Anthrax Vaccine -- posts by Meryl Nass, M.D.: Coated and uncoated spores may look alike

Sunday, January 18, 2009

Coated and uncoated spores may look alike

Pictures of coated and uncoated spores are here, courtesy of Anonymous.

57 comments:

Anonymous said...: 1. Note, Anonymous, that the communications from the scientist who did the controlled study confirm that only the silanizing solution was used in the slurry before drying. Silica -- nanoparticles or not -- was not used. The same scientist first suggested to me -- or, rather, reported that it had been suggested to him -- that the Silicon Signature could have resulted from silanizing of the glassware. That typically would be done, I believe, when the glassware is used for sticky substances. Wet bacillus spore preps typically are not sticky. Then there is the question whether use of silanized glassware would result in the Silicon Signature -- given the high spike for silicon.

2. Here are some recent filings this past month or two relating to the other "anthrax weapons suspect." Sometimes what is being said under seal is more interesting than what is spun by anonymous leakers. You just have to read the half sheets before the Court Security Officer removes it from the public file upon an inadvertent filing by defense counsel.

"282 UNDER SEAL Government's Motion for Treatment of Certain Defense Filings as Classified, filed by USA as to Ali Al-Timimi (original w/ Court Security). (tbul, ) (Entered: 01/13/2009)

281 ORDERED that pleadings 277 and 278 be and are removed from the public file and placed under seal with the Court Security Officer until further order of the Court; and, it is further ORDERED that defense counsel scrupulously adhere to the Protective Order of December 3, 2004, as further violations of the Order could result in counsel losing his clearance; and, it is further ORDERED that any half sheets prepared for the public file be presented to the Court Security Officer before being tendered to the Clerk's Office to avoid any inadvertent disclosure of classified information as to Ali Al-Timimi. Signed by District Judge Leonie M. Brinkema on 12/23/2008. (jlan, ) (Entered: 12/24/2008)

271 ORDER: For the reasons stated in open court, the defendant's Motion to Compel Discovery 269 is GRANTED IN PART; and it is hereby ORDERED that the government provide a response to the five specific inquiries in the accompanying memorandum 270 . The response must include what, if any, relevant information was generated by, or is in the possession of, the Central Intelligence Agency, the Department of Defense, the Federal Bureau of Investigation, and the National Security Agency; and it is further ORDERED that the defendant's Sealed Motion to De-Classify and Unseal the Government's Ex-Parte Filing and the Instant Motion 264 and his Motion to Compel Discovery of Undisclosed Surveillance Evidence 268 be and are DENIED; ***"

Just for fun, we need to have Rachel Maddow ask Professor Turley what it is he can't tell us. Generally. Then we someone who has a copy of the half sheets as first filed needs to upload them (to the extent lawful) the minute Barack Obama is sworn in as President and independent experts confirm he was born in the US.; January 19, 2009 at 5:34 AM
Anonymous said...: Question: The Al Qaeda manual in a section on "Poisonous Letters" instructs that a silicone sealant should be sprayed on the interior of the letter so as to avoid killing the mailman.

"Wipe the envelope from the inside with Silicone sealant," it goes on, ...
transcripts.cnn.com/TRANSCRIPTS/0111/14/tpt.00.html

Why is a hypothesis involving silanized glassware more credit-worthy than where the silanizing solution is sprayed on the interior of the envelope?; January 19, 2009 at 5:43 AM
Anonymous said...: I posted my analysis of the coated and uncoated spores in the thread about the Barry Kissin article.

This thread looks like it's just going to be a lot of messages by Ross Getman about other things that only HE wants to discuss.; January 19, 2009 at 10:18 AM
Anonymous said...: Here is an exchange between two scientists about how a spray can substitute and serve the same purpose as silanization.

"> xxxx at mbcl.rutgers.edu writes:
>
> We have found that repeated silanization of glass plates used for DNA sequencing gels eventually leads to problems in pouring the gel, i.e., the gel mix does not flow evenly and forms persistent bubbles. ***
> Has anyone else experienced similar problems? We'd appreciate any suggested alternatives to silanization that could be used on a daily basis.
>Thanks.

We coat one side of one plate with Rain-X (available from hardware stores). Other people spray their plates with Pam. In either case, it is unnecessary
to treat with silanization."

Comment:

The ingredients of Rain-X are: Ethanol/SD Alcohol 40, Ethyl sulfate, Isopropanol, Silicic acid (H4SiO4), tetraethyl ester, hydrolysis products with chlorotrimethylsilane, Polydimethylsiloxanes (Silicon oil), Siloxanes and Silicones, di-Me, hydroxy-terminated.

At the time of the Spring 2002 stories, the talk was how you can buy the additive in a hardware store.
Rather than silicon dioxide, does the Silicon Signature instead point to use of a silicon oil?

Do you think the percentage of silicon was too high to have resulted from the silanization of the envelope of use of such a spray on the interior of the envelope?

Alternatively, then, note that the Ann Arbor researchers publishing research with Ivins using virulent Ames used silicon oil as part of their research involving water-and-oil nanoemulsions

COMPOSITIONS FOR INACTIVATING PATHOGENIC MICROORGANISMS, METHODS ...
Inventor: ANNIS TED C; BAKER JAMES R; (+1), Applicant: NANOBIO CORP (BR) ... silicon oil, essential oils, water insoluble vitamins, ...

http://www.google.com/patents?hl=en&lr=&vid=USPATAPP11080629&id=zIiZAAAAEBAJ&oi=fnd&dq=%22silicon+oil%22++Hamouda+Baker; January 19, 2009 at 10:53 AM
Anonymous said...: Dr. Nass says: "Coated and uncoated spores may look alike."

The operative word is "may."

The question is: How could anyone think that the coated and uncoated spores in these images look alike?

Just look at an enlarged .jpg version HERE.

The uncoated spores are irregular and rough in shape, often with tiny points, and they look like little wads of cotton.

