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Abstract Background Plant-parasitic nematodes are economically important pests responsible for substantial losses in agriculture. Researchers focusing on plant-parasitic nematodes, especially on finding new ways of their control, often need to assess basic parameters such as their motility, viability, and reproduction. Traditionally, these assays involve visually counting juveniles and eggs under a dissecting microscope, making this investigation time-consuming and laborious. Results In this study, we established a procedure to efficiently determine the motility of two plant-parasitic nematode species, Heterodera schachtii and Ditylenchus destructor, using the WMicrotracker ONE platform. Additionally, we demonstrated that hatching of the cyst nematode H. schachtii can be evaluated using both the WMicrotracker ONE and by assessing the enzymatic activity of chitinase produced during hatching. Conclusions We present fast and straightforward protocols for studying nematode motility and hatching that allow us to draw conclusions about viability and survival. Thus, these methods are useful tools for facilitating fast and efficient evaluation in various fields of research focused on plant-parasitic nematodes.

Matjaž Fležar,1 Karina Jahnz-Rózyk,2 Gloria Enache,3 Tatiana Martynenko,4 Peter Kristufek,5 Sanda Škrinjaric-Cincar,6 Pavla Kadlecová,7 Goran Martinovic81University Clinic of Respiratory and Allergic Diseases Golnik, Golnik, Slovenia; 2Military Institute of Medicine, Department of Immunology and Allergology, Warsaw, Poland; 3Saint John Clinical Emergency Hospital, Bucharest, Romania; 4Municipal Medical Institution City Hospital, Barnaul, Russian Federation; 5SACCME, Slovak Medical University, Bratislava, Slovakia; 6Clinical Hospital Center, Osijek, Croatia; 7ADDS Sro, Jana Uhra, Brno, Czech Republic; 8Boehringer Ingelheim RCV GmbH and Co KG, Ingelheim, Vienna, AustriaPurpose: The long-acting inhaled anticholinergic agent, tiotropium, is recommended as first-line maintenance therapy for moderate to very severe Chronic Obstructive Pulmonary Disease (COPD) to improve symptoms, exercise tolerance, health status, and to reduce exacerbations. Few studies have evaluated the therapeutic efficacy of tiotropium in patients in routine clinical conditions. The current study was designed to investigate the therapeutic efficacy of tiotropium delivered via the HandiHaler® device on the health status of patients with COPD with Global initiative for chronic Obstructive Lung Disease (GOLD) disease classification 2–4 in six central and eastern European countries in a real-life clinical setting.Methods: The study was an open-label, prospective, uncontrolled, and single-arm surveillance study with three clinic visits during a 6-month observation period (baseline, and months 3 and 6). Health status was measured using the disease-specific St George's Respiratory Questionnaire (SGRQ). The primary efficacy endpoint was the mean change from baseline in SGRQ total score at the end of the 6-month observational period.Results: Patients treated with tiotropium 18 µg once daily showed statistically significant and clinically meaningful reduction (improvement) of 21.7 units in the SGRQ total score, regardless of smoking status or cardiac comorbidities at enrollment (P < 0.0001). The analysis also showed that age, treatment compliance, and GOLD disease classification were significant factors that impact the health status of patients with COPD differently.Conclusion: These results provide further support for the use of the tiotropium HandiHaler® as first-line maintenance treatment of patients with COPD with a clinician-assessed disease.Keywords: tiotropium, HandiHaler®, chronic obstructive pulmonary disease, health-related quality of life

A combination of liver and heart dysfunction worsens the prognosis of human survival. The aim of this study was to investigate whether empagliflozin (a sodium-glucose transporter-2 inhibitor) has beneficial effects not only on cardiac and renal function but also on hepatic function. Adult (6-month-old) male spontaneously hypertensive rats (SHR) were fed a high-fat diet (60% fat) for four months to induce hepatic steatosis and mild heart failure. For the last two months, the rats were treated with empagliflozin (empa, 10 mg.kg-1.day-1 in the drinking water). Renal function and oral glucose tolerance test were analyzed in control (n=8), high-fat diet (SHR+HF, n=10), and empagliflozin-treated (SHR+HF+empa, n=9) SHR throughout the study. Metabolic parameters and echocardiography were evaluated at the end of the experiment. High-fat diet feeding increased body weight and visceral adiposity, liver triglyceride and cholesterol concentrations, and worsened glucose tolerance. Although the high-fat diet did not affect renal function, it significantly worsened cardiac function in a subset of SHR rats. Empagliflozin reduced body weight gain but not visceral fat deposition. It also improved glucose sensitivity and several metabolic parameters (plasma insulin, uric acid, and HDL cholesterol). In the liver, empagliflozin reduced ectopic lipid accumulation, lipoperoxidation, inflammation and pro-inflammatory HETEs, while increasing anti-inflammatory EETs. In addition, empagliflozin improved cardiac function (systolic, diastolic and pumping) independent of blood pressure. The results of our study suggest that hepatoprotection plays a decisive role in the beneficial effects of empagliflozin in preventing the progression of cardiac dysfunction induced by high-fat diet feeding.

