATION FROM BLOOD PLASMA.”
“Here we assessed that exosomes can be isolated by SEC without significant albumin content, however, this study has limitations. The purity of exosomes isolated by SEC WAS ESTIMATED BY THE PRESENCE OF ALBUMIN ALONE. Although albumin is the most abundant serum protein, OTHER SOLUBLE PROTEINS MIGHT DISTRIBUTE DIFFERENTIALLY AND MIGHT STILL BE PRESENT IN OUR PREPARATIONS. In addition, we cannot exclude the possibility that serum proteins, especially albumin, and exosomes interact specifically, AND THEREFORE CO-ELUTE FROM SEC COLUMNS, which might explain the low efficiency of our SEC experiments.”
“In summary, here we demonstrate that SEC but not UC is suitable for the isolation of exosomes from blood without significant albumin contamination, ALTHOUGH IT’S EFFICIENCY NEEDS TO BE FURTHER IMPROVED. These results suggest that experiments on exosomes isolated by UC or SEC SHOULD BE EVALUATED CAREFULLY AND BY APPLYING APPROPRIATE CONTROLS.”
Now, before you continue to throw out recent studies claiming (but not showing) purification of exosomes, please share where exosomes were ORIGINALLY purified and isolated from the fluids, where the purified/isolated particles were characterized (biochemically, morphologically, and molecularly), and where their ascribed functions were proven. Otherwise, all you are doing is trying to show pictures of particles in TEM as proof which you should know by now is utterly ridiculous.
Can you please provide even one scientific paper that achieves your 100% purification metric on ANYTHING (could be gold, water, etc) in a vacuum in absence of all other particles?
As I stated before, I don’t think 100% purification is possible but stand to be corrected if you can show me it. Do you have a scientific paper that proves 100% purification is even possible? What you have shared is some definitions from people making theoretical statements but of course were not supported by the scientific method.
As I do not think we will get anywhere regarding debating the existence of ANYTHING especially of exosomes if we do not sort out our definitions and what is possible in reality.
I’ve already supplied definitions for purification. We both agree that 100% purification can not be achieved. Why are you still focused on this? Something being “pure” has a specific meaning, i.e. being free of contaminants, pollutants, foreign elements, anything that debases it, etc. Isolation also has a specific meaning, i.e. separated from everything else:
“In chemistry, a pure substance is an element or compound MADE UP OF ONE TYPE OF PARTICLE. Pure substances have a fixed structure with definite properties. Purity is the measurement for how pure a substance is. For instance, for a substance to be considered pure IT MUST BE MADE UP OF ONLY ONE ELEMENT OR COMPOUND. For gold to be pure it must have only gold atoms contained in it. Likewise, for water to be considered pure, it must have only water compounds.”
“In Chemistry, the concept of pure substances is that they MUST HAVE A PURITY OF 100% TO BE CLASSIFIED AS PURE. This means that they can only contain the same elements or compounds throughout. However, in reality only some substances can be considered pure. Most pure substances such as table salt have a purity rating of about 99.9%.”
“In order to be a pure substance, the substance MUST CONTAIN ONLY ONE TYPE OF ELEMENT OR COMPOUND. For example, gold only contains gold elements. If there is another substance present, then it is considered a mixture.”
It is obvious that, when getting down to the size of nanoparticles, this purification and isolation can not take place. This is admitted by the researchers. Thus we are left with degrees of purity, which is an oxymoron as something is either pure or impure. What is an acceptable degree of purity to you? 99.99%? Have you seen this claim with exosomes? How about 80%? 50%? How many impurities should be allowed?
When dealing with discovering a new entity, in order to characterize and study it, the entity must be in a purified/isolated state. If this can not be achieved, then the entity can not be studied.
Why is this purification and isolation important?
Basic concepts of separations
“This section is concerned with separations of the smallest subdivisions of matter, such as atoms, molecules, and minute particles (sand, minerals, bacteria, etc.). Such processes start with a sample in a mixed state (composed of more than one substance) and transform it into new samples, each of which—in the ideal case—CONSISTS OF A SINGLE SUBSTANCE. Separation methods, then, can be defined as processes that change the relative amounts of substances in a mixture. In chemical methods, one may start with a completely homogeneous mixture (a solution) or a heterogeneous sample (e.g., solid plus liquid); in the act of separation, some particles are either partially OR TOTALLY REMOVED FROM THE SAMPLE.
Reasons for making separations
There are two general reasons for performing separations on mixtures. First, THE MIXTURE MAY CONTAIN SOME SUBSTANCE THAT SHOULD BE ISOLATED FROM THE REST OF THE MIXTURE: this process of isolating and thus removing substances considered to be contaminants is called purification. For example, in the manufacture of synthetic drugs, mixtures containing variable proportions of several compounds usually arise. The removal of the desired drug from the rest of the mixture is important if the product is to have uniform potency and is to be free of other components that may be dangerous to the body.
The second reason for performing separations IS TO ALTER THE COMPOSITION OF A SAMPLE SO THAT ONE OR MORE OF THE COMPONENTS CAN BE ANALYZED. For example, the analysis of air pollutants to assess the quality of the air is of great interest, yet many of the pollutants are at a concentration too low for direct analysis, even with the most sensitive devices. Pollutants can be collected by passing samples of air through a tube containing an adsorbent material. By this process the pollutants are concentrated to a level such that straightforward analysis and monitoring can take place. In a second example, SEVERAL IMPURITIES IN A SAMPLE MAY INTERFERE WITH THE ANALYSIS OF THE SUBSTANCE OF PRIMARY INTEREST. Thus, in the analysis of trace concentrations of metals in rivers, organic substances can cause erroneous results. THESE INTERFERENCES MUST BE REMOVED PRIOR TO THE ANALYSIS.”
As researchers admit that what are claimed to be exosomes can not be isolated from particles of the same size/density (or smaller), there are potentially millions of other known and unknown substances within the mixture that may be the cause of any observed effect.
Beyond the lack of purification and isolation, the heavy alterations done to the sample could be the cause of the particles seen, whether from the increased forces applied from centrifugation breaking bigger substances into smaller ones, filtration breaking the bigger particles into even smaller particles, or the prepartion process for TEM imaging (fixing, dehydrating, staining, embedding) causing artefacts in the images.
Now, moving along, can you please provide the ORIGINAL paper for the complete purification/isolation and characterization of the particles assumed to be exosomes and show how they were discovered using the scientific method?
Rather than write more here I put a response at
I feel it speaks to what runs beneath our capacity to entangle love in conflict. For no one engages in anything utterly valueless or meaningless to them.
You’re not reading for comprehension. You are reading for confirmation.
The proteins in the exosomes have lettered and numbered names because they are recognized informational proteins, hormones, or whatever, as established by PCR. Mullis, being the inventor of this cataloging method, would concur (appeal to authority). The other remaining molecules that are unavoidably present, such as albumin, do not cross-reference with the bread of cataloged exosomal contents.
The truth always requires not throwing out the baby with the bathwater. And the truth often requires triangulation, even in normal, everyday life, and the beautiful thing about that is that you can’t wrap it up and put a bow on it. Because in 3D it’s a pyramid and the tighter you tie the bow the faster it will just keep slipping off. The all-seeing eye at the top of the pyramid is for them is a symbol of mastery.
You avoided my questions to you. I take it you can not answer them?
Regarding your current comment, how were these exosome proteins discovered? Where is the ORIGINAL purification, isolation, and characterization of the particles claimed to be exosomes? Where were the functions of these particles established? This obviously must come first before “exosome proteins” can be identified.
I’m sorry, friend, but the answers to all your questions have been made abundantly clear already. There’s no turning back the clock.
No, they were not. I’m sorry you can not see this but if you can not provide the ORIGINAL proof for the purification/isolation, characterization, validation, etc. of the particles claimed to be exosomes, you are chasing the same unicorns virologists are chasing.
I cant see where the basis is for your statement?
Why not link to the answers instead of a condescending dismissal affecting apology?
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Easy, brian. I’m not being affectatious, condescending, or dismissive, even though Mike is asking (rhetorical) questions that with total certainty will have zero bearing on his already fixed position. I’ve been discussing the intelligence behind horizontal gene transfer and exosomes for a couple months here. And I’m not about to do it again in this discussion in the face of the implacable force that is the tautological nature that is Mike’s ‘flat earthist’ denialism, and given the additional info Michael has presented. When Mike gets argumentatively backed into a corner he doesn’t like he goes into masking mode which is when I usually say to him that unless he disagrees I think the conversation has hit a dead end.
Loved your comment over at your place btw.
How have I been backed into a corner? I’m asking for the foundational evidence used to claim the existence of exosomes. This should include purified/isolated particles that have been characterized. The study should adhere to the scientific method which requires a valid independent variable. Presumably, the purified/isolated particles could be used to determine cause and effect yet that is seemingly impossible for exosomes. All I have been shown is images of particles of varying “purity” that have been ascribed unproven functions. Do you have better evidence Reante?
Mike you having claimed that I am hunting unicorns just like virologists may just be shadow play on your part insofar as you have made a personal religion of ‘science’ just like they have made for themselves a collectivist one. You’re the only one who can know that for sure. WADR.
You can not provide direct proof that exosomes exist and function as claimed. How else should I describe it rather than hunting unicorns? Have you even looked into the foundational evidence for exosomes before believing in the concept?
