CASE CLOSED … what really happened in the 2001 anthrax attacks?

Archive for June 30th, 2009

* questions to NAS regarding scientific aspects of the NAS review of FBI science used in investigating the 2001 anthrax attacks; questions posed by CASE CLOSED blog participants

Posted by DXer on June 30, 2009

why the FBI failed to solve the 2001 anthrax caseCASE CLOSED

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questions to NAS regarding scientific aspects

of the NAS review of FBI science

used in investigating the 2001 anthrax attacks;

questions posed by CASE CLOSED blog participants

NAS Publications

NAS Publications

***

The following email was sent (6-30-09) to Mr. William Kearney, Deputy Executive Director & Director of Media Relations, Office of News & Public Information, National Academy of Sciences, 2101 Constitution Ave. NW #182, Washington, DC 20418

***

BILL … here are the questions submitted to the CASE CLOSED blog related to scientific aspects of the NAS-FBI study. A couple of questions stray a bit into procedural aspects, but most are straight science. The questions reflect the varied interests of CASE CLOSED blog participants; collectively I think they add up to a very impressive and rather daunting agenda … LEW

Dear Mr. Kearney,

Let me repeat that I, along with the many readers and contributors to the CASE CLOSED blog, truly appreciate that the National Academy of Sciences (NAS) has undertaken the difficult task of reviewing scientific approaches and conclusions of the FBI investigation of the 2001 anthrax attacks.

In addition to procedural questions related to this review already submitted, there are scientific questions presented below, also submitted, as were the procedural questions, by contributors to the CASE CLOSED blog.

We understand that the list is long and that the study team has yet to be formulated, so we are surely not expecting a speedy response. However, we hope that you will distribute these questions to the study team as it comes into being, and that serious consideration will be given to answering each of these questions as your study proceeds.

Here are the questions …

1. Will the NAS study team evaluate the scientific basis for the FBI’s explanation that the White House staff was taking the anthrax drug Ciprofloxacin prior to the anthrax mailings?

2. Will the NAS study team comment on aspects of the FBI investigation if it is deemed that the investigative methods clearly impacted the scientific analysis?

3. Will the NAS study team provide definitive documentation regarding the science and methodology that allowed the FBI to exclude as suspects the perhaps 100 to 300 other scientists (other than Dr. Ivins) who had isolates from flask 1029?

4. Will the NAS study team be obtaining and evaluating the internal laboratory reports written by the Armed Forces Institute of Pathology (AFIP) on the SEM and EDX analysis of the Daschle and New York Post powders?

5. Will the NAS study team be limited to sources of scientific data provided by the FBI or will the NAS study team have unlimited access to whatever scientific data it deems necessary to perform its tasks?

6. Will the NAS study team address the implications of

6a. anthrax in the letters to the Senators being composed of pure spores, one trillion in a gram

6b. particles of said anthrax milled down (by some method) to the size of one or two spores?

7. Will the NAS study team verify whether anthrax spores in the 2001 mailings were treated post-production to make them more friable?

8. Will the NAS study team verify whether anthrax spores in the 2001 mailings were “weaponized?”

9. Will the NAS study team evaluate if it was feasible for mail sorting machines to have assisted in the pulverization and aerosolization of the BA spores in the postal facility?

10. Will the NAS study team seek to provide sufficient proof of its independent investigation of these scientific issues to dispel an understandable skepticism that it is an inherent conflict of interest for NAS to be taking money from the FBI, the very agency whose science is being evaluated, rather than being directly funded by the Congress?

11. Will the NAS study team permit any scientists employed at Battelle-managed labs to be part of the project team? Would such participation constitute a conflict of interest under NAS policy?

12. Will the NAS study team determine if b. subtilis genetically identical to the contaminant present in the letters mailed to New York was also isolated from the AMI building in Florida?

13. Will the NAS study team consider and scientifically evaluate hypotheses other than those which comprise the “official” theory of the FBI?

14. Will the NAS study team evaluate the scientific methods used by the FBI to reach their stated conclusion that the powder sent to Senators Daschle, Leahy and other recipients contained no special additives and was not weaponized?

14a. did the FBI use accepted scientific method to reach that conclusion?

14b. if the FBI prepared a powder of anthrax or anthrax simulant that acted the same as the powder in the Hart building?

