Funded projects under the Grant Programme for SAS PhD students in 2022

Ladybirds are important predators of aphids and coccids on trees. A current research of arboricolous ladybirds within Europe focuses mainly on ladybirds associated with deciduous trees, to a lesser extent on coniferous trees. Ladybird research within the proposed doctoral grant will be carried out on Norway spruce in towns and forests in Slovakia. It will provide original knowledge about vertical distribution of ladybirds in the presence of the invasive Harlequin ladybird along the gradient of the occurrence of Norway spruce from warmer lowlands to cool timber line area. It will contribute to better understanding of altitudinal segregation of native ladybird species and the fast spreading non-native Harlequin ladybird as a basis for the future research of impacts of climate change on insect communities and ecosystems in Central Europe.

The aim of this project is a detailed karyological study of Acanthocephalus anguillae (Acanthocephala: Echinorhynchidae) from two biotopes with different levels of anthropogenic pollution. Populations of the studied parasite from the heavily industrially polluted Zemplínska Šírava and from the Palcmanská Maša water reservoir, which is part of the European network of protected areas NATURA 2000, will be investigated by classical and molecular cytogenetic techniques. The karyotype will be characterized in detail and the number and localization of ribosomal and histone H3 genes on the chromosomes will be determined. For the first time, the RepeatExplorer software will be used to identify and characterize repetitive sequences in the genome of A. anguillae in order to obtain new suitable chromosomal markers. We expect that this new methodological approach and the mapping of individual markers on chromosome in both populations will provide valuable information on the possible impact of environmental pollution on the chromosomes of the parasite under study.

The present project will aim to characterize the biochemical and antifungal properties of chitinase from the carnivorous plant Drosera binata, where this hydrolase is an important part of the digestive processes. The cDNA of the respective chitinase will be introduced into the bacterial expression system and, after induction of expression, the recombinant protein will be isolated with affinity chromatography. Subsequently, we determine the thermal stability of chitinase in different conditions and after that, we’ll define optimal temperature and pH. We will also test its antifungal properties and compare them with chitinase from Drosera rotundifolia, which was a subject of research in the previous period. We expect that characterization of hydrolases involved in digestion processes will not only clarify the process of digestion in carnivorous plants, but may reveal their potential use in plant biotechnology especially in strengthening the defence of economically important crops against phytopathogens, in the production of pharmacologically important chito-oligomers, or even in enzymatical degradation of chitinous waste.

Composition of complex soil fungal communities is most frequently assessed by molecular methods including high-throughput Illumina sequencing using short DNA fragments as molecular markers (metabarcoding). Majority of Illumina based fungal metabarcoding studies rely on usage of either ITS1 or ITS2 partition of the Internal Transcribed Spacer (ITS) region of nrDNA. Advantages in efficiency of amplification from substrates and well-defined barcoding gap put ITS region as the primary fungal barcoding marker, however it is not unproblematic. One of these problems is that ITS based libraries suffer from diversity overestimation due to sequence copy polymorphism and unequal sequence length. The main goal of this research is to compare performances of both ITS1 and ITS2 partitions with two protein-coding low-copy regions of nDNA (ef1-a and rpb2) on artificial fungal mock communities with defined composition. Mock communities will be constructed of three unrelated agaric (Agaricomycotina) lineages of closely related species (genera Hodophilus, Dermoloma and Russula) and sequenced on Illumina platform using selected molecular markers. The sequencing data will be analyzed using most common types of bioinformatics tools and verified with reference dataset resulting from Sanger sequencing of fruiting bodies of input species. Although the ITS region is an obvious choice for characterization of general fungal assemblages, due to rising reference sequence databases for rpb2 and ef1-a markers, this project seeks to evaluate them as alternative metabarcoding markers useful for studies of target sequencing of defined groups.

Climate change is one of the main global problems, the consequences of which, such as changes in temperature and precipitation regimes, are very likely to worsen in the coming decades. In connection with climate change, the overall management of the country is also changing, including ski resorts, which cope with the problem of insufficient natural snow cover by artificial snowmaking. The effects of the operation of ski resorts on the landscape are closely connected with the fragmentation and degradation of the landscape, particularly the disturbance of the topsoil layer which is an important source of nutrients for plants. Management of artificially snowed ski slopes, therefore, has a great influence on the chemical and physical properties of underlying soils. The main goal of this project is to study how artificial snowing is changing soil properties and to supplement the ongoing vegetation research focusing on the impact of ski resorts’ operation on plant communities. We plan to perform physical and chemical analyses of soils from both the artificially and naturally snowed ski slopes within 30 selected Slovak ski resorts. In combination with a vegetation survey, this approach could help discover new connections between the disturbance coming from the operation of ski resorts and its impact on the environment, mainly the soil.

Symbiotic relationships between vascular plants and endophytic mycorrhizal fungi are thought to be one of the most important factors in vascular plant survival and adaptation to the land environment. This is even more remarkable in habitats with challenging climatic and edaphic conditions. The proposed study will investigate the mycorrhizal colonization in the root system of the endemic Daphne arbuscula Čelak (Thymelaeaceae) that inhabits climatically extreme rocky habitats in the Western Carpathians. We will determine the diversity of all groups of endophytic fungi associated with the root system of D. arbuscula using light microscopy and amplicon sequencing of nuclear ribosomal DNA. We will also investigate the possibility of differentiating endophytic fungal assemblages between populations from ecologically distinct sites, specifically warm and dry versus cold and humid microenvironments. The findings of this investigation will shed more light on the diversity and composition of symbiotic assemblages in D. arbuscula root systems, which may be one of the key mechanisms allowing D. arbuscula to survive in the climatically extreme environment. The outcomes of this project will contribute to improving conservation strategies for this endangered species and its natural habitat.

