Friday, March 7, 2014

Conflict and Disease: Syria's Wild Polio 1 Outbreak and European Public Health Security

Update: "For the first time ever, the World Health Organization on Monday declared the spread of polio an international public health emergency that could grow in the next few months and unravel the nearly three-decade effort to eradicate the crippling disease. The agency described the current polio outbreaks across at least 10 countries in Asia, Africa, and the Middle East as an "extraordinary event" that required a coordinated international response. It identified Pakistan, Syria and Cameroon as having allowed the virus to spread beyond their borders, and recommended that those three governments require citizens to obtain a certificate proving they have been vaccinated for polio before traveling abroad." See: http://www.foxnews.com/health/2014/05/05/who-declares-polio-international-public-health-emergency/?intcmp=latestnews






"Detection and control of emerging infectious diseases in conflict situations are major challenges due to multiple risk factors known to enhance emergence and transmission of infectious diseases. These include inadequate surveillance and response systems, destroyed infrastructure, collapsed health systems and distribution of disease control programs, and infection control practices even more inadequate than those in resource-poor settings, as well as ongoing insecurity and poor coordination among humanitarian agencies."  Michelle Gayer, Dominique Legros, Pierre-Formenty and Maire A. Connonlly seehttp://wwwnc.cdc.gov/eid/article/13/11/06-1093_article.htm*** 


In 1980, the World Health Organization (WHO) declared smallpox, the first disease in human history, eradicated. Smallpox is the only disease to have been eradicated and only two repositories the Center for Disease Control and Prevention in Atlanta, Georgia and Vector in Novosibirsk, Russia maintain live viral strain collections.Wild Polio Virus, was the next scheduled disease to face eradication. Unfortunately, the recent polio outbreak in Syria serves as a reminder that conflict and war severely impact vaccination campaigns and eradication strategies leaving the most vulnerable open to the spread of this horrific disease. While ample polio vaccine exists, other diseases such as smallpox, avian flu, highly pathogenic strains of Category A agents, typically considered suitable for weaponization are not maintained in significant global stockpiles to contain a pandemic outbreak and in some instances not even an epidemic. It takes nearly ten years and around one billion US dollars to bring a vaccine onto the market. While war and disease often go hand in hand and conflict disease is not a new topic, changes in the demographics of war, terrorism and emerging disease pose a critical nexus that until very recently was not fully appreciated. The recently reported outbreak of Wild Polio Virus 1 (WPV1), serves as a reminder of the effect of war and terrorism on emerging and re-emerging highly pathogenic disease. The transmissible Polio virus had long been eradicated in all but three countries: Pakistan, Afghanistan and Nigeria. Its re-emergence in Syria, the likely result of Pakistani fighters entering Syria to fight against the regime of Bashar al Assad, is among several potentially life threatening diseases facing not only Syrian populations and refugee camps, but due to modern air travel and high numbers of fighters returning to Europe, the global community as a whole.see: http://www.redorbit.com/news/health/1113001691/polio-virus-confirmed-syria-origin-pakistan-threats-111313  


polio vaccine


In a November 11th report, the WHO, under the direction of the United Nations, confirmed that genetic sequencing showed the isolated viruses were most closely related to environmental samples pulled from Egypt in December 2012. Interestingly enough, those samples had in fact been linked to the poliovirus circulating in Pakistan. see: http://www.redorbit.com/news/health/1113001691/polio-virus-confirmed-syria-origin-pakistan-threats-111313/   


Highly pathogenic and transmissible diseases do not respect national boarder, any outbreak of a contagious pathogenic agent, given modern air travel, presents health risks to the wider global community. In protracted and post conflict situations, populations may have high rates of illness and mortality due to breakdown of health systems, flight of trained staff, failure of existing disease control programs and destruction of infrastructure. These populations may be more vulnerable to infection and disease because of high levels of malnutrition, low vaccine coverage, or long term stress. See: http://wwwnc.cdc.gov/eid/article/13/11/06-   Moreover, with significant numbers of fighters returning to Europe from conflict in Syria, communities once considered to have relatively high herd immunity, due to previous vaccination, are likely to see outbreaks of polio, a disease recently scheduled for and nearly eradicated by WHO. WPV3 has possibly been eradicated, WPV2 was eradicated in 1999, but WPV1 remains endemic in countries with poor public health infrastructures. See:  http://www.polioeradication.org/tabid/488/iid/336/Default.aspx#sthash.U3cWLlcw.dpuf Chris Maher, Senior Scientist for Polio Eradication at the World Health Organization made the following observation published in the Global Polio Eradication Initiative website:http://www.polioeradication.org/tabid/488/iid/336/Default.aspx#sthash.U3cWLlcw.dpuf 

"The danger with WPV3 is that it is less virulent than WPV1, Maher explained. It causes cases at a rate of approximately 1 in 2,000 infections, compared with 1 in 200 infections for WPV1. Causing fewer cases is a good thing of course, but it also means that the virus can transmit more widely and longer, without being detected. "It's a sneaky virus, in that sense, so we have to be cautious not to let it surprise us." Maher said. The other challenge is that the last known remaining WPV3 reservoirs (Khyber Agency in Pakistan, and Bomo and Yobe states in Northern Nigeria), are areas where access is compromised due to insecurity. Undetected circulation therefore cannot be fully ruled out. Efforts are ongoing to address these and other challenges, as part of national emergency action plans being implemented in both countries. see:  http://www.polioeradication.org/tabid/488/iid/336/Default.aspx#sthash.U3cWLlcw.dpu 

"It is vital that we work in areas which offer opportunities beyond war zones as well to contain highly pathogenic diseases at the earliest identification such as refugee camps and neighboring nations, which often witness the influx of large numbers of refugees. In the case of WPV1 outbreak in Syria, Europeans are at greater risk due to the numbers of fighters returning to Europe. while this may seem to be a straight forward counter terrorism issue, it is also a European public health issue with potentially global health consequences. Increasing public health infrastructures in nations such as Turkey and Lebanon will likely help reduce risk of spread, it is not nearly enough." http://www.polioeradication.org/tabid/488/iid/336/Default.aspx#sthash.U3cWLlcw.dpu 

