Laboratory of Bacterial and Tumor Resistance
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DOC. ING. JITKA VIKTOROVÁ, PH.D.
Group Leader
During her Master's studies she was dedicated to the preparation of transgenic plants useful for phytoremediation, her PhD was dedicated to the recombinant production of substances for medical purposes under the supervision of Prof. Martina Macková and Prof. Tomáš Macek. She spent her postdoc in the group of Prof. Tomáš Ruml, where she was involved in the introduction of high throughput methods for bioprospecting. She is now trying to apply her broad methodological background and collaborative skills to establish a scientific group focused on solving the antibiotic crisis by finding new antibiotic and chemotherapeutic adjuvants.
„The future belongs to those who believe in the beauty of their dreams“ Eleanor Roosevelt
ING. BÁRA KŘÍŽKOVSKÁ, Ph.D.
Thesis topic: Inhibitions of efflux pumps in resistant bacterial strains
The main focus of her PhD studies was to search for compounds capable of inhibiting efflux pumps in resistant bacterial strains using semi-automated methods with EtBr as a fluorescent substrate. She is also interested in measuring inhibition of bacterial biofilm formation and antibacterial testing in general.
Internships and courses:
Postgraduate course Antimicrobial Susceptibility Testing with EUCAST Criteria and Methods in Tallinn, Estonia.
21st ESCMID Sumer School in Seville, Spain
Internship at Universidad de Magallanes in Punta Arenas, Chile
Internship at the University of Szeged, Hungary
Methods: Antibacterial activity testing; bacterial adhesion inhibition testing; measurement of bacterial efflux pump inhibition (EtBr accumulation and direct efflux); cytotoxicity detection on cell monolayers; genotoxicity measurement (HPRT or γH2AX).
Thesis topic: Mechanisms of bacterial antibiotic resistance and their modulation
My dissertation focuses on a deep understanding of the bacterial mechanisms of antibiotic resistance and the search for possible ways to modulate these mechanisms. My main goal is to identify and characterize inhibitors of particular bacterial resistance mechanisms that could be used in adjuvant therapy.
Specifically, I am focusing on staphylococcal infections, which pose a significant clinical problem due to their ability to develop resistance to commonly used antibiotics. I use a wide range of microbiological and molecular biological methods in my work.
My long-term goal is to contribute to the development of effective strategies to combat antibiotic resistance. By identifying new inhibitors that can be used in combination with existing antibiotics, I aim to improve the treatment of resistant infections and reduce their incidence.
Methods: antimicrobial activity and sensitization testing using microdilution, nucleic acid isolation, polymerase chain reaction (PCR) and quantitative PCR (qPCR) for detection and quantification of specific genes and for gene expression analysis.
Methods: antimicrobial activity and sensitization testing using microdilution, nucleic acid isolation, polymerase chain reaction (PCR) and quantitative PCR (qPCR) for detection and quantification of specific genes and for gene expression analysis.
ING. ONDŘEJ STRNAD
Thesis topic: Determination of anti-inflammatory activity of phytocannabinoids and their potential in the treatment of chronic inflammatory diseases
During my PhD studies, I have specialized in inflammatory and stress responses of cells. The main focus of my research is atherosclerosis, which I study as an inflammatory disease of the vascular wall. My research focuses on potential anti-inflammatory drugs, particularly phytocannabinoids. In addition, I am studying the NFκB and Nrf2 signalling pathways, which are crucial for regulating cellular responses to stress and inflammation.
Internships and courses: Internship at Universidad de Magallanes v Punta Arenas, Chile
Methods: ELISA, reporter assays, transfection, Western blot, preparation of immunoliposomes for tumour cell targeting
ING. TOMÁŠ NEJEDLÝ
Thesis topic: Introduction of new models for testing multiple drug resistance in cancer
In our laboratory, I am dedicated to the search for new agents capable of suppressing the resistance phenotype of cancer cells and thus increasing their sensitivity to chemotherapy. Selection of active compounds is performed on a monolayer of cells from a collection of drug-resistant tumour lines. The mechanism of action is then studied by monitoring the activity of transmembrane efflux pumps (P-gp, BCRP, MRP1 ...), expression of genes of signalling pathways involved in drug resistance (ARE, KEAP1) or cellular response to oxidative stress. Furthermore, I am working on the preparation of advanced 3D cell models that allow to study drug resistance in an environment mimicking in vivo conditions. In addition to spheroids consisting of a single cell line, I also introduce tumour models cultured together with components of the microenvironment (fibroblasts, adipocytes, endothelial cells, macrophages...). Among the most advanced models under development for the study of tumour drug resistance is the preparation of organoids derived from biopsies of cancer tissues from cancer patients.
