CBMTS III: A Summary
by Dr. Rudolf Portmann
AC Laboratorium Spiez, Switzerland
At CBMTS III ninety-eight scientists from 29 countries participated
and presented papers in the areas of defense against chemical
and biological warfare and chemical and bioterrorism. The presentations
took place in a total of 16 sessions, including the three poster
Symposium Chair: Dr. Brian Davey
Symposium Co-chair: Dr. Barbara Price
Sector Chemical (Sessions: 1,2,
4, 5, 8, 9, 10, 11,
Chair: Dr. Rudolf Portmann
Co-Chair: Dr. David Moore
Sector Biological (Sessions: 3, 6,
7, 12, 14,
Chair: Dr. Barbara Price
Co-chair: Prof. Ji-Sheng Chen
Session 1 - OP Treatment: State of the
Chair: Peter Eyer; Co-chair: Jiri Bajgar
This session was devoted to the practical question of treatment
of OP intoxication as well as the effects of low dose exposure
to OP compounds. Dr Elsa Reiner gave an introductory overview
of the current knowledge about research topics concerned with
treatment of OP intoxication. Dr. Balali-Mood presented the results
of a study aimed at investigation the effects of extra sodium
bicarbonate infusion (alkalinization to pH 7.47) in OP-poisoned
patients which all received atropine. There was the suggestion
of some benefits, but this was not statistically significant.
Due to the potential low cost of bicarbonate therapy if it is
successful, the study will be extended with even higher alkalinization.
Dr. Bajgar presented a study using the erythrocyte-AChE inhibition
to identify the underlying nerve agent. Dr. Worek presented results
indicating that if an oxime substantially reactivates the BuChE
(serumcholinesterase), this will lead to a re-inhibition of the
AChE (acetylcholinesterase). Dr. Scott presented a highly advanced
approach to follow subtle behavioral and electrophysiological
changes upon low-dose exposure towards OP in marmosets. It appears
that a single sign-free dose exposure of non-human primates to
sarin was without any lasting significant effect.
Session 2 - Posters Animal Treatment/ChE
Chair: Elsa Reiner; Co-chair: Atilla Hincal
This first poster session included treatment studies of OP intoxicated
animals (Dr. Antonijevic, Dr. Dorandeu). The authors gave a brief
platform presentation, followed by an extremely lively discussion
in the poster area. This format seems to be very stimulating
for further discussions and collaborations.
Session 3 - Bioterrorism: National Approach
Chair: Murray Hamilton; Co-chair: Lotfali Haghighi
This session was dedicated to the national approach against chemical
and biological terrorism. Here overviews of national readiness
and response planning were presented Dr. Erasmus, South Africa,
Dr. Harwood, Canada, Dr. Treadwell, USA, Dr. Spies, USA and Dr.
Robertson, Australia. Col. Khan from Pakistan opened the Session
with a presentation detailing characteristics of modern terrorists
as they have moved away from large organizations towards smaller
but more focussed groups.
Session 4 - OP Treatment Studies
Chair: Leah Scott; Co-chair: Florin Paul
This session's theme concerned the studies of treatment of OP-intoxications.
Dr. Phillipens presented a summary of guinea pig studies investigating
whether stressors are influencing the effects of pretreatment
using physostigmine and the effectiveness of such pretreatment
in protecting against soman. Stressors appear to induce adverse
behavioral effects in pretreated animals and that protective
efficacy was compromised. Dr. Zhou described in vitro studies
involving cholinolytic drugs and oximes. Unlike the oximes, toxogonin
and Hl-6, these compounds did not by themselves reactivate AChE.
But the results confirmed that, if added at the start of ageing,
reversible ChE inhibitors increased the portion of AChE reactivatible
by the oximes. Dr. Tonkopii discussed the effectiveness of a
series of familiar and less familiar carbamates as pretreatment
for nerve agent poisoning in vivo. Interestingly, some of the
carbamates were more toxic than the nerve agents and others made
the nerve agent mixture more toxic.
