Current projects

Abstract:

The host immune system plays a critical role in the development and progression of sepsis, but the mechanisms that lead to cellular dysfunction, organ failure and death remain poorly defined. Peripheral blood monocytes are a
major source of the effector molecules that are closely correlated with sepsis development and clinical outcomes. We aim to dissect the signalling pathways and transcription factor networks that control monocyte function in human sepsis.

We will also use a newly-developed 3D organoid model to assess how patient monocyte signalling is influenced by lung tissue, which is the most common site of infection in sepsis and serves as a reservoir of monocytes for
release into the blood. Given that sepsis-linked changes in monocyte function alter the metabolite content of blood, we will also use a metabolomic approach to identify novel biomarkers within the volatile fraction of patient serum. This project will better define the molecular determinants of monocyte activity in human sepsis and assist the identification of new prognostic biomarkers for use in the clinic.

Abstract:

A tumour cell uses both genetic and protein weapons in its development. Gaining a greater understanding of these lethal mechanisms is a key step towards developing novel and more effective treatments. Because the metal ion metabolism of a tumour cell is not fully understood, we will address the challenge of explaining the mechanisms of how a tumour cell copes both with essential metal ions and platinum based drugs. The metal-based mechanisms help a tumour to grow on one side and to protect itself against commonly used metal-based drugs. On the other side, the exact description of these mechanisms, which are being associated with multi-drug resistance occurrence and failure of a treatment, still remains unclear. We will reveal the mechanism of the as yet not understood biochemical and molecularly-biological relationships and correlations between metal ions and proteins in a tumour development revealing the way how to suppress the growth and development of a tumour and to markedly enhance the effectiveness of a treatment.

To achieve this goal, we will focus on metallothionein and its interactions with essential metals and metal-containing anticancer drugs (cisplatin, carboplatin, and oxaliplatin). Their actions will be monitored both in vitro and in vivo. For this purpose, we will optimize electrochemical, mass spectrometric and immune-based methods. Based on processing of data obtained, new carcinogenetic pathways will be sought on cell level and proved by genetic modifications of target genes. The discovered processes and the pathways found will then be tested on two animal experimental models mice bearing breast tumours (MCF-7 and 4T1) and MeLiM minipigs bearing melanomas.

The precise description of the tumour related pathways coping with metal ions based on metallothioneins will direct new highly effective treatment strategies. Moreover, the discovery of new carcinogenetic pathways will open a window for understanding of cancer formation and development.

Abstract:

Increasing chemical pollution seriously compromises the health of ecosystems and humans worldwide. Hazardous compounds, such as polycyclic aromatic hydrocarbons, heavy metals and emerging pollutants contaminate soils/sediments, ground and surface waters. To prevent/minimise the risks associated with the accumulations of these chemicals in the environment it is key to establish low-cost/green methodologies for the treatment and redevelopment of contaminated areas. Several physico-chemical methods have been explored to remove pollutants in the environment, but these are complex, energy consuming or expensive. The exploitation of the capability of bacteria, fungi and phototrophs to transform toxic contaminants into harmless end-products, can lead instead to cheap and sustainable bioremediation alternatives.

GREENER proposes the development of innovative, efficient and low-cost hybrid solutions that integrate bioremediation technologies with bio-electrochemical systems (BES). BES, such as microbial fuel cells, break down organic contaminants
through the action of electroactive bacteria while generating electrical current. We will investigate the synergetic effect of different bioremediation strategies and demonstrate effective pollutants removal in water and soil/sediments, while generating side products of interest, such as bioelectricity. The type and entity of contamination, along with the specific physico-chemical/microbial characteristics of the environment to be depolluted, will feed into a decision-making toolbox. The latter will allow the establishment of ad hoc integrated solutions, which will take into account effectiveness of biodegradation, costs, environmental risks and social aspects. Fundamental research will be performed at lab-scale, while pilot-tests will be used to proof the scaling-up feasibility for field applications. Environmental benefits and risks, compared to standard remediation approaches, including energy efficiency, will be investigated.

Abstract:

Genus Bactrocera Macquart (Diptera: Tephritidae) belongs to the family of true fruit flies and contains over 500 species occurring in South-east Asian and Pacific regions. The genus Zeugodacus currently includes 192 species. Most species within this genus are restricted to the Oriental and Australasian Regions, with a few species reaching into the eastern Palearctic in China and Japan, except for Z. cucurbitae which was introduced into other parts of the world.