The coated spores look as smooth as pearls, and some near the top of the image even seem to be fused together with the coating filling in the areas between the spores.; January 19, 2009 at 11:28 AM
Anonymous said...: Ed Lake missed the point entirely. The two pictures are completely different. The uncoated spores are imaged under excellent lighting conditions. The light in the coated spores images is poor. In addition, the untreated spores are well focused, but the treated spores are out of focus.

Ed Lake claims that the coated spores look as smooth as pearls; but that is misleading. An objective and accurate assessment would be -- the poor lighting and lack of focus in the images of the coated spores make it impossible to access whether they would appear the same as uncoated spores if comparably focused and well lit images of the uncoated spores were made.

Dr. Nass did indeed say "Coated and uncoated spores may look alike." Dr. Nass was entirely correct.

Ed Lake, on the other hand, did manage to get one thing correct -- The operative word is indeed "may". Ed just didn't understand what that meant.; January 19, 2009 at 5:58 PM
Anonymous said...: Naturally Ed Lake missed the point about the pictures. It's obvious that the coated spore picture is poorer quality. That's not the point however.
The point of the comparison was to demonstrate that the coated spores in this case are NOT coated with silica nanoparticles - which makes them look like sugar coated donuts. Sandia made a big point of the lack of the sugar coated donut-look on the mailed spores - they used this as proof that the mailed spores were not treated with silica or other silicon agents.

However, the polymerized glass coated spores clearly show that they look more like uncoated spores when treated like this. It is a known FACT that the exosporium on the spores is penetrated by small molecules - the polymerized glass coating thus forms BENEATH the exosporium - exactly as Sandia observed. The total % silicon would quickly confirm the presence of an artificial silicon additive.; January 19, 2009 at 9:30 PM
Anonymous said...: I saw it reported that Bruce Ivins told his supervisor that the powders were very difficult to get into water. Ivins had to do this when he did the spore counts. Basically, you take a small piece of powder, weigh it out, then add it to a flask of water and get the spores to disperse thoughout the water, making a slurry. Then you can take dilutions of this. Ivins said it was difficult to get the spores to go into the water. That meant the spores were hydrophobic (they repel water) -- it is my lay understanding that this is what the silicon oil would do.; January 20, 2009 at 5:21 AM
Anonymous said...: Replecote is oil based and requires removal of the oil from the prep by a volatile solvent that does not kill the spores. Hydrophobic solvents could penetrate a spore.

Ed is correct that there presently is no reason to think Dr. Ivins knew this or used this method -- although others do.; January 20, 2009 at 8:44 AM
Anonymous said...: Anonymous wrote: "Dr. Nass did indeed say "Coated and uncoated spores may look alike." Dr. Nass was entirely correct."

So, this is all about word games? You post pictures of uncoated and coated spores that clearly DO NOT look alike, and you say they MAY look alike if the pictures were clearer???

That's your idea of evidence? Of logic?

The images indicate that they will NOT look alike, they'll look totally different.

An in another thread you wrote:

"2) It is a FACT that these spores look identical to uncoated spores."

And then you show us pictures which indicate just the opposite. They do NOT look identical to uncoated spores at all.

And, you change your wording to say, "they look more like uncoated spores when treated like this."

The point is: The spores may look more like uncoated spores than spores coated with fumed silica, but they still do not look like uncoated spores in any way.

Playing word games by claiming one thing, then when that is shown to be total nonsense, claiming you actually said something else is just plain silly.; January 20, 2009 at 10:00 AM
Anonymous said...: Here's an accurate enlargement of the coated and uncoated spore images. Ed Lake's "enlarged .jpg version" includes various distortions, intentional or unintentional. In particular, Ed Lake's new pictures have a different aspect ratio than the originals -- Lake's pictures are compressed horizontally as compared to the originals. Ed Lake's new pictures also feature a substantial loss of detail as compared to the originals.

Loss of detail in digital enlargement occurs for various reasons. The most common problems include (i) enlarging a copy of the original saved in a "lossy compression" format ( - http://en.wikipedia.org/wiki/Lossy_compression - ), with the result that the "enlargements" include a compounded loss of detail; and (ii) failure to use an appropriate enlargement "filter", the net result being that details in the original become smeared in the enlargement.

In the attached, I saved each of the originals from the .ppt file in bitmap format; used a "bicubic" filter to enlarge each image to the same height as the images in Ed Lake's "enlarged .jpg version", without changing the aspect ratio of the originals; copied the enlargements to an empty background image; then saved the composite as a .jpg file using a compression value of 5%.

It is apparent that the coated spores cannot accurately be characterized as "smooth as pearls". Further no basis is seen for concluding any of the spores are "fused together" (the uncoated spores, on the other hand, do appear "fused together" at various points, but without additional imaging one simply cannot say).; January 20, 2009 at 12:34 PM
Anonymous said...: Anonymous wrote: "Lake's pictures are compressed horizontally as compared to the originals."

Hmmm. SOMEONE's images are distorted. My version is just a straight screen capture from the Powerpoint file. It's exactly what I see on my computer screen when I use Powerpoint, and then doubled in size horizontally AND vertically.

I don't have another copy of the uncoated image, but I have another .jpg copy of the image of the coated spores that I got from somewhere else. Strangely, it's a bit wider and a bit narrower vertically than the Powerpoint version.

My guess is that whoever created the Powerpoint file distorted one or both of the images to make them the exact same size. Or maybe Powerpoints does that automatically somehow.

Anonymous also wrote: "It is apparent that the coated spores cannot accurately be characterized as "smooth as pearls". Further no basis is seen for concluding any of the spores are "fused together" (the uncoated spores, on the other hand, do appear "fused together" at various points, but without additional imaging one simply cannot say)."

Both images show spores which might be described as "fused together," but the Repelcoat spores are fused together in a very different way. They're fused together to the point where they look like two doorknobs inside a sock.

Click HERE to see what I mean.

I agree that better resolution is needed, but you are the one who attempted to use these images to prove some point about similarity, even though there is no similarity whatsoever - except maybe in size.; January 20, 2009 at 5:15 PM
Anonymous said...: I noticed something in the new "accurate enlargement" posted by Anonymous. There is a BIG difference in the size of the uncoated and coated spores, even though the scales are identical

To verify it, I copied the scales and placed them one above the other on the version HERE. They match exactly.

