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The aim of the study is to investigate whether the mental stress resulting from the assignment of military professionals affects the parameters of metabolic syndrome (MetS) as a significant risk factor for cardio-vascular diseases.The study retrospectively analyses the data obtained during extended annual preventive examinations of professional soldiers in 2012-2016. The dataset was divided into two cohorts. The first "performance" cohort included soldiers from combat units and paratroopers (aiborne). The second cohort consisted of "others", i.e. soldiers from non-combat units or airborne. The psychological burden was determined by the soldiers' assignment to a combat or airborne units.In the years 2012-2016, almost all compared parameters were better in the "performance" group. Exceptions were a higher prevalence of overweight (according to body mass index) and sporadically higher glycaemia and alanine aminotrasferase (ALT) levels. When comparing years 2012 and 2016 of the “performace” group, statistically significant better values prevailed in relation to the MetS in 2016. The exceptions were ALT and uric acid levels.The outcome of this study suggests a better status of MetS risk factors and MetS-related factors in the "performance" cohort, and psychological stress did not augment MetS manifestations in these professionals. This study has confirmed the necessity of waist circumference measurement to eliminate overestimation of obesity in individuals with developed musculature. To confirm the hepatic origin of ALT, the medical history should include an inquiry about exercise habits. Subsequently, the effect of chronic stress may also be considered for higher liver enzymes. Screening for hypercholesterolaemia and hyperuricaemia has important clinical relevance for the prevention of cardiovascular diseases in younger population, particularly in those aged

Alcoholism (Alcohol dependence) is considered a serial health problem because of tendency for con-sumption more alcohol, control losing and the physical dependence development. This cycle in most often featured by periods of craving, abstinence, and relapse. The cholinergic, adrenergic, dopaminergic, glutamatergic, GABAergic, serotonergic, peptidergic, and different neurotransmitter systems in the brain are affected by alcohol consumption. The development of alcohol dependence is attributed to neuro-adaptations within the extended amygdala and mesocorticolimbic systems. Principally, the glutamatergic neurotransmission variations that resulted by alcohol consumption, lead to researches recommended in a focus on normalization of glutamatergic neurotransmission and glutamatergic receptors as a targeting therapy. Glutamatergic receptors are classified to ionotropic glutamate receptors (AMPA, Kainate, and NMDA) and metabotropic receptors. Sodium dependent excitatory amino acid transporters (EAATs) and vesicular glutamate transporters (VGLUTs). Additionally, cysteine-glutamate antiporter which regulates cysteine -glutamate exchange at the synapse. At least 90% of extracellular glutamate displace particularly by GLT1. Ceftriaxone, GPI-1046 and MS-153, upregulate expression of glutamate transporter 1 (GLT1) in mesocorticolimbic brain region.

A representative survey was conducted among personnel of the Czech Armed Forces to assess various aspects of perceived work and service conditions. ....

The Military Health Service of the Czech Army can support the Integrated Rescue System in the Czech Republic in dealing with emergency and crisis in cases where the basic components cannot ensure the execution of rescue work and clean-up operation by their own forces. The aim of the article is to evaluate the real possibilities of deploying the forces and resources of the Military Health Service to support the integrated rescue system from the point of view of valid legal and internal regulations and to analyze their availability. It was found that the fundamental conditions for the support of the Integrated Rescue System are regulated in the current legal regulations, mainly for the forces and resources of the military health service, which have the character of the other components of the Integrated Rescue System. Dedicated forces and resources of the Military Health Service have a set time of preparation for deployment of 5 - 72 hours. For the use of forces and means within the framework of other assistance, the essential prerequisites for their deployment are established, but no requirements are determined for the capabilities or availability of these forces.

Rheumatoid arthritis (RA) is a chronic multisystem illness that affects millions of individuals. The primary goal of RA therapy is to improve patients' quality of life by reducing pain severity, preserving or improving functional capability, and decreasing disability. The significance of resistin in the pathophysiology of RA has been explored in recent years, although its role is unclear as it is largely produced by macrophages. In this review, we have analyzed 10 studies from the PubMed site that demonstrate a relationship between resistin levels and the severity of RA.