According to your small religious method, you must not believe that sperm look like tadpoles, and
you must believe that IVF doesn’t exist. If it’s in vitro then to you it’s not real, and THAT is a profoundly broken understanding of reality borne of fear and denial. You refuse to talk about anything that you do believe so as to avoid having your contradictory beliefs exposed. Well you can’t hide them from people who can still deduce the truth.
You use are using foundational and ORIGINAL as if they have some added value when in actuality they have no value the way that you are using them. SEEING accurately is the ORIGIN of science and every day is a new day for science. Experiments are merely confirmational. Or not.
Let it be known that the Q-anon flat earthism of naysaying whole swaths of reality in light of obvious evidence to the contrary is easy, convenience food borne of a maximal mental fragility separation trauma. It’s a foodborne illness. Intellectual GAPS.
Collapse scares the pants off of people in various ways. Denial has many pathways. Religions come in many guises.
The (deduced) idea is to honor those of our ancestors that died on their feet more than we honor those that bore us that were bred by people farmers. Because they have much more to offer us.
You continually avoid answering my questions. Your sperm comment is ridiculous. Obviously sperm can be seen in a microscope in an alive state. What does that have to do with exosomes which can only be seen in a dead state in TEM after heavy alterations from the culturing (in many cases), ultracentrifugation, filtration, fixing, dehydrating, staining, and embedding processes?
The foundational evidence obviously matters. This is where the original evidence claiming the existence of the particles and their supposed functioning comes from which all other studies are built off of. Why would that not matter? What evidence did you see that proves to you that exosomes exist inside us in the state they are presented in TEM and that the particles function as claimed?
I’m not avoiding anything. I’m just not playing your masking game that you are playing so as to save face. All your questions have been answered sufficiently and my feeling is that most of the readership here recognizes that.
You attempt to mask PCR by fixating on TEM. PCR are biochemical eyes. I realize that you are unable to accept that at present.
So you rely on PCR as evidence? I’m guessing you mean the theoretical genome. Please share where exosomes were purified and isolated away from everything else in order to make PCR results and the genome relevant.
None of my questions have been answered by you or anyone for that matter. You avoid sharing any evidence that you have seen that proves exosomes exist and function as claimed. It is a belief on your part not based on direct evidence.
Strawman. I don’t mean the “theoretical genome” as in another of Mike’s flat earth unicorns. I mean the patterning of chemical compounds. PCR is capable of patterning the body’s chemical compounds.
You have a good day friend and I will too.
And how does this prove exosomes and their functioning?
Is it not the fact that PCR can only detect the primers that it is set to detect, which are short code fragments of proteins that in themselves do not constitute anything in entirety, such that they can only ‘see’ what you already know, so as to set unique identifying primers to ascertain for example sequence ONLY found in chickens (if such a thing exists)? This quite apart from artefacts of the exponential sample doubling by which Mullis said you can find ANYTHING if you run it enough times?
You HAVE to first KNOW what you are looking for. This means fining it empirically which is what we mean by isolate. Else you find a source of billion dollar patents that is simply snot with GoF applied
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No, they can synthesize whatever primer they want. They can synthesize something that doesn’t exist and amplify it 100X and get no result.
When Mullis said “anything” he was talking about abusing the tool for the purpose of diagnostical medical fraud. Taken out of that context, “anything” is a testament to the profound power of PCR.
PCR is the applied science of the Spiegelman’s Monster experiments I mentioned soon after I arrived here, which are the canonical self-assembling RNA ‘conscious resonance’ experiments from the 70s (?) that Tom Cowan refers to. PCR tests are living cultures of genetically modified, self-assembling DNA. They are growing DNAs in real time, at the speed of bacteria (because they’re using bacterial polymerase) so that the computers can calculate the relative population of the target DNA they’re interested in. Relative population heralds the relevance of the target in any specific sample.
With exosomes we’re talking about intelligently packaged particles of a very specific range of size and with a ‘genetic’ cargo found in ‘clean’ bodily fluids and legitimately isolated. If the cargo is nucleic acids then it is ‘genetic’ because that is the word chosen to mean informational. If someone wants to use, say, akashic (memory) instead genetic that’s fine, so long as the function doesn’t change.
Mapping the genetic chemical compounds of the body requires a tool that can read compounds, a tool that can make compounds, a mass production facility, supercomputers, and mass human intelligence. The human intelligence provides the process of elimination framework. Choose a target length of DNA and make a primer for it. Take samples, get results. Take samples, get results. Take samples, get results. Take samples, get results. Compare results. Pattern forms. Reduce the amplification. Increase the amplification. Do the same for another target. And another. Compare results. Pattern deepens. Targets start coming together in silico all “watched over by machines of loving grace.”
They map the informational network underpinning Life by synthetically growing the puzzle pieces such that they can see them in very specific contexts, contextualize the context, and contextualize the context in ever greater context until the picture forms.
That’s where we’re at. Believe it or deny it, your choice.
“With exosomes we’re talking about intelligently packaged particles of a very specific range of size and with a ‘genetic’ cargo found in ‘clean’ bodily fluids and LEGITIMATELY ISOLATED.”
Shall we look at the definition of isolation again?
“to separate from another substance so as to obtain pure or in a free state”
Can you show where exosomes were separated from everything else in a pure or free state?
You mean they can synthesis short strands of DNA that don’t find any match in nature?
Like I can write a made up word that doesn’t appear in any book ever written?
I see that you are enthusiastic from your current sense of possibility.
I don’t trust it at all or rather it doesn’t offer me anything that I want.
The capacity for consciousness to generate results in the world that confirm it is natural to the desire to have the experience. There are a huge number of non reproducible experimental results. Consciousness as a hidden variable that can never be removed.
Computer modelling (applied math) is what post-truth ‘science’ runs. Modelling outcomes according to input parameters.
What I read is ‘All the king’s horses and all the king’s men putting Humpty together again’.
Reverse engineering according to ‘ideal’ outcomes. What could possibly go right?
I recognise your current choice, perhaps you can accept that my withholding acceptance of its claims is not a denial of your own. Inheritances are evident. Even as a result of what people see on tv. My sense is that memory is stored in field effects not in DNA, but as I say I keep brackets around anything that hasn’t my full acceptance yet there are functions of inheritance as the very fabric of space and time.
Yes but they don’t work as primers so they can’t do anything with them. I was just using that as an example in response to your previous comment.
I’m a enthusiastic seeker and seer but this dark science holds no possibility for me. It’ll all largely be washed away in a few short years anyway.
The exosomal topic holds no possibility for me either. The interconnectedness is already evident. But the truth of it still matters. The exosomal research is already being used to allopathic dark ends. If the vaxxxes are what they say they are then they are a practice in deleterious synthetic exosome bombing.
The DNA *is* the field effect. The anti- materialist, ‘quantum’ (or whatever) reactionary swing away from materialism has no more basis in Reason than the idea that matter exists. The holograph is the field effect. The appearance of the hologram — ‘what’ it is — is the localized effect that consciousness imposes on the energy field. That’s the field effect. Why would there be anything else, as if that’s not enough?
The planet earth is the collective, spherical effect of the collective consciousnesses applied to the local field.
“I’m a enthusiastic seeker and seer but this dark science holds no possibility for me. It’ll all largely be washed away in a few short years anyway.”
Well yes, everything changes, but the fact remains that what we give sets the measure of our receiving. therefore to know or be certain in our own purpose and desire is the find a grounded reference point of recognising and appreciation ourselves in each other and everything.
For while we have a temporary or time-bound experience of a life, we each have a unique perspective within the Infinity of being that partakes of it – knowingly or not.
The nature of our will is that it is free to accept reality or not, in time. But what we accept will either know itself free or at some point recognise it has been phished or mis-directed to seek freedom from self-conflicted premise, that automatically redefines will and freedom to its own framing. So here the experience of ‘being lied to’ but also the awakening responsibility for what we give authority or credibility to. And as a result sharing or growing the culture of such a renewal of the mind we thought to have lost to lies and denials that are never wholly attributable to others or to an ‘othered’ terrain, but are an expression OF terrain in unique expression. This likewise automatically restores Cause to it total Event, rather than casting out to a fragmentation of conflicted causes seeking dominance or even the right to be.
Sure. Mike’s cybernetic, fallacious use of repetition would make Edward Bernays proud. Flat-earth-mind zombiehood is just another mental phenotype of the obedient ‘sheep’-like behavior of the mainstream that shoots itself up with these shots on command. The total Event is a separation trauma that has cut to the bone over and over again for generations. It’s the serial raping of man by civilization. So the stunted psychologies of some grown men go into the flat earth because ‘nothing can get them there.’ In the schizoid flat earth they get have their cake and eat it, too; maintain the spoils of a no-responsibility mainstream industrial lifestyle and at the same time completely psychologically divorce themselves from it. Obviously that’s a common function of Christianity too hence the big overlap between the two (along with biblical material they draw from.) When Mike goes into flat-earth zombie masking mode just like his zombie flat-earth mentor Jordan Grant does, it is that same protective, glazed-over, ‘you-can’t-hurt-me if-i’m not-here’ shell that little boys and girls go into when they are being serially raped. But in Mike’s case it made me angry because he’s a grown man and the ‘raping’ is simply not of the same caliber as child rape, and part of what is contributing to Mike’s zombie shell mode is lust for excessive power — aka what Clarifire said: being an influencer. Of course, that’s consistent with victims of child rape too. They alternate between shell mode and some form of power abuse as was imprinted upon them by rapist. I don’t mean to make too much of it — zombie shell rape victim modes are just extreme masking modes — but the parallel is obvious. I obviously have a lot of experience with wielding Reason during argumentation but but all the same Mike’s extreme masking in light of the two very straightforward and reasonable isolation papers Michael presented was shockingly irresponsible. If you put yourself up on a soapbox as a blogger, your responsibility to the truth only increases otherwise you effectively become one of them. They love disinformation even if it’s antiestablishmentarian. They love it because they hate the truth more than anything, and it kills the truth for the marginal seeker, which is what Mike just did, WADR to him.