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Thursday, March 6, 2014

3D Vaccine Printing: Countering Furutre Terrorist Weapons





In 2000, when Craig Venter, among other notable scientists made impressive breakthroughs in synthetic biology, it took some time before the non-proliferation community expressed any public concern. In fact it wasn't until The National Science Advisory Board for Biosecurity, a panel tasked with advising on science policy and the publication of what could be considered dual-use or sensitive research, was instituted that such advances were reviewed. Steven Block from Stanford Block Lab, warned early on about the potential for synthetic biology to be used for 'black' biology, noting in an article by Marc Schwarz: 


"If anthrax, smallpox and other 'conventional' biological agents aren't frightening enough, Block also raises the specter of 'black biology' a shadowy science in which microorganisms are genetically engineered for the sole purpose of creating novel weapons of terror. The idea that anybody can brew this stuff in their garage vastly overstates the case," he said, "but any technology that can be used to insert genes into DNA can be used for either good or bad." Block points out that genetic maps of deadly viruses, bacteria and other microorganisms are widely available in the public domain. Any scientist bent on destruction could use this information to attempt to clone extremely virulent strains of bacteria and viruses, Block contends. He notes that there are plenty of underpaid microbiologists in the world who might be eager to work for unscrupulous clients to produce incurable 'designer diseases, such as penicillin resistant anthrax or 'stealth viruses' that infect the host but remain silent until activated by some external trigger. Anthrax spores are easy to produce and can remain viable for more than 100 years if kept dry and out of direct sunlight. Their long shelf life makes them 'well suited to weaponization in a device that can deliver a widespread aerosol," Block notes. See: http://news.stanford.edu/news/2001/january17/bioterror-117.html



With concern over advances in the life sciences including synthetic biology, came attempts by NSABB to formulate Codes of Conduct. Unfortunately, terrorists and scientists working in state WMD programs which sponsor terrorist organizations such as Hezbollah, do not tend to adhere to Codes of Conduct defined by Western institutions or even international UN arms control treaties such as the Biological and Toxin Weapon Convention (BTWC). While synthetic biology and advances made by Craig Venter's team offers us incredible opportunities, the potential for mis-use, remains an issue. National Security sectors struggle to keep pace with bio-defence research to counter rapid advances in the life sciences. See: http://www.opbw.org  


While breakthroughs in synthetic biology offer impressive advancements and opportunities in the life sciences there remains concern that this technology could outstrip our ability to counter nefarious applications. The traditional, one bug one drug approach to countering the threat of biological weapons is no longer a viable model for bio-defence. This approach lags substantially behind current biological weapon development, which terrorist states may select to develop, using synthetic applications. In the future the Pentagon's Defence Advanced Research Projects Agency (DARPA) Blue Angle Program, launched in 2009, which has sought to manufacture vaccines in an accelerated time frame, previously not imagined. DARPA has worked to develop methods for rapidly producing vaccines. Until recently it took about ten years and up to one billion USD to produce one bio-defence drug. In July 2012 DARPA announded they had successfully manufactured 10 million doses of influenza vaccine in one month. See:http://www.darpa.mil/Our_Work/DSO/Programs/H1N1_Acsceleration_(BLUE_ANGEL).aspx. http://rt.com/usa/future-vaccine-darpa-research-255/   More promising is Venter's assertion that we will be able to 3D bio-print vaccines. This cutting edge research will likely revolutionize how was counter both naturally occurring and potentially deliberate disease.
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Wednesday, March 5, 2014

Ancient Strains: pithovirus sibericum and smallpox


In February 2012, Scientists from the Institute of Cell Biophysics announced they had regrown plants stored in permafrost by squirrels over 30,000 years ago. The institute raised plants of the silene stenophylla. Writing in the Proceedings of the National Academy of Sciences (PNAS), they note this is the oldest plant material so far to have been brought to life." See: http://www.bbc.com/news/science-environment-17100574
Exciting as this research and discovery was, more concerning perhaps is the recent discovery of pithovirus sibericum, not in an of itself a concern, but with implications for the re-emergence of other more harmful viruses such as ancient smallpox virus, the only human disease ever declared eradicated.

smallpox (variola major)
Recently a French scientific team from the National Center for Scientific Research (CNRS) presented their study on pithovirus sibericum. The Abstract of their study published in the: Proceedings of the National Academy of Sciences, available here:http://www.pnas.org/content/early/2014/02/26/1320670111 states: 

The largest known DNA viruses infect Acanthamoeba and belong to two markedly different families. The Megaviridae exhibit pseudo-icosahedral virions up to 0.7 μm in diameter and adenine–thymine (AT)-rich genomes of up to 1.25 Mb encoding a thousand proteins. Like their Mimivirus prototype discovered 10 y ago, they entirely replicate within cytoplasmic virion factories. In contrast, the recently discovered Pandoraviruses exhibit larger amphora-shaped virions 1 μm in length and guanine–cytosine-rich genomes up to 2.8 Mb long encoding up to 2,500 proteins. Their replication involves the host nucleus. Whereas the Megaviridae share some general features with the previously described icosahedral large DNA viruses, the Pandoraviruses appear unrelated to them. Here we report the discovery of a third type of giant virus combining an even larger pandoravirus-like particle 1.5 μm in length with a surprisingly smaller 600 kb AT-rich genome, a gene content more similar to Iridoviruses and Marseillevirus, and a fully cytoplasmic replication reminiscent of the Megaviridae. This suggests that pandoravirus-like particles may be associated with a variety of virus families more diverse than previously envisioned. This giant virus, named Pithovirus sibericum, was isolated from a >30,000-y-old radiocarbon-dated sample when we initiated a survey of the virome of Siberian permafrost. The revival of such an ancestral amoeba-infecting virus used as a safe indicator of the possible presence of pathogenic DNA viruses, suggests that the thawing of permafrost either from global warming or industrial exploitation of circumpolar regions might not be exempt from future threats to human or animal health.



