Unfinished Business: Efforts to Define Dual-Use Research of Bioterrorism Concern

Abstract

Biotechnological research poses a special security problem because of the duality between beneficial use and misuse. In order to find a balance between regulating potentially dangerous research and assuring scientific advancement, a number of assessments have tried to define which types of research are especially open to misuse and should therefore be considered dual-use research of special concern requiring rigorous oversight. So far, there has been no common understanding of what such activities are. Here we present a review of 27 assessments focusing on biological dual-use issues published between 1997 and 2008. Dual-use research activities identified by these assessments as being of special concern were compiled and compared. Moreover, from these 27 assessments, the primary research publications explicitly identified as examples of concerning research activities were extracted and analyzed. We extracted a core list of 11 activities of special concern and show that this list does not match with the reasons why primary research publications were identified as being of special concern. Additionally, we note that the 11 activities identified are not easily conducted or replicated, and therefore the likelihood of their being used in a high-tech mass casualty bioterrorism event should be reevaluated.

The authors looked at 27 assessments focusing on biological dual-use issues published between 1997 and 2008 and extracted a core list of 11 activities of special concern. They note that the activities identified are not easily conducted or replicated, and therefore the likelihood of their being used in a high-tech mass casualty bioterrorism event should be reevaluated.

Biotechnological research poses a special security problem because of the duality between beneficial use and misuse. All activities in modern life sciences are dual use to a certain degree. Controlling them all would not only pose an enormous administrative burden but would also place unacceptable restrictions on the research itself and would hamper scientific and medical progress. In order to find a balance between regulating potentially dangerous research and assuring scientific advancement, various assessments have tried to define which types of research are especially open to misuse and should therefore be considered dual-use research of special concern requiring rigorous oversight.^1–3

Lost in the extended discussion, however, has been a common understanding of what exactly we need to be afraid of.⁴ As a consequence, approaches to preventing and preparing for bioterrorism vary widely. An essential starting point for rectifying this situation would be to identify an internationally harmonized understanding of what research causes most concern. The U.S. National Science Advisory Board for Biosecurity (NSABB) calls such research “dual-use research of concern” (DURC) and defines it as:

Research that, based on current understanding, can be reasonably anticipated to provide knowledge, products, or technologies that could be directly misapplied by others to pose a threat to public health and safety, agricultural crops and other plants, animals, the environment, or materiel.³^(p17)

This definition is so general, however, that it does not focus on oversight procedures on specific research activities. To further a common understanding of which dual-use research activities are of special concern, we reviewed what different authors have considered to be such activities, analyzing whether there is a baseline understanding of dangerous research. We assess the different usages of the term “dual-use research of concern” with the aim of both identifying and bridging the discourse gap surrounding biosecurity policy discussions.

Methodology

For this review, we analyzed all assessments focusing on biological dual-use issues published before 2009.* The assessments were identified by a literature search and by interviewing biosecurity experts.⁵ To ensure that views from outside Europe and the United States were not missed, we contacted experts in the leading biotechnology countries of all relevant geographical subregions as defined by the UN: Australia, Brazil, Cuba, Egypt, India, Israel, Japan, Kenya, Mexico, Nigeria, Russia, Singapore, and South Africa.⁶ Twenty-seven assessments could be identified.^1–3,7–30 Most were authored by U.S. (8) or European (13) experts or expert groups. Authors came from government and nongovernmental institutions and had diverse backgrounds including military, health, and academia.

Dual-use research activities of special concern mentioned by these assessments were identified and compiled into a table to find similarities and differences between the assessments. Most of the assessments focus on the life sciences in general. In such cases, we list in the table all activities mentioned. A few assessments exclusively deal with specific topics, such as synthetic biology. In such cases, we list only the specific activity addressed in the assessment. We assume that authors of such assessments had conducted a threat evaluation that revealed the topic addressed to be one of special concern. In addition, we identified the primary research publications that are explicitly mentioned in the 27 assessments as examples of research of special concern.

Results

All of the assessments share the view that all life science activities are dual use to a certain degree and that there needs to be a core area of activities defined that has a greater potential for misuse than the rest in order to focus oversight and control measures.