We’re all adults here. No hard feelings.
I think you need to take a break Reante. Your accusations against me are getting excessive. I want to allow free and open discussion here but you are now attacking myself and my friend Dr. Jordan Grant. We can have discussions without resorting to ad hominem attacks. If you think I am against truth, you are mistaken. Just because your truth does not align with mine does not make me against truth any more than it does you. I have a different set of standards as to what I require as evidence which is based on logic and the scientific method. I require direct proof to be convinced that what I am being sold is true or not. You do not seem to need it.
If I repeat myself, it is because both you and Micheal failed over and over again to address anything I said. I looked at the “isolation” papers and pointed out the flaws. If you disagree with my assessment, please feel free to counter. I’m happy to debate it.
Your ‘direct proof ‘ is a flat-earth parlor trick you play on yourself and everyone else. Your children, above all, deserve better, because that self-protective parlor trick is a short-term patch that serves a coping mechanism, and the future belongs to those children being raised by Reason -based, long-term thinkers because the future belongs to those who have the best grasp of Reality. Twas ever thus.
I appreciate you having me Mike. You have a good heart. And thanks to Clarifire and Brian. Take care.
You continue to hide behind ad hominem attacks rather than answering anything I said directly. I am happy to have a discussion around the evidence presented and you are free to counter but you do not seem to want to have that discussion.
But apart from all this pettiness – what’s the strongest argument for exosomal functions via extrusions or breakdown products?
I can theorise that cell function and even breakdown carries information and resources that serve other cells. The nature of the information may be ‘packets’ of proteins, water memory, enzymatic catalysts, or indeed nano-unicorns, but I don’t know specific mechanisms for certain. But I am open to the idea of something like it as I see life as one in many and many in one – rather than as war upon itself.
I can hold an idea in brackets – as awaiting further clarification as to what is speaks of and where it belongs. I now see viruses belonging to the mind that thinks them, for the purpose they derive from doing so. Not to the living energy-expression of body.
The world is a screen onto which we project, but can be a living relationship extended and shared in.
Why do you defend the idea of exosomes as beyond question for you?
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It’s not pettiness brian. It’s life. We’re animals. That you call it pettiness makes you an animal too.
Everything’s ok, friend. It’s just another fork in the road.
Reducing to personal put downs is petty or locking down to trivia set large so as to block the issue. You can believe you are an animal, but the fact you have that choice proves otherwise!
In ego death, brian, put downs don’t exist. In ego, ‘put downs’ are looked-for in order to mask the message coming one’s way.
Sometimes I bring heat exactly in order to give the receiver the excuse he’s already looking for. Heat signifies my having given up on the person, in the moment at least. And obviously it’s also a reflection of a sense of betrayal – to the truth. Not to me, but to the truth.
We are biological animals, brian. Mammals. Pack animals. There’s no choice there. If you repress that instead of (hopefully) harnessing it constructively, well, there goes Reality again, which doesn’t bode well at all for the sustainable existence of the species.
The ego – as a part of your own thought – will never die. But as the idea of a separated self, never truly lived.
I can see put-downs as the strategy that they are to serve the purpose given them. But You are right – they are nothings to the unsusceptible.
Not unlike the idea of contagion.
Why are you asking me why I defend the idea of exosomes as beyond questioning? Why *would* you ask me that?
Mike is not questioning exosomes. He’s shitting all over them, which means he’s shitting on honesty.
I’m asking for the direct proof for the existence of exosomes and their function. You can not provide it. As I said, this is a belief for you not based on sound scientific evidence. It is OK to believe in exosomes without proof if you want. However, I require actual direct evidence proving they exist and function as claimed.
Like anyone questioning viruses is shitting all over people who get sick and die?
Give me a break!
You are CHOOSING to take offence needlessly.
An honourable way might be to say that at the moment you haven a means to articulate what you feel or intuit to be valuable, pertinent and worthy, so you will come back on it after further reflection and research.
If the Tower of Babel collapses as I see it is and must, there is the task of establishing true and truly workable foundation from which to live. Not as good intentions or ideals – but workable as in holding integrity of function under all conditions.
Much has been built on or with false presumptions by which we suffer a destructive result.
Particularly where invested models that fail are propped up with fudged data and complex extensions by which to save the appearances.
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“Like anyone questioning viruses is shitting all over people who get sick and die?
Give me a break!
You are CHOOSING to take offence needlessly.”
I’m choosing to take offense to be sure, because Mike’s conflating exosomes with viruses and belittling Michael (and myself by extension) is a dishonest and telling thing to do, and I take offense at that. YMMV.
Nobody has the means against stonewalling, brian, whether the stonewalling is conscious or just shadow play. Apparently you don’t see the stonewalling so you think I’m just engaging in shadow play myself and Mike is the one on the up and up. That’s fine. Just another he said she said in a relativistic world of pack animals living a multikulti existence. What else is new? 🙂
In what way was I belittling Michael (and you by extension)?
I don’t seem to be the only one “conflating exosomes with viruses.” Dr. James Hildreth also seems to agree:
Hildreth now proposes that “THE VIRUS IS FULLY AN EXOSOME IN EVERY SENSE OF THE WORD.”
So do the authors of this article:
“Not long ago, EVs were considered to be “cellular dust” or garbage and did not attract much attention. However, it has recently been found that EVs can have important biological functions and that in both structural and functional aspects THEY RESEMBLE VIRUSES. This resemblance becomes even more evident with EVs produced by cells productively infected with viruses. SUCH EVs CONTAIN VIRAL PROTEINS AND PARTS OF VIRAL GENETIC MATERIAL. In this article, WE EMPHASIZE THE SIMILARITY BETWEEN EVs AND VIRUSES, in particular retroviruses. Moreover, we emphasize that in the specific case of virus-infected cells, IT IS ALMOST IMPOSSIBLE TO DISTINGUISH EVs FROM (noninfectious) VIRUSES AND TO SEPARATE THEM.”
This article also seems to conflate them:
“The similar structural and physicochemical properties of exosomes and viruses, facilitate the entry, biogenesis, and multiplication of the viruses in the host cells 16. VIRUSES AND EXOSOMES SHARE VARIOUS COMMON FEATURES SUCH AS SIZE, STRUCTURE, BIOCHEMICAL COMPOSITION, AND MECHANISMS OF BIOMOLECULAR TRANSPORT WITHIN THE CELLS (20, 21).”
I can find more examples but I think these should suffice.
“So once again, where is the foundational proof for the existence of exosomes?”
Where is the foundational proof for the existence of ANYTHING?
The logic problem is this:
A. Mike requires 100% purification for proof of existence of a ‘thing’
B. It is impossible to 100% purify any ‘thing’
C. Therefore, it is impossible to prove existence of any ‘thing’.
Based on the above logic, there is nothing I can provide that satisfies A for anything including chickens, rats, gold, water, etc.
The problem is your invention of 100% purification and your requirement for 100% purification in order to prove existence of something. As long as you maintain that, I don’t see this discussion going anywhere. Because if I provide scientific papers on the purification of chickens (as the Myth Busters pointed out today in their latest awesome documentary), you will not accept it because it is not 100% purified.
Your chicken example is a ridiculous comparison. First, we know chickens exist as we can actually see them. Second, you can isolate chickens from other animals/things easily. Is that seriously the best analogy you could come up with?
If you are stuck on 100% purity, please share any example where 99.99% purity was achieved for exosomes. Please show how this was determined. If that is not acceptable to you, what is an acceptable purity level? If other particles/substances with the same size/density/morphology exist in the sample, how are the exosomes distinguished and differentiated in TEM? Where was this shown in the original publications claiming the existence of exosomes?
We are dealing with invisible particles which we can not observe in the natural environment. For all we know, these particles do not exist in humans at all and are nothing more than a creation from the processes the samples go through. Or, as Dr. Lanka and others have pointed out, they are nothing but cellular debris from dying cells.
You continue to avoid looking at the foundational evidence for the existence of exosomes. Don’t worry. I looked at it for you and will be writing up an article about it. Feel free to respond then if you want but it seems rather clear to me you are unable to grasp the fact that in order to have a valid independent variable to study utilizing the scientific method, the thing (i.e. your presumed cause; purified/isolated particles assumed to be exosomes) must be proven to exist by themselves first. This is so that they can be varied, manipulated, characterized, studied, etc. As with virology, exosome research does not have the thing they claim to be studying. They have a mixture of things from which they pick out certain particles that fit their preconceived ideas of what the thing is. As always, the concept came before the “evidence.”
Mike, you finally stated that 100% purification is impossible for any particle, molecule, animal, etc. You also claim that complete purification is necessary for proving the existence of a thing.
Based on those to points, in your world model, it is impossible to prove the existence of anything.
And this breaks any argumentation for the existence of viruses. The pro-virus people will say, well by your own measure, you cannot prove the existence of anything. Therefore, your work is simply invalid. And they would be correct.