Session 5 - Medical Treatment
Chair: Mahdi Balali-Mood; Co-chair: Cornelis Erasmus
Questions on the treatment of convulsion, mustard and heavy metal
intoxication were discussed. Dr. Rump first pointed out the side
effects of benzodiazepines, and in particular, diazepam as anti-convulsant,
and then introduced imidazenil as the ideal drug for the management
of OP-induced convulsions. He concluded that imidazenil, in a
dose of 2 mg/kg, blocked soman induced convulsions and increased
the anti-lethal effectiveness of atropine and HI-6 in soman poisoned
mice. Dr Szinicz pointed out that the role of inflammatory cytokines
released after sulfur mustard (SM) injury is not well known.
The data indicate HaCaT cells to be an appropriate model for
the study of SM effects in keratinocytes compared to SCL II cells.
According to Dr. Jaiswal no satisfactory prophylactic for the
treatment regimen has evolved for sulfur mustard. He introduced
Amifostine and its analogs as prophylactic agents for SM poisoning.
Amifostine and one of its analogs, DRDE-07, gave significant
protection. Dr. Wormser carried out a prolonged study on the
therapeutic effects of iodine at 15, 30, 45, and 60 minutes after
SM exposure on guinea pig skin. Gross pathology analysis showed
a strong protective effect at intervals of 15 and 30 minutes,
and to a lesser extent at 45 and 60 minutes. Dr. Mircioiu presented
a paper on using chelators or "decorporators" as a
possible medical treatment for poisoning by radioactive metals.
Using pharmacokinetic theory he explored a bicompartmental pharmacokinetic
model, that appeared to work, in spite of the fact that the ion
cannot leave the central compartment. Ca, Zn and Cu complexes
of EDTA were among the "decorporators" studied.
Session 6 - Toxins
Chair: Leo Laughlin; Co-chair: Liaquat Ali Khan
Toxins were the theme. Dr. Laughlin discussed selected US BW
Agent detection technologies from an historical perspective.
The initial problems identified in the methods discussed still
persist: sensitivity, time for detection / identification, and
specificity of the methodology. Dr. Chen from China provided
an assessment of toxins that clearly indicated that that there
are far more toxins of concern than are currently listed in the
Chemical Weapons or Biological & Toxins Weapons Conventions.
He described a number of factors to assess the threat from toxins.
Dr. Hamilton explained how botulinum toxins inhibited acetylcholine
release by cleaving presynaptic membrane proteins necessary for
exocytosis of the neurotransmitter acetylcholine. A specific
library was prepared by combinatorial synthesis that contained
a repeating sequence (probably the binding recognition site)
that did inhibit botulinum toxin.
Session 7 - Bio/Toxin: Threat and Detection
Chair: Paul Russel; Co-chair: Sven-Åke Pearson
Main theme: identifying infectious diseases and toxins as candidate
biological warfare agents and techniques to assist diagnosis
of both natural disease outbreaks and biological warfare agent
outbreaks. In the first instance criteria based on biological
and physical properties of pathogens and toxins such as inactivity,
lethality, transmissibility, disseminability and weaponisation
were discussed by Dr Bokan as tools for identifying candidate
biological warfare agents against human, animal and plant targets.
A scoring mechanism was used to create a threat list which largely
concurred with agents listed in the BTWC. The use of rapid PCR
and light cycler were mentioned by Dr Schütz as a rapid
means of diagnosing a disease outbreak. The use of genetic techniques
for rapid identification and epidemiology was discussed using
Swiss studies of army training areas as an example. The dual
use of these techniques was highlighted and the fact that recombinant
gene research had increased dramatically over the last 15 years.
The best means of preventing less ethical research would be through
the BTWC and honesty in such fora as the CBMTS meetings. Dr Taleski
discussed direct epidemiological studies as a means of identifying
a disease outbreak due to the dissemination of a biological weapon.