Of the 27 assessments, 22 specifically identify research activities of concern. Two general ways of defining such activities were identified: an agent-based approach and an activity-based approach. Five of the assessments define research of concern only according to the agent used.^{13,17,27–29} These assessments consider all activities per se to be of concern when a certain dangerous agent is used. Ten assessments define research of concern exclusively through activities and technologies used or through the aims and outcomes of the research.^{3,8–11,14,15,18,20,21} Another 7 assessments define research of concern by both the activities conducted and the agents used.^{1,2,16,22–24,26}

The assessments that refer to research of concern mention 54 different types of activities.³¹ About half of these are not specific research activities but whole technology branches like synthetic biology or gene therapy. Taken together, the research activities referred to reflect the whole spectrum of biotechnological research. Excluding research of concern defined by an agent-based approach, we identified a core set of 11 research activities that were frequently mentioned as having high potential for misuse:

• Conferring resistance to therapeutically useful antibiotics or antiviral agents (mentioned in 13 assessments);

• Demonstrating how to render a vaccine ineffective (11);

• Enabling the evasion of diagnostic or detection modalities (11);

• Altering the host range of a pathogen (10);

• Enhancing the virulence of a pathogen (10);

• Increasing the transmissibility of a pathogen (9);

• Enhancing dissemination of a pathogen by powder or aerosol (8);

• Synthetic creation of pathogens (8);

• Increasing environmental stability (7);

• Enabling the weaponization of a biological agent or toxin (7); and

• Rendering a nonpathogen virulent (7).

Most of these activities were first mentioned in 2002 by Zilinskas and colleagues and later became widely known through the Fink Report.†^1,2 The remaining 43 types of research of concern were only mentioned by 1 or 2 assessments. Besides this core area of 11 activities, from which only 4 were mentioned by half or more assessments, there is almost no agreement about what research activities are of special concern. This reflects the feeling that most activities can be misused but some more readily than others.

Primary research publications that are of concern are specifically referred to in 14 of the 27 assessments, yet the overall number of cited publications is rather small—just 15.^32–46 The most frequently cited publication is the “mousepox paper” by Jackson et al (2001),³³ which is mentioned by all 14 assessments. Table 1 shows the percentage of assessments in which each publication is referred to.

Table 1.

Publications of Concern as Mentioned in the Assessments Reviewed

Publication of Concern	Number of Assessments	Percentage
Jackson RJ, Ramsay AJ, Christensen CD, Beaton S, Hall DF, Ramshaw IA. Expression of mouse interleukin-4 by a recombinant ectromelia virus suppresses cytolytic lymphocyte responses and overcomes genetic resistance to mousepox. J Virol 2001;75(3):1205–1210.	14/14	100
Cello J, Paul AV, Wimmer E. Chemical synthesis of poliovirus cDNA: generation of infectious virus in the absence of natural template. Science 2002;297(5583):1016–1018.	11/13	84
Tumpey TM, Basler CF, Aquilar PV, et al. Characterization of the reconstructed 1918 Spanish influenza pandemic virus. Science 2005;310(5745):77–80.	6/9	66
Taubenberger JK, Reid AH, Lourens RM, Wang R, Jin G, Fanning TG. Characterization of the 1918 influenza virus polymerase genes. Nature 2005;437(7060):889–893.	5/9	55
Gibbs MJ, Armstrong JS, Gibbs AJ. Recombination in the hemagglutinin gene of the 1918 “Spanish flu.” Science 2001;293(5536):1842–1845.	6/13	46
Rosengard AM, Liu Y, Nie Z, Jimenez R. Variola virus immune evasion design: expression of a highly efficient inhibitor of human complement. Proc Natl Acad Sci U S A 2002;99(13):8808–8813.	5/13	38
Wein LM, Liu Y. Analyzing a bioterror attack on the food supply: the case of botulinum toxin in milk. Proc Natl Acad Sci U S A 2005;102(28):9984–9989.	2/9	22
Hatta M, Gao P, Halfmann P, Kawaoka Y. Molecular basis for high virulence of Hong Kong H5N1 influenza A viruses. Science 2001;293(5536):1840–1842.	2/13	15
Wang XG, Zhang GH, Liu CX, Zhang YH, Xiao CZ, Fang RX. Purified cholera toxin B subunit from transgenic tobacco plants possesses authentic antigenicity. Biotechnol Bioengin 2001;72(4):490–494.	2/13	15
Domi A, Moss B. Cloning the vaccinia virus genome as a bacterial artificial chromosome in Escherichia coli and recovery of infectious virus in mammalian cells. Proc Natl Acad Sci U S A 2002;99(19):12415–12420.	1/12	8
Seo SH, Hoffmann E, Webster RG. Lethal H5N1 influenza viruses escape host anti-viral cytokine responses. Nat Med 2002;8(9):950–954.	1/12	8
Smith HO, Hutchison CA 3rd, Pfannkoch C, Venter CG. Generating a synthetic genome by whole genome assembly: ΦX174 bacteriophage from synthetic oligonucleotides. Proc Natl Acad Sci U S A 2003;100:15440–15445.	1/12	8
Read TD, Peterson SN, Tourasse N, et al. The genome sequence of Bacillus anthracis Ames and comparison to closely related bacteria. Nature 2003;423(6935):81–86.	1/12	8
Kolkman JA, Stemmer WP. Directed evolution of proteins by exon shuffling. Nature Biotechnol 2001;19(5):423–428.	1/14	7
Arora N, Leppa SH. Fusions of anthrax toxin lethal factor with shiga toxin and diphtheria toxin enzymatic domains are toxic to mammalian cells. Infect Immun 1994;62(11):4955–4961.	1/14	7