14c. if, in reaching its conclusion, did the FBI used equipment similar to what was available to their suspect Dr Bruce Ivins inside Fort Detrick where they claim the powder was manufactured?

15. Will the NAS study team determine whether, if the FBI did prepare a powder or powder stimulant in this simple manner, it also re-aerosolize (in other words after initially falling on surfaces, then formed a secondary aerosol) under common office activities? (ref. Secondary Aerosolization of Viable Bacillus anthracis Spores in a Contaminated US Senate Office. Christopher P. Weis, PhD; Anthony J. Intrepido, MS, CIH; Aubrey K. Miller, MD, MPH; Patricia G. Cowin, MS, CIH; Mark A. Durno, BS; Joan S. Gebhardt, PhD; Robert Bull, PhD; JAMA. 2002;288:2853-2858; http://jama.ama-assn.org/cgi/content/abstract/288/22/2853).

16. Will the NAS study team determine if more than 80% of the B anthracis particles collected on stationary monitors fell within an alveolar respirable size range of 0.95 to 3.5 µm? (ref. Secondary Aerosolization of Viable Bacillus anthracis Spores in a Contaminated US Senate Office. Christopher P. Weis, PhD; Anthony J. Intrepido, MS, CIH; Aubrey K. Miller, MD, MPH; Patricia G. Cowin, MS, CIH; Mark A. Durno, BS; Joan S. Gebhardt, PhD; Robert Bull, PhD; JAMA. 2002;288:2853-2858; http://jama.ama-assn.org/cgi/content/abstract/288/22/2853).

17. Will the NAS study team determine if the amounts of silica detected by the Armed Forces Institute of Pathology (AFIP) in their analysis of both SEMs and EDX spectra of the Daschle and New York Post powders (AFIP detected the elements silicon and oxygen in both powders; their reports show that there was more than one order of magnitude more silica in the New York Post powder compared with the Daschle powder) can be explained as being “naturally occurring” as the FBI claim?

18. Will the NAS study team determine if either the FBI or Sandia correctly concluded that the silica found in the spores sent to senators Daschle and Leahy could have been added as a molecular monomer rather than as already formed solid fumed silica nanoparticles? The FBI and Sandia national labs announced last year that the silica found in the spores sent to senators Daschle and Leahy consisted of a continuous coating of a phase of SiOx underneath the exosporium. Since this was underneath the exosporium they concluded that the spores were not “weaponized” – since weaponization usually involves forming a coating of fumed silica nanoparticles (silica particles with individual particles in the size range 10-50nm) which is located OUTSIDE the exosporium.

19. Will the NAS study team determine if the FBI attempted and/or succeeded in re-creating the silicon signature found in the Daschle, Leahy and New York Post powders? More specifically, was the FBI able to recreate the identical continuous phase of SiOx under the exosporium? Was the quantity of SiOx the same or similar? What form of silicon was added to the spores if they did try to do this? Was it added during spore preparation or after spore preparation?

20. Will NAS determine if and how the FBI ruled out that the spores were treated with a monomer of a siloxane compound (for example Dimethyldichlorosilane (trade name Repelcote)) that penetrated the exosporium in its monomer form and then polymerized on the spore coat in situ? When Dimethyldichlorosilane polymerizes it loses HCl and forms a polymer of SiOx.  ( A 1961 study shows that even large molecules can penetrate the highly porous exosporium (ref. Gerhardt, P. & Black, S. H. . J. Bact. 82, 750–759 (1961)).

21. Will the NAS study team determine if the FBI attempted and/or succeeded with reverse engineering with polymerized glass techniques as described above?

22. Will the NAS study team evaluate the reported FBI results which indicated (a) that the spore powder on the Leahy letter contained 14,479 ppm of silicon (1.4%), (b) that the spore powder on the New York Post letter was found to have silicon present in the sample; however, due to the limited amount of material, a reliable quantitative measurement was not possible, and (c) that insufficient quantities of spore powder on both the Daschle and Brokaw letters precluded analysis of those samples?

23. Will the NAS study team determine if the FBI used Inductively Coupled Plasma (ICP) spectroscopy to obtain the result of 14,479 ppm of silicon in the Leahy powder? How much sample was needed for this in total? Was the sample dissolved in solution first using standard ICP protocols? How much of the solution was used to introduce into the ICP nebulizer? Were standards used for calibration per normal ICP protocol?