The proposed work focuses on photo-induced atom transfer radical polymerization (photo-ATRP) and its subsequent application in the discipline of surface engineering via surface-initiated photo ATRP (SI-Photo ATRP). Al2O3 is widely known for its low cost, non-toxicity and is the most biocompatible material for clinical use. Hence, surfaces deposited with Al2O3 nanolayers via atomic layer deposition (ALD) will be used for surface modification. We plan to synthesize functional polymer brushes using a renewable monomer, α-methylene-γ-butyrolactone (MBL). MBL consists of a five-member lactone ring and an exocyclic carbon−carbon double bond, which will be used for SI-Photo ATRP. Post-polymerization functionalization of the lactone ring via ring-opening will be performed using various amines/ acids. This is followed by an investigation of the reactivity of the end-capped functional groups towards the attachment of biomolecules such as Biotin (protein). The interaction of these proteins and their affinity towards ligands such as avidin/streptavidin is a major field of interest in the biomolecules community. These synthesized polymer brushes attached with biomolecules will be characterized using techniques such as VASE, WCA, AFM, GPC etc.

The wetting study of a molten MoNbTaVW high entropy alloy (HEA) on (MoNbTaVW)C high entropy ceramics (HEC) is the first step of introducing a new metal filler for joining of these HEC ceramics, since the wettability plays an important role in the processes and properties of joining of ceramics with fillers. Furthermore, high entropy carbides have remarkable properties leading to applications in the areas of aerospace field owing to their outstanding mechanical properties and stability at high temperatures. Due to the intrinsic brittleness of ceramics, it is difficult to obtain HEC bulk ceramics with large and complex shape by sintering and machining. In order to further extend the application potential of HEC ceramics, the development of a practical jointing technology is desirable. Despite the fact that HEAs have perfect potential for joining of ceramics, they have not yet been used as the filler to join HEC. Therefore, this project will focus on the study of high temperature interactions and wettability between HEC and HEA (consisting of the same metal elements) to introduce a new filler for joining of HEC ceramics.

In the last decade, advanced hybrid materials based on the clay minerals and polymers have been widely studied due to their unique properties. Clay-polymer hybrids gained considerable attention from researchers since the pioneering research of the Toyota research team on montmorillonite-filled Nylon 6 in 1980s. Polymer materials have some limitations, such as low mechanical strength, high solvent sensitivity, and low heat resistance. Clay minerals are used to overcome these limitations and enhance the performance of polymers. Clay-polymer hybrids provide remarkably improved mechanical, thermal, and gas barrier properties. Moreover, the use of clay-polymer hybrids in industrial and environmental applications are increasing due to their intrinsic properties. Molecular simulations can give a deep insight into clay polymer hybrids, considering the limitation of experimental techniques and the exponential growth of computer performance. They can add and supply information obtained experimentally about the structure and properties of these materials on an atomic scale. For example, density functional theory (DFT) method can give reliable information about the interactions of clay with the polymer matrix and its properties. In this research, we will perform a detailed study of the interactions of different clay minerals (saponite/ beidellite/hectorite) and polymer (2-methyl-2-oxazoline) using a DFT method involving dispersive corrections (D3 scheme). Further, we will investigate the mechanical properties of these hybrids. This study elucidates their stability using computational method saving chemical compounds for synthesis and time of measurement of their mechanical properties.

Non-pathogenic members of the genus Corynebacterium are widely used as industrial producers in many biotechnological processes. The bacterial strain [Brevibacterium] flavum CCM 251 has been used as a producer of the essential amino acid L-lysine. A serious and still common problem in many biotechnological laboratories and factories is infection of bacterial cultures by bacteriophages. Bacteriophages are intracellular parasites that require a bacterial host cell for multiplication and proliferation, which often results in bacterial cell lysis and loss of production. We have previously isolated and characterized corynephage BFK20, which causes lysis of [B]. flavum CCM 251. During phage infection, the primary function of the phage tail is the formation of the stable phage-host connection to enable transfer of phage DNA into the host cell. Infection begins with phages attaching to one or more receptors on the surface of their bacterial host. Studying the mechanism by which the bacteriophage BFK20 infects the bacterial cell will contribute to clarifying the first steps of phage infection. Understanding how phages that target Gram-positive bacteria recognize their host and adsorb to the cell surface using specific cell wall receptors is crucial for developing novel detection and biocontrol tools, and for evaluating the efficacy of anti-phage mechanisms in fermentation production. The main objectives of this project are i) bioinformatic analysis of potential [B]. flavum CCM 251 receptors ii) provide supporting evidence of BFK20 infection mechanism.

Microorganisms can be used as single-cell factories for production of various bioactive compounds, including punicic acid originated from pomegranate. Punicic acid belongs to conjugated fatty acids with a wide range of beneficial nutraceutical properties. Recently, we constructed recombinant yeast Schizosaccharomyces pombe strain with the highest level of punicic acid reported for transgenic organisms. In the presented project we aim to study yet uncharacterized isoforms of long-chain acyl-coenzyme A synthetases (LACSs) from pomegranate, their substrate specificity and their involvement in the process of lipid and punicic acid accumulation. We will focus on the examination of the punicic acid turnover in yeast cell to maximize its production in the recombinant strain. The obtained results have a great potential for biotechnological production of punicic acid as well as other valuable fatty acids by oleaginous microorganisms.

