ESR12: Toxicology of logic gates and evaluation of logic gates in an in-vitro blood-brain barrier model and ex-vivo

at Centre of Experimental Medicine at the Slovak Academy of Sciences/Slovakia
Institute of Experimental Pharmacology
Dr. Mojmir Mach (mojmir.mach@savba.sk)

Candidates with a background in cell biology, toxicology, biochemistry, chemistry, molecular biology or related fields are invited to apply for a PhD position in the department of Cell Pharmacology and Developmental Toxicology (head: Dr. Mojmír Mach) at the Institute of Experimental Pharmacology and Toxicology, Centre of Experimental Medicine, Slovak Academy of Sciences.

The nitric oxide radical represents one of the most versatile mediators not only in the vascular, but also in the nervous and immune systems. The wide range of physiological and pathological effects are partially induced by the reactivity of the molecule, which is able to modify biomolecules including proteins, lipids and nucleic acids. However, its availability and biological function may be hampered by reactivity with other reactive species, such as superoxide anion (O2•-) or H2O2, which play also significant roles in physiological and pathological processes. Furthermore, these reactions give rise to the products with the highly cytotoxic impacts on the cells. Thus, the development of the novel probes allowing simultaneous detection of intracellular NO along with other associated reactive species, especially at the cellular level, is important.

The topic of PhD study will focus on evaluation of cytotoxicity and applicability of novel fluorescent reporters designed for simultaneous multicolour detection of reactive nitrogen (RNS) and reactive oxygen species (ROS) at the cellular level. The cytotoxicity will be evaluated by standard assays (such as MTT, neutral red uptake assay or cell cycle analysis) in the cellular models (microglia, fibroblasts, endothelial cells). The probes specificity will be evaluated in cell and cell-free systems exposed to different ROS and RNS as well as in the systems simultaneously generating these species (such as LPS-stimulated microglia, as the brain resident macrophages) using fluorescence-based methods. The probe applicability will be validated in such cellular models as microglia or brain microvascular endothelial cells. The methods based on novel molecular logics will be compared with standard methods of detection of RNS and ROS and confronted with biological consequences and signalling pathways of RNS and ROS generation (such as the expression of iNOS protein or proteins involved in regulation of the endothelial function).

Secondments: study of metabolomic approaches to assess cytotoxicity of sensors in in-vitro cell cultures (one month at University of Leiden/Netherlands, evaluation of microfluidic artery models as experimental platform for pharmacological research (2 months at the company Mimetas/Netherlands), and study of signalling pathways of endothelial dysfunction (2 months at Jagiellonian University of Krakow /Poland).

Requirements: Master/Engineering degree in Biochemistry, Pharmacology, Chemistry or related sciences. Good knowledge and interest in both experimental and theoretical work. Interest in the field of cell biology is required. Knowledge in the field of ROS-sensitive probes are great assets. As an equal opportunity employer IEPT CEM SAS is committed toincrease the percentage of female scientists and therefore especiallyencourages them to apply

Planned start date: 1st June 2019

Application deadline: 31st March 2019

Contact: Applications should be sent to Dr. Mojmir Mach at mojmir.mach@savba.sk