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Restored role of the grey wolf in ecological networks of newly recolonized areas can be studied via surveys of parasite communities of this predator. As helminths circulating in multi-host systems, the tapeworms directly reflect wolves’ diet, while some species are also important from the One Health perspective. The Czech experienced centuries of wolves’ absence, however, now it is situated on the crossroad of recolonising wolves’ populations, which is opening questions of their role in ecological networks in this area and thus in sylvatic cycles of heteroxenous parasites. Five wolf carcasses from this area were obtained and genetic affinity to a particular population was inspected. Tapeworms isolated from wolves’ intestines during necropsies were molecularly identified based on sequences of COI marker. Three wolf haplotypes (w1, w2, w14) correspond with the dominance of haplogroup 1 (w1, w2) within Central European lowland population and haplogroup 2 (w14) within the Carpathian population. Two Taenia spp. were revealed: T. krabbei in Central European population wolves and T. hydatigena in an individual from Carpathian population. The results serve as a base for future monitoring and studies of the recolonising wolf population and its impact on ecosystems in the studied area to contribute to the hypothesis about differentiation of parasite communities in particular wolf population and higher parasite diversity and richness in established populations in comparison to newly settled ones.

[Image: see text] DC-SIGN (dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin) is a pattern recognition receptor expressed on immune cells and involved in the recognition of carbohydrate signatures present on various pathogens, including HIV, Ebola, and SARS-CoV-2. Therefore, developing inhibitors blocking the carbohydrate-binding site of DC-SIGN could generate a valuable tool to investigate the role of this receptor in several infectious diseases. Herein, we performed a fragment-based ligand design using 4-quinolone as a scaffold. We synthesized a library of 61 compounds, performed a screening against DC-SIGN using an STD reporter assay, and validated these data using protein-based (1)H–(15)N HSQC NMR. Based on the structure–activity relationship data, we demonstrate that ethoxycarbonyl or dimethylaminocarbonyl in position 2 or 3 is favorable for the DC-SIGN binding activity, especially in combination with fluorine, ethoxycarbonyl, or dimethylaminocarbonyl in position 7 or 8. Furthermore, we demonstrate that these quinolones can allosterically modulate the carbohydrate binding site, which offers an alternative approach toward this challenging protein target.

Diethylnitrosamine is well known for its toxic and carcinogenic properties affecting mainly liver and kidneys. Capsaicin has been proven in previous years as a promising protective agent against many health problems affecting modern people. In this study we used diethylnitrosamine induced mouse experimental model of liver and kidney damage to assess the potential chemopreventive effect of capsaicin in vivo. Fifty female ICR mice were randomly divided into five groups intraperitoneally administered 1% ethanol solution in controls, capsaicin to assess its toxicity, diethylnitrosamine alone, and diethylnitrosamine and capsaicin in combination in different manners in two groups. After 14 weeks all mice were sacrificed, complete necropsy was performed and liver and kidneys were used for further examination. Slides of both organs stained with haematoxylin and eosin were histologically evaluated and immunohistochemical detection of proliferating cell nuclear antigen and glutamine synthetase in the liver tissue was performed. Histological evaluation of the liver and kidneys revealed toxic damage of diethylnitrosamine treated animals, whereas mice that received the combination of the substances showed milder lesions. Proliferating cell nuclear antigen expression was lower in diethylnitrosamine treated animals compared to the control and capsaicin groups, pointing to a disruption of the proliferative activity of hepatocytes in the juvenile liver. Glutamine synthetase expression did not differ between the groups, indicating that no tumours were induced by any of the substances used in our study. In conclusion, our experiment demonstrated the toxic properties of diethylnitrosamine in mice liver and kidneys, with the promising beneficial effect of capsaicin.