Lynch syndrome is the most common hereditary cancer syndrome with a prevalence between 1/100 and 1/1000. It is caused by mutations in DNA mismatch repair genes. Lynch syndrome is associated with elevated predisposition mainly to colorectal and endometrial cancer. Colorectal cancer (CRC) is one of the most prevalent cancers in Slovakia and worldwide. Early diagnosis, which requires reliable, specific, and sensitive biomarkers, allows for faster therapy administration and reduction in mortality. Tumors are known to significantly alter protein glycosylation. Glycomic analysis as a part of the multi-omics approach is a modern method of searching for new biomarkers of various diseases, cancers included. The proposed project focuses on the identification, quantification, and monitoring of potential glycobiomarkers in the blood plasma of patients with CRC before and after colorectal resection. Experimental work is based on N-glycoprofiling using modern analytical methods of mass spectrometry and liquid chromatography with fluorescence detection. The outcome of this project will be the identification of potential glycobiomarkers of CRC and Lynch syndrome, as well as the evaluation of their potential use in diagnostics and disease monitoring.

The growth and production of plants, their yield and harvest, is largely affected by the climatic conditions. The drought is a natural phenomenon that might cause wide range of problems in agriculture. The drying of flowers and their sexual organs, and the deterioration of pollination result in lower yield, especially of cereals. During drought, plants lose the tissue flexibility and are more susceptible to mechanical damage. Furthermore, the amount of harmful reactive oxygen species in plants increases and the uptake and distribution of nutrients is inhibited. This contributes to the decrease in the photosynthetic activity. Previous studies reported that the extracts of some yeast strains have properties of plant biostimulants that are effective in the amelioration of stress caused by drought, heavy metals, high temperature, pests and pathogens. The project is focused on the selection of a suitable strain of yeast, which after direct application on maize grains (Zea mays L.) will have a positive effect on the growth of maize plants exposed to environmental stress - drought. The knowledge that will be obtained from the solution of the project has a current character and potential for use in plant production.

A method that uses the fluorescence of genetically modified bacteria is coming to the fore for the detection of infectious agents and the testing of antimicrobial procedures. These bacteria were transfected with a plasmid carrying a green fluorescent protein (GFP), which illuminated with a suitable wavelength allows monitoring the viability of these pathogens based on their fluorescence emission. By using affordable and technically undemanding in vitro models, the culturability of these pathogens can be monitored even before the in vivo testing itself. However, in vitro models cannot accurately reproduce the complex microenvironment and disease processes at the organ level. Therefore, expensive and time-consuming animal tests are often implemented, raising ethical issues. In accordance with the 3R rules, it is therefore necessary to search for new predictive screening methods. A suitable alternative to animal in vivo models is the avian chorioallantoic membrane (CAM). The CAM is a densely vascularized membrane formed extraembryonally during the development of the avian embryo. It provides a simple, rapid and low-cost in vivo model in the field of microbiology for testing the invasive capacity of pathogens, for preliminary screening of pathogenicity and interactions of inoculated microbes, genetic regulation of infection and protective effects of antimicrobial agents. The main goal of the project will be to test the culturability of GFP E. coli in in vitro conditions and then create an in vivo model of the invasion capacity of these bacteria using the Japanese quail CAM, where changes at the level of gene expression and tissue structure will be evaluated.

Congenital disorders of glycosylation (CDG) are a quite rapidly growing group of rare inherited metabolic disorders caused by defects in the complex glycosylation pathway of proteins. So far, >130 types of CDG diseases have been described, which affect individuals already in the newborn age. The disorders are accompanied by several clinical symptoms that may differ among individual CDG subtypes, what makes them difficult to diagnose. The gold standard for the diagnosis is the testing of serum transferrin, a glycosylated protein that transports iron in the bloodstream. The screening test of transferrin is a suitable method for detecting a patient with a suspected disorder in glycosylation and for distinguishing the two basic types of CDG diseases, but the will not provide any information about the specific area of the pathway where the disorder is located. CDG can also be diagnosed through molecular-genetic methods, such as whole-exome sequencing.. A suitable method is also an analytical method - mass spectrometry (MS), which is faster and less demanding. As part of this project, mass spectrometry (in our case MALDI TOF MS) is used to diagnose the N‑glycoprofile of fibroblasts and blood serum of suspected CDG patients, thanks to which we can obtain the patient's N-glycoprofile and observe changes related to the glycosylation pathway and other secondary causes. The results of the analysis play an important role not only in the understanding of protein glycosylation, but also in the search and characterization of the so-called glycobiomarkers for specific forms of CDG, but also in monitoring glycome changes during disease, in monitoring the enzymatic activity of glycosylation enzymes and possible repair of mutations.

Plant viral vectors are a perspective tool for rapid, efficient, and affordable production of recombinant proteins in plants. Proteolytic degradation of expressed proteins in plant cells or in tissue homogenates during the downstream processing of plant biomass may cause serious problems. Our recently published results demonstrated that our plum pox virus-based expression vector pAD-agro enabled production of foreign proteins of different size, structure and origin. However, their accumulation was in several cases significantly reduced due to their low stability in planta. Current strategies to increase product yields rely on the co-expression of plant protease inhibitors, silencing of protease-encoding genes, or targeting of proteins to the subcellular compartments with a lower activity of proteolytic enzymes. The aim of the presented project is to increase the yields of foreign proteins expressed in plants using the pAD-agro vector by targeting their transport to the nucleus, apoplast (secretion pathway), or their retention in the endoplasmic reticulum. The C-terminal fragment of SARS-CoV-2 nucleoprotein will be used as model polypeptide, as its detectable levels, yet significant proteolytic degradation were observed in previous experiments.

The increasing prevalence of obesity is associated with numerous human diseases, including breast and colorectal cancer. Adipose tissue, consisting of adipocytes, produces various types of adipokines. Apart from their physiological role, adipokines are involved in carcinogenesis. Adipokine leptin affects the growth and metastatic spread of various tumours. Another adipokine, adiponectin, is linked to the inhibition of tumour formation. While leptin levels are elevated in obese individuals compared to lean ones, adiponectin levels decline. Along with increased secretion of adipokines, adipose tissue is also involved in the development of hypoxia, a state of low oxygen concentration, limited nutrient supply, and the formation of an acidic extracellular environment, which worsens the prognosis of various cancers. Carbonic anhydrase IX (CA IX) is one of the most potent hypoxia-induced proteins. Its expression correlates with poor prognosis and resistance to therapy. The aim of the proposed project is to investigate the effect of adipokines on the expression of CA IX in tumour cells during normoxic and hypoxic conditions.