Background: The second-leading source of neoplasm-related death and a primary factor in gastrointestinal cancer, colorectal cancer (CCR) affects both genders globally. Poor eating behaviours, tobacco, an intestinal inflammatory disorder, swellings, inherited characteristics, and the elderly all increase the threat of acquiring this malignancy. The illness is more hostile in patients detected at earlier ages, although 90% of patients with colorectal tumours are older than 50, with a median oldness of 64 years. American Cancer Association estimates that it caused more than 49,700 fatalities in 2015.Objectives: Study the correlation of midkine with carcinoembryonic antigen (CEA), liver function tests, and white blood cell count in patients with colorectal carcinoma.Methods: The serum midkine and CEA of all subjects were measured by the ELISA technique, Liver enzymes were measured by colourimetric methods and neutrophils, and lymphocytes were measured by an Electrical Impedance Cell Counting method (automated machine).Conclusion: The study results of the correlation between serum midkine and other parameters in colorectal carcinoma patients show a significant positive correlation of midkine with CEA, liver enzyme, neutrophils, and lymphocytes.

Introduction: Olive (Olea uropeae) is a traditional plant containing oleuropein and hydroxytyrosol, which are useful and used empirically for treating diabetes mellitus.Objective: To review the potential of oleuropein and hydroxytyrosol as an evidence base for diabetes potential treatment and safety.Methods: This chapter summarizes several studies available on Pubmed and Google Scholar regarding the characteristic method and extraction method as well as the effectiveness and toxicity of oleuropein and hydroxytyrosol in vitro and in vivo.Result: Oleuropein and hydroxytyrosol are effective antihyperglycemics for treating T2D. They can reduce body weight, basal glycemia, and insulin resistance by stopping the liver from making glucose and stopping the body from absorbing glucose. Several studies have shown that both isolates can control glycemic levels equivalent to free fatty acids and are safe to use.Conclusion: Oleuropein and hydroxytyrosol are extracted by several methods and can be used as potential anti-diabetics with obesity risk factors. Evidence shows that both isolates are safe for both acute and chronic use.

Background: Low back pain (LBP) is one of the common musckloskeletal diseases and usual treated by epidural steroid injection (ESI). ESIs improve patients' quality of life, reduce lumbar radicular pain, and postpone spinal surgery. The mechanism of improvement  is yet unscertain, perhaps involve type α collagen (COL2α) for bone maintenance, hence, we sought to investigate the role of injected steroids in bone healing focusing on the role of COL2α.Methods: All patients in this research were diagnosed by specialists based on their histories and clinical features and associated diseaeses or compiling therapy. Serum samples collected from LBP patients and control group for comparisons.Results: The present study found a significant (<0.0001) increase in the concentration of COL2α in patients with LBP after injection with ESI treatment compared with patients before injection and healthy individuals.Conclusion: ESI helps LBP sufferers by boosting COL2α, which repairs damaged tissues.

This topic is probably the most important in the field of military healthcare in the long term, because care of this nature is the very reason why the whole sector was created. Effective forwarded surgical care is a crucial factor in reducing preventable battlefield casualties, known and developed since the historical beginnings of military healthcare. This highly demanding medical discipline is an absolutely essential component of the modern war effort, in which it takes on a whole new dimension thanks to unimaginable technological, medical and military-tactical progress. The changed character of the world's main battlefields seems to be crucial at present, with the return of conventional warfare accompanied by attacks on health workers. We stand before the next stage of challenges related primarily to the mobility and tactical capabilities of professionals providing this specific type of care. In the text, we have come out of the historical context that led to the evolution of this care into today's form, which we have analyzed in detail on the sample of the most advanced medical support of armies in the world. This example shows us the breadth and variability of these elements with regard to the various eventualities of their use. On this basis, we present the concept of systems providing this kind of care from both the global and the Czech Republic's point of view. At the same time, for understandable reasons, we pay close attention to the specifics of this type of care in Czech conditions.

Extreme physical exercise is potentially harmful or even lethal activity. It is associated with several compensatory mechanisms focused on increasing functional capacity. Hormonal and metabolic pathways rearrange to maintain glucose supply in working tissue. Cardiovascular system substantially increases cardiac output and redistributes blood flow in working muscles. Respiratory system increases tidal volume and respiratory rate. Immune system and other systems are also affected.If capacity of these mechanisms is exceeded, organism can be severely damaged. Frequently observed disturbances include hypoglycemia, mineral level disorders, bronchoconstriction, immune system impairment etc. More severe disturbances include arrhythmias, myocardial stunning, hypoxemia, rhabdomyolysis and many others.Early diagnosis and treatment of these disturbances is essential for minimizing the health impairment. Unfortunately, in military settings are diagnostic and therapeutic options often limited. Moreover, physiological or benign processes can mimic life threatening conditions. Without complex understanding of participating pathophysiological mechanisms an interpretation of clinical findings might be confusing and treatment inefficient or even deleterious.