In multiple reports they have stated that, "The discovery of Pithovirus shows how incomplete our understanding of microscopic biodiversity is when it comes to exploring new environments. And the potentially bad news: the reemergence of other viruses is no longer the domain of science fiction. These include viruses that were long considered eradicated such as smallpox, whose replication process is similar to that of pithovirus. This is an indication that viruses pathogenic for human or animals might also be preserved in old permafrost layers, including some that have caused planet wide epidemics in the past, said Jean Michel Claverie, one of the study's co-authors." Prof. Claverie warns that exposing deep layers could expose new viral threats. He has been quoted as saying that 'ancient strains of the smallpox virus, which was declared eradicated 30 years ago, could pose a risk. "If it is true that these viruses survive in the same way those amoeba viruses survive, then smallpox is not eradicated from the planet--only the surface." "By going deeper we may reactive the possibility that smallpox could become a disease of humans in modern times." See: http://www.bbc.com/news/science-environment-26387276

In a February, 2014 report entitled: Poxvirus Viability and Signatures in Historical Relics, published in Emerging Infectious Diseases (see:http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3901489/), the authors Andrea M. McCollum, Yu Li and CDC orthopox expert Inger K. Damon, suggest that "Although it has been >30 years since the eradication of smallpox, the unearthing of well preserved tissue material in which the virus may reside has called into question the viability of variola virus decades or centuries after its original occurrence. Experimental data to address the long-term stability and viability of the virus are limited. There are several instances of well preserved corpses and tissue that have been examined for poxvirus viability and viral DNA. These historical specimens cause concern for potential exposures, and each situation should be approached cautiously and independently with the available information. Nevertheless, these specimens provide information on the history of a major disease and vaccination against it. 

www.vaccines.mil
Both Prof. Claverie and the authors of the EID study, one of whom I had the pleasure of meeting at a major smallpox bio-security conference in Geneve, bring up valid points with regard to the potential risk of re-emergence and exposure to the only successfully eradicated human disease. But eradication may now be as Prof. Claverie states "not eradication from the planet --only the surface." Maintaining our Strategic National Stockpiles and continuing research on smallpox remains a vital aspect of bio-defence and US national security. For further reading I recommend: Orthopox Targets for the Development of New Anti-viral Agents published by NIH (See: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3773844/)
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Sunday, March 2, 2014

Bad Bugs: Micro Air Vehicles as a Delivery System for Biological Weapons

Authors note: In a recent search for articles on 'smart dust' I came across this on the Ministry of Defence website for Singapore. It closely mirrored one of my recent publications, so I thought I should share it here instead. It appears Mindef Singapore has updated a version of a previous paper I published. Their statementfound at: http://www.mindef.gov.sg/imindef/publications/pointer/journals/2007/v33n4/Emerging_Capabilities_-_A_Precision_Weapon_of_Mass_Destruction.print.html?Status=1

As somewhat of a stickler for attribution, I've included the MINDEF Singapore link


"With the rapid development and convergence of the three technologies; biotechnology, nano-sensors and unmanned systems, a new employment concept for biological warfare could emerge. Biological weapons could be the future precision weapon while retaining its capability for mass destruction. The precision would be two-fold in that it would be released at the most favorable conditions to ensure success while inflicting casualties confined to a specific target group. Unmanned "Smart Dust" sensors occupying the battlefield space would allow real-time, undetectable reconnaissance of targets and weather conditions of the area. Coupled with the delivery system of miniature unmanned vehicles with their genetically modified biological payloads, a timely attack with a high probability of success could be executed. The biological payloads would inflict casualties on a certain target group, preventing fratricide as they would be modified to act on a gene predominantly present within a particular population. Compared to current kinetic precision weapons, the price tag would be substantially lower, thus allowing once excluded parties or nations to acquire such a weapon. Its destructive effects would be precise and yet also much more widespread than kinetic precision weapons due to its infection-proliferation-infect cycle which can only be broken by quarantine. Collateral damage or fratricide often associated with both asymmetric warfare and precision warfare would be reduced or even negated. " Mindef Singapore


Micro Air Vehicles or MAV's may be modified to be a highly complimentary deployment platform for weaponized biological warfare agents. In contrast to chemical weapons for which a significant payload is required and volumes which would not, baring perhaps toxins be well suited for this delivery platform, BW would be.  A 2011, Popular Science article by Clay Dillow, entitled: Bug like Robotic Drones Becoming More Bug-Like With Bulging Eyes and Tiny Sensing Hairs (see: http://www.popsci.com/technology/article/2011-10/big-robotic-drones-become-more-bug-bulging-eyes-and-tiny-sensing-hairs) noted: 

"Micro air vehicles or MAVs, make for a tantalizing option for intelligence and surveillance agencies looking to surreptitiously gather information or deliver surveillance devices without being seen. But MAVs usually modeled after small birds or insects--are notoriously unstable in flight and difficult to maneuver in cluttered environments. So the Pentagon is handing out research contracts to make the DoD's little robotic bugs more stable by making them more bug-like. Specifically, the DoD wants big bulging bug eyes and hairy wings for its MAVs. The main problem with MAVs has to do with the way they respond (or don't respond) to dynamic environments--things like shifting or gusting winds, moving bodies, and other variables that have to be accounted for in real time. MAVs are tiny, so there's  not a lot of space for computing assets or sensor payloads, and that leads to a sort of intractable problem: how can engineers make these things smaller and more capable while also adding increased situational awareness and better in flight processing?"  