Background The range of the ornate dog tick Dermacentorreticulatus is rapidly expanding in Europe. This tick species is the vector of canine babesiosis, caused by Babesiacanis, and also plays a role in the transmission of Theileriaequi and Babesiacaballi in equids. Methods The geographic range of D.reticulatus in the Czech Republic was re-assessed, and an up-to-date distribution map is presented based on material and data obtained during a nationwide citizen science campaign. Received and flagged individuals of D.reticulatus were also analysed for the presence of B.canis DNA. Results In striking contrast to historical records, D.reticulatus was found in all regions of the Czech Republic, with most reports coming from the southeast and northwest of the country. Between February 2018 and June 2021, the project team received 558 photo reports of ticks and 250 packages containing ticks. Of the former, 71.1% were identified as Dermacentor sp. with the remainder identified as Ixodes sp., Haemaphysalis sp., Argas sp. or Hyalomma sp. The majority of specimens in the subset of ticks that were received (N = 610) were D.reticulatus (N = 568, 93.7%), followed by Ixodesricinus and Hyalomma spp. A total of 783 adult D.reticulatus, either received (568) or collected by flagging (215), were tested for the presence of B.canis DNA using species-specific nested PCR targeting part of the 18S rRNA gene; B.canis DNA was demonstrated in 22 samples (2.81%). Conclusions The continuous spread of D.reticulatus in the Czech Republic was documented in this study. In addition, DNA of B.canis was also detected in a number of ticks, suggesting the establishment of B.canis in the Czech Republic. These results suggest that veterinarians need to consider the possibility of canine babesiosis even in dogs without a history of travel. Graphical Abstract

Due to the recent recovery of brown bear populations in Central Europe, information about their ascarid parasite, Baylisascaris transfuga is necessary as the parasite represents a part of natural ecological networks. B. transfuga can lead to larva migrans syndrome in accidental hosts, but its zoonotic potential has not been confirmed. The resent study compares development of larva migrans in infected mice inoculated with two infectious doses (ID 200 and ID 2000) of B. transfuga embryonated eggs, and the clinical manifestation to evaluate the pathogenicity of the larvae. Histopathology revealed that the liver was the most severely infected organ. The moderately infected organs included lung, brain, skeletal muscles and jejunum and the less infected ones were the eyes, heart, kidneys and spleen. The high pathogenicity of B. transfuga to mice was reflected in high mortality (33,3%) after infection, with mortality increasing with higher infectious dose. The results extend the knowledge of the interaction of B. transfuga and its aberrant hosts and contribute to the understanding of the epidemiology and transmission of this bears roundworm.

Hyalomma marginatum and Hyalomma rufipes are important vectors of Crimean-Congo Hemorrhagic Fever Virus (CCHFV) in North Africa and Southern Europe. They are occasionally also reported from Central and Western Europe where they are likely introduced from their natural range by migratory birds. In this study, we report findings and molecular identification of adults and one nymph of H. marginatum and H. rufipes, primarily from horses from different regions of the Czech Republic. While the number of the reported ticks is small, this is likely to be an underrepresentation of the actual number. Due to their vector competence for CCHFV and potential expansion into new areas with a changing climate, surveillance programs in Europe are warranted.

Diethylnitrosamine (DEN) is proven to be toxic to kidneys and liver and to act as a potent carcinogen mainly in liver. Capsaicin (CAP) is an alkaloid produced by Capsicum genus plants and is considered to be a protective agent against toxicity and carcinogenicity of many substances including DEN. The aim of this study was to assess the toxicity of DEN and CAP in liver and kidneys in mice. The experiment started after two weeks of acclimatisation and was conducted according to the Czech animal welfare protection legal guidelines. At the end of the experiment the mice were sacrificed and the toxicity of DEN and CAP in liver and kidneys were analysed. The histopathological examination of the liver revealed multifocal lymphoplasmacytic reaction in parenchyma in DEN treated group. CAP used as both preventive and therapeutic agent caused reduction in number and extent of lesions. In CAP group mitotic figures were found indicating xenobiotic-induced hepatotoxicity or regenerative changes. In the kidneys DEN revealed also multifocal lymphoplasmacytic reaction that has been mitigated by CAP. Moreover histopathological observation of the kidney in DEN group has revealed granular dystrophy of the renal tubules which has not been presented in CAP treated mice. Levels of ALT, AST activity, total protein and albumin concentration was not statistically different among control and experimental groups. In this study mild protective effect of CAP on DEN-induced hepatotoxicity and nephrotoxicity was shown only in histopathological changes. The toxicity of CAP itself is questionable and further studies should be performed to verify its chemopreventive potential.