Purpose: Among other reasons, the deteriorating global security situation and dangers associated with nuclear weapons have increased the need for deeper knowledge of the basic mechanisms involving the human immune system and ionizing radiation (IR). We conducted a review as to the effects of IR on thymic tissue, and particularly on the development of thymocytes and the T lymphocytes population in peripheral blood.Existing knowledge on this topic is based in part on national registers that store records concerning irradiated people. The majority of studies in this area, however, are based on experimental animal models. The main open question in this subject area regards the delayed effects of IR on thymus tissue, development of thymocytes, and subsequent impact on the immune system. Findings acquired to date on effects of IR are contributing to emerging fields such as immunotherapy, the objective of which is to support or activate natural immunity response.Methods: Recent research articles were reviewed regarding the influence of IR on thymus tissue and thymocytes development.Results: Differentiation and proliferation of thymocytes constitute a complex and sensitive process that is partially altered after irradiation, as are, too, the mechanisms for movement of early (derived from bone marrow) and derived (thymus derivatives) precursors. Disruption of these processes may lead to alteration of immune system function.Conclusions: Low doses (<200 mGy) may lead to changes in or disruption of functions of the thymus, thymocytes, and mechanisms of the immune system. The extent of IR’s influence is dependent not only on the individual’s radiosensitivity but also on his or her sex and age. With increasing absorbed IR dose, the risk of damage to thymus tissue and thymocytes in the organism rises and the extent of damage increases.

Cynomorium coccineum and Cynomorium songaricum are unusual parasitic plants in the family Cynomoriaceae characterized by their unusual appearance and way of life. Due to their special habit and extremely rare occurrence, in the past, various magical properties have been attributed to these plants. The plant Cynomorium coccineum, which is found in the Mediterranean, has been used in Europe since the Middle Ages in folk medicine under the name fungus maletensis (maltese fungus). It was used for the same purpose in Arabic cultures under the name tarthuth. The Asian species (Cynomorium songaricum), sometimes considered a subspecies of C. coccineum, is still used in traditional Chinese medicine under the name Suo Yang. Recent studies show that the plant has a number of beneficial effects on the body and there is a real possibility that the substances present in Suo Yang or substances derived therefrom will be used in the future for therapeutic purposes. Of particular interest are their effects on sexual function. Preclinical experiments in male rats showed an increase in fertility after administration of Cynomorium extract. In these tests, an increase in the weight of the testes and an increase in the number of spermatozoa and their viability were observed. Cynomorium plants offer a new approach to the treatment of human infertility, which is currently a major problem.

Neonicotinoid insecticide acetamiprid is a part of relatively frequently used commercial formulations such as Mospilan 20SP or newly developed Carnadine, Kestrel and others. The popularity of these insecticides lies in their ease of handling and high efficacy in treating canola, corn, potatoes and apple trees. However, the possibility that pesticide products could also affect non-target organisms still needs to be taken into account. For this reason, the aim of our work was to assess the possible cytotoxic and genotoxic effects of acetamiprid-based insecticide Mospilan 20SP, which is one of the well-known commercially available formulations. Conventional tests such as chromosomal aberration assay and comet assays (alkaline and neutral) were employed under in vitro conditions using cultures of bovine whole blood cells and isolated bovine lymphocytes. The insecticide formulation was dissolved in pure water to prepare quantities of 5, 10, 25 and 50 μg.mL^-1 for analysis after 2 h, 24 h and 48 h of exposure, respectively. After 24 h of exposure, values of mitotic indices showed a decrease in mitotic activity of cells (p<0,05; p<0,01) depending on increasing concentrations. The 24 h exposure to pesticides also showed an increase in chromatide and chromosome breaks in a dose-dependent manner for the insecticide under study (p<0.05; p<0.01) compared to the negative control. Alkaline and neutral comet tests revealed the potential of acetamiprid-based formulation to induce DNA damage in bovine cells.