I have stated previously that 100% purification of these particles was not possible. They admit this in numerous instances. This is not a new statement from me if you reread my comments. However, this is the corner virologist and exosome researchers have backed themselves into trying to claim functions for nanoparticles that can not be observed in a purified/isolated state and without heavy alterations in order to be imaged. They have created a fictional narrative for dead particles which can not be shown to exist as presented in living organisms. You have bought into their fiction without asking for validation.
So once again, where is the foundational proof for the existence of exosomes? Where were the particles assumed to be exosomes purified and isolated from everything else in order to be studied and characterized? Where were the functions of these entities proven?
Do you have this evidence or can we finally skip to the part where you admit that all they are doing is pointing at the same particles in TEM images and declaring functions to them just as virologists do?
Another great article. We question the virologists and they have no answers that make much sense or are believable even to the average person who tries to think for himself.
Why can’t “modern” science send a mini-robot or microscope into the body to hunt for virus particles? We have heard much about the possible mechanical gizmos in the mRNA gene therapy injections and how they want to reinvent humanity into some artificial beings using robotics.
There must be some kind of biological solution they could make up that could be injected into the body and that would somehow attach itself to a virus, like a marker. It then could be observed somehow.
Strange that bacteria, fungi and other matter can be found and isolated and why not a virus? If viruses cannot be proven to exist in an uncultured state, it makes one wonder what other nasty things they claim are out there that can’t proven to exist. These medical bozos could make anything and pass it on as a “plague” of some kind. If all it takes is there say-so, how can we prove otherwise?
There are so many articles and videos done by the presumed good guys (those doctors questioning the covid narrative and mRNA gene therapies) but everything is formulated on the acceptance of viruses. So, this renders most of these sources highly questionable since the basis for their observations and conclusions (viruses) might not exist.
If ever had any trust in the FDA, CDC, AMA or big pharma or even my doctor, those horses have left the stable and are long gone. So when someone refers to FDA or CDC guidelines, I know they are stepping in a giant pile of cowflop.
Orthopoxvirus…can’t wait for the media to go wild with that term.
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Thanks Tom! The more we use logic to question the lie, the more it unravels. Virology has had well over 100 years to show “viruses” exist. The fact that they can not do so when challenged repeatedly is very telling. 😉
I enjoyed this https://www.bitchute.com/video/1xW7NfOXOoIs/?_kx=4U-EIWJvmU9f8YsR14QbuOgqcMHEEXF9EBhCVNDbesI%3D.TVKZzX from tom Cowan though I wish he would stop going on about the ice woman. Some great points;
why does no-one get two or more viruses at the same time- they could be in different organs?!
Going to a restaurant and ordering turmeric, when you mean curry, the same as saying you have virus in a cell culture
I really like the analogy of the radio, we know that EM radiation causes the sound to come out of it- but there is no point taking the radio apart and looking for the cause inside it cos it isn’t there!
An answer to what are ‘mock’ cultures, they were not adding a sample from a sick person or antibiotics and were not starving the cells.
Finally, they have made us forget everything we already know. We can’t explain it but we know that cats can tell when we need to get up for a pee, without us moving, from three doors away.
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Thanks Jo. I will take this opportunity to note in Jeffrey’s general direction that TC explicitly says viruses do not exist during his opening statement. That viruses are a misconception.
Unfortunately, TC goes on to just dismiss all cell particles as only being “garbage” that only occurs during cell “breakdown.” Talk about a bold (and unreasonable) statement, to suggest that particles aren’t generated by cells other than when they disintegrate.
yes I see. There are likely to be functions for exosomes etc you mean?
Yeah exosomes being the elephant in the room but also all manner of things. Membranes are called membranes for a reason. Maybe TC would respond, “well, yes of course, I’m just talking about in vitro dynamics” but my response to that would be, “then you need to be explicit about that and you also need to say that in vivo intercellular dynamics would include lots of cell particles all the time, otherwise it’s misleading.” But then again I can’t really recall him talking about in vivo particle generation.
Dr. Cowan most likely is referring to the particles seen in EM images after cell culturing. These particles are most likely nothing but cellular debris created through the culture, fixation, dehydration, staining, and embedding processes. There is no evidence that these oarticles are ever within a human body in the state they are observed in EM imaging. They can not observe these particles without heavy alterations and dead imaging.
Thanks for the link Jo! I love Dr. Cowan’s work and I will definitely give it a watch!
How were exosomes proven to exist?
The cell debris may be said to exist as an empirical observation – assuming it is not an artefact of the means for observation – as with electron microscopy (Re H Hillman’s critiques).
But the name implies extracellular exchange or function.
The modelling and mapping out of body function has a lot of reliance on in vitro experiments and dissections of the dead, and the captive. Along with techniques that are invasive that carry their own capacity to influence results. But not least of which is the basis from which we already presume to model our self and world, and operate from as ‘normal’ or presumed self-evident. Which is to say we cannot start with a blank slate, but only use such tools as we have already developed, as part of an ongoing development of modelling, imaging and theorising or theologising our sense of self each other and world. The key fact being that the image of reality is never the reality. But resonances found helpful become mapped in as part of explorative focus and experience.
I sense the nature of cells in reality is not at all as our frozen snapshots have led us to model – along with all the fudge that then attaches to or is imagined within. The physics of water as self-structuring according to various terrain attributes is opening a much more complex and alive symbiosis than a pathological model rising from a species in trauma from which it has yet to waken. The way of which calls a discernment of felt resonance or kinship in our being, rather than confirmations to boost an invested thinking as a private sense of control set over life. Or at least Gaining the illusion Of such a Function by the lens of narrative distortions given priority.
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Good to have you back brian. It’s true that we have been talking of cells in a static way. Combining what Clarifire and I talked about — how the intelligent body is still essentially a collective consciousness of single-celled eukaryotes — with what is known about cell migration, we can move towards a more realistic understanding of a self-structuring fluid dynamics holography. Check out the cell migration field if you haven’t. Migration dynamics also relates to cell force dynamics (muscle power) which shares Hillman-type views on the protein folding that generates muscle power. Muscle proteins have alternating hydrophobic and hydrophilic electrostatic/hydrostatic qualities, using the fluid dynamics of water chemistry to fold-unfold in order to generate power. Other muscle proteins like Titin, which is the largest protein in the body, perform the structural elasticity function of muscle fiber.
(Here’s an article on the water-based electro-thermodynamics of muscle power:
I’ve read Mae Wan Ho on the manifold complex of ‘water and dust’ or indeed water & proteins, solutes enzymes etc. (among others).
Complexity can seem chaotic, but my sense is to abide with apparent chaos to the recognition of patterns, relationships and meanings.
Simple but partial views can be more comfortable, but like a kindergarten can become very limiting if clung to instead of moving on from.
I have read of the alternating states not just as phase changes in the folding/unfolding of proteins, but as enzymatic processes within cytoplasm operating or indeed oscillating at billions of cycles per second.
My sense of actual cutting edge biology (as distinct from Research™spun out of biolabs or dark defence operations) is that it expands our view in ways that make our current models either false, partial or grossly over simplified.
But electromagnetic or charge field communication and exchange is universally fundamental at all scales, and the ‘waters’ serve as medium of ionic exchange that self-structures what we call matter.
I will look at your proffered link – thanks.
Ah maybe you don’t need to look at the link. I forgot to look up Mae Wan Ho.
Do you think RNA is just the first solenoid ever invented?
I suspect the organising field via structured waters provides the organic helical patterning of proteins, and that the information or ‘informing’ principle is a much more living two way process of exchange than the genetic dogma that embodies a material determinism, just as virology embodies a pathological mindset.
One of the points I keep in mind is that the assigning of authoritative faith in such thinking, takes on the signature vibration they embody.
If you believe you have no freedom, you will not use it to question your belief!
Such is the power of belief as a means to persist an idea as a default – until and unless it is brought to awareness as an idea, rather than taken as ‘self-evident’ – as in ‘of course viruses are real!’.
Beliefs can be be-lived as private realities seeking social reinforcement – with consequences of perception and response as adaptations or developments of the belief as a social identity complex. Or social masking complex.
Our symbols at best – point us toward a living relational terrain.
Transformers are innate to life. Our inventions are attempts to mimic or recreate a life we dissociated from by the wish or belief to make a self in our own image as if to regain or restore wholeness from a fragmented or split perspective. As in Humpty Dumpty.
That part about freedom and belief is excellent.
I was thinking that proteins take charges really well and that consciousness needs a center of gravity within the collective field from which to work. The solenoidal, helical protein formations generating a torus dynamic that hold the space of an individualized, informational field of consciousness as it fans out from one end of the helix and returns into the other end. I know this is ‘new age’ iconography.
I have a hydraulic valve I’m troubleshooting so solenoids are on my mind but I haven’t traced all the wiring yet and the problem is probably a wire shorting to ground somewhere.
The phase change of a protein folding as I understand (Gilbert Ling) is by a molecular transmutation of its vicinal waters according to various factors (terrain conditions) that are of course an electro-chemical bonding/unbonding.
The torus as a symbol may be associated with anything, but the balance point of any such interactive field effect holds the nature of the still point of a turning world.
Though I sense this is relevant to vital or biological organisation – as the bio-field of nested fields within a larger embrace, I also sense our attempts to hack it, are actively undermining our ‘life support’ and consciousness.
There is much research into synthetic proteins, enzymes and nanotechnological interventions, some of which is ubiquitous and unregulated, some running under stealth. Disconnection from our ‘field and its expression’ in exchange for surveillance and control. What does it profit a man to gain a world if it costs his (feeling awareness) as Soul?
What is the character limit for a comment? Do you accept email?