24. Will the NAS study team determine if, when the FBI in their April 17, 2009 response above stated that the New York Post sample also contained silicon, that this was or was not an ICP result? How much total sample was used? Was the sample prepared in solution first?  How much solution was placed in the ICP nebulizer? What were the results that the FBI stated were unreliable?

25. Will the NAS study team draw its own conclusion as to whether the quantitative silicon result for the New York Post powder is unreliable?

26. Will the NAS study team determine if the FBI science labs used accepted scientific methods to demonstrate that it was possible to manufacture 5-10g of anthrax powder in the time frame of 12-24 hours total work performed over a period of 2-3 weeks as they claim Dr. Bruce Ivins did at Fort Detrick?

27. Will the NAS study team determine how many runs would be needed in 3 litre preparation flasks? How many spores per ml of liquid in each flask? How much centrifugation would be needed to concentrate the powder to one trillion spores per gram? And whether the material could have been dried using only the equipment available at Detrick?

28. Will the NAS study team determine if the FBI performed these runs in order to scientifically prove their theories are sound?

29. Will the NAS study team determine if Detrick had access to the proprietary azeotropic drying technology used by Dugway and described in an April 2008 joint publication between Dugway and the CDC which reported on the simulated opening of a letter filled with spores? And if Detrick had an exclusionary ball mill separated by increasingly finer mesh screens? (ref. Development of an Aerosol System for Uniformly Depositing Bacillus Anthracis Spore Particles on Surfaces, Aerosol Science and Technology, Volume 42, Issue 3 March 2008 , pages 159 – 172 ; Paul A. Baron,1 Cherie F. Estill,1 Gregory J. Deye,1 Misty J. Hein,1Jeremy K. Beard,2 Lloyd D. Larsen,2 and Gregory E. Dahlstrom2; (http://pdfserve.informaworld.com/560588 790515467.pdf)

29a. Will NAS determine why the authors of this paper chose to simulate the envelope containing the Daschle powder being opened in the Hart building by using a powder especially prepared for aerosolization instead of simply drying without any special equipment as the FBI claim?

30. Will the NAS study team determine, if Detrick did not have the equipment used in the Dugway/CDC report, what basis there was (or not) for the FBI theory that a product like the Daschle or Leahy powders could be made without this equipment?

31. Will the NAS study team determine if the FBI demonstrated that they could make an identical product without such equipment that yielded powder particles that formed secondary aerosols as was shown by the EPA to take place in the Hart building?

32. Will the NAS study team determine the significance of the Sandia report that, out of 200 samples they tested for the FBI, none matched the silicon signature of the mailed spores?

33. Will the NAS study team seek independent information regarding the silica content of the New York Post powder from AFIP, since the FBI refused to share that information with Sandia?

34. Will the NAS study team evaluate the significance of the WMD Chief’s admission, under the DARPA-funded cutting edge patent, that there would be large amounts of silica that then were removed from the surface through repeated centrifugation or an air chamber, which would lead one to expect trillion spore concentration and large amounts of silica absorbed in the coat and in debris in the first batch and then pure spores in the second batch after repeated centrifugation.

35. Will the NAS study team determine if the 1070 samples examined by the morphotype analysis accurately reflects the actual number of anthrax samples held in the U.S. in the 2001 timeframe?

36. Will the NAS study team evaluate whether the process of self-submission of anthrax samples by labs and individuals (per the FBI methodology) was a reliable representation of the Ames samples that may or may not have existed prior to 2001?

37. Will the NAS study team determine the ancesteral linkage between RMR1029 and the tested samples that were positive for 2 of 4 or 3 of 4 of the morphotype markers?

38. Will the NAS study team report on the expected links between labs that contributed precursor spore preps that were used to make RMR1029?

39. Will the NAS study team determine why there is a discrepancy between the number of labs known to have samples of RMR1029 prior to September 2001 and the two labs reported to test positive for RMR1029 by the testing methodology?

40. Will the NAS study team determine why the FBI did not use the labs known to have RMR1029 plus some self-generated samples of RMR1029 as an external proficiency test of their contract labs?

