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Perfluoroisobutene (PFIB) is a colorless toxic gas which can be absorbed into the body by inhalation. The substance irritates the respiratory tract and may be used as a potential chemical warfare agent.  Inhalation exposure may cause severe symptoms of pulmonary edema associated with wheezing, difficulty in breathing and sputum expectoration. A bluish skin color is also observed. Initially, cough and chest pain may occur. However, severe symptoms of pulmonary edema may be delayed for several hours before a rapid deterioration of health occurs. Excessive exposure may cause death.

Chiral analogous compound of methamidophos insecticide are only poorly hydrolyzed by Ca²⁺-dependent phosphotriesterases in mammals tissues including the human serum. We reported the hydrolysis of O-hexyl O-2,5-dichlorophenyl phosphoramidate (HDCP) in chicken serum. The hydrolysis of the R-(+)-HDCP isomer is strongly increased in vitro in the presence of 30-250 µM copper.  It is the opposite estereoselectivity of that showed by liver Ca²⁺-dependent activity. We name it as "antagonistic stereoselectivity". Diluted chicken serum (10 µL in 1 mL solution of 400 µM HDCP) or the equivalent amount of commercial chicken serum albumin (CSA 216 µg/mL) with 100 µM Cu²⁺, showed about 50% and 75% of R-(+)-HDCP hydrolysis after 60 and 120 min. In the same conditions other commercial serum metalloproteins with high affinity to Cu²⁺ (cuproproteins) as human serum ceruloplasmin or horse kidney metallothionein did not showed significant Cu²⁺-dependent hydrolysis. Moreover, other divalent cations (Zn²⁺, Fe²⁺, Ca²⁺, Mn²⁺ and Mg²⁺) did not showed this activation. The results confirm that the CSA is the protein responsible of "antagonistic stereoselectivity" that had been observed in the chicken serum. The effect of copper on the hydrolysis of HDCP by other animal albumins is shown in this work.

Alzheimer’s disease (AD) is a complex and progressive neurodegenerative disorder. The available therapy is limited to the symptomatic treatment and its efficacy remains unsatisfactory ^[1]. In view of the prevalence and expected increase in the incidence of AD, the development of an effective therapy is crucial for public health. Due to the multifactorial etiology of this disease, the multi-target-directed ligand (MTDL) approach is a promising method in search for new drugs for AD. Aiming at developing new MTDLs, this project consists on the development of new multifunctional agents, which will act simultaneously on the different players in AD pathology. The project aims at developing MTDLs by combining an AChE inhibitory activity with an alpha-7 nAChR activation ^[2].

Background: The occurrence of retroperitoneal bleeding and the manner of detection was retrospectively evaluated in patients with life-threatening traumatic bleeding.Methods: The cohort included patients who died in the Trauma Centre of the University Hospital Hradec Kralove in Czech Republic between 2008 and 2012 due to traumatic hemorrhagic shock. Findings of retroperitoneal bleeding and the findings found for life (i.e., CT, FAST, pre-operative findings) were compared.Results: During the five-year period, deaths due to the post-mortem diagnosis of traumatic hemorrhagic shock were recorded in 75 patients, 26 of which (35%) were verified by post-mortem autopsy to have hemodynamically significant bleeding into the retroperitoneum (HSBR) from 31 sources. HSBR was identified for life in 10 patients with HSBR (38.5%). Sensitivity was 55% in CT angiography and 36% in laparotomy without previous CT. The sensitivity of laparotomy with surgical exploration of the retroperi-toneum was 67%. A predisposing factor for hemodynamically significant bleeding into the retroperitoneum, which may escape the surgeon’s attention, is high-energy blunt trauma to the trunk.Conclusions: In the acute stage of treatment of patients with life-threatening bleeding due to high-energy blunt trauma, the surgeon has to decide whether the patient’s condition allows CT and whether hematoma of the retroperitoneum should be revised surgically. However, in the present cohort few patients with HSBR underwent surgical exploration of the retroperitoneum because the hematoma was ascribed to the known injury of the pelvis and spine.Level of evidence: Prognostic study, level III

N-Methyl-D-aspartate (NMDA) receptor belongs to the group of glutamate receptors, which are further divided into ionotropic and metabotropic. It affects synaptic plasticity and the development of neuronal synapsis in CNS. Ionotropic NMDA receptors are activated by glutamate, thereby flowing positively charged ions through the membrane along its concentration gradient. However, glutamate overload leads to excitotoxicity, due to high levels of Ca²⁺, which leads to cell death assocciated with neurodegenerative diseases. NMDA antagonists like dizocilpine reduce intracellular concentration of Ca²⁺ by modulating permeability of NMDA receptor channel. Dizocilpine act as a non-competitive NMDA receptor antagonist with anticonvulsant and anesthetic properties. Its therapeutic use in humans is limited due its numerous side effects, but it is experimentally used as an animal model of schizophrenia.