Bactrocera dorsalis, B. carambolae, B. oleae and Zeugodacus cucurbitae are considered one the most destructive pests of agricultural crops worldwide. Despite their economic importance, little is known about the production of cuticular hydrocarbons in both sexes of Bactorcera spp. and Zeugodacus. Investigating the chemical ecology of this important pests may allow shedding light on mate choice mechanisms, adding useful information to improve behavior-based control strategies. In this research we propose to ivestigate in detail the epicuticular composition of Bactrocera spp. and Z. cucurbitae males and females using two-dimensional gas chromatography coupled to mass spectrometric detection and multivariance factorial analysis together with behavioral assays.

This study will add basic knowledge to the chemical ecology of the metioned fruit flies, pointing out the potential implications for Integrated Pest Management, with special reference to the development of behavior-based control tools and new tephritid lures.

Abstract:

Organophosphorus compounds are produced as chemical warfare agents or used as insecticides and cause life endangering intoxications. For the treatment of intoxication, acetylcholinesterase reactivators (oximes) are used as causal antidotes.

Currently, clinically used or promising experimental reactivators (so-called quaternary reactivators) have major
limitation in a very limited penetration into the central nervous system, where intoxication and irreversible changes in nerve tissue also occur. For this reason, biocompatible delivery systems are developed to encapsulate the reactivator´s molecules, transfer them to the central nervous system, and then release for reactivation of organophosphorus intoxication. This project focuses on acetylcholinesterase reactivators encapsulated with apoferritin and preclinical research on their bioavailability in the central nervous system.

Abstract:

Paper-based microfluidic analytical devices (μ-PADs) significantly advance the options of rapid and low-cost portable analysis. Instrumentation-free readout based on distance-based detection enhances the value of μ-PADs due to their extremely low cost, and simplicity of use without any instrumentation required. Selective reagents have to be immobilised on the μ-PAD to achieve formation of an accurate reaction boundary allowing distance-based detection, but so far only very limited range of immobilised reagent have been used. Herein we propose new ways of reagent immobilisation, thus broadening the range of reagents that can be used. We propose to utilise electrostatic interaction on μ-PADs with paper substrate modified to carry ionic charges, and explore other approaches including paper-embedded nano- or microparticles. Finally, integrated sample preparation compatible with distance-based detection μ-PADs will be studied for the determination of selected (bio)analytes in samples with various types of matrices.

Abstract:

The proposed project deals with the optimisation and utilisation of the diffusive gradient in thin film technique (DGT) for the determination of bioavailable mercury species (Hg2+, CH3Hg+, C2H5Hg+, C6H5Hg+) in contaminated soil and aquatic environment (in the locality of Jedová hora – Brdy, Czech Republic), and with the assessing the capability of the diffusive gradient in thin film (DGT) technique to predict the bioavailability of mercury for agricultural crops and for aquatic plants. Furthermore, the influence of acid rain and soil parameters on the transport of mobile mercury forms from soil to plants will be also observed, and the contamination of the
Záskalská aquatic ecosystem, which is located at the foothills of Jedová hora, will be evaluated. The obtained results will serve to better understand the mercury bioaccumulation in similarly mercury-contaminated areas.

Abstract:

The project is focused on a detailed study of zinc-dependent signalization and its relation to expression of sub/isoforms of metallothioneins in selected subtypes of breast cancer. Research plan is a cooperation effort, connecting expertise of Prof. Haase (Technical University in Berlin) in analysis and biochemistry of zinc and zinc-dependent signalization with expertise of Prof. Adam (Mendel University in Brno) in metallomics and metalloproteomics with special emphasis on metallothioneins. In the project, we will study the influence of zinc-depletion and supplementation (in vitro and in vivo) on signal transductions, invassiveness and susceptibility to cytostatics using advanced analytical and molecular biology methods. Proposed study will enhance our knowledge on relation between zinc, metallothioneins and behavior of various subtypes of breast cancer, which is characterized by a steadily increasing incidence. The obtained data will form the basis for the use of zinc-dependent molecules as prognostic biomarkers for breast cancer.