I then copied spores from the coated image and placed them next to spores on the uncoated image.

Clearly, the coated spores are MUCH larger.

It appears that we're looking at something equivalent to peanuts and candy-coated peanuts. The "candy-coated peanuts" are not only LARGER, occasionally two or more "peanuts" will be coated together as one double-size item.

The idea that the coated and uncoated spores "may" look alike is becoming more and more absurd. Clearly, they would NOT look like under high resolution.; January 20, 2009 at 5:47 PM
Anonymous said...: -> Ed Lake

We know beyond any doubt that the images were not made on the same instrument. We don't know whether the images were made on instruments having identical focal lengths or whether the image settings were otherwise identical. It takes only a smidgen of knowledge of photography to know that these and similar factors result in significantly different image sizes at identical magnifications.

And nothing in your comments explains why you posted images with significantly diminished resolution and distorted aspect ratios that just happened to support your arguments.; January 20, 2009 at 6:39 PM
Anonymous said...: Anonymous wrote: "And nothing in your comments explains why you posted images with significantly diminished resolution and distorted aspect ratios that just happened to support your arguments."

I posted comments about this, but either the comments didn't get to Dr. Nass or she chose not to post them.

I don't know where the distortions come from, but they APPEAR to come from you. I'll double check the numbers, and get back to you.

But, right now it appears that your images of the COATED spores are 675x505x24 (when not doubled in size).

BUT, the original .jpg picture of the Repelcoat COATED spores that has been circulated is 712x484x24. So, it's not only wider, it's also narrower.

The difference doesn't have any impact on my arguments. Any way you look at it, the coated spores are CLEARLY COATED. The difference in the images is as clear as when looking a peanut and an M&M candy coated peanut.; January 21, 2009 at 10:15 AM
Anonymous said...: Anonymous wrote: "And nothing in your comments explains why you posted images with significantly diminished resolution and distorted aspect ratios that just happened to support your arguments."

I posted comments about this, but either the comments didn't get to Dr. Nass or she chose not to post them.

I don't know where the distortions come from, but they APPEAR to come from you. I'll double check the numbers, and get back to you.

But, right now it appears that your images of the COATED spores are 675x505x24 (when not doubled in size).

BUT, the original .jpg picture of the Repelcoat COATED spores that has been circulated is 712x484x24. So, it's not only wider, it's also narrower.

The difference doesn't have any impact on my arguments. Any way you look at it, the coated spores are CLEARLY COATED. The difference in the images is as clear as when looking a peanut and an M&M candy coated peanut.; January 21, 2009 at 10:16 AM
Anonymous said...: Hmmm. There may be a glitch in the software for this site.

I posted the message about the image sizes, but I didn't get back the screen with the notification at the top that I have to wait for the moderator to read it. So, I did a BACK, retrieved the message I had posted and posted it again. The second time I got the moderator message.

So, I think the explanation of the difference in sizes that I posted yesterday may have been caught by the glitch.; January 21, 2009 at 10:23 AM
Anonymous said...: Okay, I just repeated all the steps for turning a PowerPoint image into a .jpg image. I displayed the Powerpoint image on my computer, and I did a screen capture of it. I then extracted only the RepelCoat image from the screen and saved it as a separate file. It measures: 678x508x24.

I then cut out only the Repelcoat image from the double-size image I put on my site, and it measures: 1350x1015x24

Half size would be: 675x508x24

That's nearly identical to the Powerpoint file ratio.

Then I cut the RepelCoat image out of Anonymous's second try, where he posted a double size .jpg image. That image is 1486x1015x24

Half size is 735x508x24.

So, Anonymous posted a different size image the first time. OR, different software changes the size of a Powerpoint image.

Again, the RepelCoat .jpg file I received from other sources is: 712x484x24.

That's close to the .jpg file from Anonymous. .9687075 difference in width, .9527559 difference in height.

The conclusion is that Anonymous either distorted the image size when he created the Powerpoint file, or the Powerpoint software did it for him. I just used what he provided.

And, as stated before, the difference in size doesn't change my argument in any way whatsoever. It is still abundantly clear that the difference between the coated and uncoated spores is as great as the difference between a shelled peanut and M&M candy coated peanut.; January 21, 2009 at 11:31 AM
Anonymous said...: Anonymous,

Can you pick out the differences between the objects in the two pictures located HERE?; January 21, 2009 at 12:12 PM
Anonymous said...: Hmmm.

I received an email this morning advising me that the "Anonymous" who accused me of altering the pictures HE altered is not the same "Anonymous" posting to another thread.

This 2nd (or is it 20th) "Anonymous" also apparently didn't realize that if the images were BOTH distorted equally, comparisons would still be valid.

I still find it totally bizarre that someone would post two pictures and claim that the spores in the two pictures MAY look alike, when they look nothing alike. What is the point?

Are we supposed to believe they look alike even though they clearly do not look alike?; January 22, 2009 at 9:28 AM
Anonymous said...: Hmmm.

I received an email this morning advising me that the "Anonymous" who accused me of altering the pictures HE altered is not the same "Anonymous" posting to another thread.

This 2nd (or is it 20th) "Anonymous" also apparently didn't realize that if the images were BOTH distorted equally, comparisons would still be valid.

I still find it totally bizarre that someone would post two pictures and claim that the spores in the two pictures MAY look alike, when they look nothing alike. What is the point?

Are we supposed to believe they look alike even though they clearly do not look alike?; January 22, 2009 at 9:29 AM
Anonymous said...: Unlike the situation with peanuts and M&M candies, anthrax spores cannot be viewed without the aid of analytical imaging devices. It is therefore imperative to carefully examine all of the detail revealed by the imaging device in order to evaluate the samples.

Blurred enlargements that obfuscate analytical information isn't a tool that would be used in objective scientific evaluation. Blaming myriads of others and other causes for one's own mistakes also reflects a lack of objectivity.