Cellular senescence is an important homeostasis maintenance process that regulates the elimination of redundant, damaged, and potentially malignant cells. Since its initial discovery in 1961, senescence has been characterized as a stable state of permanent growth arrest occurring due to the finite proliferative capacity of replication-competent cells. Recent ground-breaking discoveries showed, however, that even post-mitotic fully differentiated cells, including neurons, can develop a senescence-like phenotype. It was found that during natural aging but also under various stress conditions, neurons express elevated levels of multiple senescence markers (e.g. increased senescence-associated β galactosidase activity, upregulated synthesis of cyclin-dependent kinase inhibitors p16INK4a and p21Waf-1/Cip1, production of various inflammatory molecules). Although senescence functions primarily as a tumor-suppressor mechanism, research has shown that the accumulation of senescent cells and attributes associated with the senescent phenotype are among the adverse effects of aging and can significantly influence the onset and progression of various age-related pathologies, including neurodegeneration. Increased expression of senescence-associated genes was detected in multiple brain regions and cell types of animal models and patients with neurodegenerative diseases, such as Alzheimer's and Parkinson's disease. However, it is not clear yet whether elevated levels of these features specifically in neurons are involved in the pathogenesis of chronic conditions and if senescent-like neurons could represent new diagnostic and therapeutic targets in neurodegenerative diseases. Thus, the aim of our research is to analyze senescence induction and features of the resulting phenotype in human neuronal cells and subsequently evaluate their role in neurodegeneration.

The occurrence of new (sub)variant-driven infection waves across the globe indicates that the coronavirus disease 2019 (COVID-19) pandemic is not over. Continuous emergence of new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants can be anticipated. Easy to perform rapid assays able to detect the neutralising activity of the human sera even before the variant is highly abundant in the population seem to be an effective tool for evaluating the current serological immunity in humans. Since the beginning of the COVID-19 pandemic, numerous SARS-CoV-2 variants have evolved. A new variant characterised every pandemic wave. Recently, a new omicron sub-variant BA.2.75 caused concern because of its additional mutations and rapid spread in India. However, the accessibility of the specific virus variant is initially very limited and often delayed to the timepoint when the variant is already prevalent globally. Therefore, other options have been sought to perform neutralisation assays (NA) even before the clinical samples with the new variant are available in a given region. An approach called "pseudotyping" has already been shown in previous studies. It is considered a solid and reliable substitute for the live virus for testing neutralisation activity because of its safety and versatility. Our project will be based on a vesicular stomatitis virus (VSV) pseudotyped virus system containing spike (S) protein from desired SARS-CoV-2 variants, such as BA.2.75. In the project's first part, several variant-specific SARS-CoV-2 pseudotypes will be generated and NAs established. The second part will validate the assays by comparing the pseudotypes with their wild-type counterparts. Finally, the established assays will be employed in a serological study performed on a panel of human sera collected after the fourth vaccination dose to evaluate the improvement of the humoral immunity against new variants not included in the vaccine. Altogether, this project will combine highly advanced and innovative molecular biology and virology techniques to allow rapid evaluation of neutralising antibody response against new emerging SARS CoV 2 variants.

Traumatic spinal cord injury is one of the most serious diseases of the nervous system, which results in partial or complete loss of motor, sensory and autonomic functions. Despite the intensive efforts of scientific community, there is still no clinically approved and effective form of therapy for the regeneration and functional regeneration of the spinal cord after injury. The scientific community focuses on the use of neuroprotective (minimizing the extent of damage) and neuroregenerative (inducing effective spinal cord regeneration) therapeutic strategies. The aim of the project will be to explore the possibilities of regenerative therapy based on the combination of methods of electrical stimulation, exercise and supportive pharmacological treatment.

Pancreatic ductal adenocarcinoma (PDAC) represents the majority of all pancreatic cancers, and it is estimated to become the second-leading cause of cancer-related deaths by 2030. PDAC is characterized by extensive desmoplastic stroma containing cancer-associated fibroblasts (CAFs), which, together with hypovascularization, reduces the penetration of chemotherapeutics into the tumor tissue. Moreover, PDAC's aggressive nature can be attributed to early local invasion, high metastatic potential, and resistance to conventional treatment. A combination of epigenetic drugs with other therapeutic approaches seems promising to overcome the chemoresistance of tumor cells and improve the survival rate in solid tumors. We have shown that DNA methyltransferase inhibitor decitabine was able to reactivate aberrant gene expression in human PDAC cell lines. In the proposed project, we aim to test the combination of epigenetic drug decitabine with standard chemotherapy gemcitabine in vivo, using an advanced xenograft mouse model. Co-injection of CAFs with cancer cells will allow us to mimic the human PDAC microenvironment more reliably than standard models.

Testicular germ cell tumours (TGCTs) do not show a significant number of mutations, but the development of advanced next-generation sequencing (NGS) technologies has enabled the identification of new mutations, variants and markers in nuclear and mitochondrial DNA also in this malignancy. Mutations alter gene function, contributing to the progression of malignancy and resistance to treatment. Accumulated mutations can have a significant impact on therapeutic response. Several risk loci have been identified in TGCT, but the alterations contributing to the development of the chemoresistance phenotype require further characterization. To identify genetic changes that may relate to different treatment responses, we will compare TGCT cell lines resistant/sensitive to cisplatin (CDDP) using the NGS of nuclear and mitochondrial DNA. Our results might contribute to the discovery of genetic changes associated with CDDP resistance and thus potential new biomarkers for its early detection in TGCT patients.