Dr Paul, as an example, used anthrax's capabilities as a biological
weapon to show how the epidemiology would apply. The epidemiological
curve, with casualty numbers and fatalities were presented and
the current medical countermeasures to anthrax were shown. The
role of macrophages in viral hemorrhagic fevers (VHFs) was presented
by Dr. Ryabchikova, indicating that most viruses causing VHF
are able to multiply within these cells. Hemorrhagic rash, and
bleeding were seen in the baboon model.
Session 8 - Risk Assessment/Data Bank/Mass
Chair: Gurayten Ozyurt; Co-chair: Julie Tremblay-Lutter
Risk assessment, data banks and mass casualty management were
the overall theme of this session. Dr. Hincal pointed out, that
Drug and Poison information Centers may well provide the informational
links between medical institutions, military groups, police forces,
civil defense personnel and public transportation systems in
mass casualty management. Dr. Hughart introduced the developing
database, consisting of information about a chemical disaster,
likely to be encountered by first responders (military men or
women, disaster assessment teams, police officers, firemen and
emergency medical technicians). It has key words as DISASTER,
ASSESSMENT and ASSISTANCE. The understanding of the long-term
toxic effects of environmental toxicants and example data sets
from DYNCORP Project covering civilian and military risk management
were presented by Dr. Rao. Dr. Kosnett presented the key services
Poison Centers can provide in the event of CB incident: a) centralized
communication to hospital emergency department b) assistance
of antidote and related emergency medical supplies, c) communication
of information to the general public either through the media
or coming telephone calls. As pointed out by Dr. Mathenge, industrial
accidents are becoming a common feature in the process of industrial
development. The proactive action that would prevent the hazards
of these factories brought by natural and human induction, was
introduced. Dr. Hincal introduced the Mass Casualty Management
programs and preparations for Pharmaceutical Industry, which
is mainly located in Istanbul, Turkey, and their risk management
Session 9 - Riot control and Detection
Chair: Richard Gordon; Co-chair: Urs Brodbeck
Questions on detection, direct or retrospective, and riot control
agents were discussed by Dr. Davey, Dr. Noort, Dr. Juruli, Dr.
Surovtsev and Dr. Wicki. The overall message from this Session
was that both innocent control of civilians (such as pepper spray
or the events that occurred at Tbilisi), as well as more threatening
events (such as the Japan Tokyo incident that used impure sarin),
require techniques to identify the toxic chemical agents used.
Furthermore, a better understanding of the mechanism of action
of the agents would provide a means to detect the agents better,
and determine those agents that are indeed toxic. Most important,
the standardization of the methodology to identify post-exposure
to agents (biomedical samples) would provide the means to better
enforce the CWC. A result of the discussions was that the estimation
of cholinesterase activity is a useful indication of pesticide
intoxication in occupational health and therapeutic monitoring
of various neurological diseases.
Session 10 - Cholinesterase Determination
Chair: Uri Wormser; Co-chair: Valerii Tonkopii
As discussed, determination of ChE, is a crucial factor in detection
and monitoring exposure of humans to organophosphates and carbamates.
Much effort has been invested in developing appropriate analysis
that can simply and reliably determine ChE. The papers presented
in this Session discussed the recent advances in this area. Dr.
Eyer presented an improved method that, in addition of being
easily performed, gives reliable results down to 3% residual
activity. Dr. Feaster demonstrated a new and accurate method
for simultaneous determination of blood concentrations of both
AChE and BuChE. This method was successfully employed in screening
320 blood samples in one four-hour time interval. Dr. Portmann
discussed the reasons for the relatively high variations in BuChE
levels obtained in different laboratories. He discussed several
important technical improvements, such as the use of detergent
and the loss of hydrocyanic acid in the reagent mixture. Dr.
Simeon-Rudolf described a simple method for AChE and BuChE using
a colorimetric method and DTNB for thiol groups determination.