This is apparently being adjusted for according to a Wikipedia piece which contends: 

"A new trend in the MAV community is to take inspiration from flying insects or birds to achieve unprecedented flight capabilities. Biological systems are not only interesting to MAV engineers for their use of unsteady aerodynamics with flapping wings; they are increasingly inspiring engineers for other aspects such as distributed sensing and acting, sensor fusion and information processing. Various symposium bringing together biologists and aerial roboticists have been held with increasing frequency since 2000.Although there are currently no true MAVs (i.e., truly micro scaled flyers) in existence, DARPA has attempted a program to develop even smaller Nano Air Vehicles (NAVs) with a wingspan of 7.5 centimeters.[17] However, no NAVs meeting DARPA's original program specification were forthcoming until 2009 when AeroVironment demonstrated a controlled hovering of DARPA's flapping-wing NAV.[18]http://en.wikipedia.org/wiki/Micro_air_vehicleBeyond the difficulties in developing MAVs, few designs adequately address control issues. The MAVs' small size makes teleoperation impractical because a ground station pilot cannot see it beyond 100 meters. An onboard camera allowing the ground pilot to stabilize and navigate the craft was first demonstrated in the Aerovironment Black Widow, but truly micro air vehicles cannot carry onboard transmitters powerful enough to allow for teleoperation. For this reason, some researchers have focused on fully autonomous MAV flight. One such device, which has been designed from its inception as a fully autonomous MAV, is the biologically-inspired Entomopter originally developed at the Georgia Institute of Technology under a DARPA contract by Robert C. Michelson (19).


Clostridium Botulium 
Although one might consider that the lack of space on the very small MAV's could inhibit its use as an effective platform for BW deployment, considering that something like modified botulinum toxin or a synthetic version might be aerosolized beyond the 0.1-0.3mcm  and then merging this with swarm technology could well prove to be a formidable biological weapon deployment platform in a theatre of combat or even in civilian population centers. I will make the point that we are way beyond silica coatings which make anthrax weaponization almost obsolete. While my preference within the field of Category A highly pathogenic agents and toxins has always been those which are highly virulent and transmissible, such as smallpox, one could imagine anthrax or botulinum toxin deployed in this way could be effective and could result in high kill ratios. Its an interesting concept, the scenarios one can imagine are almost limitless. Note: 'Some contemporary analysis discount the potential of botulinum toxin as a bioweapon because of constraints in concentrating and stabilizing the toxin for aerosol dissemination. However these analysis pertain to military uses of botulinum toxin to immobilize an opponent (William C. Patrick unpublished data, 1998). In contrast, deliberate release of botulinum toxin in a civilian population would be able to cause substantial disruption and distress. For example, it is estimated that a point source aerosol release of botulinum toxin could incapacitate or kill 10% of persons within 0.5 km downwind (William C. Patrick, unpublished data, 1998) see: http://www.bt.cdc.gov/agent/botulism/botulismconsensus.pdf  Just as a personal note and to be complete, Botulinum Type H has recently been discovered in the feces of a child suffering from botulism. The toxin's DNA hasn't been released to the public as it has no antidote, which clearly makes it a good BW candidate. See http://www.natureworldnews.com/articles/4442/20131015/botulinum-toxin-type-h-deadliest-known-antidote-discovered.htm

Countering BW MAV Payloads Converged with Swarming  (To be Continued in Part II)

Intro: No doubt there is already research being done to assess the risks such technology pose, should it be used offensively against our own forces and civilian populations.
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Thursday, February 27, 2014

Virus Like Particles Applications for Bio-Defence Vaccine Delivery





Virus Like particles (VLPs) are a promising vaccine platform due to safety and efficiency. When we consider specific issues related to bio-defence vaccine development and of course non-bio-defence related vaccine R and D, VLP's would appear to be a highly promising technology. Bio-defence vaccines, particularly those which are perceived as less likely agents for development, testing and use for bio-terrorism and/or warfare,  have typically faced funding issues. BioShield and BARDA have attempted to level the playing field, but there remain significant concerns related to such investment. Its difficult to develop vaccines for diseases or therepeutic counter-measures for diseases which in some instances, have never occurred on a large scale or at all. As a bio-defence specialist and risk analyst, in my view, these vaccines and medical counter-measures are a critical aspect of public health protection. Threat reduction, on the security policy side is the perview of social sciences and the intelligence community, and although useful in countering the threat of bio-terrorism, it is not a replacement for the continued need to ensure public health security at the therapeutic level. The bio-defence community would be remiss in their responsibilities if they did not do this. Vaccines are costly and investment, partiuclarly in early stage or by the time a company is entering the "Valley of Death", is vital to production and ultimately public health security. 

Aside from reducing safety issues related to some vaccines and medical counter measures, VLPs may make investments in bio-defence vaccines and orphan drug research and development, a far more secure bet in terms of efficacy, safety, manufacturing costs, and timeline delivery. Investment in diseases which occurr naturally and have the potential to result in pandemic outbreaks with severe consequences to public health security, have typically been of more economic and investment interest than diseases which may or may not ever occur. It is the later group of vaccines, which is likely to benefit considerably from advanced VLP technology. One can imagine that even a product like VIG (Vaccinia Immune Globulin) for example, developed in the 1960's to treat those counter-indicated for smallpox vaccine, might be an interesting candidate. Still other neglected diseases could well benefit from advances in VLP research and development.

Monday, February 24, 2014

DARPA's 7-Day Bio-Defence and the Future of Synthetic Vaccines

US military personnel don special biohazard gear during a training exercise designed to simulate a biological weapon attack. The Department of Defense and other agencies routinely hold training sessions throughout the country as part of a domestic bioterrorism preparedness program. 'The challenge is to integrate these forces to mount an effective response under various attack scenarios," says Prof. Steven Block. Courtsey: US Navy 

Author's note: In 2007, I attended a bio-defence briefing delivered by a DARPA scientist. Since then I have always thought of DARPA as the 'bugs on the wall folks' and indeed they have successfully produced robotic bugs, but more significantly, they discussed advanced bio-defence technologies which we were not allowed to take notes on or photograph the slides. It was extremely exciting and I left with a sense of awe which DARPA tends to inspire. Six years later, they have successfully manufactured 10 million doses of vaccine within in a month. For pharmaceutical companies who invest up and over a billion per drug and which including research and development takes about ten years to bring onto the market, this was a phenomenal feat. Here we are on the threshold of major breakthroughs in vaccine research, development and production, even manufacturing technologies and the future of bio-defence couldn't look brighter. While industry of course continues to work on live attenuated vaccine production is the future a synthetic one?