Nerve agents (NAs) belong to family of organophosphorus inhibitors (OPIs) of acetylcholinesterase (AChE) enzyme. Although the use of highly toxic OPIs as chemical warfare agents is prohibited, they have been misused several times and thus still represents an emerging threat. For these reasons, the development of novel prophylactic agents and therapeutic intervention against NAs is still up-to-date and of high importance. Prophylactic administration of antidotes not only increases body's resistance to the effects of NAa, e.g. when operating at a contaminated environment after a terrorist attack, but also potentiates the subsequent antidotal therapy after exposure. Currently, pharmacological prophylaxis can be delivered either by protection of AChE against irreversible Inhibition, administration of commonly used antidotes in advance (i.e. oxime reactivators) or by the use of bioscavengers. The aim of the recent review is to highlight the current trends in prophylaxis and outline breakthroughs in prophylaxis based on reversible cholinesterase inhibitors like huperzine A or acridine derivatives.

The acute toxicity of OPNA results from irreversible inhibition of AChE (EC 3.1.1.7), a key enzyme in neurotransmission, via the formation of a covalent P–O bond at the catalytic serine. Inhibition of AChE leads to the accumulation of acetylcholine neurotransmitter (ACh) in the synaptic cleft causing among other symptoms, seizures and respiratory arrest leading to death.  The current urgency treatment of OPNA poisoning is based on the administration of a cocktail of three components: an antimuscarinic agent (e.g. atropine), an anticonvulsant drug (e.g. diazepam) and mono or bispyridinium AChE reactivator (e.g. pralidoxime, obidoxime, trimedoxime). The high nucleophilicity of these alpha-nucleophiles allows the displacement of the phosphyl group from the catalytic serine, yielding to the restoration of AChE activity.  However, reactivation of central AChE is inefficient due to the fact that positively charged pyridiniums poorly cross the brain blood barrier (BBB). Moreover pyridinium(s) oximes exhibit a quite narrow spectrum of reactivation. Despite decades of research in this field, there are no efficient and general broad-spectrum reactivators for OP-inhibited AChE.  In this context, we have developed families of new uncharged reactivators of OP-inhibited acetylcholinesterase and/or OP-inhibited butyrylcholinesterase with the potential to cross the BBB. Three new families of uncharged reactivators display in vitro reactivation potencies towards VX-, tabun- and paraoxon-inhibited human AChE that are superior to those of the mono- and bis-pyridinium aldoximes (e.g. 2-PAM, HI-6, obidoxime, HLö-7, TMB-4) which include those currently used in the armed forces.

The interest and the increasing need for communication, exchange of experience and materials, training in diagnostics and research methods has led to the idea of creating an international scientific society focusing on tularemia. Tularemia meetings focus on fundamental, clinical, and applied research regarding any aspect of Francisella tularensis.  Research uncovers not only the biology of this fascinating bacterium, but also provides the basis for future development of treatments, preventive interventions, and disease diagnosis. Under the umbrella and with active participation of WHO, a group of scientists has met to develop this idea.

Hemothorax is a frequent clinical situation often associated with chest injury or with iatrogenic lesions. Spontaneous hemothorax is uncommon and among its cause may include coagulation disorders, pleural, pulmonary or vascular pathology. Diagnostics is based on radiography or ultrasound and thoracentesis which may be also therapeutic solution. The majority of hemothoraxes can be managed non-operatively but hemodynamic instability, the volume of evacuated blood and persisting blood loss or persisting hemothorax require surgery. A surgical approach may vary from open thoracotomy to rapidly developing minimally invasive methods - video-assisted thoracoscopic surgery (VATS) and videothoracoscopy (VTS).