Dr. Brodbeck compared AChE and BuChE levels in blood taken from
capillary (finger) and from venous blood. He also tested the
effect of different detergent on these enzymatic activities.
The Session was an important methodological step towards a commonly
accepted methods that may be applied in practice.
Session 11 - Decontamination
Chair: Devendra Jaiswal; Co-chair: Slavomir Rump
Decontamination plays a vital role in defence against toxic chemicals
including chemical warfare agents. It is a process of removal
and conversion of the toxic chemicals into non-toxic or harmless
substances by physical, chemical or biochemical methods, depending
on the nature and type of contamination and surface on which
the decontaminant is dispersed. The research & development
in NBC defence obviously includes decontamination as an important
area of work. The Session gave a wide spectrum of information
on new developments on materials, methods and technologies for
decontamination. The decontamination of mass casualties after
exposure to harmful liquid chemicals can be quite important in
several situations particularly in an industrial accident as
was highlighted by Dr.Persson. The casualties with traumatic
injuries and exposure to toxic chemicals will create problems
with regard to transportation, decontamination and medical treatment.
The polyurethane foam linked mammalian cholinesterase for decontamination
and detection of OP nerve agents was an interesting development
reported by Dr. Gordon. It involves a bioscavenger approach to
the protection against OP toxicity. A sponge product containing
ChEs, oximes and polyurethane was developed for removal and decontamination
of OP compounds from medically important surfaces such as skin
and wounds. The bioscavengers can also act as a biosensor for
detection of OPs. Maj. Eaton discussed large-scale mass decontamination
and special event planning criteria for special international
events based on Canadian Aqueous System for Chemical, Biological
and Radiological Decontamination (CASCAD) system and Canadian
model. Recent and planned research to extend and further validate
RSDL as a broad spectrum personal decontamination system for
BCW agent decontamination had been studied by Dr. Philip O'Dell.
The Reactive Skin Decontamination Lotion (RSDL) has been fielded
in a number of countries for C agent protection of personnel.
It is also useful for BW agent decontamination and for riot agent
removal. Mr. Earl Laurie presented the useful development of
new Chemical and Biological (CB) moulded gloves with far better
tear resistance and physical properties He highlighted the general
composition of the material used in such gloves and the advantages
of the new product.
Session 12 - B-Agent Protection and Treatment
Chair: Elena Ryabchikova; Co-chair: Kim Lindsay
This session was devoted to discussion of B-agent protection
and treatment. Only a small part of this huge field may be discussed
in any meeting, and the reports presented in this Session were
related to several events. Dr. Moore presented new promising
biocidal resin Triosyn, characterized by high potential of microorganisms
destruction and simplicity and convenience in practical application.
The resin represents new approach to usage of iodine abilities
to kill various microorganisms. Dr. Ryabchikova presented current
situation of smallpox researches, and proposed plan of variola
virus studies in SRC VB "Vector" (Russia). Dr. Haghighi's
topic was related to analysis of the infections among Iranian
troops during Iran-Iraq war. Isolated agents were identified
and their biological properties were examined. Resistance of
the selected microorganisms to antibiotics was evaluated. Dr.
Russel's presentation gave an overview of plague vaccine history,
current situation and main directions for future. Plague is still
dangerous disease requiring attention of epidemiologists and
development of new modern vaccines. Dr. Melling, who presented
common principles of biodefense vaccinology, completed the Session.
Biodefense vaccines are a special kind of vaccines, however,
they should correspond to requirements for vaccines destined
for public health. The report summarized general problems of
biodefense vaccinology. Taken together, the Session paid attention
to the most significant problems of B-agent protection and treatment.