Arguments about m&ms and peanuts cannot change the actual information revealed in the images: (i) each image includes some spores that are larger or smaller than some of the spores in the other image; (ii) objective examination of the coated spore images reveals what appears to be a rough and dimpled surface in some cases; (iii) instances of spores partially superimposed vertically above and below each other are found in both slides, but one cannot properly infer 'fusion' of the superimposed spores since the superimposition prevents determination of whether the hidden portions of the spores are even in contact.

Mere arguments cannot change the facts. The Scandia imaging scientists have now backed away from their original statements of no additives, and very high quantitative silicon uptake from natural sources as the explanation for the high silicon levels in the attack spores.

The silicon content of the attack spores is a significant anomaly that needs further investigation.; January 22, 2009 at 10:11 AM
Anonymous said...: The military scientist whose lab did the anthrax simulant experiment (resulting in the nice picture above) advises me that if you run an infrared spectroscopy on them they contain a large peak of silica, but no silica particles are to be seen. He says they "float like a butterfly and sting like a bee." They look very different from the ones in the journal article by the Dugway authors that used silica particles using the method historically used since the 1960s.

The Sandia scientists oversimplified in referring to only spores using the traditional Dugway method as "weaponized." (At the very least, the term is not meaningful unless expressly defined). (Treated to aid dispersability does not cut it as a definition). The spores pictured above are BETTER than the ones Sandia calls "weaponized."

At the end of the day, we should credit the FBI WMD's head conclusion that it could have been in the culture medium. Moreover, the Sandia scientists did excellent work at precisely identifying the location of the silicon and deserve our thanks. Their work will prove critical to the investigation. (It it in no way, based on what has been disclosed, supports a theory that Ivins is guilty).

Everyone should just avoid overbroad conclusions based on whether they edit Wikipedia, use screen names, or anonymously leaves a young woman half his age a gift card for Lands End.

Everyone should read the DARPA-funded research that is not so secret at all. In 2005, they were even funding a new and improved windshield wiper fluid.
Those of us who are non-scientists should stick to making new articles available or consulting with scientists and reporting what they say.; January 22, 2009 at 11:16 AM
Anonymous said...: Anonymouse wrote: "The military scientist whose lab did the anthrax simulant experiment (resulting in the nice picture above) advises me that if you run an infrared spectroscopy on them they contain a large peak of silica, but no silica particles are to be seen."

There are no candy "particles" visible on the M&M candy coated peanuts either. It's one solid coating of candy. That's what the Repelcoat image looks like, and no one is questioning that such spores would show a silicon peak. The questions are: Why should anyone care? What reason is there to believe this coating process has anything to do with the attack anthrax.; January 22, 2009 at 11:47 AM
Anonymous said...: An "Anonymous" wrote: "The Scandia imaging scientists have now backed away from their original statements of no additives, and very high quantitative silicon uptake from natural sources as the explanation for the high silicon levels in the attack spores."

I just contacted a scientist at Sandia (NOT Scandia), and he says what you say is UNTRUE. They have NOT "backed away" from their original statements, etc.

An "Anonymous" also wrote: "Blurred enlargements that obfuscate analytical information isn't a tool that would be used in objective scientific evaluation."

THEN WHY DID YOU POST THE BLURRED ENLARGMENTS?

This is my second try at responding to the comments by "Anonymous."; January 22, 2009 at 12:15 PM
Anonymous said...: There is another possible explaination for the silicon contained within the spore coat.

It could be the result of silicic acid being introduced during the spore concentration / purification process.

Silicic acid is a common contaminant of a very common compound occasionally used in some spore purification procedures.

It is very unlikely that such procedures were in use at USAMRID, however.; January 22, 2009 at 3:51 PM
Anonymous said...: Anonymous wrote: "It is a known FACT that the exosporium on the spores is penetrated by small molecules - the polymerized glass coating thus forms BENEATH the exosporium - exactly as Sandia observed."

This makes no sense whatsoever.

First: What polymer consists of "small molecules?" The definition of polymer is that it is a LARGE molecule.

Second: The idea that these molecules (large or small) will penetrate the exosporium means nothing without an explanation of why ONLY the spore coat will be coated?

Here's an analogy: Let's say that instead of a spore with its exosporium we have a baseball covered with a loose-fitting gauze cover.

The claim is that the polymer will penetrate the gauze cover and coat the baseball.

Fine. No problem. But why won't the gauze cover also be coated?

Bascially, you're talking about dipping the gauze-covered baseball in a bucket of acrylic paint. The paint penetrates the gauze and coats the baseball. But, the gauze will also be coated.

The tests done at Sandia show that the exosporium is NOT coated. NO silicon was detected in the exosporium. The silicon was concentrated INSIDE the spore coat, not ON the spore coat.

So, the idea that this polymer coating process somehow "penetrated" the exosporium without coating the exosporium seems to be total nonsense. How is that accomplished? What magic kept the polymer coating from also coating the exosporium?; January 22, 2009 at 5:09 PM
Anonymous said...: BugMaster wrote: "It could be the result of silicic acid being introduced during the spore concentration / purification process."

I think that will introduce the same problem as with putting a layer of polymerized silicon on the spore coat without also coating the exosporium.

Any process of applying silicon AFTER the spore is formed will either coat the exosporium or BOTH the exosporium AND the spore coat.

And Sandia can PROVE that the exosporium contained NO silicon. The silicon was INSIDE the spore coat.; January 22, 2009 at 5:18 PM
Anonymous said...: Blurred enlargenments are posted on Ed Lake's web page (http://www.anthraxinvestigation.com/CoatedUncoatedSporesBIG.jpg) and referenced here (at Dr. Nass's site) by Ed Lake to support Lake's assertions made here (at Dr. Nass's site):

"The question is: How could anyone think that the coated and uncoated spores in these images look alike?

Just look at an enlarged .jpg version HERE.

The uncoated spores are irregular and rough in shape, often with tiny points, and they look like little wads of cotton.

The coated spores look as smooth as pearls, and some near the top of the image even seem to be fused together with the coating filling in the areas between the spores."

Lake now shouts a bizarre claim that someone else posted the blurred enlargements "THEN WHY DID YOU POST THE BLURRED ENLARGMENTS?"