Abstract Rotations of schoolchildren were considered as a non-pharmacological intervention in the COVID-19 pandemic. This study investigates the impact of different rotation and testing schedules.We built an agent-based model of interactions among pupils and teachers based on a survey in an elementary school in Prague, Czechia. This model contains 624 schoolchildren and 55 teachers and about 27 thousands social contacts in 10 layers. The layers reflect different types of contacts (classroom, cafeteria, etc.) in the survey. On this multi-graph structure we run a modified SEIR model of covid-19 infection. The parameters of the model are calibrated on data from the outbreak in the Czech Republic in spring 2020. Weekly rotations of in-class and distance learning are an effective preventative measure in schools reducing the spread of covid-19 by 75–81% . Antigen testing twice a week or PCR once a week significantly reduces infections even when using tests with a lower sensitivity. The structure of social contacts between pupils and teachers strongly influences the transmission. While the density of contact graphs for older pupils is 1.5 times higher than for younger pupils, the teachers’ network is an order of magnitude denser. Teachers moreover act as bridges between groups of children, responsible for 14–18% of infections in the secondary school compared to 8–11% in the primary school. Weekly rotations with regular testing are a highly effective non-pharmacological intervention for the prevention of covid-19 spread in schools and a way to keep schools open during an epidemic.

Abstract Late Oligocene (ca 25 Ma) volcano‐sedimentary successions exposed on the western periphery of the Doupovské Hory Volcanic Complex reveal a complex sedimentation history influenced in various ways by decay of the alkali basalt volcanic edifice. Weathering of the volcanic rocks supplied abundant reactants that promoted carbonate precipitation in the peripheral palaeolakes—as evidenced by strongly non‐radiogenic 87Sr/86Sr values (0.7038–0.7041). On the other hand, the sediments of the initial shallow lake became deformed by the bulldozing effect of a debris avalanche. The debris flow and avalanche deposits filled up the original depression, modified the basin morphology and shifted the peripheral lacustrine setting further away from the volcano. At this stage, surface water influx from the surrounding granites conferred a more radiogenic character (87Sr/86Sr values 0.7046–0.7049) to the calcrete deposits. Fossil assemblages as well as limestone textures suggest significant seasonal water‐level fluctuations, possibly reflecting the alternating rainy and dry‐seasons of a prevalently humid Central‐European Late Oligocene climate. The seasonal drying out of the ponds resulted in significant 18O enrichments. Although the ca 0‰ δ13C values might suggest mixing of atmospheric and volcanic CO2 during carbonate precipitation, no active volcanic conduits of relevant age are known in the close vicinity. The lower δ13C values are likely a result of mantle degassing through rift faults, a phenomenon observed in the magmatically extinct Ohře Rift until present. This paper demonstrates that limestones derived from weathered alkaline basalts are characterised by highly non‐radiogenic Sr isotopic ratios (87Sr/86Sr ca 0.704), suggesting a magmatic origin for the Ca within these carbonates. Contrary to the notion of carbonatites being present when highly non‐radiogenic Sr isotopes are found, these results show that Sr isotopes in carbonates formed in alkali basalt‐sourced environments only reveal the source of the Sr (and Ca) ions, not necessarily the presence of carbonatite.