Session 13 - Poster 2: Chem Poisoning,
Chair: Keith Vesely; Co-chair: Michael McMillan
This poster session was devoted to chemical poisoning, treatment,
warning and decontamination. Topics included: antidote delivery
systems; effects of low-level inhaled sarin; development of adequate
warning systems and risk assessment procedures in and around
chemical demilitarization sites; present and future treatments
for chemical warfare agents; and examination of mycotoxin levels
in feed and animal livers. Dr. Pita explained the functioning,
manufacturing process and technical characteristics of Spanish
Armed Forces autoinjectors. New directions will include: replacing
1 mL syringes with 2 mL syringes; introducing HI-6; research
and development of combination atropine, oxime and anticonvulsant;
and possibility of formulating autoinjectors with other drugs.
Dr. Bajgar reported on a study examining the effects of sublethal
concentrations of inhaled sarin as measured by AChE activity
in rats. Results suggested that at low concentrations, AChE in
blood is "titrated" by sarin. AChE in the blood appears
to be is a very sensitive marker for diagnosis of inhalation
exposure to low concentrations of sarin. Dr. Kassa described
the long-term effects of low-level sarin inhalation in rats.
Non-convulsive symptomatic levels of sarin can cause subtle long-term
signs of neurotoxicity and immunotoxicity. These sarin-induced
long-term signs of neurotoxicity may be caused by mechanism(s)
separate from the cholinergic nervous system. Dr. Petrov described
the need to replace obsolete warning systems to properly warn
personnel at chemical warfare storage facilities, as well as
surrounding populations in Russia. Dr. Juruli evaluated the health
risk of the "old military waste" in Georgia. To prevent
damage to human health, the national legal instruments related
to chemical management must be improved. Dr. Saxena presented
work establishing a relationship between ligand-induced acceleration
of AChE reactivation and phosphoryloxime inhibition of the reactivated
enzyme. Results suggest that ChE, oximes and organophosphorus
hydrolases can work in tandem to hydrolyze or inactivate organophosphorus
compounds in vitro and in vivo. Dr. Kalantari presented a study
designed to determine the extent of contamination of mixed feed
by mycotoxin and the subsequent presence of the toxin in livers
of poultry fed the contaminated feed. Results showed aflatoxin
contamination but at levels that were safe. Dr. Ozyurt described
the role of serum ChE and the Namba classification in diagnosing
and managing organophosphorus intoxication in humans. Clinical
appearance was found to be more important than only BuChE levels
in determining when to cease antidotal therapy.
Session 14 - Poster 3: B-Agents and Terrorism
Chair: Lisa Rotz; Co-chair: Maarten Nieuwenhuizen
This poster session addressed a variety of subjects from policy
approaches to combating chemical terrorism to specific methods
for detection and early warning for chemical and biological agents.
In addition, research projects that evaluated countermeasures
for botulinum toxin B and T-2 mycotoxin were presented. Dr. Huber
described the Swiss approach to protect from chemical terrorism.
The observation was made that the problems and approaches for
responding to chemical terrorism seem to be similar for various
countries. Discussion occurred regarding the necessity of each
country developing their own solutions, or could a common response
template be developed that may then be used by several countries.
Dr. Schürch discussed research regarding the development
of a detection method for biological agents and toxins using
monoclonal antibodies. The authors used a semisynthetic method
to develop antibody libraries against F. tulariensis. The antibody
libraries created using the animal model and the semisynthetic
method were compared and appeared to be similar in their specificity
for F. tularensis antigens. Buforin I (a peptide from the stomach
of Asian frog) was found to be a competitive inhibitor of botulinum
toxin B by Dr. Gordon who evaluated several peptides that contained
the QF cleavage site. Buforin I functioned as a competitive,
dose-dependent inhibitor for botulinum B toxin activity but not
as a substrate. A method for assessing the risk of different
agrochemicals for use as a terrorist weapon utilizing an analysis
of their availability, volatility, and human and animal toxicity
was presented by Dr. Yousefi. Ratings of high, medium, or low
for each category were assigned to the agrochemicals currently
available for industry use in South Africa and this scoring system
was used to determine which chemicals present the greatest risk.