Abstract:

Projekt využívá multidisciplinárního přístupu pro návrh, vývoj a testování pokročilých nanomateriálů, především na bázi selenu, ale i ostatních kovů, polokovů a biomakromolekulárních materiálů, jako alternativ antibiotik. Vyvíjené nanomateriály budou aplikovány do tří oblastí s cílem akcelerovat jejich využití v zemědělské praxi. Budou testovány tak, aby došlo k validaci jejich aplikačního potenciálu, a to primárně ve veterinární medicíně a rostlinolékařství, kde bakteriální resistence přináší velké socioekonomické problémy. Projekt samotný obsahuje čtyři provázané výzkumné záměry:
• VZ1 Pokročilé nanomateriály a využití proteinových klecí pro jejich cílený transport
• VZ2 Využití pokročilých nanomateriálů v léčbě mastitid u hospodářských zvířat
• VZ3 Využití pokročilých nanomateriálů pro zajištění sterility a antimikrobiality krycích materiálů v zemědělské praxi
• VZ4 Využití pokročilých nanomateriálů k ošetření rostlin vůči bakteriálním kmenům Xanthomonas campestris pv. Campestris
Hlavním výstupem projektu budou na straně jedné publikace ve vysoce impaktovaných ISI indexovaných časopisech, kam budou směřovat veškeré prezentace výzkumné aktivity výzkumného záměru VZ1, a na straně druhé tři mezinárodní patentové přihlášky, které budou výstupem VZ2, VZ3 a VZ4. Konkrétně se bude jednat o výzkum a vývoj intramamární injekce jako ochrany vůči mastitidám (VZ2), sterilního krycího materiálu s antimikrobiálními vlastnostmi (VZ3) a prostředku pro ochranu rostlin (VZ4). Všechny tři výstupy směřují do oblasti zemědělské praxe, a jejich využití bude významným socioekonomickým přínosem v tomto odvětví.

Abstract:

The aim of this project is treatment of gastro waste (GW) into a solid carbonaceous product for material utilization by drying, pretreatment with the addition of additives for the pyrolysis process. The first is focused on defining energy in dried GW, as well as pyrolysis products: biochar, pyrolysis oil and syngas. The second direction focuses on the use of biochar in agriculture. The third direction focuses on biochar as a filter medium for removing pollutants from WW. Part of the project is design, production and testing of a functional sample: drying equipment, pyrolysis and filtration units. The project will determine which direction will be acceptable from the point of view of legislation, technical solution and economy. This goal will be achieved at the end of this project.

Abstract:

The aim of the project is to support the functional diversity of soil organisms by applying stable organic matter. This is also achieved by finding methodological procedures and designing equipment for the conditioning (activation) of biochar and digestate. This will eliminate their negative impacts on the soil and improve the
ecological and economic aspects of agricultural production. The project will result in improved soil properties,
supporting functional diversity, promoting fertility and also returning carbon back into the soil. The project
will achieve certification of methodologies of the investigated technologies, design of devices for activation of
org. matter and verification of the result in semi-operation. The objectives of the project will be achieved in
relation to the field.

Abstract:

In the project, the partner’s workplace will have the task of designing and preparing magnetic particles with a defined size in the order of 100-2000 nm and defined coverage by organic molecules enabling subsequent conjugation of antigens and thus use in in-vitro immunodiagnostics. The particles will be developed with an emphasis on high stability when stored in solution (according to Testline’s requirements) and also with an emphasis on low production cost, so that production costs after all related inputs are competitive compared to globally sold analog products.

Abstract:

The aim of the presented project is the development of laboratory tests for the diagnosis of inflammation, sepsis and cardiovascular diseases. The diagnostic tests will operate on the principle of chemiluminescent immunoassay and will be compatible with an automated random access platform. The project also includes the development of a new Point-of-care testing (POCT) analyzer.

Abstract:

The thin film diffusion gradient (DGT) technique will be used to measure bioavailable forms of arsenic in contaminated soils, both in France and in the Czech Republic. The main goal of this project is to compare the DGT technique with different traditional methods of assessing the bioavailability of arsenic in soils with different physicochemical properties and also to investigate the effectiveness of these methods in predicting the absorption of arsenic by organisms. For this prediction, the conditions of use of the newly developed sorption gel Lewatit FO 36 will be optimized. The accumulation rate and the sorption capacity of the gel will be monitored. Simple and sequential extraction procedures will be optimized for fractional analysis of arsenic in soil samples. Furthermore, the influence of soil parameters (eg pH, TOC – total organic carbon, Fe and P content) on the transport of mobile forms of arsenic will be studied. Correlations between soil parameters and the content of mobile forms of arsenic will be monitored. Furthermore, the influence of soil properties in different types of soil samples (agricultural, urban and forest) will be observed.