Current pharmacotherapy for Alzheimer's disease (AD) involves compounds aimed at increasing the levels of acetylcholine in the brain through inhibition of AChE. These drugs, known as acetylcholinesterase inhibitors, have been shown to improve cognition and global functions but have little impact on improving the eventual progression of the disease. However, there are evidences that other cholinesterases such as butyrylcholinesterase (BuChE) can play an important role in cholinergic function in the brain, and the long-suspected non-cholinergic actions of acetylcholinesterase, mainly the interference with the beta-amyloid protein cascade, have recently driven a profound revolution in cholinesterase drug research [1-2]. We will present our journey from dual binding site AChE inhibitors as potent beta-amyloid modulators to the more recent serie of indolylpiperidines hybrids with an unexpected and very potent hBuChE inhibition. Experimental and computational studies have revealed the chameleon behavior of these molecules able to change their bioactive conformation depending on the cholinesterase binding site. Based on the potent activity of these compounds targeting BuChE, the low cellular toxicity and the in vivo target engagement, we can propose these indolylpiperidine derivatives as valuable tools for the study of the role of BuChE in AD and probably as potential drugs candidates for its future pharmacotherapy.

Alzheimer’s disease (AD) is a broadly spread neurodegenerative disorder of ageing population manifesting itself in progressing loss of cognitive functions down to total demolition of intellect and disability. Profound synaptic dysfunction contributes to early loss of short-term memory in Alzheimer’s disease. Here we show the protective effects against amyloid-induced synaptic toxicity of C-35, a potent reversible inhibitor of acetylcholinesterase (AChE). Crystal structure of the complex between human AChE and C-35 revealed tight contacts of ligand along the enzyme active site gorge. Molecular dynamics simulations indicated that the external flexible part of the ligand establishes multiple transient interactions with the enzyme peripheral anionic site. Thus, C-35 is a dual binding site inhibitor of AChE. In amyloid-transgenic mice, C-35, when administered after disease onset, reversed synapse loss, decreased the number of amyloid plaques and restored learning and memory. When administration of C-35 and the clinically relevant AChE dual inhibitor donepezil was terminated three weeks after the trial started, animals, that were receiving C-35 showed a much better ability to learn than those who received physiological saline or donepezil. Our results provide evidence that C-35 has a more pronounced Alzheimer’s disease-modifying action than donepezil.

The following thesis focuses on the issues regarding catering and food and its impact on health and life satisfaction of coalition soldiers during their deployment on a military operation in the Middle East area. Obviously, food is a part of our lifestyle and it is certainly an area affecting our life satisfaction, all the more affecting the life satisfaction of soldiers specifically in an armed conflict environment where they are exposed to higher stress conditions. Furthermore, life satisfaction is one of the variables influencing the work itself. And work influences our lifestyle and vice versa. It can be also said that life satisfaction influenced by the working environment also affects the health of soldiers. It turns out that work satisfaction has a very strong and lasting relationship with life satisfaction, as work is a central factor for most people, influencing their identity Firstly, it was necessary to find out and describe information from the area of catering of troops in field conditions and fighting food rations which can be provided to deployed soldiers. The authors’ fundamen-tal research question is how the catering and food in the military operational environment affects the life satisfaction and health of deployed soldiers. To obtain the data, a standardized method, Life satisfaction question-naire (authors: J. Fahrenberg, M. Myrtek, J. Schumacher and E. Brähler, 2001), was used. Food and catering play a very high role in our lifestyle, affects our health and furthermore, the quality of life in any situation. In life satisfaction, Danish soldiers have higher values, followed by American soldiers, and British men are the least satisfied during the foreign operation. The reasons for differences and their clarification corespondes with the last published “Happines Index” which is investigated annually by the United Nations agency.

Chitosan is low toxic, non-immunogenic and biodegradable natural biopolymer. As the deacetylated form of chitin, it is extensively used in medicine, agriculture and industry for easy production, biocompatibility and antimicrobial activity. Chitosan is also widely studied as the main structural unit of nanomaterials. Numerous studies have shown the strong antimicrobial activity of chitosan including its efficacy as an adjuvant for construction of new vaccines. In this overview, we present the basic information on chitosan nanopar-ticles, their preparation, purification, and  characterization as well as the new data on their utilization for the construction of targetted drugs based on nanotechnologies.

