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Biomedical subjects

M Koukal

Publications and source records attributed to M Koukal.

11 recordsLinked to original sources

Phagocytosis of synthetic particles in earthworms. Effect of antigenic stimulation and opsonization.

We examined the phagocytic activity of coelomocytes in earthworms Lumbricus terrestris. The synthetic polymeric HEMA particles were used in this study. The phagocytic capacity was increased after the protein administration or preincubation of HEMA particles with coelomic fluid. Furthermore, we isolated and characterized the proteins adsorbed on the surface of the particles which have opsonin-like properties.

Animals↗

Monoclonal antibodies against pig transferrin. Blocking and binding activity.

Monoclonal mouse anti-pig transferrin antibodies PTF-01, PTF-02 and PTF-03 and anti-human transferrin antibody HTF-14 detect transferrin coupled with Sepharose particles in an indirect immunofluorescence test. Only the PTF-03 antibody can be used for immunofluorescence detection of pig transferrin bound to specific receptors on the plasma membrane. The binding of iodinated pig transferrin to PK cells was studied. It could be blocked by non-labelled transferrin in excess, by pig serum or by anti-pig transferrin monoclonal antibodies. PTF-03 expressed the lowest blocking activity among the antibodies tested.

Animals↗

Polyester film as an advantageous tool for light and ultrastructural examination of single cells infected with viruses.

The use of marked transparent polyester film as a support of cell monolayer, smears or sections (cryostat or paraffin), makes possible to examine the same cell preselected in the light microscope by electron microscope (EM). Consequently, this technique enables to use histological or immunohistological investigations (using immunofluorescence or peroxidase (Px) labelling) by light microscopy before examination in EM. As example we used herpes simplex virus (HSV) type 1-infected cells grown on the above mentioned polyester film. Visualization of the antigen with hyperimmune serum and Px-labelled antiserum was followed by examination of selected cells in EM, where viral particles were clearly visible.

Cells, Cultured↗

Selective decontamination, induced colonization resistance and connected immunological changes in piglets.

14-d-old conventional piglets were picked from normal piggery, washed with disinfectants, placed into isolators suitable for germfree work, fed a sterile diet and treated with peroral antibiotics (nalidixic acid, kanamycin, and nystatin). Beginning with day 5 or 7, Enterobacteriaceae were not found in feces. The absence of these bacteria was proved by inoculation of germfree newborn piglets with caecal content. In selectively decontaminated piglets, the white blood cell count in blood had fallen to 6 X 10(9)/L; this decrease was due to an extremely low number of granulocytes (to 0.8 X 10(9)/L). On day 35, IgG-positive cells almost disappeared from the spleen, whereas IgA cells were found in an unusually great amount. Corresponding changes in serum levels were established. The colonization resistance effect in Enterobacteriaceae-deprived piglets was confirmed; settling of introduced various E. coli strains did not occur or was delayed.

Animals↗

Phages in eukaryotic cells.

Urine from a patient after kidney transplantation added to a human embryonic lung culture caused the appearance of dark spots shown to contain cells with phages resembling group A, as well as bodies resembling Chlamydiae.

Chlamydia↗

[Use of colloidal gold in ultrastructural cytochemistry].

Colloid gold stabilized by macromolecules with various binding characteristics proved to be usable in transmission electron microscopy: Nucleic acids localization was studied in tissue cultures after embedding by techniques autoimmune serum-protein A--gold and RNase--gold. Technique with serum autoantibodies against double-stranded DNA resulted in marking condensed chromatin. Further experiments with ultrastructural localization of other autoimmune components seemed to be perspective and of diagnostic importance. Technique with RNase--gold complex marked especially ribosomes, nucleolus and interchromatin nuclear spaces. Herpes virus, rotavirus and enterovirus were identified in negative contrast by the technique antivirus serum--protein A--gold. A higher warrant of evidence achieved by the method may be used in virologic diagnosis. A small amount of tubulin was found in isolated permeabilized nuclei of Xenopus laevis by a direct immunocytochemical method with complex monoclonal IgG against tubulin-gold. Binding of complex of triiodothyronine-bovine serum albumin--gold on the cytoplasmic membrane of LEP cells in a short term tissue culture showed a possibility of tracing non-peptidic hormones binding on specific receptors.

Animals↗