The date of the assessment and the publication was taken into account in determining the percentage. The total number first refers to all assessments that mention a certain publication and secondly to the number of assessments for which the publication was theoretically available. If the assessment and the paper were published within the same year, we assumed that the paper was not available except in cases where it was explicitly mentioned.

It is noteworthy that, with the exception of the paper by Cello et al,³⁸ all papers cited by more than half of the assessments are directly or indirectly related to work with extinct or eradicated agents with extremely negative public health impacts.

Discussion and Conclusions

While a broad definition of dual use is used in all the assessments reviewed here—which can be summarized as the tangible and intangible aspects of life science research that have the potential to be used for benevolent and malevolent purposes—this is insufficient to develop concrete oversight measures. Although all the assessments agreed that adding a subcategory—such as what the NSABB calls “dual-use research of concern” (DURC)—would be of value, this is not sufficiently defined in any of the assessments. Instead, we note that the term dual-use research appears to be used almost interchangeably with the term dual-use research of concern. The problem, therefore, is the lack of a consistent way for deciding what is and what is not “of concern.” Without this, it is hardly possible to decide what research in practice needs tighter oversight and what does not.

The outcome of our review can be summarized as follows: There is almost no agreement on what dual-use research of special concern is, but there are 2 different non-exclusive approaches in trying to define it. And what is theoretically considered to be of concern differs considerably from what is in practice identified as research of concern.

Our review shows that, taken together, the activities mentioned in the assessments cover almost all areas of life science research. Trying to focus on a core area, however, we identified 11 activities that are mentioned by at least a third of the assessments as being of concern. This core area overlaps to a large degree with the experiments of concern first described by Zilinskas et al. and in the Fink Report.

The publication of the Fink Report in 2004 influenced all later publications about dual-use issues in the life sciences. It is by far the most widely cited assessment, and its list of activities of concern is copied in many of the later assessments. This makes the 7 experiments of concern mentioned in the Fink Report part of the core area of research of concern identified through our review. The fact that the Fink list is obviously reproduced in a number of later assessments might lead one to question the independence and added value of these assessments. But as all the assessments, quite naturally, are influenced by earlier writing, in their entirety they represent the debate around dual-use research in the global expert community.

However, the core area identified here includes 3 activities not mentioned in the Fink Report: enhancing dissemination of a pathogen by powder or aerosol, increasing environmental stability (both activities would, however, fit under the broader Fink criterion “enabling the weaponization of a biological agent or toxin”), and the synthetic creation of a pathogen.‡

Even if there is no universal agreement within the assessments reviewed about what dual-use research of concern is, some general starting points can be deducted. All assessments use 1 or both of 2 different approaches to identify it: agent-based and/or activity-based. The agent-based approach defines research of concern on the basis of the agent involved. The most frequently mentioned agents in this context are the pathogens causing anthrax, botulism, plague, smallpox, and viral hemorrhagic fevers, but the agent lists differ from assessment to assessment both in terms of length and agents covered. The activity-based approach defines research of concern on the basis of the aims and results of a research activity. This approach assumes that some activities are more useful in the creation of a biological weapon than others. The majority of assessments reviewed apply the activity-based approach, but the lists of activities differ from assessment to assessment both in terms of length and level of detail.