ABSTRACT: Objectives: This study aimed to characterise compounds with activity against carbapenemase-expressing Gram-negative bacteria and nematodes and evaluate their cytotoxicity to non-cancerous human cells. Methods: The antimicrobial activity and toxicity of a series of phenyl-substituted urea derivatives were evaluated using broth microdilution, chitinase, and resazurin reduction assays. Results: The effects of different substitutions present on the nitrogen atoms of the urea backbone were investigated. Several compounds were active against Staphylococcus aureus and Escherichia coli control strains. Specifically, derivatives 7b, 11b, and 67d exhibited antimicrobial activity against Klebsiella pneumoniae 16, a carbapenemase-producing Enterobacteriaceae species, with minimum inhibitory concentration (MIC) values of 100, 50, and 72 µM (32, 64, and 32 mg/L), respectively. In addition, the MICs obtained against a multidrug-resistant E. coli strain were 100, 50, and 36 µM (32, 16, and 16 mg/L) for the same compounds, respectively. Furthermore, the urea derivatives 18b, 29b, 50c, 51c, 52c, 55c–59c, and 62c were very active towards the nematode Caenorhabditis elegans. Conclusions: Testing on non-cancerous human cell lines suggested that some of the compounds have the potential to affect bacteria, especially helminths, with limited cytotoxicity to humans. Given the simplicity of synthesis for this class of compounds and their potency against Gram-negative, carbapenemase-expressing K. pneumoniae, aryl ureas possessing the 3,5-dichloro-phenyl group certainly warrant further investigation to exploit their selectivity.

Giardia intestinalis, a cosmopolitan gastrointestinal protist, is detected mainly in patients with clinical giardiasis in high-income countries. In contrast, there is very little information on the presence of Giardia in asymptomatic individuals. Therefore, the aim of this study was to determine the presence and prevalence of Giardia in gut-healthy volunteers in the Czech Republic and to perform a comparative evaluation of different diagnostic methods, since Giardia diagnostics is complicated. Our results confirmed that the qPCR method is the most sensitive method for detecting Giardia and revealed a prevalence of 7% (22/296) in asymptomatic individuals. In most cases, the colonization intensity ranged from 10−1–101. A conventional PCR protocol targeting the TPI gene was used to identify the assemblages. However, this protocol had limited sensitivity for Giardia amplification, effectively detecting colonization above an intensity of 104. In addition, Giardia was detected in 19% of the animals, which were closely associated with the study participants. However, due to methodological limitations, zoonotic transmission could not be clearly confirmed. Notably, contact with animals proved to be the only factor that had a significant impact on the incidence of Giardia in gut-healthy humans.

This article deals with the implementation of automated guided vehicles (AGVs) in a selected company. The aim is to analyse the use of AGVs in our country and abroad and to provide information about the use of AGVs in other countries and operations other than ours. The result of the analysis was a literature review, which points out the individual advantages and disadvantages of the use of AGVs in companies. Within the review we also address the issue of AMR vehicles, due to the modernization of existing AGVs in the company, or the replacement of AMRs with AGVs in general. Our aim is to show why AGVs can replace human work. This is mainly because of the continuous increase in the wages of employees, because of safety, but also because of the modernization of the selected company. The company has positive experience of AGVs in other sites. We wanted to point out a higher form of automation, and how it would be possible to use AMR vehicles for the same work as AGVs. In the company, we have identified jobs where we would like to introduce AGVs or AMR vehicles. Consequently, we chose the AGV from CEIT operated by magnetic tape and the AMR from SEER as an example. Based on studies, the demand for AGVs is expected to increase by up to 17% in 2019–2024. Therefore, the company is looking into the issue of the implementation of AGVs at multiple sites. The question which remains is the economic return and the possibility of investing in the automation of processes in the company, which we discuss in more detail in the conclusion of the article and in the research. The article describes the exact processes for AGVs, their workload, and also the routes for AGVs, such as loading/unloading points, stopping points, checkpoints, junctions with other AGVs, charging stations, and field elements, as well as their speed, frequency and the possibility of collision with other AGVs. Our research shows that by applying the new technology, the company will save a large amount of money on employee wages. The purchase of two AGVs will cost the company EUR 49,000, while the original technology used in the company cost EUR 79,200 annually. The payback period for such an investment is 8 months. The benefits of implementing AGVs are evaluated in the last section of this paper, where both the economic and time requirements of the different proposals are included. This section also includes recommendations for improving specific parts of the enterprise.