Photo: www.csc.com
Current vaccine production based on inactivated  viruses (live attenuated vaccines) has been successful in reducing significant disease burden associated with major epidemics of the 19th and 20th centuries. However, a major draw back has been the lengthy research and development phase, the significant investment costs and the inability to respond rapidly to changing strains. Synthetic vaccines may overcome many of the more traditionally based production obstacles. In 2012 DARPA announced it's Blue Angel Program. On their site they lay out the problem quite concisely with regard to responding to pandemics of the future. Such pandemics may or may not be natural and may or may not be caused by natural pathogens, viruses and toxins. Consider fighting a synthetically produced outbreak. The DARPA site states:

"The 2009 Army Posture Statement, cites a World Health Organization estimate of between 20 and 50 percent of the world's population being effected if a pandemic were to emerge. WHO forecasts 'it may be six to nine months before a vaccine for a pandemic virus strain becomes available." In a separate report on pandemic influenza, the WHO described several challenges to producing sufficient volumes of vaccine using current, egg based protein-production technology, including the likihood that two doses per person could be required due to the absence of pre-existing immunity. In short, the potential for a pandemic exists and current technological limitations on defensive measures put the health and readiness of U.S. military forces at risk. A technological solution to increase the speed and adaptability of vaccine production is urgently needed to match the broad biological threat. DARPA's Blue Angel Program seeks to demonstrate a flexible and agile capability for the Department of Defence to rapidly react to and neutralize any natural or intentional pandemic disease. Building on a previous DARPA program, Accelerated Manufacture of Pharmaceuticals, Blue Angel targets new ways of producing large amounts of high quality, vaccine grade protein in less than three months in response to emerging and novel biological threats. One of the research avenues explores plant made proteins for candidate vaccine production. "Vaccinating susceptible populations during the initial stage of a pandemic is critical to containment," said Dr. Alan Magill, DARPA program manager. "We're looking at plant based solutions to vaccine production as a more rapid and efficient alternative to the standard egg-based technologies, and the research is very promising." In a recent milestone development under Blue Angel, researchers at Medicago Inc. produced more than 10 million doses (as defined in an animal model) of an H1N1 influenza vaccine candidate based on virus-like particles (VLP) in one month. Production adhered to Phase 1 appropriate current good manufacturing practices. The work was part of a 'rapid fire' test that ran from March 25, 2012 to April 24, 2012, at a facility in Durham, NC. A third party laboratory tested the production lots to confirm the immunogenicity of the vaccine candidate. Testing confirmed that a single dose of the H1N1 VLP influenza vaccine candidate induced protective levels of hemagglutinin antibodies in an animal model when combined with a standard aluminum adjuvant. The equivalent dose required to protect humans from natural disease can only be determined by future, prospective clinical trials. 

DARPA's 7 Day Bio-Defence 

Photo: DARPA

In May, 2013 Medicago Inc. announced it had successfully produced a VLP vaccine candidate for the H7N9 virus responsible for an influenza outbreak in China. Medicago's future in VLP vaccines couldn't look brighter especially given their success with rapid vaccine production and their work with DARPA. 
An additional, although slightly different project which I believe will make significant strides in bio-defence counter-measures is DARPA's 7-Day Bio-Defence project, again, worth considering in terms of how future vaccines will be manufactured and how investment in this technology could well shift. As stated on their site:

"Military readiness and national security depends on the health and well being of military service members. The Department of Defence's (DoD) cumulative investment in personnel comprises the second largest share of the total defence budget. As such DoD seeks advances in health care to ensure war-fighters can operate at peak performance. Research into natural and synthetic pathogens, and treatments against them is one plank of ensuring military readiness in the face of accidental and offensive biological threats to both war-fighters and the supply chain supporting them. In this context, the 7-Day Biodefence program will seek to develop novel technologies focused on preventing infection by any emerging pathogen, sustaining survival once infected, and building immunity. In recent years, global surveillance networks have determined an increase in the frequency and diversity with which new infectious micro-organisms are emerging. While this increase is due in part to improved reporting, multiple examples demonstrate it is also promulgated by changes in natural systems and possibly human activity. The potential biological threat breaks down into two primary categories: 

1. Exposure to natural pathogens that are the result of: increased human-animal interface; increased population densities and co-location of vulnerable species with pathogen reservoirs; climate change, particularly affecting migration and spread of vectors; and narrowing of genetic diversity among food-animal stocks. 

2. Exposure to synthetic and highly diverse pathogens that have become easier to produce as bio-medical and genetic-engineering technologies proliferate internationally; such that pathogens could be used by adversaries for offensive purposes in a direct attack on war-fighters for covert sabotage of the agricultural industry that supports war fighters. 

Together, these emerging threat agents challenge current medical countermeasures. Today's research and development cycle for countermeasures is ill-equipped for rapid response to emerging biological threats. In response to the unspecified potential threat from emerging pathogens, the goal of the 7-Day Biodefence program is to develop innovative approaches to counter pathogens without regard to their exact nature. The methods being explored do not require prior knowledge of the pathogen and are broadly applicable to multiple, unrelated infectious agents. The program consists of four technical areas invstigating novel technologies to: 1. prevent infection; 2. sustain survival; 3. provide transient immunity; and 4. create persistent immunity. See: http://www.darpa.mil/Our_Work/DSO/Programs/7-Day_Biodefense.aspx

Advances in synthetic vaccine manufacturing, VLP's and even 3D bio-printing will significantly change our concept of bio-defence and the manufacturing of counter-measures. The incorporation of these counter-measures will additionally change how we approach threat reduction and possibly remove many of the traditional concerns at the technical level. This could increase interest in areas which have typically been an after thought.
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Sunday, February 23, 2014

Manufacturing of Recombinant Botulinum Neurotoxin Vaccine

PHOTO: http://iai.asm.org/

As the  Medical Countermeasure System Joint Vaccine Acquisition Program (MCS/JVAP), the advanced developer for the Department of Defence (DoD) responsible  for developing, producing and stockpiling FDA licensed vaccines to protect the Warfighter from biological agents puts out a Request for Information a requirement to develop and license a pre-exposure prophylaxis recombinant botulim neurotoxin serotype A and B vaccine, scientists, Jason Barash and Stephen Arnon at California Department of Public Health announced they had discovered the first new form of botulinum toxin in over forty years.  Moreover, the scientist published two reports describing their work online in the Journal of Infectious Diseases. As with previous study which could provide a recipe for would be bio-weaponeers, the genetic sequences were withheld.See: http://www.npr.org/blogs/health/2013/10/11/230957188/why-scientists-held-back-details-on-a-unique-botulinum-toxin