Restored role of the grey wolf in ecological networks of newly recolonized areas can be studied via surveys of parasite communities of this predator. As helminths circulating in multi-host systems, the tapeworms directly reflect wolves' diet, while some species are also important from the One Health perspective. The Czech experienced centuries of wolves' absence, however, now it is situated on the crossroad of recolonising wolves' populations, which is opening questions of their role in ecological networks in this area and thus in sylvatic cycles of heteroxenous parasites.Five wolf carcasses from this area were obtained and genetic affinity to a particular population was inspected. Tapeworms isolated from wolves' intestines during necropsies were molecularly identified based on sequences of COI marker.Three wolf haplotypes (w1, w2, w14) correspond with the dominance of haplogroup 1 (w1, w2) within Central European lowland population and haplogroup 2 (w14) within the Carpathian population. Two Taenia spp. were revealed: T. krabbei in Central European population wolves and T. hydatigena in an individual from Carpathian population.The results serve as a base for future monitoring and studies of the recolonising wolf population and its impact on ecosystems in the studied area to contribute to the hypothesis about differentiation of parasite communities in particular wolf population and higher parasite diversity and richness in established populations in comparison to newly settled ones.

Nanocrystalline mixed oxides of Ti, Zn, Al and Fe were prepared by a homogeneous hydrolysis of sulphates with urea at temperature of 100 °C in an aqueous solution. The prepared samples were characterized by BET and BJH measurements, an X-ray powder diffraction and scanning electron microscopy. These oxides were taken for an experimental evaluation of their reactivity with yperite (2,2′-dichloroethyl sulphide), soman (3,3-dimethyl-2-butyl methylphosphonofluoridate) and matter VX (O-ethyl S-2-(diisopropylamino)ethyl methylphosphonothionate). An excellent activity in decomposition of chemical warfare agents was observed in these materials (conversion degree higher then 96%/h).

Background Protein exchange kinetics correlate with the level of chromatin condensation and, in many cases, with the level of transcription. We used fluorescence recovery after photobleaching (FRAP) to analyse the kinetics of 18 proteins and determine the relationships between nuclear arrangement, protein molecular weight, global transcription level, and recovery kinetics. In particular, we studied heterochromatin-specific heterochromatin protein 1β (HP1β) B lymphoma Mo-MLV insertion region 1 (BMI1), and telomeric-repeat binding factor 1 (TRF1) proteins, and nucleolus-related proteins, upstream binding factor (UBF) and RNA polymerase I large subunit (RPA194). We considered whether the trajectories and kinetics of particular proteins change in response to histone hyperacetylation by histone deacetylase (HDAC) inhibitors or after suppression of transcription by actinomycin D. Results We show that protein dynamics are influenced by many factors and events, including nuclear pattern and transcription activity. A slower recovery after photobleaching was found when proteins, such as HP1β, BMI1, TRF1, and others accumulated at specific foci. In identical cells, proteins that were evenly dispersed throughout the nucleoplasm recovered more rapidly. Distinct trajectories for HP1β, BMI1, and TRF1 were observed after hyperacetylation or suppression of transcription. The relationship between protein trajectory and transcription level was confirmed for telomeric protein TRF1, but not for HP1β or BMI1 proteins. Moreover, heterogeneity of foci movement was especially observed when we made distinctions between centrally and peripherally positioned foci. Conclusion Based on our results, we propose that protein kinetics are likely influenced by several factors, including chromatin condensation, differentiation, local protein density, protein binding efficiency, and nuclear pattern. These factors and events likely cooperate to dictate the mobility of particular proteins.