Abstract:

The general goal of this project, which is further divided into sub-objectives, is to use modern technologies for delivery of ribonucleoproteins (RNP) and RNA to selected model organisms in order to influence the expression of desired genes and precision modifications without integration of foreign recombinant DNA into the host genome. The aim will be to assess the possibility of using the method of electroporation and the application of nanoparticles with surface treatments for the adsorption of nucleic acids. Complexes of nanoparticles and biomolecules will be further investigated in terms of prospective application for transport into cells derived from selected plant systems. The result of the project will be the creation of a robust methodology for the delivery of the required biomolecules to cells for minor modifications in the host genome regardless of the species.

An integral goal of the project will be to deepen the cooperation of Mendel University in Brno with the Krakow University of Agriculture within this and other potential projects. It will include efforts to institutionalize the technical infrastructure, while emphasizing the sharing of scientific experience and knowledge. Emphasis will also be placed on the international direction of members of research teams, including the modernization and improvement of teaching and social innovation. The aim of the project is, in addition to the stated goals, also the education of young doctoral students towards interinstitutional cooperation and internationalization in science and research. Emphasis will also be placed on the general development of education in the field of modern approaches to gene technologies, which can be considered as safe alternatives to classical approaches.

Abstract:

The proposed project seeks to contribute to scientific knowledge using modern advanced scientific methods (mass spectrometry with ambient techniques DART / DESI-MSI, scanning laser confocal microscopy) and should bring new insight into the issue of nanoparticles on plants. It focuses on the study of penetration of nanoparticles into tissues, their transport and possibly the oxidative stress caused by them and changes in the metabolome. Exposure of sown peppers to titanium dioxide nanoparticles has not been studied. Data from the literature differ on whether the mentioned nanoparticles have a biostimulating effect or, on the contrary, act as stressors. It will therefore be interesting to study their effect on another plant species.

The expected output of the project will be the publication of results in impacted professional journals, which will contribute to increasing the competitiveness and prestige of the participating workplaces. Also, deepening cooperation and establishing new contacts between Czech and Polish partners, especially in the education and internationalization of young researchers, will certainly bring positive results in terms of work and social and will become the basis for future cooperation.

Abstact:

Advances in understanding of cancer biology have only slowly been translated into improvements in cancer care. Among the main reasons belongs the lack of selective transport of anticancer drugs towards tumor tissue. With these limitations in mind, and in attempts to apply Paul Ehrlich´s “magic bullet” concept, liposomes and polymeric drugs were developed in 1960s. Recent developments in nanotechnologies are expected to improve drug delivery, primarily through enabling the drug solubilization, protection from degradation and decreasing the side effects. Nanomaterials can be functionalized with biomolecules, enabling to target specific cells within certain tissues or even specific organelles. However, only little improvements have been achieved in case of chemoresistant diseases, which is most likely due to inefficiency of cargo. Therefore, we propose to combine cutting-edge smart bionanotechnologies based on biocompatible protein nanocages – ferritins, with a heterogeneous cargo of short interfering RNA (to make cells vulnerable) and cytostatic drugs (a killing ingredient).

Abstract:

The proposed production process is a guide for the production of paramagnetic particles of defined size and defined surface modification enabling efficient separation of nucleic acids from a solution of a body fluid sample (eg blood, blood serum, plasma, sputum, etc.). The paramagnetic particles can be advantageously used for the pre-separation and pre-concentration step before the actual analysis by PCR or by another molecular diagnostic method.

Abstract:

The project will use ferritin nanocells prepared by molecular cloning and protein engineering techniques. To achieve active targeting of selected molecules overexpressed on neuroblastoma cell membranes, the surface of ferritin cages will be modified with short biomimetic peptides derived from ligands with natural affinity for these molecules, such as conotoxins, neurotrophins, neurotrophic factors and the target molecules themselves. The sequence of these peptides will be designed using in silico molecular dynamics and docking techniques. Clinically used cytostatics with high toxicity to non-target cells (such as doxorubicin) and experimental cytotoxic molecules (such as ellipticine) will be delivered. Modern biophysical and biochemical methods will be used for complex characterization of prepared nanocarriers. Much attention will be paid to specific toxicity to neuroblastoma target cells comparable to that of the free drug, while ensuring the protection of non-target cells. Ideally, the results obtained could serve as a basis for the design of selective drug nanocarriers with high translational potential for oncological nanotherapy.