The recent discovery of Botulinum Type H, the most lethal of botulinum neurotoxins, with no known antidote is likely to concern bio-defence specialists. The previously known seven serotypes: A,B,C1,C2,D,E,F and G produced by the bacterium clostridium botilinum, block acetylcholine, a neurotransmitter which then leads to muscle paralysis. According to New Scientist, this is the first time a genetic sequence has been withheld from publication over security concerns. Patients are usually treated with monoclonal antibodies, immune proteins which react to the specific toxin type. In a March, 2013 report,  Medscape notes: 
In an abstract (Botulinum neurotoxin vaccines: past present, and future),  published by PubMed back in 2007, Smith, LA and Rusnak JM detail that 'In the early 1930's, a formalin-inactivated toxoid against botulium neurotoxin was first tested in humans. In 1965, a pentavalent botulinum toxiod (PBT) received Investigational New Drug (IND) status under the Centers for Disease Control's IND 161 (for at risk workers), and in 1991 under the United States Army's Office of the Surgeon General IND 3723 (for military deployment). This PBT vaccine has been shown to be safe with over 20,000 injections given to date, and continues to be used in at risk individuals. During the past decade, recombinant DNA technology has been employed to develop second generation vaccines to prevent botulism. Recombinant subunit vaccines utilizing the receptor-binding domains of botulinum neurtoxin (BoNT) have been shown to be safe and efficacious in protecting animal models against BoNT serotypes A, B, C1, D, E and F. In 2004, the first recombinant subunit vaccine [rBV A/B(Pichia pastoris) vaccine] was tested in humans during a phase 1 clinical trial. Results from that study demonstrated that the recombinant bivalent vaccine was safe and well tolerated at all dosage levels tested and stimulated serotype specific neutralizing antibodies among the majority of vaccine recipients. See: http://www.ncbi.nlm.nih.gov/pubmed/18197811

PHIL Image 12052
Clostridium innocuum bacteria
"The US Food and Drug Administration (FDA) last Friday approved the first botulism antitoxin to neutralize all 7 known botulinum nerve serotypes valuable versatility for a drug in the nation's emergency medicine cabinet against a bioterrorist attack. The heptavalent botulism antitoxin (BAT,Cangene) had been available on an investigational basis from the Centers for Disease Control and Prevention (CDC). Cangene began supplying doses of BAT to the US Strategic National Stockpile in 2007 under a 427 million contract with the Department of Health and Human Services, according to a company press release. The CDC will distribute the stockpile antitoxin." See: http://www.medscape.com/viewarticle/781389

I would agree with Cangene's statement to the effect that "Currently, no specific, licensed therapies are available to treat all seven known serotypes" and "Given the inability to rapidly detect and identify botulinum neurotoxin (BoNT) serotypes in emergency situations, there is a critical need for a single effective treatment against all BoNT serotypes." See: http://www.raps.org/focus-online/news/news-article-view/article/2849/could-cangenes-botulism-antitoxin-become-the-seventh-product-approved-under-the.aspx

As a defence specialist, I would suggest that it is wise to recall in 1995, Iraq admitted it had produced 19,000 liters of botulinum toxin (See: http://www.fas.org/spp/starwars/crs/98042705_npo.html). Botulinum is listed by CDC and WHO as a Category A agent. It can and has been weaponized and poses a risk as bio-warfare agent.  In a 2005 Proceedings of the National Academy of Sciences (PNAS) announced it would publish a paper that presented a mathematical model of the possible consequences of deliberate botulinum toxin contamination of the US milk supply. The paper was authored by Dr. Lawrence Wein of Stanford University Graduate School of Business and Yifan Liu a graduate student. An overview is available here: http://www.nytimes.com/2005/05/30/opinion/30wein.html?pagewanted=all 
While I sat in on the ethics meetings to decide if Wein and Liu's study should be published in full, the first time such a meeting convened due to security concerns, botulinum and its potential use a weapon came to the foreground. Protection against such toxins is a vital aspect of national security. 
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Jill Bellamy is an internationally recognized expert on biological warfare and defence. She has formerly advised NATO and for the past seventeen years has represented a number of bio-pharmaceutical and government clients working on procurement strategy between NATO MS and Washington DC. Her articles have appeared in the National Review, The Wall Street Journal, The Washington Post, The Sunday Times of London, Le Temps, Le Monde and the Jerusalem Post among other publications. She is a CBRN SME with the U.S. Department of Defence, Chemical, Biological, Radiological and Nuclear Defence Information Analysis Center and CEO of Warfare Technology Analytics, a private consultancy based in the Netherlands. She is an Associate Fellow with the Henry Jackson Society, UK.

up-coming: Merging Social Network Analysis with DARPA's Bio-Intelligence Chips (BIC)

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Emerging Technologies: Bio-hacking and the future of bio-terrorism




For some time concern has been raised over 3D and 4D technologies, (with synthetic biology the emerging technological forerunner of these concerns and the NSABB playing watchdog), with regard to how inherent de-skilling may reduce the technical threshold which inhibits most would-be weaponeers from developing and deploying a weaponized biological agent capable of mass destruction. At the somewhat more extreme end, bio-hacking could reduce barriers which are perhaps better left in place.  Bio-hacking  was put on the map in 2013 when molecular biologist Ellen Jorgensen delivered a TED talk about Genespace, the DIY science lab she opened in New York in late 2010. See: http://new.ted.com/talks/ellen_jorgensen_biohacking_you_can_do_it_too  


The lab Jorgensen oversees is one of approximately 45 DIY international science groups,of  more than twenty in the US. While some of these labs are rather extreme in their goals, emerging technology such as 3D bioprinting could theoretically reduce the knowledge needed to develop synthetic weapons. So far several of the bio-hacking groups seem to be content with using themselves in experiments and implanting magnets but coding life for the masses and or the non-scientific community, could become a lot easier in a relatively short period of time. 