Even though silicon is frequent compound in soil, its use in plant nutrient media is rare. Based on known physiological role and up to now performed studies it seems that silicon has a good potential to improve growth characteristics of in vitro cultivated plants. Before practical application, however, it is always necessary to assess the optimal conditions of application with regard to the fact that plant reactions to different chemicals added to nutritional media can vary on the species or even cultivar level. The presented study evaluate effects of potassium silicate used in in vitro cultivation media on growth parameters of Prunus persica × Prunus davidiana ‘Cadaman,‘ Prunus × amygdalopersica ‘GF 677‘ and Corylus avellana ‘Tonda di Giffoni‘ genotypes, which are frequently subject of commercial in vitro multiplication. In fact, four different concentrations of potassium silicate was added to the multiplication media, control medium was left silicon-free. Three different characteristics were observed during cultivation – number of new shoots per explant, weight of a new plant and length of new shoots, from which number of new shoots per explant was considered the most important factor. In all cases a positive effects of potassium silicate on the condition and other growth parameters of treated cultures were observed. In ‘Cadaman‘ culture significant growth changes appeared on media with 20 mg.l-1 potassium silicate, which can be recommended for future applications. For ‘GF 677‘ the best results were obtained on media with 2 mg.l-1 which can be recommended for improving condition and number of shoots on new plants. For hazelnut genotype ‘Tonda di Giffoni‘ best results were obtained on media with 10 mg.l-1 of potassium silicate, but also use of 5 mg.l-1 of potassium silicate significantly improved growth parameters. Generally, presented study provides important and practically useful insights into the practical use of silicon in cultivation media designated for commercial in vitro micropropagation.

Dientamoeba fragilis is a cosmopolitan intestinal protist colonizing the human gut with varying prevalence depending on the cohort studied and the diagnostic methods used. Its role in human health remains unclear mainly due to the very sporadic number of cross-sectional studies in gut-healthy populations. The main objective of this study was to expand knowledge of the epidemiology of D. fragilis in gut-healthy humans and their animals. A total of 296 stool samples from humans and 135 samples from 18 animal species were analyzed. Using qPCR, a prevalence of 24% was found in humans in contrast to conventional PCR (7%). In humans, several factors were found to influence the prevalence of D. fragilis. A more frequent occurrence of D. fragilis was associated with living in a village, traveling outside Europe and contact with farm animals. In addition, co-infection with Blastocystis spp. was observed in nearly half of the colonized humans. In animals, D. fragilis was detected in 13% of samples from eight species using qPCR. Our molecular phylogenies demonstrate a more frequent occurrence of Genotype 1 in gut-healthy humans and also revealed a likely a new protist species/lineage in rabbits related to D. fragilis and other related organisms.

Gliflozins (sodium-glucose transporter-2 inhibitors) exhibited renoprotective effects not only in diabetic but also in non-diabetic patients with chronic kidney disease (CKD). Controversial results were reported in experimental non-diabetic models of CKD. Therefore, we examined empagliflozin effects in three CKD models, namely, in fawn-hooded hypertensive (FHH) rats, uninephrectomized salt-loaded (UNX + HS) rats, and in rats with Goldblatt hypertension (two-kidney, one-clip 2K1C) that were either untreated or treated with empagliflozin (10 mg/kg/day) for eight weeks. Plethysmography blood pressure (BP) was recorded weekly, and renal parameters (proteinuria, plasma urea, creatinine clearance, and sodium excretion) were analyzed three times during the experiment. At the end of the study, blood pressure was also measured directly. Markers of oxidative stress (TBARS) and inflammation (MCP-1) were analyzed in kidney and plasma, respectively. Body weight and visceral adiposity were reduced by empagliflozin in FHH rats, without a significant effect on BP. Experimentally induced CKD (UNX + HS and 2K1C) was associated with a substantial increase in BP and relative heart and kidney weights. Empagliflozin influenced neither visceral adiposity nor BP in these two models. Although empagliflozin increased sodium excretion, suggesting effective SGLT-2 inhibition, it did not affect diuresis in any experimental model. Unexpectedly, empagliflozin did not provide renoprotection because proteinuria, plasma urea, and plasma creatinine were not lowered by empagliflozin treatment in all three CKD models. In line with these results, empagliflozin treatment did not decrease TBARS or MCP-1 levels in either model. In conclusion, empagliflozin did not provide the expected beneficial effects on kidney function in experimental models of CKD.