The efficacy of multiple treatment regimens utilizing corticosteroids
in rats exposed to T-2 mycotoxin was evaluated by Dr. Jovanovic.
The dose of corticosteroid but not the route of administration
produced significant differences in the protective index. A novel
strategy using synthetic nucleic acids combine with conventional
immunoassays for biological agent detection was described by
Dr. Filer. The poster presentation was a description of a Multiplexed
Detection System that would serve as a bridge between two distinct
assay technologies in an attempt to ameliorate the limits endemic
to current immuno-diagnostic systems.
Session 15 - Bioscavenger
Chair: Bhupendra Doctor; Co-chair: Yongxiang Zhang
The session addressed bioscavengers, research on the development
of stoichiometric bioscavengers such as cholinesterases (ChEs)
and catalytic bioscavengers such as OPAA and OPH was presented.
Dr. Doctor demonstrated the use of three different types of ChEs
(HuS BChE, Eq BChE, and FBS AchE) in protection against OP intoxication
in four animal species (rats, mice, guinea pigs, and rhesus monkeys).
He was followed by Dr. Broomfield, who described the kinetics
of nerve agent hydrolysis by human OPAA hydrolase and demonstrated
that despite the high Km, this enzyme can catalyze the rapid
hydrolysis of soman and sarin, and to a lesser extent VX. Dr.
Cheng described his studies on the isolation of an OPAA hydrolase
from Alteromonas, its characterization as a prolidase, and the
cloning and large-scale production of recombinant enzyme. He
also described the formulation of recombinant enzyme in different
matrices for possible use in chemical agent decontamination,
personnel protection, and detoxification. Dr. Saxena discussed
her attempt at improving the bioscavenging property of Mo AChE
by site directed mutagenesis studies and demonstrated that the
E202Q AChE was 2- to 3-fold more efficient than wild-type AChE
in detoxifying soman and sarin compared to wild-type AChE. Dr.
Way demonstrated the use of recombinant OPH encapsulated into
sterically stabilized liposomes (SL) in the presence of 2-PAM
and atropine, for the protection of mice against a 1000 LD50
of paraoxon. In similar studies, Dr. Petrikovics demonstrated
the use of recombinant OPAA encapsulated into sterically stabilized
liposomes (SL) in the presence of 2-PAM and atropine, for the
protection of mice against DFP.
Session 16 - Terror
Chair: Ladislaw Szinicz; Co-chair: Andrew Robertson
Session 16 was devoted to terrorism. Various presentations from
other Sessions (3, 7, 14)
already stressed the disastrous results that may arise from terrorist
or non-state use of chemical and biological warfare (CBW) agents.
Mitigating the impact of a CBW attack requires timely planning,
properly trained personnel, rapid detection methods, protection
and medical equipment, pharmaceuticals, dedicated facilities
and a rapid, and well trained, emergency response organization.
Any reaction to a CBW terrorist attack will be infused with high
levels of anxiety, emotion and psychological pressure. Dr. Santana
noted that intensive communication at different levels is inevitable
in any incident response and stressed the importance of the early
"hardening" of any disaster communication system. Intensive
education and training, coupled with appropriate equipment, is
also important. Col. Centonze outlined the particular CBW protection
and detection training which is provided by the Swiss Government
and emphasized the benefits of cooperation between civil and
military authorities. Dr. Lisa Rotz addressed the evaluation
and prioritization of potential biological agents against objective
criteria to ensure appropriate allocation of the limited funding
by public health authorities. By identifying priority agents,
public health programs can be targeted appropriately. Dr. Lindsey
stressed the importance of the availability of standardized laboratory
protocols during a CBW attack and outlined the approach CDC had
taken to make this information available using the internet.
Dr. Treadwell, using the Seattle World Trade Organization meeting
as an example, emphasized the need for surveillance programs
for early recognition of characteristic symptom clusters, which
may be the first indication of terrorist use of biological agents.
The daunting task of improving protection against CBW terrorist
attack will be more cost-effective with continued national and