The nerve agents of the A-series are relatively recent chemical weapons with no antidote available yet (1). Once inside the human body, those chemicals act similarly to the classic nerve agents, by binding to the catalytic residue Serine 203 (Ser203) of human acetylcholinesterase (HssAChE) and thus preventing the proper function of this enzyme. However, there is no experimental evidence yet if the current antidotes for intoxication by nerve agents are also capable of restoring AChE inhibited by the nerve agents of the A-series. In order to launch some light on this issue, we used computational techniques (molecular docking, molecular dynamics and MM-PBSA interaction energy calculations) to assess the performances of the four currently available commercial oximes (2-PAM, HI-6, obidoxime and trimedoxime) when in contact with HssAChE inhibited by the agent A-242 (2). Based on the near-attack conformation (NAC) criterion, our results suggest that the commercial oximes would have limited efficacy to reactivate the enzyme since they are not able to properly approach the adduct Ser203-A-242. Among those oximes, trimedoxime seems to be the most promising, since it showed lower values of energy in the MM-PBSA calculations, a higher stability inside the catalytic anionic center (CAS) of HssAChE and was able to adopt a position closer to the NAC that could enable the reactivation mechanism.

The Cytomegalovirus (CMV) pass through placenta is connected with consequences in prenatal as well as postnatal phase of development. Newborns can be asymptomatic, can have mild or even life-threating symptoms with serious long-term sequels. All CMV infected newborns are at risk of long-term neurodevelopmental problems and sensorineural hearing problems.
The aim of the work: To analyse prenatal and postnatal characteristics, clinical and laboratory findings, results of investigations in the group of 11 newborns with congenital CMV infection, who were hospitalized at Neonatal Department of Intensive Medicine between January 1st 2012 and March 31st, 2022 included.
Results: Prenatal foetal sonography revealed in patient 5 and 8 positive calcifications in brain; in patient 6, 9 and 11 isolated ventriculomegaly was found. Neurological examination was clinically negative in patient 1 and 10, changes of muscular tonicity and spontaneous activity was confirmed in the rest of group. In patient 5 and 10 one-sided positivity of otoacoustic emissions was confirmed. Chorioretinitis with bilateral negative otoacoustic emissions was confirmed in patient 5. Clinical status of patient 11 was complicated by pneumonitis. Three patients were treated with antiviral drugs orally, and 11 newborns had combination of intravenous and oral form of treatment.
Conclusion: The results of analysis will contribute to a society-wide solution of prevention. Monitoring of the frequency of CMV infection in population with education of population can decreased the number of affected newborns.

Since 2002, no clinical candidate against Alzheimer’s disease has reached the market; hence, an effective therapy is urgently needed. We followed the so-called “multitarget directed ligand” approach and designed 36 novel tacrine phenothiazine heterodimers which were in vitro evaluated for their anticholinesterase properties. The assessment of the structure−activity relationships of such derivatives highlighted compound 1dC as a potent and selective acetylcholinesterase inhibitor with IC₅₀ = 8 nM and 1aA as a potent butyrylcholinesterase inhibitor with IC₅₀ = 15 nM. Selected hybrids, namely, 1aC, 1bC, 1cC, 1dC, and 2dC, showed a significant inhibitory activity toward τ(306−336) peptide aggregation with percent inhibition ranging from 50.5 to 62.1%. Likewise, 1dC and 2dC exerted a remarkable ability to inhibit self-induced Aβ₁₋₄₂ aggregation. Notwithstanding, in vitro studies displayed cytotoxicity toward HepG2 cells and cerebellar granule neurons; no pathophysiological abnormality was observed when 1dC was administered to mice at 14 mg/kg (i.p.). 1dC was also able to permeate to the CNS as shown by in vitro and in vivo models. The maximum brain concentration was close to the IC₅₀ value for acetylcholinesterase inhibition with a relatively slow elimination half-time. 1dC showed an acceptable safety and good pharmacokinetic properties and a multifunctional biological profile.