The primary research publications that are mentioned in half or more of the assessments reviewed as representing research of concern deal either with extinct agents (refs 33, 44, 45), the synthetic creation of viruses (ref 38), or rendering a vaccine ineffective (again, ref 33), which is a small subset of the activities identified as core areas of research of concern. In fact, it is questionable whether the paper by Jackson et al would have become so prominent if the agent used had not been such a close relative of smallpox. Interestingly, these 4 papers are also the papers that provoked wider controversial discussions of their proliferation potential. At this point the review reveals a significant mismatch between what is theoretically considered to be an activity of special concern and what is empirically considered to be one when looking at research publications. It seems to us that in practice, special concern is created if easily transmissible and highly pathogenic extinct or eradicated agents—such as those causing smallpox or 1918 influenza—are used in research. Even if the assessments do use an activity-based approach, the final decision whether a research project is considered to be of concern is made primarily on the basis of the agent used.

Also interesting is the fact that the total number of publications mentioned in the assessments we reviewed as being of concern is less than the total number of assessments (15 versus 27), and much less than the numerous activities of concern that the 27 assessments identified (15 versus 54). One explanation could be that there simply are not many publications that have caused severe security concerns in the past—in other words, that research of concern is a very recent phenomenon. This explanation is supported by a 2006 survey that revealed that scientific journals that do screen publications for biosecurity issues have in the past considered only very few publications to be problematic.⁴⁷ All of these publications were finally published with no or only minor changes.

Obviously, scientists are very cautious in classifying research as being of special concern, either because the awareness of security aspects is still not completely internalized in the life science community, or because scientists know how difficult the reproduction of research is and therefore how limited the advantage for terrorists would be. An additional explanation why the number of publications is small and only the most prominent and widely known examples were mentioned in the assessments reviewed could be that many of the assessments were authored by nonscientists for nonscientific audiences and have to rely on examples that are illustrating the problem with glaring clarity and are still easily understandable. It remains a mystery, however, why certain publications with extreme misuse potential—for example, Pomerantsev et al,⁴⁸ describing the construction of a vaccine-resistant anthrax strain—are mentioned in none of the assessments. Even if this paper was widely discussed after its publication, this discussion is not reflected in the assessments reviewed.

Whatever the explanation for the low number of publications is, it is likely that this number will increase in the future. The increasing amount of research activity globally will quite naturally lead to a proportionate increase in the amount of research with a high misuse potential, unless we actively look for alternatives.

Most of the 11 activities in the core area we identified are carried out as a means to reach a certain end with no immediate security concern, such as to examine the pathway of infection. In such cases, the activities of concern are published in the methodology sections of papers, are known in detail only to a limited number of scientists, and can rarely be identified by scanning titles and abstracts of papers. Arguably, such “transient” use of activities of concern is less dangerous than activities with the explicit and sole intent to undermine vaccine effectiveness, produce antibiotic resistance, or increase environmental stability.⁴⁹ Even if a low visibility reduces the likelihood of such activities being misused, they should be addressed by oversight mechanisms. This could, for instance, mean focusing more on the rationale for using activities of concern instead of other methodologies, and more seriously looking for alternative, low-concern means of reaching the same aim. It would be desirable if research calls elaborating such alternatives would be put out for tender.

Although “dual-use research of concern” remains diffuse, the core area of 11 activities of concern can be used as a starting point to elaborate a more agreed-on list of such activities. The list will need to be reduced or expanded and regularly updated and would benefit from widespread international consultation. Elaborating a more agreed-on list will be very demanding not only because of the fast pace at which new research technologies are appearing, but also because judgment on the likelihood of misuse varies widely.

A recently published study by the authors showed that most activities usually considered to be of concern require a substantial amount of expertise and equipment.⁵⁰ They are not easily conducted or replicated, and therefore the likelihood of a high-tech bioterrorism event and the ease with which mass casualties can be caused, should be reevaluated.

In general, the misuse potential of the core area of activities is a long-term concern. In contrast, there might be activities that can directly be misused and therefore cause short-term concern. The focus of attention has thus far been too much on the high-tech, potentially very serious, but nevertheless long-term risks of dual-use research. Clarifying what activities are of immediate concern remains a most urgent requirement.