Boykov and Kolmogorov showed that it is possible to find globally minimal contours and surfaces via graph cuts by embedding an appropriate metric approximation into the graph edge weights and derived the requisite formulas for Euclidean and Riemannian metrics [3]. In [9] we have proposed an improved Euclidean metric approximation that is invariant under (horizontal and vertical) mirroring, applicable to grids with anisotropic resolution and with a smaller approximation error. In this paper, we extend our method to general Riemannian metrics that are essential for graph cut based image segmentation or stereo matching. It is achieved by the introduction of a transformation reducing the Riemannian case to the Euclidean one and adjusting the formulas from [9] to be able to cope with non-orthogonal grids. We demonstrate that the proposed method yields smaller approximation errors than the previous approaches both in theory and practice.

The graph cut framework presents a popular energy minimization tool. In order to be able to minimize contour length dependent energy terms an appropriate metric approximation has to be embedded into the graph such that the cost of every cut approximates the length of a corresponding contour under a given metric. Formulas giving a good approximation have been introduced by Boykov and Kolmogorov for both Euclidean and Riemannian metrics. In this paper, we improve their method and obtain a better approximation in case of the Euclidean metric. In our approach, we combine the well-known Cauchy-Crofton formulas with Voronoi diagrams theory to devise a general method with straightforward extension from 2D to 3D space. Our edge weight formulas are invariant to mirroring and directly applicable to grids with anisotropic node spacing. Finally, we show that our approach yields smaller metrication errors in both the isotropic and anisotropic case and demonstrate an application of the derived formulas to biomedical image segmentation.

Implicit active contours are widely employed in image processing and related areas. Their implementation using the level set framework brings several advantages over parametric snakes. In particular, a parameterization independence, topological flexibility, and straightforward extension into higher dimensions have led to their popularity. On the other hand, a numerical solution of associated partial differential equations (PDEs) is very time-consuming, especially for large 3D images. In this paper, we modify a fast level set-like algorithm by Nilsson and Heyden [14] intended for tracking gradient-based active contours in order to obtain a fast algorithm for tracking region-based active contours driven by the Chan-Vese model. The potential of the proposed algorithm and its comparison with two other fast methods minimizing the Chan-Vese model are demonstrated on both synthetic and real image data.

An accurate localization of a cell nucleus boundary is inevitable for any further quantitative analysis of various subnuclear structures within the cell nucleus. In this paper, we present a novel approach to the cell nucleus segmentation in fluorescence microscope images exploiting the level set framework. The proposed method works in two phases. In the first phase, the image foreground is separated from the background using a fast level set-like algorithm by Nilsson and Heyden [1]. A binary mask of isolated cell nuclei as well as their clusters is obtained as a result of the first phase. A fast topology-preserving level set-like algorithm by Maska and Matula [2] is applied in the second phase to delineate individual cell nuclei within the clusters. The potential of the new method is demonstrated on images of DAPI-stained nuclei of a lung cancer cell line A549 and promyelocytic leukemia cell line HL60.

Methods based on combinatorial graph cut algorithms received a lot of attention in the recent years for their robustness as well as reasonable computational demands. These methods are built upon an underlying Maximum a Posteriori estimation of Markov Random Fields and are suitable to solve accurately many different problems in image analysis, including image segmentation. In this paper we present a two-stage graph cut based model for segmentation of touching cell nuclei in fluorescence microscopy images. In the first stage voxels with very high probability of being foreground or background are found and separated by a boundary with a minimal geodesic length. In the second stage the obtained clusters are split into isolated cells by combining image gradient information and incorporated a priori knowledge about the shape of the nuclei. Moreover, these two qualities can be easily balanced using a single user parameter. Preliminary tests on real data show promising results of the method.