Human acetylcholinesterase (hAChE) and butyrylcholinesterase (hBChE) are related enzymes. hAChE plays a key role in neurotransmission and is the target of organophosphorus nerve agents. hBChE is good a natural stoichiometric scavenger of nerve agents, preventing their diffusion to the central and peripheral nervous system where they inhibit hAChE. hAChE and hBChE display different specificities for substrates and ligands due to differences in the number of aromatic residues lining the active site gorge. These aromatic residues are essential for the binding of quaternary and aromatic ligands. Some molecules containing quaternary and/or aromatic moieties form supramolecular structures by chelating Zinc. The nature of these molecules suggested that they could have affinity for the aromatic residues in the active site gorge of human cholinesterases. It was confirmed by determining their inhibition properties. A key question was whether these supramolecular ligands bind to human cholinesterases as their Zn-complex or monomeric form? The X-ray structures of two supramolecular complexes binding to the gorge of the hAChE and the hBChE reported herein showed that either cases are possible. These structural data on two new types of ligand can be used to design original cholinesterases inhibitors or reactivators.

Current symptomatic treatment has only limited clinical efficacy and minute effect on progression of Alzheimer’s disease. The research focus has thus shifted from single targets towards multifunctional ligands targeting several pathological processes of the disease [1, 2]. A potent picomolar selective inhibitor of human butyrylcholinesterase [3] was used as the starting point to develop a new series of multifunctional ligands. A focused library of derivatives was designed and synthesized that showed both butyrylcholinesterase inhibition and good antioxidant activity comparable to natural antioxidants. The crystal structure of compound 11 in complex with butyrylcholinesterase revealed the molecular basis for its low nanomolar inhibition of butyrylcholinesterase (K_i = 1.09 ±0.12 nM). In addition, compounds 8 and 11 show metal-chelating properties as determined by the UV-Vis titrations, and reduce the redox activity of chelated Cu²⁺ ions in a Cu-ascorbate redox system. Compounds 8 and 11 decrease intracellular levels of reactive oxygen species, and are not substrates of the active efflux transport system, as determined in Caco2 cells. Compound 11 also protects neuroblastoma SH-SY5Y cells from toxic Aβ_1–42 species. These data indicate that compounds 8 and 11 are promising multifunctional lead ligands for treatment of Alzheimer’s disease.

Currently, the best bioscavenger candidate against nerve agent intoxication is human butyrylcholinesterase (BChE). However, the effective dose cost, estimated to about 200 milligrams of pure enzyme, remains challenging despite the production and purification progresses realized these last years. A strategy for reducing dosage and cost would be to turn this scavenging protein into a nerve agent hydrolyzing enzyme, a catalytic bioscavenger. Up to now, screening of large mutant libraries has been hindered by the restricted eukaryotic expression of active BChE. Here we present the successful prokaryotic expression of an active human BChE variant designed with PROSS, a sequence- and structure-based algorithm for the soluble prokaryotic expression of difficult proteins. The protein is easily purified with two simple chromatographic steps. Despite 47 point mutations, the enzyme presents similar enzymatic parameters than the wild-type enzyme and its active site gorge structure is identical to that of the native enzyme produced in eukaryotic systems as determined by X-ray crystallography. These data validate the prokaryotic expression of human BChE which will greatly facilitate the screening of variants with nerve agent hydrolytic properties. We have initiated animal studies to assess the protein potency (immunogenicity, pharmacokinetic and bioscavenger efficiency) and will study the production of the tetramer form. On the other hand, we are currently developing high-throughput protocols for the prokaryotic expression, purification and screening of nerve agent hydrolysis.

The best-known function of acetylcholinesterase (AChE) is the hydrolysis of the neurotransmitter acetylcholine, however we are increasingly aware of the multifunctionality of this enzyme [1]. The non-hydrolytic functions of AChE are driven by allosteric sites as the peripheral allosteric site (PAS) responsible for amyloidosis in Alzheimer’s disease through interaction with β-amyloid peptide. We would like to show our work about the identification and characterization of new allosteric sites in AChE, using computational tools. This study has allowed us to identify allosteric inhibitors by virtual screening using our in-house MBC chemical library [2] guided by structure-based and fragment hotspot strategies. The identified compounds were also screened for in vitro inhibition of AChE and three of them were observed to be active. Further experimental (kinetic) and computational (molecular dynamics) studies have been performed to verify the allosteric activity. Thus, new compounds have been developed as allosteric modulators that may be valuable pharmacological tools in the study of non-cholinergic functions of AChE.