41. Will the NAS study team determine if the FBI used any type of proficiency testing for their genetic analysis?

42. Will the NAS study team evaluate the statistical probability of finding 4 positive morphotypes in any Ames sample, in any of the 32 or so precursor Ames strains that made up RMR1029, or in repeat analysis of RMR1029 itself?

43. Will the NAS study team determine the probability of losing one or more of the morphotypes from progeny/descendent samples derived from RMR1029? The number of generations occurring before the sample becomes negative equals 3 or fewer morphs according to the FBI.

44. Will the NAS study team determine and report on the precision, accuracy, linearity, sensitivity, specificity, reproducibility, and upper and lower limits of quantitation data values for the four morph assays?  Will the NAS study team determine if the FBI validated parallelism, robustness and ruggedness of the assays?

45. Will the NAS study team determine the reasonableness of the claim by the FBIs subcontractors that they had no issues with cross contamination and false positives with the morph analyses, and indicate if such a claim was substantiated by any data?

46. Will the NAS study team determine if the B. subtilis contaminant found in the first batch of letters was genetically identical to any forensic evidence collected from any lab?

47. Will the NAS study team determine if the B. subtilis contaminant found in the first batch of letters was tested against strains from Dugway proving grounds?

48. Will the NAS study team determine if the FBI’s scientific analysis of the silicon content was sufficient to support their claim that exognenous silicon was not added?  In particular, Will the NAS study team determine if the FBI performed titration experiments to show that their analysis was sufficiently sensitive to detect added silcon compounds?

49. Will the NAS study team determine if the FBI’s conclusion of “no added silicon” is supported by scientific evidence?

50. Will the NAS study team determine what analyses were completed to substantiate the FBI claim that none of the spore preps were milled or otherwise processed?

51. Will the NAS study team determine what data was used to substantiate the claim that mail sorting equipment imparted physical characteristics to the spore preps?

52. Will the NAS study team determine, given that both sets of letters were handled by mail sorting equipment, why is there such a discrepancy between their physical states (factors other than purity and concentration)?

53. Will the NAS study team evaluate the FBI “theory” that Ottillie Lundgren and Kathy Nguyen died from “cross contamination,” and, if so, why there were not many more casualties along the east coast from such “cross contamination”?

54. Will the NAS study team determine if the same genotype of subtilis was discovered in each of the hundreds of swab samples taken from the AMI building by the FBI during 2 visits to this building?

55. Will the NAS study team determine if any of the swab samples taken from the New York Post (NYP) building and other contaminated buildings associated with the NYP powder and NBC powder showed the presence of the subtilis genotype found in the raw powders?

56. Will the NAS study team evaluate the implications of the following scenario: (a) if subtilis was found in swab samples from the NYP and NBC buildings but not the AMI building, is it reasonable to conclude that the powder sent to the AMI building (the envelope for which was not recovered) did not contain the subtilis genotype? (b) if this is the case, does this cast doubt on the FBI theory that only 2 batches of anthrax powder were prepared from RMR-1029 by the perpetrator(s)? (c) If only 2 batches were prepared, does this suggest a different timeline for the production than the FBI theory that the powder sent to AMI must have been the same batch as the Daschle/Leahy powder, which, according to the FBI theory, was manufactured after the media powders had been sent?

57. Will the NAS study team determine if hydrophobicity testing of the Leahy and/or Daschle powder was undertaken? This can be performed using a simple phase transfer test where a small sample of powder is shaken in a tube containing water and a non-polar solven such as hexane – the test measures what fraction of the powder sample goes in to the water phase.

57a. If this test was performed what was the degree of hydrophobicity of the Daschle and/or Leahy powder?

57b. If the degree of hydrophobicity was close to 100% what is the theory behind this?

57c. Could a coating of hydrophobic polymerized glass on the spore coat underneath the exosporium in a dry powder (where the exosprorium will have collapsed) cause the spores to be unusually hydrophobic?

57d. Would this also enhance the powder’s ability to form an aerosol since it will inhibit clumping of spores due to water-bridging phenomenon?

58. Will the NAS study team evaluate the origin of the tin detected in letters to the New York Post and to Senators Daschle and Leahy, and perhaps in other letters as well?

59. Will the NAS study team evaluate the significance of absence of fiber evidence?

60. Will the NAS study team the validity of printing defects on the envelopes as establishing where the envelopes were purchased?

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