Internships and courses:
Internship at Universidad de Magallanes in Punta Arenas, Chile
Training school in 3D bioprinting and modelling of pathophysiological multicellular systems, Davos, Switzerland
Pan-European Educational Platform on Multidrug Resistant Tumours and Personalised Cancer Treatment (PANDORA)
Methods: Cytotoxicity testing of agents (cell monolayer, 3D cell models); Modulation of multidrug resistance in tumours (cell monolayer, 3D cell models); Fluorescent substrate accumulation; Solubilization of transmembrane proteins SMA nanodiscs; RT-qPCR; CAA (Cellular Antioxidant Activity); ORAC (Oxygen Radical Absorbance Capacity); Preparation of spheroids (hanging drop, low-attachment plates, coating wells); Preparation of Patient Derived Organoids (PDOs)
ING. ANNA LUDVÍKOVÁ
Thesis topic: Mechanisms of antibiotic resistance in Pseudomonas aeruginosa and possibilities of their modulation
I am dealing with antibiotic-resistant gram-negative bacteria of the species Pseudomonas aeruginosa and testing potential adjuvants on them. At the same time, I am cloning single genes causing antibiotic resistance into Escherichia coli. These modified E. coli strains are then used to test antimicrobials and potential adjuvants or to isolate the relevant enzymes and determine their activity.
Methods: PPCR, agarose gel electrophoresis, nucleic acid isolation, protein isolation and purification, antimicrobial activity testing and sensitization
ING. EMÍLIE KUČEROVÁ
Thesis topic: In vitro preparation of the blood-brain barrier for prediction of potential drug transfer
I am involved in the preparation of an in vitro model of the blood-brain barrier by co-culturing three human cell lines. This model will be further refined with cells resistant to selected antiepileptic drugs and used to develop potential antiepileptic drugs effective in patients with drug-resistant epilepsy. Furthermore, I am working on the preparation of an in vitro intestinal barrier model consisting of Goblet cells theirs function is mucus production in addition to standard epithelial cells.
Methods: work with cell lines and preparation of human barrier models, measurement of transmembrane resistance, cytotoxicity assays, permeability assays, PCR, STR analysis in the cell genome
ING. PETRA TŘEŠŇÁKOVÁ
Thesis topic: Testing substances with the potential for use in the dermocosmetic industry
I am involved in the preparation of in vitro skin models that can be used for cytotoxicity testing of skin cells, or that may reveal anti-cancer potential. I am also investigating the potential antimicrobial activity of these compounds against common skin pathogens. I also study inflammatory processes in skin cells and investigate the possible antioxidant activity of the tested substances. In addition, I am analysing the effect of these substances on wound healing.
Methods: cytotoxicity testing, antimicrobial activity testing, CAA (Cellular Antioxidant Activity), ELISA, testing the effect of substances on wound healing
ING. JAN ŠPAČEK
Thesis topic: The use of nanopore sequencing to suppress antibiotic resistance in bacteria
During my Master's studies, I introduced a method to identify compounds capable of suppressing the resistance phenotype of bacteria and thus restoring the efficacy of conventional antibiotics. I also learned how to work with the nanopore sequencing method, including evaluating and working with the results. It is this method that I would like to use in my PhD studies to study resistance genes in bacteria, including their transcription, and then use the information to gain a deeper understanding of antibiotic resistance.
Methods: genome and transcriptome sequencing of bacteria using Oxford Nanopore technology, isolation and transformation of bacterial plasmids, determination of minimum inhibitory concentration of antibiotics by microdilution method, method for identification of antibiotic adjuvants
BC. JAN JANOUŠEK
Lab Technician
I take care of the running of the laboratory, making sure that the things needed for the work are ready and replenished. I order materials, service instruments and other matters necessary for the running of the laboratory. I also take care of preparing, shipping and receiving samples. I passage cells and perform cytotoxicity tests using various cell lines.