In recent years, the most mentioned nerve agents by the public are A–agents, so-called Novichoks. The official chemical structures of these new organophosphates have never been published. The possible structures were introduced by Vil Mirzayanov in 2009 (1), however, they do not correspond with structures published by Hoenig (2).
One type of Novichok was evaluated in our laboratory and its physical-chemical and biological properties were compared to well-known structures of nerve agents such as sarin or VX. Inhibition kinetics of human acetylcholinesterase and butyrylcholinesterase was determined. Further, the ability of standard oxime nerve agent antidotes to reactivate both inhibited cholinesterases was assessed. Cytotoxicity of Novichok compound was evaluated using human origin cell lines, including normal human lung fibroblasts (NHLF) and neuroblastoma cell line (SH-SY5Y), which undergo differentiation to mature neurons.
The research findings are initial dates in our research of fourth-generation nerve agents and should be useful towards the development of effective antidotes and possible subsequent therapy.

Three preservatives  used as ingredients in cosmetics and other consumer products (triclosan , triclocarban and resorcinol) were evaluated for their genotoxic/mutagenic potential by means of a set of in vitro alternative methods, namely Ames Test (MPF Test, Xenometrix, OECD TG 471) with strains TA 98, TA 100, TA 1535 and TA 1537, Comet assay on HaCat cell line (non-tumor human keratinocytes) and Mammalian chromosome aberration test (OECD TG 473) using human peripheral lymphocytes. In the chromosome aberration test all three chemicals were positive in the highest tested concentrations. Similarly, in the Comet assay  the percentage of DNA in tail was significantly increased in the highest concentrations. Moreover, the genotoxic effects  were clearly dependent on the increasing concentration and duration of exposure. The Ames Test revealed positivity only for resorcinol using strain TA 1537, detecting frameshift mutations. The positive results were recorded for extremely high concentrations which under foreseeable conditions could not be present in the human body, however, the results justify the need to regulate and limit the use of these preservatives in final products.

Every medical device that is in contact with human body must be subjected to series of biological tests during the risk assessment process. Cytotoxicity testing belongs to the group of endpoints for biological evaluation of medical devices and its whole process is defined in ISO 10993-5 “Biological evaluation of medical devices – Part 5: Tests for in vitro cytotoxicity”. In a short time, it provides an initial information about toxicity, which serves as a good indicator of general toxic properties and thus it can reduce the number of in vivo models required for subsequent toxicity analyzes. There are different approaches to cytotoxicity testing, either direct contact toxicity evaluation or elution methods. In our study, we focused on the latter approach, namely on optimizing the preparation of the material extracts in accordance with recommendations from ISO 10993-12 “Biological evaluation of medical devices – Part 12: Sample preparation and reference materials”. Based on the specific properties of tested materials, we selected volume of solvent used per surface area of test samples. Material samples were extracted in appropriate cell culture medium supplemented with serum. The materials were extracted at 37 °C for 24 hours with continuous circulation of sterile material immersed in the extraction medium in sealed tubes. In addition, we further extended the analyzes to dose ranging cytotoxicity evaluation to determine the level at which cytotoxicity no longer occurs.

The aim of the presented work was to evaluate the decontamination efficacy of newly developed decontamination agents based on surfactants with hydroxyl functional group.
Sarin and VX were selected as representative nerve agents (NA). The ability to accelerate the catalytic hydrolysis of the NA was assessed by the pH value measurement using an automatic burette. The hydrolysis of NA in a contaminated solution leads to a decline of pH value due to the formation of O-alkyl methylphosphonic acids (MPAs). The ability of tested surfactants to penetrate through the skin and their rinsing effectivity was observed using modified Franz-type of glass diffusion cells and ex vivo dermatomed porcine skin as a model membrane. The amounts of surfactants penetrated into the receptor fluid after application on the skin surface were determined by HPLC with fluorescent detection. The solutions of surfactants were used to rinse the contaminated skin. Simultaneously, the same solution was contaminated by corresponding amount of NA to simulate the situation when 100 % of a contaminant is rinsed off. The MPAs in the rinsing fluid and NA penetrated into the receptor fluid were determined by HPLC-MS. The decontamination efficacy of the tested solutions was expressed in % of an agent washed away by the rinsing fluid.
All of tested newly synthesized compounds had better hydrolytic activity compared to the commercially available surfactant. Their penetration through the skin was not observed. The rinsing efficacy of all tested solutions was comparable.