Protists are a normal component of mammalian intestinal ecosystems that live alongside, and interact with, bacterial microbiota. Blastocystis, one of the most common intestinal eukaryotes, is reported as a pathogen that causes inflammation and disease, though health consequences likely vary depending on host health, the gut ecosystem, and genetic diversity. Accumulating evidence suggests that Blastocystis is by and large commensal. Blastocystis is more common in healthy individuals than those with immune mediated diseases such as Inflammatory Bowel Diseases (IBD). Blastocystis presence is also associated with altered composition and higher richness of the bacterial gut microbiota. It is not clear whether Blastocystis directly promotes a healthy gut and microbiome or is more likely to colonize and persist in a healthy gut environment. We test this hypothesis by measuring the effect of Blastocystis ST3 colonization on the health and microbiota in a rat experimental model of intestinal inflammation using the haptenizing agent dinitrobenzene sulfonic acid (DNBS). We experimentally colonized rats with Blastocystis ST3 obtained from a healthy, asymptomatic human donor and then induced colitis after 3 weeks (short term exposure experiment) or after 13 weeks (long term exposure experiment) and compared these colonized rats to a colitis-only control group. Across experiments Blastocystis ST3 colonization alters microbiome composition, but not richness, and induces only mild gut inflammation but no clinical symptoms. Our results showed no effect of short-term exposure to Blastocystis ST3 on gut inflammation following colitis induction. In contrast, long-term Blastocystis exposure appears to promote a faster recovery from colitis. There was a significant reduction in inflammatory markers, pathology 2 days after colitis induction in the colonized group, and clinical scores also improved in this group. Blastocystis colonization resulted in a significant reduction in tumor necrosis factor alpha (TNFα) and IL-1β relative gene expression, while expression of IFNγ and IL17re/17C were elevated. We obtained similar results in a previous pilot study. We further found that bacterial richness rebounded in rats colonized by Blastocystis ST3. These results suggest that Blastocystis sp. may alter the gut ecosystem in a protective manner and promote faster recovery from disturbance.

The article considers a possible improvement of road vehicle safety by using eCall – a system which initiates an emergency call in case of traffic accident. A possible way of better description of a frontal impact accident of a vehicle is examined and enriched by the information from the onboard e-call unit. In this article, we analyze results of frontal crash tests with different types of barriers and overlapping area and look for the correlation between the individual vehicle and collision parameters in order to provide a better description of the severity of the accident by the eCall system. The relation among the selected parameters is described using the correlation analysis.

The ciliary ultrastructure can be damaged in various situations. Such changes include primary defects found in primary ciliary dyskinesia (PCD) and secondary defects developing in secondary ciliary dyskinesia (SCD). PCD is a genetic disease resulting from impaired ciliary motility causing chronic disease of the respiratory tract. SCD is an acquired condition that can be caused, for example, by respiratory infection or exposure to tobacco smoke. The diagnosis of these diseases is a complex process with many diagnostic methods, including the evaluation of ciliary ultrastructure using transmission electron microscopy (the golden standard of examination). Our goal was to create a program capable of automatic quantitative analysis of the ciliary ultrastructure, determining the ratio of primary and secondary defects, as well as analysis of the mutual orientation of cilia in the ciliary border. PCD Quant, a program developed for the automatic quantitative analysis of cilia, cannot yet be used as a stand-alone method for evaluation and provides limited assistance in classifying primary and secondary defect classes and evaluating central pair angle deviations. Nevertheless, we see great potential for the future in automatic analysis of the ciliary ultrastructure.