Hi George, there is no character limit to comments as far as I know. You can definitely email if you want at email@example.com.
“I have read of the alternating states not just as phase changes in the folding/unfolding of proteins, but as enzymatic processes within cytoplasm operating or indeed oscillating at billions of cycles per second.”
When I read this from you yesterday, about enzymatic oscillation, the first thing that came to mind was Hillman’s visual representation of the cell from one of his videos, which was just a nucleus with no detail and a cytoplasm with a bunch of flecks in it. I had been thinking of oscillations as only circular. I noodled around a bit just now to begin to understand what you mean and I found that enzymatic oscillation modeling is toroidal. And interlinked pairs of cross-regulating enzymatic toroids are modeled to account for functional complexity.
Check out this link. On the right hand side you can page between the three models. The first model details the particulars of the schematic. Note the role of mRNA.
It took me a while to understand how they were using the term “negative feedback” until I realized it means homeostatic regulation. In enzymatic oscillation, negative feedback maintains that center of conscious gravity (my animist home cosmology holds that gravity is based on consciousness) in the universal, entropic tendency towards ‘chaos.’ The solenoid effect.
When I looked up “cellular pool” i found that it’s referring to the the cell waters in which chemicals that form true solutions take on a crystalline structure. Nice. If we take these two dimensional models and flesh them out, then the cellular pool is obviously surrounding these these oscillations and the 3D oscillations that generate the “metabolic species” required for crystallizing the waters such that the toroidal consciousness has a structural fundamental that it can work with in order to hold up the space (holding the space) of the volumetric, crystalline waters, if you follow me. Is that right? Is this the holography of the nucleolus?
Are similar ‘muscle’ oscillations happening out in the cytoplasm? Is everything in the cytoplasm just Hillman’s flecks, with the flecks being the centers of gravity of the oscillations? And, say, mitochondrial bodies just being more ancient groupings of symbiotic oscillation? Is this the object modeling beyond object modeling?
Yes I follow your gist.
Whether centres of gravity of zero-points (infinity points) within polarised expression.
I sense a series of stepping down of energy expression from total synchronicity or ‘always’ to partial realms of expression that at heart are not polarised exclusion but a whole in all its parts.
The instrument of our perception is itself a result of a polarised exclusion or split mind, in which a ‘consciousness’ is projected AS IF into its own imaged modelling. Which is also a means of oscillations or charge cycles that can maintain a specific pattern of focus like an eddy in the river. My sense of intuited appreciation is the recognition of the total flow of river as one with it ‘eddies’. To embrace the field through the particular rather than focus in objects as reflections of disconnected objects.
I may be wandering off here. But at some level this is a willingness to be aligned by a ‘field communication’ or Conscious Terrain, rather than double down in a sense of conflicting levels, orders, meanings, minds. Symbiotic Consciousness as a felt quality not just as a codified insight or belief set in symbol.
The Universe exhibits charge separations as well as discharge pathways both. I see seemingly closed entropic systems within open ‘system’ that may seem ‘chaos’ at the threshold of our ‘consciousness’.
Yeah thanks so much brian.
Reblogged this on Snooze 2 Awaken and commented:
Enjoy yet another devastating (to the pseudoscience of virology, that is) by Mike Stone at ViroLIEgy …
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Thank again! 😁
I’ve been looking forward to George’s lengthy comment. Are you able to get that posted and piecemeal if necessary? Thanks.
As brian has also confirmed, these exosome papers presented by Michael S fall into a different category from the stupid virus papers. The truth dictates that not giving a militant inch to viroliegy is the same thing as *giving* a circumspect inch to the straightforward, “extended sensing” of new and greatly improved exosomal research, and it can only be a regressive and reactionary form of ‘Terrain’ orthodoxy that would refuse to give that circumspect inch. Orthodoxy is where ego, opinion, and structural power meet.
Hi Reante, I’m not sure what comment from George you are referring to. I’m sorry, I’ve been on vacation the last two weeks so it has been a bit difficult keeping up.
Hey MIke 🙂
I see you since saw George’s query and replied. Thanks!
No apologies, please. Way to knock out a blogworthy comment during vacation.
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Well, That is a LOT of comments! Great article and glad I read it. So basically it said bla bla bla, the scientist have NO idea what they are looking at and are only speculating. Though it seems that they maybe closer to the truth on Exomes, than on viruses, just based on what you wrote and other information I have gathered, though that is just my opinion.
And, just in case you wanted to know, I found a tiny typo “there is no need for the vitus” to enter into the equation”. I am guessing that is supposed to say “virus”?
Great job once again!!! It certainly adds to the understanding of our current “science” model, which is sooo out of wack. Is there anything in science we can trust these days? I am really starting to wonder.
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Thanks! I corrected the typo and appreciate the kind words and support. 🙂
Mike wrote: “First, we know chickens exist as we can actually see them”
Are you suggesting that purification is not needed in order to prove existence for things?
If that is the case, you have introduced additional methods for proving the existence of things. What are the methods that you will accept for proving existence?
While my offering to yours and others posts here is in the ongoing theme of the existence or not of various scientifically claimed particles as functions assigned them, its scope is contextual not competing in an attempt to convince or discredit others so as to maintain my own convictions unquestioned.
I put it at:
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“That communication occurs between the various parts of an organism or living complex is redundant – for that is the life. The actual nature of such communications is complex and subtle such as to elude – in my view – any strictly materially based consideration.”
Redundancies are SOP in all lifeforms. Exosomes aren’t strictly material. And conscious resonance (fieldwork) isn’t strictly energetic. Both are symbiotic holography. To say exosomes are strictly material is to say the whole body is strictly material.
‘Bodies’ are major points of emphasis wouldn’t you say? Communications amplified to bodily status/form are communicative points of emphasis. Redundancies. Packaging the damn things up is a point of emphasis. That they contain genetic material is a point of emphasis.
It seems clear to me that this is a belief for you not based on any direct evidence otherwise you would share it.
Mike for such a nice guy you’re true colors are showing. Your use of fallacious repetition is one of their favorite tools.
My “true colors?” I do not like what you are implying. I have repeatedly asked for validation of exosomes which you have consistently evaded. It’s fine if you don’t have the evidence. I doubt you or anyone else does. It’s OK to say “I don’t have any paper showing the purification/isolation of exosomes nor proof of their functioning.” As I said, if you want to believe in exosomes without direct evidence, that is fine. It is not good enough for me.
I appreciate you walking back the insult to Michael and myself.
I have yet to hear how I insulted either of you.
The terrain is an interaction.
If Mike gives emphasis to empirical verification as proof of claim, I look for a correspondence within my own connection with life for a willingness or openness to communication within what is being thought or said – not merely the social coding.
Are genetic traits coded materially or is that a premature attempt to pitch a claim on behalf of a materialist reductionism (what a mouthful!) so as to persist in vitro (in the ego frame of life set in pieces).
I don’t know what you are saying.
It is so, that what matters to us become what we mind or give focus to at such a level as to internalise as if to seem to be a mind or think/thing of its own.
‘Empirical verification’ *is* materialist reductionism. Reason comes from cumulative observation of cause and effect *plus patterning.* Mike shat on the deep patterning because he’s previously invested in exosomes not existing. And Dr. Tom is invested in exosomes not existing.
Of course genetic traits are coded ‘materially’ (holographically). We’re holographs How else would they be coded? Why is it that genes are the ONLY functional component involved in reproduction? Egg and sperm are receptacles for nucleic acids. They have no metabolisms.
Genes existed before cells existed. They built Life. To not believe that is to not believe in primordial Life. Reason tells me that the primordial origins of Life are true to the patterning of Reality. YMMV, but if it does then naturally I would consider you to be wrong unless you can come up with a better Reasoning process.
Usually the first person to shit on another is the person with the worse reasoning process. Masking mode. Truth tellers don’t have anything hide.
We do not need to purify chickens to know they exist. We can easily observe chickens with our own eyes and in nature. We can take a chicken, remove it from other chickens/animals, and study it. Chickens are not invisible nanoparticles contained within an unpurified sample mixed with millions of identical particles. I’m not sure why you are doubling down on your chicken analogy. I would drop it and move on as it’s pretty bad.
“Chickens are not invisible nanoparticles contained within an unpurified sample mixed with millions of identical particles”
This is a prime example of intellectual dishonesty that bears no reflection on the experiments Michael presented.
Masking mode turned all the way up leads back around to bare nakedness.
You are not making sense. Chickens exist. We can see them without having to search through unpurified sludge to find them. Please explain how isolating chickens hold any relevance to exosomes.
Hi Mike, I cannot answer your question regarding purification until we sort out your definition of existence.
I will repeat my question as this is foundational:
“What about Gases? Does Oxygen exist? You cannot see oxygen, so what are the methods for proving existence Mike uses for O2?”
That is a cop-out. It is not my definition of existence. It is logic and the scientific method that needs to be applied here.
You keep trying to procrastinate. I gave you 99.99% purity which can be established with gold. We are not discussing oxygen but since you brought it up, it can also be purified to 99+%:
“Most commercial oxygen is produced using a variation of the cryogenic distillation process originally developed in 1895. This process produces oxygen that is 99+% pure.”
Now please stop procrastinating. Do you have a paper showing exosomes purified to 99.99%?
There are many very specific proofs to the claim oxygen exists along with specific functions – as opposed to phlostogen.
The ‘tools’ for indirectly claiming the existence of these nano particulate entities or functions, are hardly in the same category as your claim of equivalence to elementary or molecular gases.