Footnotes

Acknowledgments

We would like to thank the European Centre for Disease Prevention and Control (ECDC), the Fritz Thyssen Foundation, and the German Ministry for Education and Research for providing financial support for our research, and 3 anonymous reviewers for helpful suggestions in the revision of the original manuscript.

*

The empirical research for this review was conducted in 2008 as part of a study conducted for the European Centre for Disease Prevention and Control (ECDC), Dual-use Life Science Research and Its Potential Application in Bioterrorism, Final Report, April 2009, Tender No SAU-08-768-JGab of 4 April 2008.

†

The Fink Report considers research to be of concern when it includes experiments that: (1) demonstrate how to render a vaccine ineffective; (2) confer resistance to therapeutically useful antibiotics or antiviral agents; (3) enhance the virulence of a pathogen or render a nonpathogen virulent; (4) increase transmissibility of a pathogen; (5) alter the host range of a pathogen; (6) enable the evasion of diagnostic/detection modalities; or (7) enable the weaponization of a biological agent or toxin.²

‡

In the Fink Report, “enhancing the virulence of a pathogen” and “rendering a non-pathogen virulent” are summed up into 1 activity. In our core area, we count these as 2 different activities.

References

Zilinskas

, Tucker

. Limiting the contribution of the open scientific literature to the biological weapons threat. Journal of Homeland Security, December 2002. http://www.homelandsecurity.org/journal/Search.aspx?s=tucker. 2011 September 30.

Committee on ResearchStandards and Practices to Prevent the Destructive Application of Biotechnology, National Research Council. Biotechnology Research in an Age of Terrorism [The Fink Report] Washington, DC: National Academies Press, 2004. http://www.nap.edu/catalog.php?record_id=10827. 2011 September 30.

National Science Advisory Board for Biosecurity. Proposed Framework for the Oversight of Dual Use Life Sciences Research: Strategies for Minimizing the Potential Misuse of Research Information, June 2007. http://oba.od.nih.gov/biosecurity/pdf/Framework%20for%20transmittal%200807_Sept07.pdf. 2011 September 30.

Resnik

. What is dual-use research? A response to Miller and Selgelid. Sci Eng Ethics, 2007; 13,4:523–580.

For detailed information on the identification of assessments, other methodological issues, see. Suk

, Zmorzynska

, Hunger

et al. Dual-use research and technological diffusion: reconsidering the bioterrorism threat spectrum. PLoS Pathog, 2011 Jan 13 7,1:e1001253, doi:10.1371/journal.ppat.1001253.

For detailed information on how these key countries were selected, see. Hunger

. Confidence building needs transparency. a summary of data submitted under the Biological Weapons Convention's Confidence Building Measures 1987–2003. The Sunshine Project. September 2005. http://biological-arms-control.org/publications/hunger_CBM.pdf. 2011 October 26.

Molas-Gallart

, Perry Robinson

. Assessment of Dual-Use Technologies in the Context of European Security and DefenceFinal report for the Scientific and Technological Options Assessment (STOA), European ParliamentBrighton: SPRU, 1997.

Fraser

, Dando

. Genomics and future biological weapons: the need for preventive action by the biomedical community. Nat Genet, 2001; 29,3:253–256.

Do no harm: reducing the potential for the misuse of life science research. Report of a Royal Society-Wellcome Trust meeting on 7 October 2004. http://www.wellcome.ac.uk/stellent/groups/corporatesite/@policy_communications/documents/web_document/wtx023408.pdf. 2011 September 30.

10.

McLeish

, Nightingale

. The Impact of Dual Use Controls on UK Science: Results From a Pilot StudySPRU Electronic Working Paper Series, Paper No. 132April 2005. www.sussex.ac.uk/spru/documents/sewp132. 2011 September 30.

11.

World Health Organization. Life Science Research: Opportunities and Risks for Public Health. Geneva: WHO, 2005. http://whqlibdoc.who.int/hq/2005/WHO_CDS_CSR_LYO_2005.20.pdf. 2011 September 30.

12.

Managing risks of misuse associated with grant funding activities. A joint BBSRC, MRC and Wellcome Trust policy statement. September 2005. http://www.bbsrc.ac.uk/web/FILES/Policies/misuse_of_research_joint.pdf. 2011 October 5.