Within the alpha/beta hydrolase fold superfamily of proteins, the COesterase group (carboxylesterase type B, block C, cholinesterases…) diverged from the other groups through addition of an N-terminal disulfide bond and simultaneous increase in the mean size of the protein (1). This disulfide bond creates a large loop, which is essential for the high catalytic activity of cholinesterases through formation of the upper part of the active center gorge. In some non-catalytic members of the family, the loop may be necessary for heterologous partner recognition. The shuffling of this portion of protein occurred at the time of emergence of the fungi/metazoan lineage. Homologous proteins with this N-terminal disulfide bond are absent in plants but they are found in a limited number of bacterial genomes. In prokaryotes, the genes coding for such homologous enzymes may have been acquired by horizontal transfer. However the cysteines of the first disulfide bond are often lost in bacteria. Natural expression in bacteria of CO-esterases comprising this disulfide bond may have required compensatory mutations or expression of new chaperones. This disulfide bond may also challenge expression of the eukaryote-specific cholinesterases in E. coli. Recently , catalytically active human acetylcholinesterase and butyrylcholinesterase were successfully expressed in E. coli. The key was the use of a peptidic sequence optimized through the Protein Repair One Stop Shop process, an automated structure- and sequence-based algorithm toward expression of properly folded, soluble eukaryotic proteins with an enhanced stability (2,3). Surprisingly however, the crystal structure of the optimized butyrylcholinesterase variant expressed from bacteria revealed co-existing ‘close’ and ‘open’ states of the first disulfide bond. Whether the ‘open bond’ involves two cysteines (i.e., the bond never formed) or two half-cystines (i.e., the bond properly formed, then broke during the production/analysis process) cannot be inferred from the structural data. Yet, this observation suggests that this first bond is difficult to maintain in E. coli-expressed cholinesterases.

It was with interest and pleasure that I read the contribution of Petronilho & Figueroa-Villaret in the MMSL reviewing the literature on agents for defense against chemical warfare [Petronilho & Figueroa-Villaret, 2015]. The authors briefly touch on the history of organophosphates emphasizing the pioneering contribution of Jean Louis Lassaigne, the synthesis of triethyl- phosphate (TEP) and finally the achievements of Philippe de Clermont who codeveloped the first organophosphate (OP) acetylcholinesterase inhibitor, tetraethyl pyrophosphate (TEEP). They continue by pointing out that Wilson and Ginsburg managed to reactivate OP-inhibited acetylcholinesterase using pralidoxime (2 -PAM), which reactivates the enzyme much faster than hydroxylamine. I believe that the scientists involved in organophosphorus cholinesterase inhibitor & reactivator development deserve more attention and that the colleagues' contribution contains a number of ambiguities deserving additional...

Freshwater planarians from Platyhelminthes, harboring a mammal-like cholinergic nervous system, have emerged as a promising in vivo model for investigating neurotoxicity. Moreover a large proportion of stem cells provide planarian an unconventional capacity of regeneration allowing for developmental disruption studies. Schmidtea mediterranea (Smed) was used as model for organophosphorus (OP) poisoning and for evaluating the efficacy of detoxifying enzymes. Acetylcholinesterase and butyrylcholinesterase from planarian (Smed-AChE and Smed-BChE) share 35% identity with their human counterpart (Hs-AChE and Hs-BChE). Structural predictions revealed strong similarities between planarian and human enzymes. Cholinesterase activities were detected in crude planarian homogenates after grinding and were inhibited after organophosphorus exposition. In situ Hybridization was further used to localize cholinesterases in planarians and showed two different patterns, Smed-AChE being mainly detected in cephalic ganglion and ventral nerve cords while Smed-BChE distribution was diffuse. Survival, behavior and regeneration were analyzed in whole planarian exposed to four OP [1]. The toxicity of OP degradation products generated by enzymatic hydrolysis with the robust phosphotriesterase enzyme SsoPox, from the archea Sulfolobus solfataricus [2], was further evaluated. OP were found to be highly toxic to planarians causing severe mortality and behavior disruption at sublethal concentrations as well as growth disruption during regeneration after cutting. Enzymatic decontamination drastically reduced toxicity and enhanced both mobility and development. These results underline that degradation products have a lower impact than initial organophosphorus substrates. A biotechnological application based on a filtration column incorporating detoxifying enzymes was developed to decontaminate wastewater with planarian as biosensor.