Suffice it to say that that the facts speak for themselves.; January 22, 2009 at 5:21 PM
Anonymous said...: BugMaster,

I think soluble orthosilicic acid will end up being the key to the entire silicon question. In order for the silicon to end up INSIDE the spore coat, it almost certainly had to be in some water soluable form. That means - once again - that it came from the nutrients or the water.; January 22, 2009 at 5:46 PM
Anonymous said...: In response to Ed Lake's most recent argument of the minute --

Polymers can be (and often are) formed in situ from a reactive mixture of prepolymers. Polymerization of low molecular size and weight prepolymers below the exosporium could readily produce polymers that are physically trapped beneath the exosporium, i.e., the molecular size of the polymer would prevent them from exiting back through the exosporium. Polymerized prepolymers (i.e., polymers) of the same large molecular weight and size, exterior to the exosporium, could still be susceptible to removal by liquid media.

It's neither magic nor terribly complicated.; January 22, 2009 at 5:49 PM
Anonymous said...: Anonymous wrote: "Suffice it to say that that the facts speak for themselves."

Yes, they do. Here is what YOU posted:

"Here's an accurate enlargement of the coated and uncoated spore images."

But you are only trying to avoid the real subject: the blurred images of the coated spores (original or enlarged) that you posted and your expectation that people are supposed to see what you BELIEVE is there and not what is ACTUALLY there.; January 22, 2009 at 6:00 PM
Anonymous said...: Ed:

Was it ever stated that the silicon detected in the spore coat was POLYMERIZED? I don't think so, rather, wasn't it stated that silica was detected in the spore coat?

As far as binding to the coat and not the exosporidium:

I may be mistaken, but I seem to recall from a micro lecture that the spore coat is composed of dense, basic proteins. Basic, as in the opposite of acidic. Basic proteins contain a large amount of basic amino acids, amino acids with sidechains containing an amino (NH2) group. Some of these amino groups become ionized at physiological pH, becoming NH3+. Note the positive charge, thus the term: Basic.

Silicates contain oxygen, thus have considerable negative character. They will therefore bind to the basic amino groups.

For more info about the binding of silicates to basic compounds, search for the topic "end-capping" and "HPLC column packing material".; January 22, 2009 at 6:11 PM
Anonymous said...: Ed Lake,

Your enlarged .jpg is clearly blurred compared to the accurate enlargement that I posted. My enlargement was made exactly as set forth in my post; using digital techniques commonly used for preserving detail in digital enlargements. If you cannot see or admit that your "enlarged .jpg" is blurred, that's your problem.

You also incorrectly claimed, relying on your blurred enlargements, "The coated spores look as smooth as pearls, and some near the top of the image even seem to be fused together with the coating filling in the areas between the spores."

The facts do speak for themselves.; January 22, 2009 at 7:46 PM
Anonymous said...: I'll just add this -- there are numerous other mechanisms whereby silicone polymers or oligomers could be deposited beneath the exosporium but not on the surface.

For example, oleophillic (oil loving) silicone materials could be added to aqueous media as tiny emulsified particles, or micelles. Because of their oleophillic nature they wouldn't necessarily interact with, or be deposited on, the spore surface. Nevertheless, if the micelles were sized to penetrate the exosporia, but only with difficulty, the concentration of these silicone materials could build up beneath the exosporia over a period of days. When the spores were later removed from the aqueous media, all of the water could easily exit the exosporia, but a great deal of the silicone materials would remain as the penetration rate of the micelles would prevent their rapid exit. No silicone materials would remain on the exterior of the spores because there wasn't substantial surface interaction in the first place.

There are numerous mechanisms and possibilities. BugMaster elucidates further mechanisms. We know the spores have a very high silicon content but we don't know how or why that happened - and neither does the FBI. It's an anomaly that begs further investigation.; January 22, 2009 at 7:49 PM
Anonymous said...: Someone should ask the expert on oil-and-water nanoemulsions (including silicon oil) who was working with Bruce Ivins using virulent Ames in the research funded by DARPA if this explanation about micelles, whatever they are, makes sense.

I thought micelles was a Beatles song.

I FOIAed the relevant documents but got squat.; January 22, 2009 at 9:07 PM
Anonymous said...: BugMaster wrote: "Was it ever stated that the silicon detected in the spore coat was POLYMERIZED? I don't think so, rather, wasn't it stated that silica was detected in the spore coat?"

The statements about polymers come from two different people with theories who both call themselves "Anonymous" on this thread and another thread on Dr. Nass's blog.

Silicon and Oxygen were detected in the spore coat. Some people early in the case ASSUMED that silicon and oxygen meant silica.

But there are many ways for Oxygen to be in the spores. It's the Silicon that presents the BIG questions. How did it get there? Where did it come from?

Of course, people have been arguing for SEVEN years that it MUST be from some supersophisticated weaponization process. And they continue to argue that using one theory after another as to how it got there. The latest screwball theory is about a polymerization process.

Comprendo?; January 23, 2009 at 10:19 AM
Anonymous said...: Anonymous wrote: :Your enlarged .jpg is clearly blurred compared to the accurate enlargement that I posted.

So what? The .jpg I created via a screen capture of the PowerPoint screen was the only way I could turn a .ppt file into a .jpg file. The blurring is minor, and BOTH IMAGES in the set are equally blurred, so comparisons are still valid.

In the slightly sharper image you call an "accurate enlargment," the coated spores are STILL "as smooth as pearls." Everything I said about the other image still applies.

As you say, The facts do speak for themselves.

Anonymous also wrote: "I'll just add this -- there are numerous other mechanisms whereby silicone polymers or oligomers could be deposited beneath the exosporium but not on the surface."

So you claim. But you do not explain how they would coat the spore coat and not the exosporium. You just say such things exist and you speculate that they MAY have been used.

You say: "We know the spores have a very high silicon content but we don't know how or why that happened."

We know the attack spores had a high silicon content. We don't know that it was "very high," because we don't have any solid information on what process produce what kind of silicon content.