13.

Roffey

. From bio threat reduction to cooperation in biological proliferation prevention. Background paper 4. Presented at: Conference on Strengthening European Action on WMD Non-proliferation and Disarmament: How Can Community Instruments Contribute?, December 2005.

14.

Maurer

, Lucas

, Terrell

. From Understanding to Action: Community-Based Options for Improving Safety and Security in Synthetic Biology. Berkeley, CA: University of California, Berkeley, Richard and Rhoda Goldman School of Public Policy, April 2006.

15.

National Science Advisory Board for Biosecurity. NSABB Draft Guidance Documents. July 2006.

16.

Campbell

. Empowerment and restraint in scientific communication. New developments make it easier to share information, but more difficult to deal with dual-use biology. EMBO Rep, 2006 July 7Spec NoS18–S22.

17.

World Health Organization. Biorisk Management. Laboratory Biosecurity Guidance. Geneva: World Health Organization, September 2006. http://www.who.int/csr/resources/publications/biosafety/WHO_CDS_EPR_2006_6.pdf. 2011 October 5.

18.

National Science Advisory Board for Biosecurity. Addressing Biosecurity Concerns Related to the Synthesis of Select Agents, December 2006. http://oba.od.nih.gov/biosecurity/pdf/Final_NSABB_Report_on_Synthetic_Genomics.pdf. 2011 October 5.

19.

Atlas

, Dando

. The dual-use dilemma for the life sciences: perspectives, conundrums, and global solutions. Biosecur Bioterror, 2006; 4,3:276–286.

20.

Committee on Advances in Technology and the Prevention of Their Application to Next Generation Biowarfare Threats, National Research Council. Globalization, Biosecurity, and the Future of the Life Sciences. Washington, DC: National Academies Press, 2006. http://www.nap.edu/catalog.php?record_id=11567. 2011 October 5.

21.

Miller

, Selgelid

. Ethical and philosophical consideration of the dual-use dilemma in the biological sciences. Sci Eng Ethics, 2007 Dec 13,4:523–580.

22.

Steinbruner

, Harris

, Gallagher

, Okutani

. Controlling Dangerous Pathogens. College Park, MD: Center for International and Security Studies, University of Maryland, March 2007. http://www.cissm.umd.edu/papers/display.php?id=217. 2011 October 5.

23.

Makosky

, Stephen

. Development of a Threat Assessment Framework Applicable to Dual Use BiotechnologyDefence R&D CanadaApril 2007. http://pubs.drdc.gc.ca/PDFS/unc105/p527643_A1b.pdf. 2011 October 5.

24.

Steering Committee on Issues in Biotechnological Research in an Age of Terrorism. Biotechnological Research in an Age of Terrorism. Jerusalem: Israel Academy of Science and Humanities and the Israel National Security Council, 2008. http://www.academy.ac.il/data/reports_data/48/Biotech_eng_Apr2009.pdf. 2011 October 5.

25.

Kuhlau

. Countering Bio-threats, EU Instruments for Managing Biological Materials, Technology and KnowledgeSIPRI Policy Paper No. 19Stockholm: Stockholm International Peace Research Institute, August 2007. http://books.sipri.org/files/PP/SIPRIPP19.pdf. 2011 October 5.

26.

Garfinkel

, Endy

, Epstein

, Friedman

. Synthetic Genomics: Options for GovernanceCSIS ReportsOctober 2007. http://mit.dspace.org/bitstream/handle/1721.1/39141/Synthetic%20Genomics%20Options%20for%20Governance.pdf?sequence=1. 2011 October 5.

27.

Commission of the European Communities. Green Paper on Bio-preparedness. Brussels, November 2007. http://ec.europa.eu/food/resources/gp_bio_preparedness_en.pdf. 2011 October 5.

28.

Organisation for Economic Co-operation and Development. OECD Best Practice Guidelines for Biological Resource Centres. Paris: OECD, 2007. http://www.oecd.org/dataoecd/7/13/38777417.pdf. 2011 October 5.

29.

Atlas

. Toward global harmonization for control of dual-use biothreat agents. Science and Public Policy February, 2008; 35,1:21–27.

30.

Rappert

. The benefits, risks, and threats of biotechnology. Science and Public Policy February, 2008; 35,1:37–43.