I call it black box science because – like goats entrails, only the high priest can determine its import and a priesthood is established to present some sense of unified interpretations to matters on which the turning of a world now depends – ie contractual subordination of sovereign states and laws under WHO. Or a global biosecurity state.
Hence no room for error is possible in setting the frame by which such definitions unpack to enable such consequences that are both weaponised and marketised to an anti-life agenda.
I don’t see the case as proven, so much as expressing an outdated way of looking at life as biology taken out of living context.
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“Now please stop procrastinating.”
Please keep this discussion civil. I have already provided you the reason that I am trying to understand your definition of existence. You have walked back your definition of existence from needing to be 100% purified, now to 99.99%. You also seem to have recanted on the requirement to be able to ‘see’ the substance.
What are the methods that you will accept regarding the measurement of purification?
Also, what methods do you view as acceptable for purification? For instance, you don’t seem to accept Chromatography because you have to add the thing being purified into a substance. Do you also not accept Ultra Centrifugation because you also have to add the thing being studied to a sucrose solution? Same for precipitation.
Again, not my definition of existence. The methods for purification for the particles claimed to be exosomes have already been laid out. I gave you the 99.99% purity level as you were stuck on 100% purification. Thus, you agree that the particles called exosomes can not be separated from everything else. It is obvious that in order to determine that the particles seen in EM are in fact exosomes, they must be isolated from everything first. As this can not be done, there is no guarantee that those particles have any relation to the concept of exosomes whatsoever. As the particles can only be seen dead and after heavy alterations, there is no guarantee that these particles ever exist in the body as presented in EM. As they can not be observed alive, there is no way to determine their supposed function. In other words, there is no way to prove the existence of the particles as exosomes until there is a method that separates them from all other particles and allows for them to be be seen alive.
You may find this unfair but this is what logic dictates. It is also what the scientific method dictates. Exosomes can’t even get past the first step which is to observe a natural phenomena. Exosomes can not be observed at all. As they can not be separated from everything else within the fluids, there can be no independent variable to manipulate and vary in order to prove cause and effect, once again failing the scientific method. Essentially what we have is particles created through a process involving heavy alterations that are picked as the representative of a theoretical entity. It just so happens they are the same particles chosen as “viruses” as well.
Now, as we know you do not have any paper showing 100% purification, do you have any showing 99.99%? 90%? 80%? 50%? What is the best you have? And please explain how the particles claimed to be exosomes are supposed to be distinguished from all of the other similar particles within the sample if they can not be separated from them completely. Then please explain how their functioning was determined without being able to observe them alive.
Mike Wrote: “Thus, you agree that the particles called exosomes can not be separated from everything else. ”
I agree that it is impossible to separate anything from everything else. Chicken, Gold, and O2 are just some examples.
What is the point of giving you another scientific paper when you will just go on to state that it isn’t 100% purified and the purification methods can’t possibly purify the thing because you are adding this component into the mixture (the case for Ultra Centrifguation, Chromatography, and Precipitation)?
So I again ask what are the accepted purification methods that satisfies your definition of existence?
Also, do you still demand 100% purification but you also said you will give me 99.99%? I do not understand this logic. Either you require 100% purification or not. There is no ‘give’ in science.
I have also asked you for a scientific paper that 100% purifies any substance. This paper will provide me an example so that I know what you will accept as proof of existence.
How can a chicken not be separated from everything else? We are not deconstructing the chicken at a molecular level in order to see that a chicken exists. We can see it with our own two eyes. We’ve been over this again and again so now we are just beating a dead horse.
You understand what I require. A paper that adheres to the scientific method. This requires observing a natural phenomena and detetmining cause and effect. In order to do so, there must be a valid independent variable (i.e. purified and isolated particles). Exosome and “viral” research can not provide said evidence.
So now explain to me why the current evidence is valid.
1. How can the particles picked as exosomes in EM be determined to be exosomes if there are millions of similar and/or identical particles in the sample? You realize these particles are commonly mistaken for other particles all the time in EM, correct?
2. How is the function determined if the particles are unable to be observed alive?
3. How are the particles not a creation of the purification and EM imaging process?
I have never seen nor do I believe that an exosome paper that adheres to the scientific method exists. There is no direct evidence. I am willing to look at any paper you feel proves exosomes exist and just as I have done with your other papers, I will provide my assessment. You can either agree or disagree.
The reason I used the ‘Chicken’ Reference was due to Spacebuster’s recent documentary where he explains isolation and purification – “so easy even a child can understand”. – https://odysee.com/@spacebusters:c9/Final-The-End-of-Germ-Theory:8?r=GsTgacQd7yJqHkhRTYtahmMNAv2t4NTs&t=2693
At 53minutes, he goes on to describe the isolation and purification of exosomes. He notes that Dr Andrew Kaufman personally sent him an example paperhttps://odysee.com/@spacebusters:c9/Final-The-End-of-Germ-Theory:8?r=GsTgacQd7yJqHkhRTYtahmMNAv2t4NTs&t=3207
Here is a link to the paper – https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4780996/
Note, I haven’t read the paper yet. But just skimming, it seems like all of the other exosome papers that I have read that confirm purity via western blot and other methods.
So prior to this 2016 paper, it is admitted that HIV and exosomes could not be separated:
“However, plasma of HIV-1 infected individuals CONTAINS BOTH EXOSOMES AND HIV VIRAL PARTICLES, which are similar in size and density. Thus, efficient separation of exosomes from HIV viral particles in human plasma HAS BEEN A CHALLENGE.”
This being the case, any antibody testing is utterly meaningless as both exosomes and HIV would have needed to have been properly purified/isolated and characterized biochemically first in order to determine “specific” antibodies:
“Virus particles were IDENTIFIED BY p24 ELISA. Exosomes were identified on the basis of exosome markers ACETYLCHOLINESTERASE (AChE), and the CD9, CD63, and CD45 ANTIGENS.”
This isn’t even taking into account that antibodies themselves are entirely theoretical, non-specific, and have also never been purified/isolated. One fictional entity can not be used to determine whether another exists or not nor to determine “purity.”
Also, acetylcholinesterase is not specific to exosomes and can not be utilized as a marker:
“Acetylcholinesterase (AChE) activity is found in abundance in reticulocytes and neurons and was developed as a marker of reticulocyte EVs in the 1970s. Easily, quickly, and cheaply assayed, AChE activity has more recently been proposed as a generic marker for small extracellular vesicles (sEV) or exosomes, and as a negative marker of HIV-1 virions. To evaluate these proposed uses of AChE activity, we examined data from different EV and virus isolation methods using T-lymphocytic (H9, PM1 and Jurkat) and promonocytic (U937) cell lines grown in culture conditions that differed by serum content. When EVs were isolated by differential ultracentrifugation, NO CORRELATION BETWEEN AChE ACTIVITY AND PARTICLE COUNT WAS OBSERVED. AChE activity was detected in non-conditioned medium when serum was added, and most of this activity resided in soluble fractions and could not be pelleted by centrifugation. The serum-derived pelletable AChE protein was not completely eliminated from culture medium by overnight ultracentrifugation; however, a serum “extra-depletion” protocol, in which a portion of the supernatant was left undisturbed during harvesting, achieved near-complete depletion. In conditioned medium also, only small percentages of AChE activity could be pelleted together with particles. Furthermore, no consistent enrichment of AChE activity in sEV fractions was observed. LITTLE IF ANY AChE ACTIVITY IS PRODUCED BY THE CELLS WE EXAMINED, AND THIS ACTIVITY WAS MAINLY PRESENT IN NON-VESICULAR STRUCTURES, as shown by electron microscopy. Size-exclusion chromatography and iodixanol gradient separation showed that AChE activity OVERLAPS ONLY MINIMALLY WITH EV-ENRICHED FRACTIONS. AChE activity likely betrays exposure to blood products AND NOT EV ABUNDANCE, echoing the MISEV 2014 and 2018 guidelines and other publications.”
They are also adding chemicals like iodixanol in order to “purify” their samples:
“Prepare 6% and 18% solutions of the IODIXANOL REAGENT, supplied as a 60% solution in water, by dilution in PBS.”
Iodixanol can cause apoptosis, i.e. cell death, and mitophagy, i.e. degradation of mitochondria:
“Iodixanol is a nonionic, water-soluble contrast reagent.1 IT INDUCES APOPTOSIS AND MITOPHAGY in renal epithelial cells in a rat model of contrast-induced acute kidney injury.”
On top of this, there are no EM images of purified/isolated particles whatsoever.
Hiya, I know I’m in the middle of something else again, but have a question about antibodies, which I now see as sticky globulins which probably don’t do what we’re told they do in ‘immunity’. However I have been ‘working with them’ for many years. If smooth muscle actin for example is injected into a mouse (which is I now regard as abuse and see no justification for) and a B cell taken and made into a hybridoma with a myeloma cell- the resulting production of globulins when applied to tissue sections and visualised by staining, consistently, seemingly accurately and everywhere one would expect it, stick to what we call the smooth muscle in all arteries and arterioles etc, which are not stained by adding just serum. Sticky globulins seem to be made by mammals to stick to things fairly specifically!
I know all histology itself is an artefact and only represents dead tissue, but observing completely formed breast tissue in a lymph node taken from a person with cancer cells in the breast for example, does give some information about health I think.