Some plants are as much as 20% silicon. What basis is there for claiming that 2% or even 1% silicon in a spore coat is not possible except via some supersophisticated weaponization process?

I agree that more information is needed, but I also know that MORE INFORMATION IS COMING. We just need to wait for the scientific reports to get through the peer review process and into some scientific publication.

Dreaming up one bizarre possibility after another to accuse the government of hiding some secret bioweapons program does no one any good.; January 23, 2009 at 11:00 AM
Anonymous said...: Bug Master,

David Kelly was the source in the Spring 2002, based on briefing by NATO personnel. See the articles at the time.; January 23, 2009 at 10:59 PM
Anonymous said...: If anyone has any questions regarding the controlled experiment done, they can contact expert Dr. Kiel, whose lab did the study. Having Mr. Lake, a lay person not familiar with the work, is a waste of time.; January 23, 2009 at 11:03 PM
Anonymous said...: For the sake of clarity and precision, its important to understand that Sandia National Labs' TEM images do not establish that the surface of the attack spores was free of silicon. Sandia merely claimed that the silicon was not exterior to the spores.

I've added arrows to Sandia's TEM image here, to identify some of the numerous instances in which it is clear that the silicon in the attack spores extends fully to the surface of the spores. Sandia's original SEM and TEM images can be downloaded here. Sandia's complete publication, including the images, can be viewed here.; January 24, 2009 at 10:11 AM
Anonymous said...: Anonymous wrote: "For the sake of clarity and precision, its important to understand that Sandia National Labs' TEM images do not establish that the surface of the attack spores was free of silicon. Sandia merely claimed that the silicon was not exterior to the spores."

Here is what Sandia said in another publication:

"Using highly sensitive transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM), the researchers came to a startling realization: The silicon had grown organically inside the Bacillus anthracis samples, nothing had been added to weaponize the spores. "The silicon was not on the outside of the spore," says Michael, who headed up Sandia's investigation, "but rather incorporated on the inside."

Later in the article there is this:

"Finding the Right Technology
By the time the Sandia researchers began their work in February 2002, "we had heard just like everyone else that the spores had been weaponized," says Michael, who had proposed a study of the elemental composition of any materials found growing outside the spores.

"The first step was to find the silicon. Michael was aware that FBI researchers had analyzed the samples with both scanning electron microscopy (SEM), which scans surfaces with a high-energy beam of electrons, and with energy dispersive x-ray spectroscopy (EDS), which analyzes x-rays emitted by a substance after it is hit with charged electrons. But at that point, no one had studied them with a scanning transmission electron microscope (STEM), which transmits a focused beam of electrons through a small part of a specimen to form an image and could provide compositional information by examining the spores a few nanometers (one nanometer is 40 millionths of an inch) at a time, a higher resolution than SEM could provide, Michael says.

"This enabled Sandia researchers to not only detect the presence of a foreign substance such as silica, but also to determine its location on or inside the spore. "In the FBI's mind, it was important not only that trace amount of elements were present, but also…[to determine]…where those elements were located in the sample through microanalysis," says Paul Kotula, a Sandia material scientist who studied the samples with Michael.

"The researchers could find no way that the silica could be placed inside the spore without leaving a residue on the spore's outermost layer. (They found none.) Instead, the researchers determined that the silica formed inside the spore naturally. After only a month examining the anthrax samples in March 2002, Michael and his team were convinced, contrary to other reports, that the anthrax used in the attacks had not been weaponized."

And in the roundtable discussion, Dr. Michael also stated that the locations of the silicon were very similar to a study done in 1980. They even obtained a sample from those 1980 tests and confirmed that the location of the silicon was the same -- it was incorporated into the spore coat just the way calcium gets concentrated into your bones and not into your eyeballs. That's where it works best.

If that isn't enough, I'll be glad to put you in contact with Joe Michael at Sandia, and you can ask all the questions you want. Or better yet, why not wait for his scientific articles to be published? They'll describe everything in great detail.; January 24, 2009 at 10:49 AM
Anonymous said...: Thanks Anonymous for your post discussing the Sandia pictures, and your arrows pointing out where the exosporium is missing.

Indeed, pictures of spores with and without exosporium can be viewed in this paper:

https://kb.osu.edu/dspace/bitstream/1811/31927/1/WCS_Paper_Hayes_08.pdf

I have prepared pictures of a side by side comparison of these pictures with the Sandia pictures. The Sandia pictures most clearly resemble the pictures from the paper at the link WITHOUT an exosporium. This is contrary to Sandia's claims that the exosporium is intact in the mailed spores.

I'll email my comparison pictures to Meryl and ask if she'll post them.

Finally, contrary to the repeated claims by Ed Lake (who seems to be attempting his usual tactic of deliberately misleading), that somehow polymers are large molecules and couldn't get through the exosporium, that's the whole point. When you coat spores with polymerized glass, you don't start with the polymer form - you start with the monomer.

As described at the link below, the dimethyldichlorosilane (a monomer)polymerizes into Polydimethylsiloxane (a polymer) in situ, undergoing a hydrolysis reaction - taking up water and losing HCl.

Dimethyldichlorosilane is small molecule that easily penetrates the exosporium. It then polymerizes in situ (Ed, that's Latin for "on site") into polydimethylsiloxane. The latter IS a large molecule and it forms a continuous coat on the inner spore coat. After the spores are washed and dried the polydimethylsiloxane stays there and cannot come off. Any loose remnants of exosporium would contain no polydimethylsiloxane.

http://www.silicones-science.com/chemistry_pdms.html; January 24, 2009 at 10:51 AM
Anonymous said...: An apparently different "Anonymous" wrote: "Having Mr. Lake, a lay person not familiar with the work, is a waste of time."

If I have any questions about any scientific aspects of the case, I ask the people who would know the answers, people at Sandia, the FBI and NAU. I don't ask people who are not involved with the case and who only have baseless theories.

Months ago, I contacted the person you mention to ask what his thoughts were, but all he would tell me is that he provides "alternative science based points of view" that he thinks the public should be aware of.