"But we don't smuggle plutonium. We don't supply chemical weapons. We don't build rockets. Instead we have a hobby that the FBI believes could be so dangerous that they have come up with a special programme to make sense of it. That hobby is to play with genes, proteins and bacteria in our spare time in a homemade lab we constructed from scratch. We are part of a rapidly growing community of amateur geneticists, who are often labelled biopunks, or outlaw biologists. Or, better still, in an analogy to the computer programming enthusiasts of a generation ago, some call us bio-hackers. But instead of software code, we try to tinker with DNA, the code of life. The FBI has set up the Biological Countermeasures Unit ( ) one of their goals in preventing acts of terrorism is to reach out to leading names in the field to quiz them about what they do." See: http://www.bbc.com/future/story/20130122-how-we-became-biohackers-part-1

This surely must be cutting edge bio-security, however, how close are bio-hackers to actually crossing what was considered the technological threshold to creating what might even be considered synthetic biological weapons? After 911 and the US anthrax attacks, I advised governments that mass casualty bio-terrorism was not as simple as it was being touted. In fact I, and several other scientists, focused on state warfare laboratories, considering bio-terrorism not of real world interest. Emerging technology which results in de-skilling however, may make the life of the would be bioweaponeer far easier and reduce what was always considered to be rather insurmountable technical barriers, certainly in the deployment of a mass casualty weapon. 

biohackspace.org 

What is the current view of life sciences deskilling, given the increase in DIY science? Johnathan B.Tucker, a former long time colleague, presented an excellent analysis of the issue in his paper, "Could Terrorists Exploit Synthetic Biology? published in The New Atlantis, see: http://www.thenewatlantis.com/publications/could-terrorists-exploit-synthetic-biology, although notably before bio-hacker movement emerged more openly into the media with a cohesive defined goal and group structure. Tucker, in his analysis states: 

"Member of this second school point to a contradictory trend in biotechnological development that they claim will ultimately prove stronger. They note that the evolution of many emerging technologies involves a process of de-skilling that, over time, reduces the amount of tacit knowledge required for their use. Chris Chyba of Princeton, for example, contends that as whole-genome synthesis is automated, commercialized, and 'black-boxed,' it will become more accessible to individuals with only basic scientific skills, including terrorists and other malicious actors (16).De-skilling has already occurred in several genetic-engineering techniques that have been around for more than twenty years, including gene cloning (copying foreign genes in bacteria), transfection (introducing foreign genetic material into a cell), ligation (stitching fragments of DNA together), and the polymerase chain reaction, or PCR (which makes it possible to copy any particular DNA sequence several million fold). Although one must have access to natural genetic material to use these techniques, the associated skill sets have diffused widely across the international scientific community. In fact, a few standard genetic-engineering techniques have been de-skilled to the point that they are now accessible to undergraduates and even advanced high school students, and could therefore be appropriated fairly easily by terrorist groups." See:http://www.thenewatlantis.com/publications/could-terrorists-exploit-synthetic-biology   

Gerald Epstein, of the Center for Science, Technology and Security Policy, write that whole-genome synthesis 'appears to be following a trajectory familiar to other useful techniques: Originally accessible only to a handful of top research groups working at state of the art facilities, synthesis techniques are becoming more widely available as they are refined, simplified, and improved by skilled technicians and craftsmen. Indeed, they are increasingly becoming 'commoditized,' as kits, processes, reagents, and services become available for individuals with basic lab training." (17). In 2007 Epstein and three co-authors predicted that 'ten years from now, it may be easier to synthesize almost any pathogenic virus than to obtain it through other means," although they did not imply that individuals with only basic scientific training will be among the first to acquire this capability.(18)" See:http://www.thenewatlantis.com/publications/could-terrorists-exploit-synthetic-biology 

"To date, the de-skilling of synthetic genomics has affected only a few elements of what is actually a complex, multi-step process. Practitioners of de novo viral synthesis note that the most challenging steps do not involve the synthesis of DNA fragments, which can be ordered from commercial suppliers, but the assembly of these fragments into a functional genome and the expression of the viral proteins. According to a report by the U.S. National Science Advisory Board for Biosecurity, a federal advisory committee, "The technology for synthesizing DNA is rapidly accessible, straightforward and a fundamental tool used in current biological research. In contrast, the science of constructing and expressing viruses in the laboratory is more complex and somewhat of an art. It is the laboratory procedures downstream from the actual synthesis of DNA that are the limiting steps in recovering viruses from genetic material." (19)" See: http://www.thenewatlantis.com/publications/could-terrorists-exploit-synthetic-biology 

As technology emerges which contributes to deskilling and with the advent of DIY science, we may witness rather rapid advancements which overcome the long time presumed threshold. The bio-hacking community has emerged because techniques used in molecular biology have been de-skilled and the cost has dropped. 




"A couple of decades ago, it took three years to learn how to clone and sequence a gene, and you earned a PhD in the process. Now, thanks to ready made kits you can do the same in less than three days. Specialized materials and second hand equipment are much more affordable, not to mention more available. Machines for amplifying DNA can now be purchased online, whilst enzymes and chemicals for creating, manipulating and sticking together DNA an be ordered off the shelf. The cost of sequencing DNA has plummeted , from about 100,000 for reading a million letters or base pairs, of DNA code in 2001, to around 10 cents today. See:http://www.bbc.com/future/story/20130122-how-we-became-biohackers-part-1 full review: Warfare Technology Analytics
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Monday, February 17, 2014

