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Journal of NBC Protection Corps

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Vol 3, No 2 (2019)
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ISSUES OF COMPLIANCE WITH CHEMICAL AND BIOLOGICAL WEAPONS CONVENTIONS

103-110 141
Abstract

Organophosphorus chemical agents are included in the 1st List of the Annex on Chemicals of the Convention on the Prohibition of the Development, Production, Stockpiling and Use of Chemical Weapons and on Their Destruction (Chemical Weapons Convention, CWC). For the purposes of verification of compliance with the provisions of the CWC, special methods, which are considered the most informative at determining the retrospective effects of organophosphorus toxicants on the body, are necessary. Typical long-lived biomarkers of organophosphate toxic agents are tyrosine phosphorylation products, the presence of which in biomedical samples clearly indicates the exposure to sarin, soman, tabun and V-series agents. We have elaborated methods for the synthesis and isolation of tyrosine adducts derivatives of methylphosphonic and phosphoric acids, used as reference samples. The synthesis scheme included the consecutive protection of carboxyl and amino groups of tyrosine, its O-phosphorylation by the corresponding alkylphosphonates and phosphates, the removal of protective groups with the release of corresponding O-phosphorylated tyrosine adducts. Their purification from impurities was carried out, using column chromatography (SiO2, eluent: dichloromethane/ethyl acetate 1:1). The purity of the obtained products was more than 90 %, so it was possible to involve them in further transformations with the use of catalyst without the threat of its «poisoning». Benzyl and carboxybenzyl protection of phosphorylated L-tyrosines (12–17) was removed by means of catalytic hydrogenation by molecular hydrogen under atmospheric pressure. Target adducts of phosphorylated reagents and L-tyrosin were obtained (63–82 %) in form of crystal white substances, readily soluble in water and ethanol, and poorly – in dichloromethane and acetonitrile.

CHEMICAL SECURITY AND PROTECTION AGAINST CHEMICAL TERRORISM

111-116 151
Abstract

The advanced biocatalysts based on hexahistidine-tagged organophosphorus hydrolase (His6-OPH) were recently developed for the detoxification of various organophosphorus compounds and degradation of N-acyl homoserine lactones. Due to enzyme immobilization, some of obtained biocatalysts are quite stable, easy to use and very effective/active (e.g. tens of millions of substrate solution volumes appeared to be treated with column cartridges containing immobilized His6-OPH). Recently, the possible bioengineering of different stabilized nanocomplexes of His6-OPH due to its non-covalent binding with different compounds (polymers, antioxidants, antimicrobials, etc.) was demonstrated. Firstly, it was realized by computer modeling via molecular docking. Polymers of amino acids (polyglutamic and polyasparctic acids) were established to be the most effective stabilizers of the enzyme that enabled effective preservation of the enzyme activity. Up to 100 %-retention of initial catalytic characteristics of the enzyme was reached in obtained enzymatic complexes. Such nanobiocatalysts were stabilized against inactivating effects of solvents, temperatures and were able to circulate in vivo for at least 25 hours. It appeared that different antioxidants can be applied as partners of the enzyme in the nanocomplexing. Thus, a new set of original enzymatic antidotes were developed possessing dual action: both hydrolytic activity against organophosphorus neurotoxins and improved antioxidant activity. Additionally, it was shown that different organophosphorus compounds and N-acyl homoserine lactones could be molecularly docked directly to the active centers of His6-OPH dimer, thus allowing to theoretically clarify some new prospective substrates for the enzymatic hydrolysis. It appeared that new type of nanocomplexes of the enzyme with antibiotics also can be prepared. In this case the combination of antibiotics with enzyme quenching the quorum of the pathogenic gram-negative bacteria was performed. The enzyme being stabilized by the various antibiotics (especially those containing β-lactame ring) played the role of a carrier for the antimicrobial compounds significantly improving their efficiency of the action. Such biocatalysts and/or method of their design have a great potential and can be very useful for both chemical and biological defense.

Biological Security and Protection against Biological Threats

117-136 510
Abstract

Sea anemones are well-spread everywhere in the World Ocean and represent the most ancient active poisonous organisms. Their main instrument of attack on other animals are the nematocysts – stinging organelles with the curtailed hollow thread with poisonous edge on the end. In order to attract their potential victims, they use fluorescent proteins. These proteins became a separate object of research as genetically coded markers for the observation of activity of promotors of genes. The poisonous secret of sea anemones is characterized by the presence of maximum number of peptides of various structural classes and spatial structures among the studied land and marine organisms (bees, spiders, scorpions, snakes ect.). This fact complicates the identification of sea anemones' secret and its differentiation from poisons of animals of other taxons, if the concrete source of its origin is unknown. The toxicity of some biologically active sea anemone peptides (RpI, RpIII) at intravenous administration to experimental animals is comparable with that of the most well-known and dangerous representatives of natural toxins with the similar mechanism of action (an alpha-hemolysine and tetrodotoxin), or chemical warfare agents, such as sarin and hydrogen cyanide. Based on their toxic effect, the biologically active sea anemone peptides generally can be classified as neurotoxins due to their impact on the functioning of sodium channels in the cells of the nervous system of animals. Cardiotoxic effect of sea anemone secret is caused by the specificity of interaction between its separate neurotoxins and one of the sub-types of sodium channels of muscle cells, characteristic for heart tissues. The main ways of identification of sea anemone neurotoxins in samples (for example, during the investigation of biological crimes) can be sequence by Edman`s method or tandem mass spectrometry (the analysis of fragments of toxin molecule for the establishment of its structure). Further study on the mechanisms of interaction between the sea anemone neurotoxins and the ion channels of the cells of nervous and muscular systems may result in the creation of medicines for treatment of channelopathy, as well as pluripotential antidotes, blocking the toxins, that influence on sodium channels.