Lipid metabolism is one of the most frequently altered metabolic pathways in virus-infected cells because cellular lipids play an important role in every stage of the viral life cycle. One of their functions is the storage of energy in the form of lipid droplets (LDs). During high energy demand, such as virus replication, LDs are used as a source of energy since they generate large amounts of ATP. This strategy is commonly used during replication by various viruses. The interactions between lipid metabolism of the host cell and arenavirus infection remain unclear. The proposed project aims to shed light on these processes during infection with lymphocytic choriomeningitis virus (LCMV), a prototypic model of the Arenaviridae family. We will focus on the formation and utilization of cellular LDs during LCMV infection. Detailed analysis of interactions between LCMV and cellular metabolic machinery will deepen our knowledge of LCMV replication strategy as well as contribute to the development of antiviral drugs targeting metabolic pathways essential for arenavirus propagation.

Therapy resistance is a major drawback of current cancer treatment and one of the main reasons of cancer treatment failure. Therefore, it is important to understand mechanisms which are responsible for development of resistance to anticancer agents. Although some of the mechanisms of resistance are already well-known, complexity of this phenomenon requires further studies since they may vary in individual patients and tumor types. Proposed project focuses on investigation of the changes that are related with resistance towards chemotherapeutic drugs in breast cancer using distinct cell culture models – derived cancer cell lines with acquired resistance towards chemotherapy and clinically relevant model of patient-derived cancer organoids. Furthermore, new method for cancer organoids immunofluorescence will be introduced within the project. New findings in this field will contribute to understanding of therapy resistance in breast cancer and may help to investigate biomarkers of therapy resistance along with development of strategies to overcome drug resistance.

Parkinson’s disease (PD) is one of the most common neurodegenerative diseases, characterized by the presence of protein aggregates called Lewy bodies (LB) which mainly consist of misfolded and fibrillary forms of alpha-synuclein (α-Syn). Many studies support the critical involvement of α-Syn, a small natively unstructured protein, and its aggregates in the pathogenesis and spreading of the disease. Understanding the pathological changes, diagnosis, and treatment of PD is one of the pressing issues of present-day neuroscience and requires development of robust disease models. Cell models are efficient tools that allow the quick and multifaceted analysis of the pathological changes associated with PD. This project represents a necessary step towards understanding the pathological processes underlying PD as we will create cellular models stably overexpressing α-Syn (wild type or its PD related mutated variant A53T). These newly generated cell models will then be employed to study in greater detail the intracellular events occurring as the result of overexpression first without and then with stress. We will mainly focus on the unexplored aspects like the effects on mitochondrial metabolism, reactive oxygen species (ROS) production and inflammation. Our results and subsequent analysis will contribute to establishing innovative approaches for treatment of α-Syn related pathologies in PD.

Slovakia is one of the countries with the highest mortality rate and incidence of colorectal cancer (CRC) in the world. Since current therapeutic methods (chemotherapy, surgical resection...) also include a number of undesirable effects, research focuses on substances of natural origin. Such an example is the substance isolated from Thymus vulgaris – thymol (2-isopropyl-5-methylphenol), which has proven to act against colorectal cancer cells. Its hydrophobic properties are limiting for wider applications. However, as preliminary analyzes showed, the newly synthesized hydrophilic derivative of thymol (acetic acid thymol ester), with a targeted modification of the chemical structure, acts more effectively on colorectal cancer cell cultures in 2D at much lower concentrations than thymol itself. For a comprehensive assessment of the effect of this newly synthesized derivative, the antitumor and genotoxic effect on colorectal cancer tumor cells in 3D culture will be determined. 3D cell culture ensures greater stability, while better representing real cell aggregation, morphology, and mutual cell interaction. As a result, the creation of a more complex microenvironment will be ensured, which to a greater extent describes the real conditions in vivo. By defining and summarizing the impact of the newly synthesized derivative on CRC tumor cell lines, its eventual use in the treatment process would be possible. It has the potential to facilitate progress that would lead to improvements in the prevention and treatment of colorectal cancer.

In Europe, semi-natural grasslands are among the most species-rich habitats and are essential for their ecosystem services in the form of climate change mitigation and food security. In recent decades there have been major changes in grassland use, in the form of intensification and abandonment, which have significantly impacted grassland quality. The European Union is committed to investing in the restoration and sustainable management of degraded ecosystems under the EU Biodiversity Strategy 2030. However, successful strategy implementation requires comprehensive knowledge of the parameters that reflect ecosystem quality. The intensity of grassland use is often expressed in the components - mowing frequency, grazing intensity, and the amount of fertilizer applied. However, in most agricultural regions, full-scale information on these operations is unavailable. This project aims to map the components of grassland use intensity. I will use advances in Earth observation, particularly in time series analysis and classification algorithms, along with existing field data to accomplish the goals. The project results will include a socio-economic dimension and can be used to map and conserve species-rich grasslands and better assess conservation policies in existing protected areas.

The shopping behavior of consumers is not stable in time and space, but it constantly changes under the influence of various internal and external factors. Among the important internal ones is the age of the consumers. The COVID-19 pandemic is one of the important external factors affecting the shopping behavior of consumers nowadays. The project is aimed at evaluating generational differences in the shopping behavior of consumers in Slovakia with an emphasis on changes in times of various anti-epidemic measures. It is a comparison of the shopping behavior of Baby Boomers, X, Y, and Z generation consumers.