How 3D Bioprinting would change a Wargame: Atlantic Storm




   Influenza virus Credit: Dr Paul Digard, Department of Pathology

Author's note: In 2007 I was invited to speak at a NATO instillation near Warsaw. I delivered a talk on variola major the causative agent of smallpox and highly pathogenic disease containment. At that time, the US Strategic National Stockpile was rapidly increasing its procurement of a number of vaccines and medical counter measures, including vaccine to prevent smallpox. The US strategic stockpile was comprised of vaccine from Acambis and Bavarian Nordic. As I noted in my discussion, NATO held only a virtual stockpile and one had wonder how a real response could be mounted by NATO, not just allocated to the Member States, holding their national stockpiles, but by NATO command, when they only had a virtual stockpile. Atlantic Storm, following on from Dark Winter, was in my view a conservative and realistic table top exercise. Below is the basic introduction to Atlantic Storm,  but what would happen if instead of strategic stockpiles we could use centrally located 3Dbioprinters? What if NATO had  3D bioprinters for example or even the World Health Organization? How would that change how we counter highly pathogenic and possibly deliberate outbreaks of disease? I do make a distinction between deliberate and  natural outbreaks, as a deliberate outbreak is likely to increase mortality rates. So here are the basics of Atlantic Storm:  
Atlantic Storm was a simulated bioterrorist attack which demonstrated the weakness of international public health and security systems when dealing with a sudden outbreak of highly infectious diseases. On 14 January 2005, ten heads of government from Europe and North America and the Director General of the World Health Organization (WHO; Geneva, Switzerland) were scheduled to meet for a ‘Transatlantic Security Summit' in Washington, DC, USA, to discuss the threat of international terrorism. On the eve of the meeting, news broke that citizens from several European countries appeared to have become ill with smallpox; shortly thereafter suspected smallpox cases appeared in the USA. Although the assembled leaders did not know it at the time, a radical terrorist group had obtained seed strains of Variola major—the virus causing smallpox—and deliberately released the virus in a number of main transport hubs and sites of commerce throughout Europe and North America. On 14 January, the heads of states who gathered in Washington were confronted with one of the worst nightmares imaginable: the use of contagious and deadly disease as a weapon. Full citation see:http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1369238/

Just as Atlantic Storm began, the scientific journal Nature published a paper describing a new technology that allows the rapid and accurate synthesis of long DNA segments using standard laboratory chemicals (). In October 2005, scientists at the Centers for Disease Control and Prevention (Atlanta, GA, USA) published the reconstruction of the ‘Spanish' influenza virus, which killed at least 25 million people during the winter of 1918/19 (). These and other discoveries provide researchers with better tools and knowledge to develop new medicines and vaccines against infectious diseases. However, they also make the synthesis and modification of viruses and bacteria for criminal purposes more likely. Hundreds of biological laboratories around the world already have the technical capacity to synthesize or manipulate small viruses such as polio or flu. Smallpox, with its genome of approximately 200 kb, is technically more challenging, but within the next few years, technology will undoubtedly advance to the stage where the synthesis of Variola major—based on sequence information freely available on the worldwide web—will be possible. The age of engineered biological weapons is neither science fiction nor suspense thriller—it is here today ().
For decades, NATO and other security alliances have planned their response to all kinds of military crises. Planning with that degree of rigor and strategic and operational detail is also needed to cope with potential biological threats of international consequence. Such transatlantic cooperation is also at the core of many non-proliferation programmes, such as the US Department of Defense's Cooperative Threat Reduction Program or the G8 Global Partnership. These programmes seek to reduce the threat posed by weapons of mass destruction (WMD) by detecting, deterring and interdicting illegal trafficking in such items; improving the physical security of facilities and WMD materials; destroying chemical weapons agents; preventing radiological contamination by decommissioned Russian nuclear submarines; and providing former WMD programme personnel with a decent living so they will not seek to profit from selling their knowledge to terrorist organizations or states trying to acquire WMD. However, biosecurity has often been an orphan of such programmes and must be given both higher priority and more resources commensurate to the challenge (; Dalgaard-Nielsen & Hamilton 2005). “We live in a time of new threats… What we now see is that health and security go together, so we have to combine them, and I think the lesson we should draw from this…is that we don't have the organizational structures to deal with the new threats,” commented Jan Eliasson, who acted as the Swedish Prime Minister in Atlantic Storm. Similarly, Sir Nigel Broomfield (Fig 3), who played the British Prime Minister, said after the exercise, “we have a globalized economy and globalized society, but we don't yet have globalized effective institutions to deal with the questions that come out of the globalization process.”http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1369238/

If we consider not only the time it takes to deploy a vaccine, and the US CDC estimates the time at about twelve hours, and we consider international travel, this time frame becomes critical. This photo below illustrates one aspect of what is involved in deploying medical countermeasures. If however, a state or institution such as NATO or WHO could manufacture vaccines on site, something akin to having fire hydrants strategically located throughout a city for putting out fires and easily accessible to all, containing disease would be far more efficient and effective. 


Click photo for screen-resolution image


Air Force Senior Airman Jacob Lloyd, a member of the 153rd Logistics Readiness Squadron, moves simulated vaccines onto a C-130 Hercules as part of the Strategic National Stockpile exercise in Wyoming. May 14, 2012. Airmen assigned to the Wyoming Air National Guard worked with multiple state agencies to test state health officials' abilities to receive, deliver and distribute medical supplies to various parts of the state. (Air National Guard, photo by 1st Lt. Rusty Ridley)

While it is prudent to understand that such technology can potentially be used to manufacture biological warfare agents, I believe the benefits far outweigh this possibility. In terms of Atlantic Storm, 3D bioprinting would remove policy decision making on vaccination/containment strategy, logistical issues involved in deploying a stockpile, remove transnational issues related to supply and rapidly increase our ability to respond at the first instance. One of the issues particularly with regard to smallpox vaccine availability within EU/NATO states is inconsistent stockpiles. One can imagine if there were an outbreak of smallpox in Africa, given lengthy incubation periods,  flights arriving into Schipol or Zaventem would not raise any alarm until the first cases started emerging. By this time, Belgium which holds only slightly over a 15% coverage would see citizens possibly trying to cross the boarder to the Netherlands where they hold a 100% so 1:1 stockpile. 3D bioprinting could theoretically remove all these issues possibly making bio-terrorism and warfare a less attractive option. 
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For an interactive view see: http://www.atlanticstorm.org/flash/