I don't see any way to interpret that to mean he provides definitive scientific explanations.; January 24, 2009 at 11:08 AM
Anonymous said...: One "Anonymous" wrote: "I've added arrows to Sandia's TEM image here, to identify some of the numerous instances in which it is clear that the silicon in the attack spores extends fully to the surface of the spores."

And apparently a different "Anonymous wrote: "Thanks Anonymous for your post discussing the Sandia pictures, and your arrows pointing out where the exosporium is missing."

Wow. One "Anonymous" says that the TEM images somehow show a silicon signal, and the other "Anonymous" says the images show that the spores in the TEM image are wholly or partially missing the exosporium.

I doubt that either belief is true.

Unfortunately, it's Saturday, so it may be difficult to get a response from Sandia. But, I'll try.

As far as I know, the TEM images are not STEM images and only show a cross section of spores from the attack anthrax. Nothing more, nothing less. No STEM spectography, no spores without their exosporium.

FYI, here's what a map of silicon in a spore coat looks like under an STEM. Click HERE.

That STEM image came from HERE

In the roundtable discussion, Dr. Michael said that the attack anthrax look similar except for the fact that current technology allows for a much MUCH sharper image.; January 24, 2009 at 11:35 AM
Anonymous said...: One of the Anonymi wrote: "Finally, contrary to the repeated claims by Ed Lake (who seems to be attempting his usual tactic of deliberately misleading), that somehow polymers are large molecules and couldn't get through the exosporium,..."

I made no such claim. I said just the opposite. I compared the coating process to dipping a baseball covered with gauze into a bucket of acrylic paint. I SPECIFICALLY stated that the paint would penetrate the gauze and would coat the baseball, but the paint would ALSO coat the gauze.

You also wrote: "After the spores are washed and dried the polydimethylsiloxane stays there and cannot come off. Any loose remnants of exosporium would contain no polydimethylsiloxane."

That's a claim with no basis. If the washing doesn't removed the polydimethylsiloxane, how does it remove it from the outside and inside of the exosporium?

It's a painted baseball inside a painted cover of gauze. You're claiming that washing only washes the gauze, not the baseball. NONSENSE. The paint/polydimethylsiloxane would still be detectable on the exosporium.

You need to PROVE that it would not. You cannot just make a baseless claim and expect people to believe it.; January 24, 2009 at 11:55 AM
Anonymous said...: Just in case someone is going to claim that a TEM image and an STEM image are identical, just remember that the very first TEM examination of the Daschle anthrax was at USAMRIID by Tom Geisbert. And Tom Geisbert had no way to examine spectra. He had to go to AFIP to use their EDX.

Sandia has a Scanning Transmission Electron Microscope (STEM), but the image in their article was evidently just taken using the Transmission Electron Microscope (TEM) cababilities.; January 24, 2009 at 12:07 PM
Anonymous said...: "The researchers could find no way that the silica could be placed inside the spore without leaving a residue on the spore's outermost layer. (They found none.)"

As previously noted in this thread, there are several possible ways this could have been accomplished. The researchers may just have been unaware of the actual technique(s) used, and were therefore unable to duplicate the results.

I don't think it would be socially responsible to post specifics about these possible techniques in a public forum, however.

BTW: Dimethyldichlorosilane is a highly reactive compound, releasing toxic HCL as it polymerizes. I've silonized quite a bit of glassware in the past, and when using this stuff, one has to be careful. If it was in fact used to treat the spores , it would have had to been applied in a very controlled manner to avoid killing them (or, like Ed suggests, perhaps completely covering them in polymer). I can't imagine this being accomplished easily, but I could be wrong.; January 24, 2009 at 12:25 PM
Anonymous said...: Okay. I received a response from Sandia.

First, the TEM image in the article is a STEM image. I was wrong about that. BUT, I was right in stating the image does NOT - REPEAT NOT show any spectrographic signal.

Technically, the TEM/STEM image is an "annular dark field image of the spores."

The brighter areas in the image are simply areas where the material is more dense.

Second, since the exosporium is flexible, sometimes there is a gap between the exosporium and the spore coat, and sometimes there is NO GAP. All that the other Anonymous is pointing out are areas where the exosporium is flat against the spore coat and cannot be distinguished from one another.

My Sandia contact quoted Dr. Popov:

"Popov said:. "The Sandia pictures completely agree with my expectation of partially collapsed exosporium. It may be still there but hard to detect in the dry spores."

So, summing up: BOTH Anonymi are WRONG. The TEM/STEM image shows no spectrographic signal, and the TEM/STEM image shows NO GAPS in the exosporium.; January 24, 2009 at 12:57 PM
Anonymous said...: Sandia National Labs has provided no evidence to support its assertion that silicon in the attack spores was formed naturally. To the contrary, Sandia's Paul Kotula said, "We looked at over 200 samples in our lab that were various attempts to reverse-engineer the process under which these powders were made and did not find a match." It's been over 5 months since Sandia first announced its "formed naturally" theory. To date, Sandia has provided not a single shred of scientific data in support its theory, not one. Sandia's "formed naturally" hypothesis remains as such until proven. That's how science works.

As to Sandia's explanations of its TEM image, if we take Sandia at its word that the image shows the attack spores, including their in tact exosporia, it's quite clear that the silicon content does extend fully to the surface of the exosporia as shown here.

Whether the silicon content was formed naturally, or impregnated into, the exosporia of the attack spores is unknown at this point. Sandia's "formed naturally" hypothesis hasn't even been shown to be possible, despite over 200 attempts. Future efforts by Sandia and/or others may provide answers. But at this point, Sandia's hypothesis remains unsupported by scientific evidence.; January 24, 2009 at 1:50 PM
Anonymous said...: Another comment about the exosporium:

My Sandia contact advises me that in some of the better quality images they took, the hair-like appendages can be clearly seen.

The hair-like appendages only exist when the exosporium is also present.; January 24, 2009 at 2:27 PM
Anonymous said...: I posted a long message about what Sandia told me this morning about exosporia, but for some reason it hasn't appeared.