The growing negative impact of climate change represents a global crisis that affects several areas. The impact of the climate crisis is also directly reflected in the quality and quantity of agricultural production. Longer periods of drought, lower amounts of precipitation and increasing daily temperatures, especially in the growing season of cultivated crops, significantly change the properties of agricultural soils. The preservation of suitable and stable properties of soils and their water regime for the purpose of growing crops is therefore very important and necessary. Recently, the use of organic materials, as soil additives, which can improve the hydrophysical properties of agricultural soils, has come to the fore. Biochar, which is a secondary product made by pyrolysis of organic waste, is also an organic material suitable for such use. Due to its positive properties and relatively wide use, biochar is currently becoming important in many studies. Improving soil characteristics and soil water regime by using organic material, such as biochar, can expand the possibilities of sustainable management of agricultural soils, as well as methods of growing agricultural crops.

The great interest in comets is caused by two reasons: (i) Comets presumably consist of pristine materials since the Solar-system formation; (ii) Comets comprise a large amount of water ice and organics, which were presumably supplied to Earth. Our work is aimed to search and study short-term color variations of dust in cometary comae of selected comets, which are observed at heliocentric distances larger than 3.5 AU (1 AU is the distance between the Earth and the Sun). For no clear reason, the comets almost constantly reveal outburst activity that is accompanied with ejection of considerable amount of dust, plausibly from subsurface layers of the nucleus. Analysis of the dust color allows to study of microphysical properties of cometary dust particles and its variations may, therefore, reflect changes in their chemical properties. Also, noticeable temporal variations of the color index of a comet significantly correlate with its outburst activity. This phenomenon is still poorly studied for distant comets. Using all our previous and new observations will be performed an exhaustive systematic investigation of selected distant comets that is aimed to study the chemical composition of dust in distant comets. We will use the numerical modelling of the interaction of sunlight with cometary dust, with help of Discrete Dipole Approximation (DDA). The principal advantage of the DDA approach is that it places minimum restrictions on the shape of target particles, making it possible to consider highly realistic model of particles whose shape resembles very much of the in situ findings in comets. Obtained results will improve our understanding of mechanisms responsible for fast dust color variations in distant comets and their relations with activity and chemical composition. Observed data, as well as the results of their analysis, will significantly enrich the physics of distant comets (including our knowledge about the nature of the cometary activity at a large heliocentric distance). Properties of chosen comets will be used to define possible differences between distant and close comets, i.e. their evolution, origin, etc. Consequently, we would improve our understanding of circumstances and peculiarities in forming our Solar system.

Geophysical methods are a non-invasive tool for identification of subsurface structures. They offer a broad spectrum of applications - they can be used in shallow or deep exploration of Earth. A more accurate picture of the structure of the examined area could be achieved by combining multiple geophysical methods. The Alpine-Carpathian region was chosen to solve the problem of a combination of gravimetric and geoelectric survey based on an intensive study of the deep structure of this area. Gravimetric research enabled creation of a high-quality map of complete Bouguer anomalies of Slovakia, which reflects the gravitational effect of density inhomogeneities in Earth's crust in the Alpine-Carpathian area. The results point to different structures in the northern part of Western Slovakia that should be verified by other geophysical methods. The ERT method (electrical resistivity tomography) has been chosen for the purpose of verification of anomalous gravity data. The method is able to distinguish vertical and slightly inclined boundaries of electrical resistivity distribution within the studied structures in the Male Karpaty and Povazsky Inovec mountain ranges. These results could broaden the existing knowledge of the tectonic structure of Western Slovakia.

The aim of this project is an improvement of crystal quality of β-Ga2O3 grown on SiC via liquid-injection metalorganic chemical vapor deposition (LI-MOCVD) method by use of vicinal (off-axis) 4H-SiC substrates. SiC substrates can offer greatly improved thermal performance of Ga2O3-based microelectronic devices. Apart from the ultrawide bandgap (~5 eV) and high breakdown electric field (~8 MV/cm), a significant advantage of Ga2O3 over its competitors is the availability of relatively cheap native (β-Ga2O3) substrates manufactured using bulk wafer production techniques. However, the major disadvantage of Ga2O3 (including the bulk substrates) is its low thermal conductivity (highest: 0.21-0.27 W/cm K in [010] direction, lowest: 0.11-0.14 W/cmK in [100] direction). This leads to reduced heat transport in the active device area, significant overheating, reduced device lifetime, and lower reliability. A significant improvement in the sinking of the generated waste heat in Ga2O3 devices can be achieved by epitaxial growth of Ga2O3 on a foreign substrate with high thermal conductivity, such as SiC (~5 W/cm K). In this project, we focus on the growth of high crystal quality epitaxial Ga2O3 layers on vicinal 4H-SiC substrates with various off-axis angles (e.g. 4 or 6°) using an established home-built LI-MOCVD system. We will investigate the material properties of grown layers and fabricate various microelectronic devices e.g. resistors, diodes, and transistor structures to study their electrical properties and compare them to those manufactured from Ga2O3 layers grown on the on-axis 4H-SiC substrates.

The development of hydrogen by electrolysis of water is one of the advanced ways in obtaining green energy. Today, platinum metals are used as the most effective catalysts in electrolyzers. Their high price and problematic availability cause electrolysis of water weak competitiveness of water electrolysis against fossil fuels. Therefore, development is focused on the preparation of new, cheaper, and more efficient electrocatalysts that would preserve the efficiency of platinum metals and the stability of both water decomposition reactions: hydrogen evolution reaction (HER) and oxygen evolution reaction (OER). In this presented project, attention is paid to non-platinum catalysts for both HER and OER with an emphasis on their electrocatalytic efficiency, overpotential applied in both half reactions, current density, and stability in acidic and alkaline environments. The project would also deal with the morphological and electrochemical characterization of phosphides and their application to electrolyzers with proton exchange membrane for testing and use in commercial applications.

