Výzkumný záměr Ú.Imunologie / Research Activity of D.of Immunology

Assurance of the realization of the research plan in the framework of existing spatial, material and technical conditions in the workplace of the administrator

The following Palacký University departments will participate in the project:

• Department of Immunology
• Department of Microbiology
• Department of Electron Microscopy
• Clinic of Orthopedics
• Department of Physical Chemistry
• Clinic of Pulmonary Diseases and Tuberculosis
• Clinic of Hematooncology

Individual departments have sufficient personal and technical resources necessary for specialized activities declared in the research plan.

Department of Immunology UP
The department specializes in work based on microbiology, molecular biology and biochemistry methods. For such purposes it has been fully equipped. The laboratories for molecular methods utilize equipment for work with DNA constructs and recombinant proteins. Specifically, these are: the flow-box Jouan for sterile work, PCR cycler MS Research PTC - 200, spectrophotometers Jenway 6405 UV/Vis and Spekol 11, gradient pump Beta 10, HPLC equipment, centrifuges Jouan MR 1812, Hettich mikro 22 R and Jouan KR - 25 enabling work with 0.2 – 250 ml containers. At our disposal we also have the ultracentrifuge Beckman Optima L-90K, equipment for agar electrophoresis of nucleic acids and its evaluation (UV transilluminator with a CCD camera). The laboratories contain equipment for culturing microorganisms (constant temperature shaker) and their lysis (tissue homogenizer Ultra-Turrax T 25 and ultrasound homogenizer Hielscher UP 200 S with accessories). The laboratories naturally have usual equipment such as the pH meter, electronic scales for analyses, magnetic shakers, vortexes etc. Also the microbiological laboratory is equipped with a flow-box and culture boxes. Besides, there is a 3-liter semi-continuous fermented made by MBR. The room is air-conditioned. The immunochemistry laboratory is equipped for polyacrylamide electrophoresis of proteins (protean III) and subsequent western blotting and immunodetection on membrane. The ELISA method can also be routinely performed in the laboratory. The tissue culture laboratories are fully air-conditioned and separated by a sanitary loop from the remaining part of the department. They are autonomous, i.e. use their own equipment (flow-boxes, culture boxes, inverted and normal microscopes, centrifuges) and material which is specially treated for use in tissue cultures. The methods routinely performed here involve for instance blast transformations of lymphocytes in vitro enabling to assess specific cellular immune response by incorporation of radioactively-labelled thymidine. The microscopy room is equipped with three microscopes of which particularly one is worth mentioning – the Axioskop fluorescence microscope with an Olympus CCD camera for documenting preparations visualized both by traditional staining or phase shift and by direct or indirect immunofluorescence. Cooperation has been initiated with the Department of Electron Microscopy led by RNDr. Radko Novotný.. Material storage at the department is secured by a cold room with a constant temperature of 4°C, a -80°C freezer box and Dewar flasks with liquid nitrogen pro long-term deep freezing.. The department owns autoclaves for material sterilization and equipment producing distilled and deionized water.. Each laboratory is equipped by its own sets of single-channel and multi-channel automatic pipettes which limits the risk of contamination of processed materials. The department has legislation, material and spatial conditions for work with laboratory animals – the Department for Work with Laboratory Animals accredited for keeping laboratory mice, rats, guinea-pigs and rabbits.
The Department of Immunology has been accredited by the European Federation for Immunogenetics, the laboratory areas of individual institutes comply with standards for biosafety levels BL2/3.

Department of Microbiology UP
Besides standard equipment for traditional bacteriological procedures, the institute is ready for the specific project goals as it has three separated areas for pre-PCR, PCR and post-PCR techniques which are equipped with centrifuges, a biohazard box, a PCR box, 3 common thermocyclers (Minicykler, PTC 200, Robocykler), 1 capillary cycler (RapidCycler), 1 cycler for real-time PCR (RotorGene), devices for common electrophoresis and pulsed-field gel electrophoresis, software for 2D gel analysis (GelCompar II) and a high-resolution melter HR-1 (Idaho Technology, Inc.), so far unique in the Czech Republic..