Vaccination protects the community as a whole from serious communicable diseases, such as polio, pertussis, diphteria, measles and others. The incidence of these diseases has been successfully reduced in the last decades. Switzerland with its National vaccination strategy takes part in an international effort on public health described in WHO’s 2011-2020 Global Vaccine Action Plan (1), and in the European 2015 - 2020 Vaccine Action Plan (2). However, in contrast to the Czech Republic, Switzerland doesn´t have a compulsory global vaccination schedule. It only presents the vaccination recommendations. Vaccination rates are not always high enough in Switzerland and vary from one region to another.

Alzheimer´s disease (AD) is a progressive and degenerative neurological disorder resulting in memory loss and cognitive decline. The severity of AD dementia was found to correlate with the extent of the cholinergic loss and acetylcholine (ACh) depletion. In brain synapses ACh can be hydrolyzed by two cholinesterases (ChEs), acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), which were found in neurons and glial cells as well as in AD neuritic plaques and tangles. AChE is the prevalent enzyme in the healthy brain, while BChE is considered to play a minor role in the regulation of synaptic ACh levels. However, in AD advanced stages, AChE activity is decreased while BChE activity is unchanged or even increased, making both ChEs stimulating targets for the treatment of AD.  Current AD therapy is based on AChE inhibitors, although they have very modest clinical effects in treating the symptoms of the disease and are unable to halt disease progression. Oxidative stress (OS) and mitochondrial dysfunction are also considered critical factors in AD pathogenesis. As a result, targeting mitochondrial oxidative stress (OS) in the prodromal phase of AD to slow or prevent the neurodegenerative process and restore neuronal function is thus viewed as a valid therapeutic approach. As part of our drug discovery program focused in oxidative stress-related diseases, and following a multi-target strategy, new mitochondriotropic antioxidants based on natural scaffolds acting as dual and bifunctional cholinesterase inhibitors have been developed. The results will be reported in this communication.

Drug discovery and development is a complex and expensive process. Thanks to the exponential growth of molecular data and advancement in technologies, efforts have been tremendously amplified. Among new approaches multipotent compounds are emerging as the next paradigm in drug discovery [1] and includes: (i) single drug acting on multiple targets of a unique disease pathway, or (ii) single drug acting on multiple targets pertaining to multiple disease pathways. These compounds are thought to have best beneficial effects in the treatment of complex diseases, like Alzheimer’s Disease, in which the simultaneous regulation of various pathological aspects may more efficiently interfere with the disease progression. Systematic integration of the data derived from different disciplines including computational modeling, X-ray crystallography, synthetic chemistry, in vitro / in vivo pharmacological tests, is mandatory for the selection of best-in-class compounds. In this context, we report on the key contribution of X-ray crystallography in highlighting peculiar mode of interaction of promising multi-target directed ligands, designed by combining the tacrine fragment to distinct pharmacophores i.e. juglone [2], benzofuran [3] and       L-tryptophan with a linker of a suitable length. Overall, the structural analysis highlights the molecular determinants responsible for the optimal binding of the multi-target ligands to AChE and pinpoints the utility of hybridization strategies in structure-based drug design programs. It also unveils the validity of X-ray crystallographic structures determination at certain milestones along the development of interacting inhibitory drugs based on molecular modeling studies.

Autoimmune diseases are caused by reaction of immune system against own body´s healthy structures. Diagnostics of these states require collaboration between clinical and laboratory diagnostic part. This text is focusing on diagnostics selected systemic and organ specific autoimmune diseases. It does not aim for detail description of selected diseases rather summarizing the main clues leading to right diagnostics. Also suggesting laboratory methods used in diagnostics and summarizing their benefits and drawbacks.

