Head of Laboratory

Agnieszka Ścibior, PhD, DSc

(cellbiol@kul.pl)

 

Research interests

The research area includes analysis of the effects of selected elements on the cells and mammalian organisms. It is important to know the relationship between the excess or deficiency of certain elements in the body and pathological conditions that may occur. It is also relevant to recognize the consequences of interactions between the examined metals. The question of interactions between elements is always up-to-date and it is still an important issue in toxicology. The knowledge in this research field is not only of cognitive but also of practical value. The studies are conducted in two experimental models: in vitro (on Chinese hamster ovary cells, CHO-K1 line) and in vivo (on rats).

The research area includes the following issues:

  • toxicity of selected elements and mechanisms of their toxic action;
  • consequences, character, and mechanisms of metal interactions;
  • analysis of changes in markers of oxidative stress and nephrotoxicity in body fluids/tissues under the influence of selected elements;
  • evaluation of iron metabolism indices in the conditions of exposure to some metals;
  • assessment of changes in red and white cells and in platelets under intoxication with selected metals;
  • cytometric analysis of the degree of maturation of reticulocytes;
  • analysis of alterations in the level of micro- and macroelements and their mutual proportions in biological fluids/tissues in the condition of exposure to selected metals,
  • research on the influence of some elements on the bone morphology at the microscale*,
  • analysis of the chemical composition of bone with the X-ray photoelectron spectroscopy (XPS) method*;
  • seeking the most sensitive biomarkers to assess organism reactions to some metals,
  • seeking an 'antidote' to limit the toxicity of vanadium and to alleviate its strong prooxidative properties;
  • studies on the mechanisms underlying the potential protective role of magnesium in vanadium poisoning;
  • evaluation of the cytotoxicity induced by some elements in the CHO-K1 cell line by means of the following assays: (a) the resazurin reduction test, detecting mitochondrial metabolic activity, (b) the neutral red uptake test, assessing lysosomal damage, (c) trypan blue staining, detecting plasma membrane permeability, and (d) the sulforhodamine B assay, measuring total cellular protein content;
  • analysis of cell morphology by phase-contrast microscopy* collaboration with the Laboratory of X-Ray Optics.

Keywords referring to the research area:

(a) in vivo experimental model: rats, vanadium, magnesium, vanadium-magnesium interaction, bioelements, mechanisms of toxicity, oxidative stress, lipid peroxidation, antioxidants, total antioxidant status,  biomarkers, organs/tissues, blood/urine, nephrotoxicity, urinalysis, bone morphology, bone formation markers, mineral status, haematological parameters, markers of iron status, metal transporters, surface roughness parameters, experimental toxicology, clinical chemistry.

(b) in vitro experimental model: CHO-K1 cells, vanadium, selenium, vanadium-selenium interaction, cytotoxicity, toxicity mechanism, oxidative stress,  neutral red, resazurin, trypan blue, fluorimetric measurements.

Techniques/methods:

       Microwave digestion, Atomic Absorption Spectrometry (AAS), UV-Vis Spectrophotometry, Flow cytometry, Microscopy, Potentiometry, ELISA Immunoenzymatic method.

Laboratory of Oxidative Stress is equipped with:

  • Speedwave Four Microwave Digestion System with a 12-position rotor; temperature and pressure monitoring using a contact-free sensor in each vessel with the control of the course and reading (Berghof)
  • Z-2000 Atomic Absorption Spectrometer with two atomizers (Flame and Graphite Furnace Atomization) (Hitachi)
  • Automated EasyLyte Na/K/Cl Analyzer (Medica)
  • Automated BS-120 Biochemistry Analyzer (Mindray)
  • Automated XL-640 Veterinary Analyzer (ERBA Diagnostics Mannheim GmbH)
  • ADVIA 2120i Haematology system with veterinary software (Siemens)
  • Laura Smart urine analyzer (ERBA Lachema)
  • Synergy 2 Multi-Mode Microplate Reader (BioTek) equipped with the following detection modes: UV-visible Absorbance, Fluorescence, Luminescence, and Fluorescence polarization
  • ELx50 Microplate Strip Washer (BioTek)
  • DTS-4 digital thermostatic micro plate shaker (Elmi Laboratory Equipment)
  • Flow cytometer: MACSQuant Analyzer 10 (Miltenyi Biotec)
  • Heracell 150i CO2 Incubator with copper chamber (Thermo Fisher Scientific)
  • HERAsafe KS 12, Class II Biological Safety Cabinet (Thermo Fisher Scientific)
  • SH 12/24 hematological adder
  • IX73 Inverted Microscope (Olympus) with the CellSens Imaging Software 
  • Double Beam Spectrophotometer U-2900 (Hitachi) with 2J1-0104 Temp Controller ED-5 (Hitachi) and Heating Circulator with Open Bath (Julabo)
  • XA 100 3Y.A analytical balance (Radwag)
  • PS 210 R.2 and PS 2500 3.Y precision balances (Radwag)
  • UP 50H Compact Ultrasonic Homogenizer (Hielscher Ultrasonics GmbH)
  • BioGen PRO200 Homogenizer (PRO Scientific Inc.)
  • Heraeus Megafuge 11R Refrigerated Centrifuge (Thermo Fisher Scientific)
  • HLP Spring 5R water system (Hydrolab)
  • Haake DC10-P5/U Heating Circulator with Open Bath (Thermo Fisher Scientific)
  • HeraFreeze HFU 486 Basic (Thermo Fisher Scientific)
  • LCv 4010 Freeze-Refrigerator (Liebherr)
  • TC240 Thermocenter Oven (SlavisLab)
  • Shaking WNB 14 water bath (Memmert)
  • ONE 45 oil bath (Memmert)
  • Polwater DL-100 V 717 Demineralizer (Labopol)
  • DEM-20 Distiller
  • Computer with Graphic Design Software (CorelDRAW Graphics Suite X6 PL Win Box)
  • Equipment for animal rooms: (a) Tecniplast IVC system for rats (Green Line), (b) Scantainer Z-11E ventilated cabines, (c) Tecniplast metabolic cages for rats with refrigerated collection chillers, (d) CS5 Evo Plus cage chainging station, (e) surgical table with a temperature controller (RWD Life Sciences), (f) Circus LUXO magnifying lamp with additional lenses (4x and 6x), (g) CUBIS MSU 12012S-1CE-DO precision balance
  • with a software for weighing small laboratory animals (Sartorius)