Electron Microscopy Facility UP
Department of microscopic methods MF UP Olomouc serves as service and research workplace. For this purpose it is equipped with transmission electron microscope JEOL JEM 2010 with accelerating voltage 60 – 200 kV and with high resolution ability and magnitude 50x – 800,000 x. For video recording it is equipped with special digital camera system MORADA with resolution 1941x1920 and potency of the film recording. Department is equipped with scanning electron microscope TESLA BS 340. The workplace is equipped with all instrumentation necessary for specimen preparation. The staff has longtime experience with preparing and processing assorted animal and plant samples.

Clinic of Orthopedics UP
The clinic has 72 beds used for urgent and planned care of patients with injury or diseases of kinetic apparatus. More than 2800 patients are hospitalized in the clinic during one year. Infection diseases treatment is the integral part of the medical care. Those are solved at separate septic surgery and hospitalized at detached rooms with special regimen. Diagnostic of deep periprosthetic infection is the long-term research program of the Clinic of Orthopedics Department of Microbiology, Clinic of nuclear medicine, Department of Pathology and laboratory section of the Faculty Hospital in Olomouc is. At above mentioned workplaces are investigated samples from joint tissues (joint punction, arthroplasty dross, tissue materials), blood samples and the scintigraphic investigation are performed as well. Therapeutics of these serious complication is intensively studying too. Up to 20 infected prosthesis joint replacements are treated at the clinic within one year. It presents 1/10 up to 1/5 of all revision surgery of total joint replacement.

Department of Physical Chemistry UP
For the project purposes, the department has two teaching laboratories available for selected research activities as well and five research laboratories (with the main nanomaterial laboratory located in Olomouc-Holice research centre). The research laboratories has both standard equipment required for activities common in chemistry (laboratory centrifuges, thermostats, pH-meters, a gas chromatograph, spectrophotometers) and special devices necessary for the basic project goals - potentiostats, luminometers, tensiometers, a rotation viscosimeter, EPR spectrometer, a device for studying particle size by dynamic light diffusion made by Brookhaven. A modern electron microscope JEOL 2010 is located in workplace shared with the Faculty of Medicine, Palacký University in Olomouc.

Clinic of Pulmonary Diseases and Tuberculosis UP
The clinic has 64 acute care beds, an extensive out-patient section with counselling centres including those for pulmonary inflammations and for interstitial pulmonary processes. These issues are exclusively dealt with by one of the four departments of the clinic. The bronchology department carries out more than a thousand of bronchology procedures each year including bronchoalveolar lavages, transbronchial and percutaneous lung biopsies in interstitial pulmonary processes. In cooperation with the clinics of nuclear medicine and radiology, diagnostics is complemented with the latest imaging techniques (lung HRCT, PET/CT, gallium scintigraphy and MRI). The clinic laboratories can assess specific cytokines in cooperation with biochemical, immunological and nuclear medicine laboratories. Targeted samples from lower airways are processed either in our cytological laboratory or in cooperation with microbiological laboratory specializing in modern methods for detection of typical and atypical pathogens. The bronchology department is equipped with 12 bronchoscopes of flexible as well as rigid types. The cytological laboratory has an Olympus BX 41 microscope with an ARTCAM – 300 Mi digital reader, 4 medical microscopes made by Zeiss Jena and other laboratory equipment.

Clinic of Hematooncology UP
The clinical part includes an out-patient department for 4 physicians working simultaneously, a day centre for out-patient chemotherapy, separation units with two blood cell separators, an in-patient department with 18 beds in double rooms, a 16-bed intensive hematology care unit and a transplant unit with six beds in reverse isolation with HEPA air filtration. The transplant unit has been accredited by the EBMT.

No extension of the spatial capacities will be required by the project.