Our previous studies demonstrated that chronic systemic blockade of renin-angiotensin system (RAS) lowered blood pressure (BP) of Ren-2 transgenic rats (TGR) by the attenuation of both angiotensin II-dependent and sympathetic vasoconstriction. Since systemic RAS blockade also inhibits brain RAS, we were interested which effects on these two types of vasoconstriction will have the central RAS blockade in hypertensive TGR rats. Adult male heterozygous TGR rats and their Hannover Sprague Dawley (HanSD) controls were subjected to chronic systemic or intracerebroventricular administration of either angiotensin type 1 receptor blocker losartan or direct renin inhibitor aliskiren for 4 weeks. Additional groups of TGR and HanSD rats were used for the evaluation of acute peripheral and brain effects of angiotensin II. Both chronic systemic and intracerebroventricular administrations of losartan or aliskiren normalized BP of TGR animals. BP effect of brain RAS blockade was based solely on the reduced sympathetic vasoconstriction, while systemic RAS blockade attenuated both angiotensin II-dependent and sympathetic vasoconstriction. Surprisingly, neither peripheral nor central pressor effects of acute angiotensin II administration were enhanced in TGR compared to HanSD rats. In conclusion, sympathoinhibition represents the main mechanism of BP reduction in heterozygous TGR rats subjected to chronic brain or systemic RAS blockade, while peripheral attenuation of angiotensin II-dependent vasoconstriction during systemic RAS blockade is less important. Our data suggest that the participation of angiotensin II in BP control of adult heterozygous TGR rats is shifted from peripheral vasoconstriction to central sympathoexcitation. Similar mechanisms cannot be excluded in human essential hypertension.

The problem of attributing responsibility for cyber-attacks is almost as old as cyberspace itself, yet it remains one of the most troublesome issues of that domain. It is often impossible to uncover direct evidence that would reveal the identities of the attackers. Investigators must therefore rely on other, more indirect avenues of proof. The aim of this exploratory study is to develop a basic categorisation of indirect evidence that can be used to attribute state responsibility for cyber-attacks in international relations. To do so, the article works with international legal concepts but transposes them into the analysis of international relations. The categorization of indirect proof is based on the Russian-Georgian conflict of 2008, which provides one of the richest arrays of this kind of evidence. The analysis identifies four kinds of indirect evidence: level of coordination, level of preparedness, state relations with the national hacker community, and state conception of cyber-security.

Colonization by the benign tapeworm, Hymenolepis diminuta, has been associated with a reduction in intestinal inflammation and changes in bacterial microbiota. However, the role of microbiota in the tapeworm anti-inflammatory effect is not yet clear, and the aim of this study was to determine whether disruption of the microflora during worm colonization can affect the course of intestinal inflammation. We added a phase for disrupting the intestinal microbiota using antibiotics to the experimental design for which we previously demonstrated the protective effect of H. diminuta. We monitored the immunological markers, clinical parameters, bacterial microbiota, and histological changes in the colon of rats. After a combination of colonization, antibiotics, and colitis induction, we had four differently affected experimental groups. We observed a different course of the immune response in each group, but no protective effect was found. Rats treated with colonization and antibiotics showed a strong induction of the Th2 response as well as a significant change in microbial diversity. The microbial results also revealed differences in the richness and abundance of some bacterial taxa, influenced by various factors. Our data suggest that interactions between the tapeworm and bacteria may have a major impact on its protective effect.

The aim of the paper was to determine the current state of the Turkish minority in the Greek regions of Central Macedonia and Thrace. Specifically, Xanthi (Thrace) and Thessaloniki (Central Macedonia) were of prime interest. The field research was conducted in the second largest Greek city of Thessaloniki, where historical and socially relevant sites of the Turkish minority were visited. Participant observation and semi-structured interviews were conducted at the headquarters of the Turkish Embassy. This house was the birthplace of the first President of Turkey, Mustafa Kemal Atatürk. Nowadays, this house serves as a museum and is visited mainly by Turks. Thessaloniki has well documented tourist locations around the city except for the Atatürk museum, which also has very few Greek visitors. The field research continued in Xanthi and its surroundings. In Xanthi, the field research included an initial observation followed by semi-structured interviews. These techniques were performed in the old Turkish quarter, two minority schools, a local mosque, and a traditional Turkish restaurant. The consensus of the interviewees was that they “feel” that they are Turkish, but their homeland is Greece. Similarly, they consider their native language is Turkish, while Greek is deemed essential for living. Turkish minority children attend local minority schools and in many cases commute from surrounding villages to the schools. For college, students must leave Greece for Turkey if they want to study in the Turkish language since this option is not available in Greece. The Greek language that is utilized in Greek universities is typically more difficult and specialized than the everyday Greek used by the Turkish minority and therefore this group prefers to study in Turkey or at another European university. The areas with the highest concentration of a minority Turkish population do not perceive discrimination from the majority Greek population due to their long-term cohabitation. Nonetheless, when members of this minority leave this area, they experience discriminatory behavior due with their faith and ethnicity.