137-149 423
Abstract

The researchers of the Branch Office of the Federal State Budgetary Establishment «48 Central Scientific Research Institute» of the Ministry of Defence of the Russian Federation (Kirov) organized theoretical and experimental studies on the introduction of a tangential filtration method for the separation of biological mixtures into the production of immunobiological preparations. The method of microfiltration in the tangential stream replaced the process of sedimentation at the stage of concoction of intermediate vaccines, reduced considerably the process time, and allowed to obtain suspensions from cultural liquid, substandard on an indicator of concentration of microbial cells. Along with this, the microfiltration method allowed to concentrate the cultures of Yersinia pestis of a vaccinal strain EV. In comparison with the centrifugal separation, the concentration of living microbial cells of a vaccinal strain of EV Y. pestis increased by one and a half times. The filtration in a tangential stream at the ASF-020 installation from the point of view of the production of the sporous product of anthrax vaccine STI-1 (in millions of doses), is 1.8 times more effective in comparison with the centrifugal separation. The membrane method allowed to reduce the duration of technological process. These membrane processes are used nowadays during the production of plague and anthrax vaccines, anthrax immunoglobulin, diagnostic medicines and during the sterilization of liquid nutrient mediums. This type of equipment for the sterilization of nutrient mediums can be considered as an alternative to the processes of the thermal sterilization of liquids and provides their biological and technological full functionality. Experiments on the use of the ceramic-metal filters sterilizing the air given for aeration, showed the decrease in duration of preparatory operations on 20 h and increase in the general operational opportunities of the system.

CHEMICAL AND BIOLOGICAL WEAPONS IN WARS AND CONFLICTS

150-174 905
Abstract

The Iran-Iraq war (1980–1988) was the result of the geopolitical situation in the Middle East after the Islamic revolution in Iran in 1979. Certain longstanding territorial disputes and the absence of mutually recognized state border between the rivalry countries were among the direct pretexts of the war. At the same time neither Iraq, nor Iran were ready to serious war, both did not want it in such scales, and they did not possess chemical weapons (CW). During the war, Iraq enjoyed broad international support. At the same time, revolutionary Iran turned into a pariah state. By 1983, Iraq began to suffer a defeat from Iran, which possessed considerable human resources. Because of that certain Western countries helped Iraq (on the paid basis) to start its own CW program and the industrial production of chemical agents and munitions. Gradually CW became an integral part of the offensive and defensive operations, planned by the Iraqi command. Due to the technologies, equipment and chemicals, supplied by certain foreign, mainly Western firms, Iraq was able to start the industrial production of mustard gas, tabun and sarin/cyclosarin, as well as to start the synthesis of VX. During the war, CW turned from purely defensive into offensive means of warfare. The war ended as chemical. In 1988 all the operations, which led to the end of the war, were carried out by Iraq with the use of CW. At the same time, the war revealed certain weaknesses of chemical protection means, possessed by both sides. Thus, it appeared to be impossible to sort effectively the wounded and affected by CW during the medical evacuation phase. The existing decontamination means turned out to be ineffective in case of mass arrival of the affected into hospitals. Because of that, the secondary contamination of medical stuff took place even in Western hospitals. The protective equipment against blister agents, used by NATO countries, turned out to be insufficient in case of use of «dry yperite» by the Iraqis. The accepted schemes of treatment of the affected by tabun also showed their inefficiency. As we find out, the experience of the Iran-Iraq chemical war is studied actively in the West up to now.

HISTORICAL ARCHIVE

175-193 532
Abstract

Organophosphorus compounds occupy a unique positon among all chemical warfare agents (CWA's). Since the 1930-s their high toxicity, wide range of physical-chemical properties and complex action attracted close attention of foreign military experts. In 1936 a German chemist, Dr. Gerhard Schrader, synthesized O-ethyl-dimethyl amidocyanophosphate, known as tabun, for the first time. By the beginning of World War II, more than two thousand new organophosphorus and phosphorus containing compounds were synthesized by his laboratory's stuff. Some of these compounds were selected for further study as CW agents and subsequently were adopted as weapons by the German army. In 1938 the same Gerhard Schrader have synthesized the organophosphorus compound, closed to tabun, but more toxic: О-isopropyl methyl fluorophosphate, called sarin. In 1944 the German chemist, the 1938 Nobel laureate in chemistry Richard Kuhn synthesized soman and revealed the damaging effect of organophosphorus CWA's. In 1941 the British chemist Bernard Saunders synthesized diisopropyl fluorophosphate. During World War II the industrial production of organophosphorus CWA's was organized in Germany, Great Britain and in the USA. Germany produced tabun, sarin and soman, the western allies: diisopropyl fluorophosphate. Till the end of World War II the leadership in the sphere of the development of nerve agents belonged to Nazi Germany. After the end of the war the German scientists, many of whom were devoted Nazis, continued their work under the auspices of military departments of the USA and Great Britain. Subsequently phosphorylated thiocholine esters: V-series substances (VG, VM, VR, VX, EA 3148, EA3317 agents etc.) were synthesized with their participation. The wide range of organophosphorus compounds was tested on volunteers in Porton Down (Great Britain) and in the Edgewood arsenal (USA). But after the synthesis of V-series agents the work on organophosphorus CWA's did not stop. In recent years there appeared the tendency of the transformation of real threats connected with the chemical weapons use, to propaganda sphere. The provocation which the «Novichok» agent, arranged primitively by the British intelligence, is the perfect example of such a transformation. But it does not mean that the research in the sphere of new organophosphorus CWA's in the West is stopped.

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ISSN 2587-5728 (Print)
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