This project aims the enhancement of optoelectrical and thermal properties of pn heterostructure based on n-type gallium oxide (Ga2O3) and p-type boron-doped diamond (BDD) by introducing an ultra-thin interlayers of amorphous/polycrystalline aluminum nitride (AlN) or silicon carbide (SiC). N-type Ga2O3 and p-type BDD have attracted great attention for optoelectronic device applications due to their ultra-wide bandgap (UWBG) and high breakdown electric field. However, successful fabrication of an integrated device based on Ga2O3 and BDD has been limited. This limitation arises mainly from the poor surface adherence of BDD to oxide semiconductors and harsh BDD growth conditions causing deteriorated semiconductor surfaces resulting in electronic devices with low efficiency and low reliability. Our solution for this issue is to introduce an ultra-thin AlN and SiC interlayers which can on one hand strengthen the interface between BBD and Ga2O3 layers and protect the oxide layer form deterioration during BDD deposition process, and on the other hand imply negligible changes on the optical, electrical, and thermal properties of fabricated devices.

Currently, technologies and related materials are intensively researched for energy storage. Lithium-ion batteries are one alternative. Although nowadays several materials are used as individual components for these types of batteries, global research is still focused on the synthesis and exploring new types of materials. These elements provide appropriate physical and electrochemical properties (i.e., sufficient capacity, stability, cyclability, etc.). In this way, complex oxides with a spinel structure represent a suitable alternative as anode materials for lithium-ion batteries. In addition, the modification of their composition to achieve high values of entropy (also referred to in the literature as high-entropy oxides with a spinel structure) represents one of the most researched topics in recent times. Based on it, the main goal of the project is focused on the synthesis (mainly by mechanochemical procedures) of complex oxides with spinel structure with various chemical composition. The prepared samples will be characterized by several physico-chemical methods and results will be compared with the materials described in the literature.

Shape memory microwire from Taylor UIitovsky production method is a promising final product for direct use in form of micro actuator and sensor. The promising material for this applications is Ni2FeGa alloy possesses with shape memory effect, has high production reproducibility and ferromagnetic property. The challenge is coming from the metallic core characterizations due to the micro diameter and high flexibility. Recently, we also observed that the diameter of microwire is also affecting the shape memory effect. To obtain the best statistical result of microwire diameter, observation by SEM locally and measurement by laser diffraction will be used in the whole length of microwire. Then, the crystal structures of low and high temperature phases will be studied by x-ray diffraction. In addition, to estimate the reversible martensitic transformation temperature, temperature dependence of resistivity and magnetic properties will be measured.

Frustrated magnetic systems (FMS) belong to current subject of research in the condensed matter physics. This is related with the fact, that due to the specific crystal structure and competitive exchange interactions act on the magnetic moments forces, which cannot be simultaneously satisfied by these moments. This frustration prevents magnetic ordering and leads to a degeneration of the ground state, i.e. to the formation of states with the same energy. The major achievements of FMS studies include observations of new magnetic structures or ground states, such as e.g. spin ice, quantum spin liquid, magnetic monopole analogy, etc. [1-3]. One of the reasons for our FMS study is their promising applications in field of quantum technologies. In anisotropic FMS authors [4, 5] have theoretically shown existence of “skyrmions”, the new type of magnetic vortexes with a size of a few nm, which could be suitable candidates for construction of q-bits for further development of quantum computers [6]. Considerable attention during the study of FMS with ordered magnetic moments to the two-dimensional so-called Shastry-Sutherland lattice (SSL) attracted the observation of fractional magnetization plateaus in the field dependence of magnetization [7]. Main goal of this project is to investigate the influence of pressure on properties of metallic frustrated magnetic systems (M-FMS), in particular in rare earth tetraborides TmB4, HoB4, ErB4 at low temperatures and in magnetic fields. [1] L. Balents, Nature 464 (2010) 199. [2] C. Lacroix, J. Phys. Soc. Jap. 79 (2010) 011008. [3] S. Petit et al., Nature Physics 12 (2016) 746. [4] A.O. Leonov, M. Mostovoy, Nature Commun. 6 (2016) 8275. [5] A.O. Leonov, M. Mostovoy, Nature Commun. 8 (2017) 14394. [6] Ch. Psaroudaki, Ch. Panagopoulos, Phys. Rev. Lett. 127 (2021) 067201. [7] H. Kageyama et al., Phys. Rev. Lett. 82 (1999) 3168.

The era of Covid-19 reminds us how difficult it is to treat patients with acute respiratory distress syndrome (ARDS). This project focuses on the synthesis and functionalization of magnetic nanoparticles for MRI imaging of the distribution of the drug N-acetylcysteine,or another drug, in the lungs in the aforementioned ARDS. N-acetylcysteine is a mucolytic drug commonly used in the treatment of the respiratory tract. The drug dissolves all the components that cause mucus to become viscous and thus promotes expectoration. The first step will be the preparation of a conjugate consisting of magnetic nanoparticles modified with functional groups suitable for drug conjugation. Nanoparticle functionalization and drug conjugation will be optimized and studied by physicochemical methods such as UV/VIS, SEM and TEM microscopy, Dynamic Light Scattering (DLS), Electrophoretic Light Scattering, or magnetic measurements. In the next phase, the conjugate will be analyzed by MRI and compared with the properties of commercially available MRI contrast agents. In the third step, a relevant ARDS model will be created and the prepared magnetic nanoparticles with the bound drug will be applied to the lungs. Subsequently, they will be imaged using optimized MRI techniques in order to study the spatial distribution of the drug in the lungs in ARDS. The project outputs have direct application potential for clinical practice.