Mercury exists in the environment as elemental, metallic mercury, inorganic mercury and organic mercury.  Elemental and inorganic mercury released into the atmosphere from combustion of fossil fuels, mining, smelting and various industrial activities. In the aquatic environment is mercury deposited to sediments where is transformed into the main organic form methylmercury. Mercury methylation occures in the aquatic environment due to microorganisms metabolism as sulfite reducing bacteria.  The toxicity of mercury in humans or animals depends on its chemical form. Elemental mercury is volatile, gastrointestinal absorption of inorganic mercury is between 10 - 30% range. Methylmercury is the most toxic form of mercury. In comparison with the inorganic form, methylmercury is absorbed from more than 80 % in the gastrointestinal tract, than is widely distributed to all tissues, although the largest deposition occurs in the kidney. The enterohepatic cycle results in a long half-life for this compound compared to inorganic mercury. Methylmercury is able to cross the blood-brain and the placental barriers,  mainly affects the central nervous system and is harmful to the nervous system development of the fetus.
The greatest risk of human exposure to mercury comes from food chain. Various authorities, such as United States Environmental Protection Agency (USEPA),  the Joint Expert Committee on Food additives (JECFA) of the Food Agriculture Organization (FAO), World Health Organization (WHO) and European Food Safety Authority (EFSA),  have established a reference dose for human consumption, such as Provisional Tolerable Weekly Intake (PTWI) for methylmercury.
Our study were focused on two year report from Rapid Alert System for Food and Feed (RASFF) and evaluation of  various fish species from market chain exceeded established tresholds by the Commission Regulation (EC) No. 1881/2006 of 19^th December 2006 setting maximum levels for certain contaminants in foodstuffs. According to this study, in 2017 and 2018 y., 113 cases of exceeding the Hg content limit in fish and seafood products were reported. The most commonly reported fish is swordfish (Xiphias gladius), blue shark (Prionace glauca) and mako shark (Isurus oxyrinchus).

The present study has shown the screening test of Hawthorn fruits (Crataegus spp.) using crude hydro-alcoholic extract and in vitro antimicrobial activity of extracted flavonoid which has shown more antibacterial activity than crude extract with minimal inhibitory concentration (MIC) values ranged from 5% to 2.5%. The amount of total flavonoid and antioxidant activity of Hawthorn fruits (Crataegus spp) in various concentration extracts and antioxidant activities of different concentration extracts were determined by radical scavenging by 2, 2-diphenyl-1-picrylhydrazyl (DPPH•). The output showed that the contents of flavonoid were found to be 0.386 mg quercet in equivalents (QUE/g). Dried extract displayed remarkable antioxidant activity according to (DPPH•) assays.

Bile acids (BAs) play a significant regulatory role in the pathophysiology of non-alcoholic steatohepatitis (NASH). The present study evaluates the modulation of BAs homeostasis by carvedilol, a nonselective blocker beta adrenoreceptor that is routinely used to treat cardiovascular complications accompanying NASH. NASH was induced in mice by a continuous 24-week high-fat diet (HFD) with glucose/fructose in drinking water. At week 21, individual groups of animals received carvedilol (10 mg/kg/day, p.o.) for three weeks. Biochemical and histological analysis has shown the effectiveness of a high-fat diet in inducing NASH. The spectrum of bile acid was analyzed in bile, stool, and plasma by the LC-MS method and the molecular determination of BAs-related enzymes and transporters. Carvedilol significantly increased plasma concentrations of BAs in healthy mice via downregulation of Ntcp and Bsep transporters. Carvedilol did not significantly affect net BAs plasma concentrations or BAs biliary and fecal excretions in animals with developed NASH. However, it shifted spectra of BAs toward more hydrophilic and less toxic αMCA and HCA. Carvedilol also significantly reduced liver fibrosis in NASH mice and in vitro suppressed profibrogenic response in human hepatic stellate cells. Our results suggest the hepatoprotective effect of carvedilol in NASH and support using this agent as a part of cardiovascular regimens in patients with metabolic syndrome and a high risk of NASH development.