31.

A list of all these 54 different types of research of concern is. http://www.biological-arms-control.org/projects_dualuselifescience/Research_stated_to_be_Dual_Use_Research_of_Concern.pdf. 2011 October 5.

32.

Arora

, Leppa

. Fusions of anthrax toxin lethal factor with shiga toxin and diphtheria toxin enzymatic domains are toxic to mammalian cells. Infect Immun, 1994; 62,11:4955–4961.

33.

Jackson

, Ramsay

, Christensen

, Beaton

, Hall

, Ramshaw

. Expression of mouse interleukin-4 by a recombinant ectromelia virus suppresses cytolytic lymphocyte responses and overcomes genetic resistance to mousepox. J Virol, 2001; 75,3:1205–1210.

34.

Kolkman

, Stemmer

. Directed evolution of proteins by exon shuffling. Nature Biotechnol, 2001; 19,5:423–428.

35.

Hatta

, Gao

, Halfmann

, Kawaoka

. Molecular basis for high virulence of Hong Kong H5N1 influenza A viruses. Science, 2001; 293,5536:1840–1842.

36.

Gibbs

, Armstrong

, Gibbs

. Recombination in the hemagglutinin gene of the 1918 “Spanish flu.” Science, 2001; 293,5536:1842–1845.

37.

Wang

, Zhang

, Liu

, Zhang

, Xiao

, Fang

. Purified cholera toxin B subunit from transgenic tobacco plants possesses authentic antigenicity. Biotechnol Bioeng, 2001; 72,4:490–494.

38.

Cello

, Paul

, Wimmer

. Chemical synthesis of poliovirus cDNA: generation of infectious virus in the absence of natural template. Science, 2002; 297,5583:1016–1018.

39.

Rosengard

, Liu

, Nie

, Jimenez

. Variola virus immune evasion design: expression of a highly efficient inhibitor of human complement. Proc Natl Acad Sci U S A, 2002; 99,13:8808–8813.

40.

Domi

, Moss

. Cloning the vaccinia virus genome as a bacterial artificial chromosome in Escherichia coli and recovery of infectious virus in mammalian cells. Proc Natl Acad Sci U S A, 2002; 99,19:12415–12420.

41.

Seo

, Hoffmann

, Webster

. Lethal H5N1 influenza viruses escape host anti-viral cytokine responses. Nat Med, 2002; 8,9:950–954.

42.

Smith

, Hutchison

3rd , Pfannkoch

, Venter

. Generating a synthetic genome by whole genome assembly: ΦX174 bacteriophage from synthetic oligonucleotides. Proc Natl Acad Sci U S A, 2003; 100:15440–15445.

43.

Read

, Peterson

, Tourasse

et al. The genome sequence of Bacillus anthracis Ames and comparison to closely related bacteria. Nature, 2003; 423,6935:81–86.

44.

Taubenberger

, Reid

, Lourens

, Wang

, Jin

, Fanning

. Characterization of the 1918 influenza virus polymerase genes. Nature, 2005; 437,7060:889–893.

45.

Tumpey

, Basler

, Aquilar

et al. Characterization of the reconstructed 1918 Spanish influenza pandemic virus. Science, 2005; 310,5745:77–80.

46.

Wein

, Liu

. Analyzing a bioterror attack on the food supply: the case of botulinum toxin in milk. Proc Natl Acad Sci U S A, 2005; 102,28:9984–9989.

47.

Van Aken

, Hunger

. Biosecurity policies at international life science journals. Biosecur Bioterror, 2009 Mar 7,1:61–71.

48.

Pomerantsev

, Staritsin

, Mockov

, Marinin

. Expression of cereolysine AB genes in Bacillus anthracis vaccine strain ensures protection against experimental hemolytic anthrax infection. Vaccine, 1997; 15,17-18:1846–1850.

49.

The need to take into account the willingness to make use of objects for bad ends while trying to define dual-use has been described by. Forge

. A note on the definition of “dual-use.” Sci Eng Ethics, 2010; 16,1:111–118.

50.

Suk

, Zmorzynska

, Hunger

et al. Dual-use research and technological diffusion: reconsidering the bioterrorism threat spectrum. PLoS Pathog, 2011; 7,1:e1001253, doi:10.1371/journal.ppat.1001253.