I can not speak to the effects you have seen, only to the history of antibodies and the studies I have read. Antibodies, as presented to us as a Y-shaped nanoparticle, have never been purified, isolated, and characterized. They have never been observed within living organisms. Antibodies started off as a concept first. The reactions ascribed to them are regularly non-specific and the studies using antibodies are unreproducible which has led to a reproducibility crisis. There are 6 different theories explaining their form and function. Antibody testing results are inconsistent, showing up in people who should not have them while completely missing from those who should. You can find all of the information I have gathered here:
Hiya, yes, I see that antibodies are not involved in ‘immunity’ in any way that we understand it, but it is spectacular to see the staining of different tissues with these monoclonal antibodies – I can’t believe I’m linking to wiki https://en.wikipedia.org/wiki/Immunohistochemistry but the ‘antibody’ or something else is definitely sticking to the ‘antigen’/target tissue it’s been ‘raised’ to eg actin which is then being visualised by a chromagen.
It’s hard to say what causes the staining reactions but IHC has been shown to be unreliable. You may find this article interesting where prominent immunologist Charles Saper called out IHC specifically:
“…it is often assumed that IHC has an analogous ability to identify molecular targets accurately.
Nothing could be further from the truth.
In fact, IHC methods remain as primitive, in terms of both sensitivity and specificity, as they were in the days when DNA sequencing was done by hand using sequencing gels. The fundamental principles on which antibody localization is based have not improved at all in the last two decades, and if anything, the slope occupied by IHC has become more slippery than ever.”
this applies to when gene sequences are used and the manufacturer is not clear how they were raised but ‘If the pattern of staining of the antigen is well known (e.g., antibodies against glial fibrillary acidic protein or tyrosine hydroxylase), it is reasonable to show that tissue of the type and species you are studying, when stained with your antibody, produces a pattern that is identical to that previously reported. Alternatively, it can sometimes be demonstrated that one antibody (e.g., a monoclonal antiserum against a large protein) produces a pattern of staining that is identical to another antiserum (e..g., a polyclonal serum against a synthetic peptide) that is better characterized (and does pass the adsorption test). Antisera against different parts of the same molecule which produce the same staining pattern, or can be shown to be colocalized in double label studies, provide an important strategy for establishing specificity. Similarly, comparison with the pattern of mRNA expression by using in situ hybridization histochemistry can demonstrate that the immunostaining for the protein is genuine.’
I maintain that the staining for human actin is completely genuine and that a monoclonal antibody to it was raised in an animal.
Some sticky highly specific globulins are made by mammals in response to injection with some ‘antigens’- for what purpose? for the bodies own investigation into what they are? to control them?
Of course there is some sort of chemical reaction taking place. However, this reaction is not proof of the antigen/antibody theory and as Saper and others pointed out, it is not specific and regularly unreproducible. This passage is a nice overview:
“The THEORY behind IHC is that we can use antibodies to bind to proteins in tissues, and by building up layers of secondary antibodies and coloured complexes, we can visualize the locations of these proteins with a simple light microscope. Sounds great, right? Well it’s not always straight-forward (honestly, I don’t think anything in my PhD ever was!) and as with any technique, IHC has benefits and drawbacks, some of which you can see in the table below.”
It is a theory that these antigen/antibody reactions are occuring within certain chemical reactions and seeing certain staining patterns. What actually occurs is anyone’s best guess. If one does not get the same pattern using the same antibody, there is always the built in excuse that the antibodies were not prepared properly rather than admitting that sometimes the desired chemical reaction is observed, sometimes it is not. Maybe there is use for this technique, maybe not. However, with the reactions being non-specific in many cases (such as seen in the reproducibility crisis), it is very hard to say the results show anything meaningful at all. It is all in the eye of the beholder.
I’ve done thousands of immuno runs so know exactly how to get the stain I want- by antigen retrieval (using microwaves, trypsin for exactly 30 seconds, turning round three times etc), and some antibodies are always unreliable and capricious. I’m talking about this one ‘anibody’ though- actin – which is highly reproducible, specific, robust and any other word you want to call it. A specific something is made in an animal which sticks 100 times out of 100 to what is was raised to and nothing else.
What type of antibody do you use that is 100% specific?
I feel that this will end the same as my ping pong with Mike Yeadon, https://georgiedonny.substack.com/p/what-they-really-dont-want-us-talking neither of us will be convinced otherwise.
‘Antibodies’ do ‘cross react’ in that they stick to similar things….
I am so disillusioned with science and health care that I forget I’m a published author. Please see our paper on co localisation of HMFG and HK ATPase in parietal cells in the duodenum ‘previous study of the novel antigen of epithelial membranes indicated that the mono-clonal antibodies directed against human milk fat globule(HMFG) membrane proteins cross- reacted with gastric mucosal cells. Using HMFG-1 and HMFG-2 monoclonal anti- bodies as immunocytochemical probes on human gastric body mucosa,we have shown that these antibodies are selective for the intracellular canalicular structures of parietal cels,and have confirmed this selectivity using a well characterised antibody widely used as a gastric parietal cel specific and canalicular membrane H+ translocating, K+ stimulated adenosine triphosphatase (H,K-ATPase) specific marker’ https://pubmed.ncbi.nlm.nih.gov/7490316/ and look at the pretty colourful pics.
….But it would take a lot of immuno dark arts to produce these pictures of co localised staining in the caniculi of parietal cells without a sticky thing raised in an animal to a protein kind of reaction. I’m not saying that antibody testing in a person isn’t riddled with problems but when you can see the tissue section being stained in a very ‘particular’ part of a ‘particular’ cell or structure I find it hard to just call that a chemical reaction.
Not many people know this (possibly only me and my supervisor) but anti human SMA also cross reacts with the smooth muscle in blood vessels and in the myoid cells which surround fully formed mature seminiferous tubules in the testes of both the harbour porpoise and the dolphin! Please see our paper and look at my stunning pics of SMA staining which I haven’t seen for ages. The antibody procedure does not require antigen retrieval so minimal immuno voodoo. Put antibody on the section and visualise it. The sections provide their own internal control, structures not containing smooth muscle are not stained. Again I think there must the forming of a sticky substance in an animal to a protein injected in it in order to stain ‘particular’ parts of sections of testes in such a ‘particular’ way.
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Thank you Jo for sharing your paper! I will definitely give it a read as I am genuinely curious about this. Do you know which monoclonal antibody was used? I’m just wondering, based on how monoclonal antibodies are made and the many proprietary components and secret sauce nature of their creation, if there is a way to say for certain that the substance you are referring to was actually in the animal to begin with or if it is just a creation/byproduct stemming from the process used to make the monoclonal antibodies?
HIya, the manufacturers are all in the papers. the SMA one was made by Dako. https://pubmed.ncbi.nlm.nih.gov/15379925/ I’ve never it done myself but have worked with people making antibodies and they do inject animals with antigens and their spleens harvested. For the thing to be a ‘byproduct ‘ of the process of making anitbodies it would have to be a different byproduct, of the same process, for each different antigen, (as the antibodies go on to stain very ‘particlular’ parts of particular cells), which defies belief more than the simpler explanation that cells we call B cells in an animal can make sticky globulins that stick to different antigens
Monoclonal antibodies are not taken directly from the animal and used. They are created through an artificial process of fusing mouse spleen cells with myeloma cancer cells, along with PEG and who knows what else is used as it is hidden behind proprietary reasons and trade secrets:
“Monoclonal antibodies are ARTIFICIALLY produced against a specific antigen in order to bind to their target antigens.”
This is an outline for how they are produced:
“Immunization of mice & isolation of splenocytes – Mice are immunized with an antigen and later their blood is screened for antibody production. The antibody-producing splenocytes are then isolated for in vitro hybridoma production.
Preparation of myeloma cells – Myeloma cells are immortalized cells that, once fused with spleen cells, can result in hybridoma capable of unlimited growth.
Myeloma cells are prepared for fusion.Fusion – Myeloma cells and isolated splenocytes are fused together to form hybridomas in the presence of polyehthylene glycol(PEG), which causes cell membranes to fuse.
Clone screening and picking – clones are screened and selected on the basis of antigen specificity and immunoglobulin class.
Functional characterization – Confirm, validate and characterize (e.g. ELISA) each potentially high-producing colony.
Scale up and wean – Scale up clones producing desired antibodies and wean off selection agent(s).
Expansion – Expand clones producing desired antibodies (e.g. bioreactors or large flasks).”
While they may create a certain staining pattern at times, it’s very hard to see how these lab-created substances can be claimed to have any relation to what was originally inside the mouse, especially as the very recipe used to create the product remains undisclosed in most cases.
No, ARTIFICIALLY means deliberately and on purpose in this context. Antibodies can’t be dismissed because they can be produced by injecting ‘lab’ animals (whose cells are then fused with an immortal cell to produce more antibodies, to save them the trouble of injecting the poor animal over and over again).
Because the body is more amazing, complex and wonderful than we can EVER comprehend, and that sticky globulins seem to be made by it to every single different thing it comes into contact with (piggy backed on by immunochemistry for thousands of different and very precise localisation techniques that would be flipping IMPOSSIBLE to make in a lab without using an animal,- we have a few tinctural stains!) (nor can all antibodies be dismissed by some of the techniques not working as well as others) doesn’t mean that germ theory is incorrect, or that people develop immunity, or fight ‘infections’ with them. It means that the body is more amazing, complex and wonderful than we can EVER comprehend, and that sticky globulins seem be made by it to every single different thing it comes into contact with and we need to open our minds (as always) even wider.