The ISO standard 10993 for medical device testing was accepted over three decades ago. During that period, ISO standard was changing and adapted based on the growing knowledge on adverse effects and regulatory needs. The term, medical devices, includes various products that are used in medicine, but their terapeutic effect is not acchieved by pharmacological, immunological or metabolic means. These products have different mechanical, physical and chemical properties. The procedures described in the ISO 10993 cover testing of raw materials as well as final products. However, composition of  medical devices can be quite complex, and it may sometimes be challenging to follow procedures laid down by the ISO 10993. The ISO 10993-12, reviewed in 2021, describes in detail the essentials of the sample preparation undergoing the in vivo or in vitro testing (i.e. extraction into polar and non-polar solvents). Unfortunately, there is only a little guidance for materials absorbing the extraction solvents  (ISO, 2018; ISO, 2021a; ISO, 2021b).
In our study, we tested medical devices used in dental/oral care. Among these products are adhesive pasted for dentures and cream for ulcer treatment. These formulations have shown to be highly solvent absorbing. We have performed several procedures for extracting such materials, using different ratios of the material/extractant and including/excluding the orbital shaking of the sample. The achieved extracts were subsequently tested in line with the ISO 10993-5 using 3T3 NRU assay and ISO 10993-23 using 3D reconstructed human tissue model of epidermis and another, non-keratinised epithelia, used to mimic the soft tissues in the oral cavity.
The results show that the exctract preparation procedure considerably influenced the results of the experiments. In the static conditions, the preparations showed significantly lower cytotoxic proppetries compared to the mixed extracts. Non-keratinised 3D epithelial tissue model was more sensitive to the preparations than 3D skin tissue, as expected. The testing battery helped us to predict possible side effects of the selected MD materials used in the oral cavity.

Reproductive health is challenged by contemporary lifestyles. Health issues such as menstrual problems, cancers, infertility, and sexual dysfunction have been rising (1-3). For example, nowadays, more than 20% of couples experience infertility problems (1). Recently, a growing piece of evidence supports an association between chemical exposure and reproductive disorders (2,3). Traditionally, animal models have been used to elucidate human reproductive health development, disorders, and pathologies, including mechanistic insight. Reproductive toxicity testing of chemicals for regulatory purposes also relies on them. Therefore, it is vital to address the challenges concerning the use of animal-based models for reproductive toxicology and biomedicine and provide an overview of currently available 3Rs strategies. This talk will acknowledge the urgent need for animal-free models focusing on testes and ovaries, as their proper development and functionality are critical for a lifelong healthy reproductive system and due to their sensitivity to chemicals (2,3), with some specific examples from our current studies. Then, the adverse outcome pathway (AOP) concept will be applied to summarize currently available mechanistic knowledge covering key events at all levels of the biological organization and some potential early biomarkers of reproductive impairments will be proposed. Finally, chemically-induced disruption leading to reproductive disorders and dysfunctions will be discussed.

Non-steroidal anti-inflammatory drugs (NSAIDs) are drugs with analgesic, antipyretic and anti-inflammatory effects. They are widely used in human and veterinary medicine. The most used NSAIDs include ibuprofen, diclofenac, naproxen. Due to their excessive use surface water pollution occurs. A total of 65 samples of surface water from the Elbe river basin were taken during August 2018 when the weather was constant without any significant fluctuations. The analysis was performed by means of liquid chromatography with tandem mass spectrometry (LC-MS/MS). A statistically significant negative correlation between the river flow rate in the monitored locations and the residue concentration was found for ibuprofen, naproxen, diclofenac. The most significant findings of the monitored drug residues were mostly determined in samples from small streams below larger urban settlements with a hospital or other health facilities. It was found that non-steroidal anti-inflammatory drugs can in increased concentration cause oxidative stress, damage to the liver, gills and kidneys and cause reproductive disorders in fish. They also negatively affect aquatic invertebrates, representatives of producers (phytoplankton) and consumers (zooplankton, zoobenthos) and destruents (bacteria).

Since the discovery of penicillin, antibiotic use and misuse lead to the development of bacterial strains resistant to not only antibiotics, but to common disinfecting agents as well. Monomeric quaternary ammonium compounds (QACs) – common disinfecting agents – are already rendered inefficient due to the acquired resistance. The development of novel agents is now a major component in the fight against the spread of diseases. New designs should enhance the advantages of existing substances (high efficacy, low price), while embracing biodegradability, improved solubility and lower cytotoxicity.
In recent years new, yet related class of disinfectants is on the rice. Diammonium quaternary salts – or so-called ‘Gemini’ QACs, which contain in contrast to their monomeric cousin two polar heads. In total, the molecule contains two positively charged nitrogen atoms and two alkyl chains. From this group; octenidine emerged. Despite its promising results against a broad range of resistant microorganisms, it has many shortcomings: low solubility and mild cytotoxicity. These properties need to be overcome.
In this report, we prepared novel series of octenidine derivates, which have been tested against common nosocomial bacterial strains and against biofilms, which possess unique properties in comparison to single cells. The antimicrobial activity has been evaluated against Gram-positive and Gram-negative bacterial strains. Furthermore, the substances have been tested for cytotoxicity on a mammalian cell line to evaluate one of the key safety parameters.