But, another message from Anonymous with his same argument HAS appeared:

"As to Sandia's explanations of its TEM image, if we take Sandia at its word that the image shows the attack spores, including their in tact exosporia, it's quite clear that the silicon content does extend fully to the surface of the exosporia as shown here."

So, to repeat what I wrote but which didn't show up here,

The TEM image from Sandia does NOT use STEM capabilities. It does NOT show any a spectrographic signal.

The image is an "annular dark field image of the spores," also known as a "Z contrast" image, meaning that the brightness is related to the density of the material.

So, Anonymous #1 is totally WRONG in believing that the brightness has anything to do with silicon.

Secondly, Anonymous #2 is totally WRONG in believing that the images show spores without any exosporium.

The exosporium on Bacullus anthracis is flexible, and while there may be a gap between the exosporium and the spore coat in some places, in other places the exosporium may be flat against the spore coat, which means that the TEM is unable to distinguish any boundry between them.

My contact at Sandia gave me a quote from a statement Dr. Popov made:

"Popov said: "The Sandia pictures completely agree with my expectation of partially collapsed exosporium. It may be still there but hard to detect in the dry spores."

It is ridiculous to look at a TEM images from Sandia and to ASSUME that silicon can somehow be seen when it is NOT a spectragraphic image. It is also ridiculous to assume that because the exosporium is touching the spore coat at some point, the exosporium doesn't exist at that point.

After writing that information, I posted the other information where my Sandia contact explained that some of their images of the attack spores are so clear that they can actually see the hair-like appendages which protrude from the exosporium and which do not exist if there is no exosporium.

My contact at Sandia didn't comment upon the statement about the 200 attempts at "reverse engineering," but that is just a problem with terminology. Some people use the term "reverse engineer" when no such thing took place. All that was done was to create samples for comparison. In reality, there were NO attempts at reverse engineering. That was stated very clearly by a top scientist at Dugway:

"Daniel Martin, a microbiologist in Dugway's Life Sciences Division, tells C&EN that Dugway was asked "to produce materials to see how they compared with the materials the FBI had in its possession." But, Martin says, Dugway did not reverse or back engineer the attack powder. "Back engineering implies that you know exactly what the material is and can replicate the material exactly, step by step." That isn't what Dugway did, he says.

"Instead, Martin says, Dugway used the Leahy powder as the culture starter to "produce several different preparations using different media, and different ways of drying and milling the preparation" that the FBI could use for comparison purposes. Dugway, he says, never analyzed the Leahy powder and did no comparative analyses between the preparations made and the Leahy powder."

So, again it's just bad terminology. There were NO attempts to reverse engineer the attack anthrax. Over 200 samples of anthrax were created using different preparations so that scientists could analyze the differences.

There is NO problem in creating spores that look exactly like what was in the attack anthrax. The only problem is with creating the "silicon signature" in a way that can be repeated over and over. In theory, to do that, they'd have to know with absolute certainty every step that was followed, every piece of equipment that was used, and every detail about the water, media, temperatures, etc. They don't know all those details, so they cannot "reverse engineer" the attack anthrax.

But that doesn't mean they don't know who made it and where it was made.; January 24, 2009 at 4:26 PM
Anonymous said...: The TEM image from Sandia is NOT a spectographic image. It looks nothing like a spectographic image. Yesterday, I contacted a scientist at Sandia, and he immediately confirmed that the TEM image does not show any spectrographic signals. "It is in fact an annular dark field image of the spores," the Sandia scientist advised me. "These images are actually what we call Z contrast images, meaning that the brightness of the image is related to the atomic number of the illuminated region. It is really mass thickness as well, so thicker areas will appear brighter. So, it may be that those areas are thicker, or have a higher atomic number or both. Without x-ray microanalysis not much chemical information can be reliably concluded from that image."

There is simply no way anyone can look at the TEM image and point to silicon.; January 25, 2009 at 1:58 PM
Anonymous said...: This just gets curiouser and curiouser...

According to Ed Lake's Scandia contact, one can't properly derive chemical information from the TEM images because one can't distinguish mass thickness of an imaged area from the mass of the elements in the imaged area...

But in the FBI briefing of August 18, Sandia's Dr. Michael said just the opposite....

"DR. MICHAEL: First of all, we did find we did find that the spores contained silicon and oxygen. Our quick SEM analysis, that's Scanning Electron Microscopy, we detected silicon and oxygen within the spores. Later when we had thin sections for high resolution microanalysis in the scanning transmission electron microscope we then could localize that silicon and oxygen to the spore coat, which is a layer on the spore that's within the spore itself."; January 25, 2009 at 4:37 PM
Anonymous said...: Ed Lake wrote:

"So, Anonymous #1 is totally WRONG in believing that the brightness has anything to do with silicon.

Secondly, Anonymous #2 is totally WRONG in believing that the images show spores without any exosporium."

This is wrong on both counts. z-contrast makes higher atomic number elements brighter. Silicon has a higher atomic number than carbon thus the silicon is brighter.

The published pictures of spores with and without exosporiums are completelty different. The Sandia pictures most clearly match the pictures WITHOUT exosporium.; January 25, 2009 at 4:40 PM
Anonymous said...: Anonymous wrote: "This just gets curiouser and curiouser.."

That's only because you evidently somehow believe that the single TEM image that Sandia put on their web site is the ONLY TEM image Sandia ever created.

As stated in the roundtable discussion, Sandia produced STEM images detailing the locations of the silicon in the spores. The STEM images are VERY different from what Sandia put on their site. They are MAPS of the silicon locations. Those STEM images will be part of the scientific articles that scientists at Sandia write and publish in scientific journals describing their findings in the Amerithrax case.; January 25, 2009 at 6:05 PM
Anonymous said...: In response to Lake's comment:

"That's only because you evidently somehow believe that the single TEM image that Sandia put on their web site is the ONLY TEM image Sandia ever created."

That's a thought that plainly originated in your head; and as such reflects only your limited view of the world and others.

As to the those articles that Sandia are going to write and publish in scientific journals, we are currently, and have been for five months, waiting to see them. Please pardon me for wanting to see the articles themselves instead of your predictions.; January 25, 2009 at 7:54 PM