Initiatives like #Metoo have raised important and complicated questions on the boundaries between what is wanted and what is unwanted in sexual interactions. Media space and public discourse often fail to reflect the complexities of sexual relationships which are situated in gender stereotypes and power inequalities. Despite decades of struggle for women's rights and sexual liberation, women are still more likely than men to face sexual violence. We still lack a deeper understanding of the so-called grey areas in sex. The present research focuses on the construction of boundaries in sex in the context of sexual subjectivity, which confronts the ambivalent expectations of liberated and traditional sexuality. We will use semi-structured narrative interviews and focus groups. The research participants will be middle-aged women, who are so often neglected in sexuality research. The research has the potential to provide new insights for designing prevention programmes as well as advocacy activities and policies regarding sexual violence definitions.

Testicular cancer is an oncological disease with good prognosis and a high level of healing. On the other hand, cancer and its treatment have a number of adverse effects which, both during and after treatment, have a long-term effect on the physical and psychological quality of life of patients. Research on continual changes in the quality of life of oncology patients occurs only rarely, and psychological research is mainly dominated by studies on female patients. The aim of the project is to monitor and describe short- and long-term changes in the quality of life in male patients with testicular cancer treated with chemotherapy. Continuous monitoring of the quality of life of adult patients will be carried out at different stages of treatment - shortly after diagnosis of the disease before, during and after treatment. These findings may be useful for the development of a personalised approach to the treatment of cancer, health and psychosocial care of patients during and after treatment.

The main contribution of the project is the development of identification of implications of the conceptual system of the Anthropocene, which could have an impact on the basic starting points of Philosophical anthropology. Identifying these consequences will be important for establishing a methodological starting point for the two-way penetration of knowledge from the field of natural sciences about the Anthropocene into the field of philosophical-anthropological reflection and vice versa. The framework hypothesis of the research is that the consequences of the temporal-spatial cumulation of anthropogenic influences on geological-climatic conditions on Earth are a qualitative change in these conditions and a change in the interpretive framework of the Earth's history from the Holocene to the Anthropocene. This hypothesis reflects claims from the field of natural science research that identified Man as a proxy symbol for the Anthropocene, which means, that there is a requirement to update the philosophical reflection of Man. The question about Man is the main problem precisely in Philosophical anthropology. Thus, research into the relevance and contribution of Philosophical anthropology to international interdisciplinary research on the Anthropocene becomes a challenge for the project.

The promotion of sobriety became a firm part of the communist programme in European countries in the twentieth century, and Czechoslovakia was no exception. The state regime's approach to alcoholism pointed not only to the economic and health consequences of this widespread problem but also to the ideological and symbolic meanings of anti-alcohol attitudes. Drinking alcohol was contrary to the ideal of both socialist and communist societies, which presupposed a man not only educated and aware, but also free from the unfortunate experiences of the past. Alcoholism was considered a socially dangerous relic of the past, and the struggle against alcoholism became part of the struggle for communism in Czechoslovakia. During the forty years of state socialism, the approach to drinking changed to a pragmatic solution to the most pressing problems of excessive drinking, and alcoholism ceased to be labelled as a relic of the capitalist past. This project focuses on anti-alcohol strategies and attitudes in Slovakia between 1948 and 1989. Through historical discourse analysis, it will analyse contemporary texts from professional periodicals, state campaigns, as well as the texts of laws and decrees aimed at solving the problem of alcoholism. The aim of the project is to show what the fight against alcoholism in Slovakia looked like, how the causes of alcoholism were explained, what legitimizing practices were used to present proposed solutions, and finally, what type of future appeared in the contemporary discourse.

The project focuses on research into the privileged transit traffic (transport between a point of origin and a point of destination on the territory of the same State) in which part of the route passes through the territory of another State) and its impact on the development of rail transport in the southern border territories of the Slovak Republic (1939-1945), which has lost strategic railway nodes, locomotives, wagons and railway stations following the Vienna Award. The issue of privileged transit traffic in the southern border territory of Slovakia (Bratislava District, Nitria District, Hron District, Tatra District and Šariš-Zemplín District) in the years 1938-1944 has not yet been examined in detail, either in Slovak or Hungarian historiography,. The aim of the project is to examine the state of the railway transport network in the southern territory of Slovakia within the framework of the interwar Czechoslovak Republic (before the Vienna Award) and compare it with the situation before the declaration of the Slovak State (the period between 2 November 1938 and 14 March 1939), thereby establishing the impact, following the November events, on rail transport and the consequent need for the creation of a privileged transit traffic on the southern border territories of Slovakia. Another objective of the research is the analysis of privileged transit traffic and its impact on the development of rail transport (construction of new railway lines) in the southern border territory of Slovakia in the period up to the outbreak of the Slovak National Uprising. Research into railway (transport) laws and regulations, the effects of the foreign economy and the requirements of Nazi Germany, as well as the Hungarian regulations on railways (transport) in the northern part of Hungary, is an essential element of the project’s implementation. Keywords: privileged transit traffic, Rail Transport, Slovak Republic (1939-1945), Slovak-Hungarian Relations, Kingdom of Hungary, Vienna Award.

The aim of the project is the analysis of the burial ritual of the Lusatian culture in Slovakia and comparison with neighbouring cultural-geographic regions on both sides of the Carpathian Arc during the Bronze Age and Early Iron Age. Specific aspects of the funerary ritual and its development in time and space will be analysed. Anthropological data will also be considered. Statistical analyses will be conducted to reach objective conclusions about the burial rituals of the population of the Lusatian culture. This research will allow the identification of specific features of funerary rituals in each burial ground, the presentation of similarities and differences in these rites between sites, with regard to their geographical location and chronology, and the comparison of the features of Lusatian culture funerary rituals in Slovakia with other, neighbouring, groups of this culture. This will help to better understand the funerary behaviour of the Lusatian culture in Slovakia, also as an element of beliefs, tradition, and cultural identification.