In this study, it has been investigated the oxidative status and morphological alterations in the brain of Wistar rats induced by repeated application of low doses of selected acetylcholinesterase reactivators - asoxime, obidoxime, K027, K048, K074, and K075. Each oxime (0.1 of LD50/kg im) was given 2 times per week for 4 weeks. The animals’ whole-body, organ weight, oxidative status, as well as microscopic examination of the brain, were done on day 35 of the study. Markers of oxidative stress, lipid peroxidation (malondialdehyde (MDA) and advanced oxidation protein products (AOPP)), as well as the activity of antioxidant enzymes (catalase (CAT) and superoxide dismutase (SOD)), were measured in the brain homogenates (1,2). Brain alterations were quantified by semiquantitative grading scales – brain damage score (BDS) (3). Oxidative stress parameters MDA and AOPP were significantly elevated in the K075-treated group compared to the control group (p < 0.001). SOD activity was significantly higher in the K075-treated group (p < 0.001), without differences in CAT activity. In the same groups of rats, brain injuries were significantly more severe than those observed in animals receiving only asoxime or K027 (p < 0.001). Our results can help to predict likely adverse systemic toxic effects, and target organ systems, which are crucial for establishing risk categories, as well as in dose selection of K-oximes as drug candidates. (MFVMA01/23-25).

Anthracycline (ANT) resistance represents a significant challenge in cancer therapy. Besides other mechanisms, ANT resistance is mediated by the metabolic activity of carbonyl-reducing enzymes (CREs) and/or the efflux activity of ATP-binding cassette (ABC) transporters. CREs reduce ANTs to their corresponding less potent alcohol metabolites, while ABC transporters pump ANT drugs out of cancer cells decreasing their concentrations below cytotoxic level. Among CREs, AKR1C3 is of great importance because its overexpression has been detected in many hematological and solid malignancies, similar to some ABC transporters such as ABCB1, ABCG2, and ABCC1. In the present study, we describe the effect of Bruton´s tyrosine kinase inhibitor, zanubrutinib (ZAN), on ANT carbonyl reduction catalyzed by AKR1C3 and ANT active efflux mediated by ABC transporters. Our results show potent inhibition of recombinant AKR1C3, while the interaction was confirmed at the level of intact cells as well. Subsequent experiments proved a synergistic effect of combination of ZAN with daunorubicin in cancer cells with AKR1C3 overexpression. In gene induction studies, ZAN did not affect the mRNA level of AKR1C3. Finally, using ANT accumulation assays, ZAN was found to significantly inhibit ABC transporters ABCB1 and ABCC1, suggesting additional mechanism contributing to combatting ANT resistance. In summary, our data introduce ZAN as an effective modulator targeting multiple pharmacokinetic resistance mediators. Following in vivo confirmation, our data might be translated into the therapy of ANT resistant tumors.

This presentation will introduce one-dimensional nanomaterials that we have been developing for various applications over past years. Two main types of materials will be discussed – nanofibers of inorganic oxides and nanotubes of TiO₂.
In the first part, the first comprehensive toxicity study of Al₂O₃, SiO₂, ZrO₂, TiO₂ and WO₃ nanofibers effects in cultured epithelial A549 cells will be presented. The nanofibers were produced by centrifugal spinning from suitable spinning solutions and have an average diameter in the sub-micrometer range. At first, we characterized the nanofibers for their morphological, compositional and structural properties. Then, we estimated the biological effects of nanofibers in pulmonary epithelial A549 cells comparing them with biological effects of Al₂O₃, SiO₂, ZrO₂, TiO₆ and WO₃ nanoparticles. Multiwalled carbon nanotubes (MWCNT) were used as a positive control. The cells were treated with 1, 10 and 100 µg.mL^-1 concentrations of a nanomaterial for 24 and 48 h. The dehydrogenase activity and glutathione levels were determined in cells as markers of cell injury. Experimental details and results of these investigations will be presented and discussed (1).
In the second part, the comprehensive toxicity study of TiO₂ nanotubes, prepared by anodic oxidation of Ti and followed by various post-treatments, will be presented.