Abstract:

Targeted therapy, delivering biologically active substances to tumors without interacting with the healthy cells, is the golden grail of cancer chemotherapy. Despite some therapeutics benefiting from targeted therapy have been employed in anticancer treatment, their widespread use is complicated by severe drawbacks.

The proposed project aims to establish a novel concept of smart therapy, based on active transport of enzymes for a catalytic activation of nontoxic prodrugs. In the project, advanced bio-nano delivery vehicles – ferritins for active transport of enzymes will be developed. In the project, broad spectrum of prodrugs inactivated by enzyme-sensitive protective masking moieties will be investigated. Activation kinetics of enzyme-prodrug pairs will be examined in vitro and in vivo, while a particular attention will be paid towards elimination of side-effects with a simultaneous enhancement of therapeutic efficiency.

Abstract:

The proposed production process represents the development of technology and instructions for the production of 3D printed biosensors for the detection of antibodies to SARS-COV-2 (COVID-19) virus. Decentralized detection of COVID-19 is extremely important to the global economy. 3D printing will allow decentralized instantaneous allows the preparation of sensors at the site of analysis. 3D printing composite filaments of graphene and polymer will be developed, and technology for their modification with antigen against COVID-19 antibodies will be developed. This technology will enable the preparation of sensors in thousands of pieces per day for one test site.

Abstract:

The goal of this project is to develop a unique portable system to diagnose the presence of SARS-CoV-2 virus within 20 min based on its specific RNA sequence. It will be achieved by using a microfluidic system where we load the sample potentially containing the virus of interest. The sample will be automatically processed following amplification and detection of nucleic acids, assuming the ones from the virus will be presented in the sample. Key system parameters are high sensitivity, small size and low manufacturing cost thus this proposed system would become a common tool used by mobile health workers and other security personnel of the Czech Republic. During future pandemic similar to current COVID-19 the system will be modified to quickly identify the pathogen source to be quarantine.

Abstract:

350-400 tonnes of plastic waste are produced worldwide. This amount increases every year – especially in the current pandemic situation with the usage of disposable items. PET plastics are important pollutants in the aquatic environment. It has long been known that they can form microparticles and nanoparticles that affect the health of the environment and humans. At the same time, information on the effects is incomplete, so further research is required. This project is trying to fix it, because main aim of the project will be to map the behavior of PET nanoplastics in aquatic and terrestrial environments.

Abstract:

Zinc oxide (ZnO) and zinc oxide nanoparticles (ZnO NPs) have been used in agriculture as fertilizers and feed supplements. In the European Union (EU), from June 2022, due to contamination of the environment, zinc will be allowed as a feed additive only in quantities that meet daily animal requirements. Application of zinc for extended periods of time promotes induction and spread of antibiotic resistance and virulence traits among bacteria in the digestive tract of domestic animals and the environment.

Our research is focused on biology of Escherichia coli exposed to sub-inhibitory concentrations of ZnO/ZnO NPs for 40 subcultivations. For 20 sub-culturings, E. coli will be treated with ZnO/ZnO NPs. From 20th to 40th sub-culturing, bacteria will be separated into 2 aliquots: the first aliquot will remain exposed to ZnO/ZnO NPs and the second aliquot will be without treatment. Selected sub-cultures (0th, 5th, 10th, 20th and 40th) will be exposed to antibiotics: aminoglycosides, carbapenems, cephalosporins, penicillins, sulfonamides, fluoroquinolones, and tetracyclines and minimum inhibitory concentration (MIC) will be measured to determine the effect of zinc on development of antibiotic resistance. Furthermore, the effect of zinc treatments on biology of E. coli after 40 sub-culturings will be studied by: A) genome sequencing by MiniSeq, Illumina, B) transcriptome sequencing by NextSeq 500 Illumina, C) proteome analysis by nanoflow reverse-phase liquid chromatography-mass spectrometry, and D) metabolome analysis by Agilent Technologies 6460 Triple Quad LC/MS with focus on metabolites in the Krebs cycle. Selected differentially expressed genes will be confirmed by the RT-qPCR. The obtained multiple omics data will be combined and analyzed with the goal to understand how E. coli adapts to exposure of ZnO/ZnO on genomic, transcriptomic, proteomic and metabolomic levels. PhD students will learn how to prepare, process, and analyze samples on these four omics levels in collaboration with different research groups with specific expertise at Mendelu. In addition, the principal investigator will attend an internship at the University of Koln, Germany to learn new skills in transcriptomics. The output of this project is also to publish two articles in a top peer reviewed journals.