Artificially also means in this case:
“in a way that uses an industrial process or substance, RATHER THAN BEING NATURAL”
“in a way that has been intentionally caused by people’s actions, RATHER THAN HAPPENING NATURALLY OR BY CHANCE”
I think you are misunderstanding what I meant by chemicals like PEG. My point is that we do not know what chemicals were used in the creation of the monoclonal antibodies. These remain a secret. All we know is mouse spleen, myeloma cells, and PEG. There are certain to be other substances and they are admitted to be under trade secrets.
Nothing about the creation of monoclonal antibodies is natural. Just as when virologists use all sorts of chemicals and substances when cell culturing their “viruses,” immunologists are doing the same with monoclonal antibodies. Just as “viruses” are assumed to be in the culture and causing the CPE, antibodies are assumed to be within the mixture and staining tissues. In both cases, an effect is being used as proof for the cause. The agents presumed to cause the effect are never seen. It is the exact same scenario.
Do you think you would get the same results using antibodies directly from mouse spleen or must the mouse spleen be combined unnaturally with myeloma cells and other substances first? If so, why?
We start (cruelly) by injecting a mouse with a protein we wish to locate in a section.
The mouse miraculously produces a substance that precisely locates said protein and leaves the many that surround it untouched.
Yet this can’t be (though I’m not sure why) the mouse producing (for our purposes) a highly specific substance. This observable, highly reproducible phenomena must due to some weird, wonder chemicals made in the lab by adding PEG etc?
Your arguments have totally convinced me of mine, so thank you for that!
Until next time,
The body demonstrates a quality of balanced disequilibrium – that is homeostasis as a self organising balance within change. Self-healing occurs to restore function – as its own set of functions that can be impeded or blocked. the living body will adapt, or work around to support core functions as long as workability allows – allowing there is also a psychic or spiritual element where the will to live fades.
is this so?
the exact mechanism or means by which this all occurs is the blind watchmaker’s attempt to reverse engineer the mechanisms that can be observed, interpreted or imagined and believed.
What is the root basis for the desire to understand life as mechanism?
Is it possible it is designed or intended to be lived – such that we don’t HAVE to engage in ingenious thinking and definitions as a basis to control life. But insofar as we CAN and are moved to, let it be in an appreciation and wonder of life rather than of a human as laboratory object.
so excited I didn’t put the link to the second paper https://pubmed.ncbi.nlm.nih.gov/15379925/
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If not for tragic and painful consequence, the ability to ‘construct a world’ and live as if it is true is quite an amazing feat.
The mind of its own illusion is not aware of its own ‘attack’ on reality in giving weight to wishful thinking that it wants to be true, but as a stakeholder to its belived reality naturally defends ‘its truth’ against what is now perceived to be a threat that is then ‘justifiably attacked’ on behalf of a self-convicted sense of self and world.
That conflict is not the way to establish, uncover or know anything, is because it runs from a presumption to know, that hijacks the mind of its capacity to give and receive truly or wholly.
To know – which is the nature of being, is thus masked over with derivative claims that are set indirectly, in symbols of association rather than direct witness or indeed with-ness.
Another way of referring to the isolation claim, is whether in fact we find what we say we found, or whether we find signs and traces, associations or bio-markers that themselves rest on or persist the indeterminate nature of other assumptions or modelling conjectures.
I see the same pattern in the realm of finance, whose corruptions then feed the tooling of science to protect and extend a capacity to literally make things up as the means to persist and protect invested revenue streams as if a living complex of life support for unresolved conflicts.
The encapsulation of conflicts that become toxic as a result of persistence, operates a temporary expedient that can become a loss of freedom of being or functional joy in being, to death and taxes – or sacrificial conflict management as war over life seen as threat – rather than as a realm endeavour that involves decisions, evaluations and true appreciations.
Worship of science as the arbiter of technological extensions hailed as ‘progress’ has discarded the qualities of being for the ego of becoming something ‘else’ than what we simply and always are. What an amazing feat! But at what a cost to gain a world of conflicting illusion and lose the capacity to know anything truly!
Giving and receiving are the nature of mind or thought. As we choose to accept or receive so will we give or live to others and ourself. The fact of a resonant match is not a matter of purity so much as actuality. The quality is where signal is tuned so as to ‘lose’ the noise.
If something has being, it can be truly known as part of you, as part of all. Reality is democratic in that it is for all, not for a favoured few. Indulging a-tempt to make a private agenda will conflict, but the conflict is always with ourself for reality cannot ‘attack’ itself.
In this quote, the term ‘grounded reference point’ is pertinent to human social endeavour that has lost sight of it.
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Yes, there are many assumptions that are made based on indirect evidence such as bio-markers that get put into a model and accepted as truth. They are masters of fitting these indirect markers into a model and claiming specificity yet this specificity claim rarely holds up upon further examination.
Yes – I’m a student in the willingness to recognise & release this kind of thing from my own thinking and learning to communicate with those who act from it without really knowing there are deeper choices and better outcomes!
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Mike wrote the definition of isolation is “to separate from another substance so as to obtain pure or in a free state”
Can you achieve this with anything when looking at the nanometer scale?
I argue it is impossible. The chicken example is also true that you cannot separate all of the bacteria and other particles that are on the chicken. I think Mike claims that you do not need to “isolate” or at least purify chickens in order to claim chickens exist. He said as long as you can see the thing, it can be determined to exist.
But it seems that Mike requires 100% purification for other things. So I am trying to figure out what his rules are for purification and existence of things. Why? He has asked for proof of exosome existence. However, if we do not define the rules for existence of things, than the discussion cannot progress.
Mike said that if you can see it, than it doesn’t need purification.
What about Gases? Does Oxygen exist? You cannot see oxygen, so what are the methods for proving existence Mike uses for O2? This might get us closer to understanding if it is possible to even prove gases exist.
Functional activity or process of change is a verifiable fact, but what we assign as meaning is drawn from the fact – that is a selection and a focus of interpretation according to priorities of an already established mind or structure of consciousness of intention and desire.
The nature of what a claim can be used for is proportionate to the requirement of proof.
Mapping out nanoparticulates to functions that are patented and leveraged as black box ‘science’ by mutating specialisms of unimaginable access to (or tooled) funding, is in the realm of sorcery, because no matter how naive the ‘researchers needing more research’ they are tooled to applications of such radical import as to literally seek to make life in their own image.
The ability to create or set definitions and languages as the basis of leverage over all that partakes of their attributes, is the mind’s capacity to model reality and to shape it to its own preferences – as a local or private adjustment – within the absolute limits of workability.
A dysfunctional model can be persisted in but only at cost of sacrificing to its demand as the driver of needs. Which will be the experience of countering or managing a mounting chaos.
A principle aspect of which is the seeking out of restoring lost functional wholeness – as the attempt to rejoin separated or disconnected parts in ways that suppress awareness of conflicts, in fear, pain and loss, by repackaging them to more complex definitions of support FOR a mindset as control or as tooled by suppressed and unaddressed conflict to cling to as the mask over fear of loss of control.
I realise I expand from the issue of whether exosomes have in fact been found or are only inferred to exist according to various indirect processes that are not at all a direct observation, but involve a plethora of attempts to probe the invisible via instruments or tests that are open to multiple variables, yet adopted as standard procedure within corporate and institutional biases and habits.
Archimedes postulated a lever by which to move a whole world. Technologism ‘finds’ such leverage in the basis of what we accept and define the basis of our world to be.
Yet we attack or undermine and deny ourself by putting our spanner in the works.
Many scientists assign probability to ideas. I see that as an ongoing evaluation of whether they are truly fitting and workable, but the world truly in this is everything. For who or what?
Garbage in; garbage out.
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I’m not sure why you continue to double down on your chicken analogy. Are chickens invisible nanoparticles? Do chickens need to be put through ultracentrifugation, filtration, precipitation, etc.? Do they need to be killed, fixed with glutaraldehyde, dehydrated with ethanol, stained with heavy metals, embedded in resin, and then blasted with electrons in order to be seen?
The answer is obviously no. We can observe chickens with our own eyes. We can see them in their natural habitat and study them. We can separate (i.e. isolate) chickens easily from other animals.
Exosomes are not observable in nature. They are not observable within the human body. Their functions can not be seen. The particles assumed to be exosomes can only been seen in a deadened state after heavy alterations and even then, are contained within a mixture of millions of similar and/or identical particles. If the particles claimed to be exosomes can not be separated from all of the other similar particles, can not be observed alive, and can not be seen within a living organism, then there is no proof as to their existence. Exosomes are nothing but a fictional concept.
Now, moving beyond complete purification, I gave you an inch. I asked if you had any paper showing 99.99% purification as can be achieved with gold. You have avoided this. However, even if you were to produce a paper with 99.99% purification of the particles claimed to be exosomes, this still does not prove that they carry information by delivering various effectors or signaling molecules between specific cells. This is a made-up story ascribed to particles that have also been called “viruses” and cellular debris. They are the exact same particles given different names and functions. Until these particles can be observed alive without heavy alterations, there is no direct proof that they exist and function within a living body as claimed.
Chickens don’t exist, Mike. ‘Chickens’ exist. There are more ‘not chicken’ cells in a ‘chicken’ than there are ‘chicken’ cells. Welcome to the sludge world brother.
We are not discussing breaking down chickens molecularly. We are talking about seeing the chickens existing in nature first and then separating the chickens from everything else. If you are going to study something, it must be shown to exist first before you can break it down in order to study. Neither exosomes nor “viruses” exist in nature. They must be created through numerous processes.
We do not have to purify and isolate chickens to know that they exist.
Have it your way then mike