Borrelia afzelii is one of potential etiological agents of Lyme disease in the Czech Republic. An important source of information about this microoganism is proteome analysis. It allows to understand pathogenesis of Lyme disease. The proteins were identified by 2-dimensional electrophoresis and mass spectrometry (MALDI-TOF). Overall 34 protein spots of Borrelia afzelii (strain CB43) were chosen for identification. This process was done successfully in case of 23 proteins. This number was represented by 11 different proteins. We identified some variants of OspC protein, several enzymes, protein of chemotaxis CheY and Membrane associated protein p66 precursor.

The 13^th Host Pathogen Interaction Forum was held in Slavonice, Czech Republic, between April 16 and April 19, 2018. The Forum is traditionally organized by the Faculty of Military Health Sciences, Hradec Králové, Czech Republic, in collaboration with the Czech Immunological Society in Prague, Czech Republic. The venues for our Forums are selected with respect to historically significant areas of the Czech Republic. The Forum’s reputation has grown steadily over the years such that it has earned a place on the calendars of important scientists in the field of studies on pathogen interactions with their hosts. Due to the scientific orientations of the organizers, the forums bring together scientists working mostly with intracellular (bacterial as well as viral) pathogens.

Notwithstanding clinical effectiveness evidences continue to suggest benefit from the acetylcholinesterase inhibitors (AChEIs) in alleviating Alzheimer’s disease (AD) symptoms, these drugs do not appear to delay or prevent the underlying neurodegeneration. In this context, novel prospects are offered by the strategy of developing single chemical entities able to modulate multiple targets, i.e. the multi-target-directed ligands (MTDLs). On this basis, several multifunctional AChEIs have been rationally designed with the deliberate aim of enlarging their biological profiles, beyond the ability to inhibit cholinesterases. This is because it has been recognized that a balanced simultaneous modulation of multiple targets critically intertwined in AD pathological cascade can provide a superior therapeutic and toxicological profile compared to the action of a selective AChEI.[1] Building on this founding principle, we and others have developed several series of anti-AD MTDL compounds that combine cholinesterase inhibition with anti-aggregating, anti-oxidant, and anti-neuroinflammatory properties.[2] As a further step, to explore the possibility to discover new MTDLs based on inexpensive resources, we have developed a series of MTDLs obtained by properly modifying constituents from the cashew nut shell liquid (CNSL), a waste from cashew nut processing factories.[3] Such hybrid compounds, obtained from renewable and inexpensive material, might be promising bio-based, sustainable MTDLs for AD drug discovery. Working in the field for almost 20 years, we should draw lessons from the past and try our best to chart innovative directions and hopefully address the scientific and societal challenges of neurodegenerative diseases.

Fecal microbiota therapy is going through its renaissance period. Even in ancient China, stool and its derivats were used for therapy of various diseases. Now thanks to new molecular methods a new knowledge about the intestinal microbiome and its interference with the human physiology, this method can be used for concrete therapy of disease.

Together with the protection of the sick and wounded in armed conflicts, rules of international humanitarian law (IHL) provide protection to medical facilities and personnel. Despite universal acknowledgement of these rules, their violations occur regularly. Causes of these violations are numerous and complex, however, in order to be respected, IHL rules need to be in the first place implemented within domestic legislations of individual states. Specific measures to be taken are to a certain extent left up to discretion of every state concerned. This article examines existing legislation and measures adopted in this context by the national authorities of the Czech Republic with respect to the Czech Armed Forces. A particular attention is directed at internal regulations of the Armed Forces against the wider background of Czech legal order. The article also touches upon areas of military training and criminal liability.

The chemical warfare agents and neurotoxic agents are an important threat to people all over the world, and require special attention because they are highly dangerous. Most of these agents are neurotoxic organophosphorus compounds (OP), which inhibit the enzyme acetylcholinesterase (AChE), which is responsible for controlling the transmission of nerve impulses. To be inhibited by these compounds, AChE can sometimes be reactivated using cationic oximes, which are the most used substances for this reactivation. Until today there have not been discovered agents for complete treatment of poisoning by all OPs. For this reason, the treatment of intoxicated people requires the determination of the absorbed OP, in order to select the appropriate activator, a process that usually requires long time and may cause death. Therefore, this study aims to do a review on the OPs used as chemical warfare agents and the process of inhibition and reactivation of AChE, especially to motivate the development of new agents for defense against chemical weapons, a process that is very important for protecting all humanity.