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H V Tuczek

Publications and source records attributed to H V Tuczek.

At least 19 recordsLinked to original sources
PubMed1990

[Interpretation of the estrogen receptor content of breast cancer].

The biochemical analysis of estrogen receptor (ER) content, using the DCC (dextran-coated charcoal) method, was compared with different plotting methods of the estrogen-receptor immunocytochemical assay (ER-ICA) in 80 primary breast cancers including 9 metastases under routine conditions. It was evident, that the determined content of estrogen receptors depends on the technique of measurement, as well as the microscopic organizations of the individual carcinomas and should be interpreted in respect of their content of stroma and if possible of tumour heterogeneity.

Antibodies, Monoclonal↗
PubMed1990

Application of the chromogenic reaction to conventional silver staining, the Ag-NOR staining and the silver-intensified immunogold technique.

The principle of the chromogenic reaction and the transformation of "black and white" histochemical staining results or immunohistochemical signals to coloured microscopic images is described. The chromogenic reaction was optimized and is, so far, possible with either cyan-blue or magenta-red reaction products. The application of the chromogenic reaction to conventional silver stain was optimal in the Lendrum staining resulting in red or blue stained reticulin fibres. The Ag-NOR staining of the nucleolus organizing region (NOR) could be transformed by the same reaction to coloured reaction products as well as the silver-intensified immunogold technique in immunocytochemistry.

Arthritis, Rheumatoid↗
PubMed1989

Immunocytochemical detection of sulphonylated DNA in tissue sections. An alternative to Feulgen staining of DNA.

We report here on a new sensitive and highly specific DNA staining technique which we have called sulpho-DNA staining. DNA staining is based on a sulphonylation reaction of 2'-deoxycytidine or cytidine that takes place in the 6th position of cytosine with ensuing immunodetection of the sulphonylated DNA. The specificity of DNA staining is introduced by the use of an antibody recognizing only modified DNA but not modified RNA, by recourse to an additional acid hydrolysis step which destroys RNA but not DNA. We describe here the optimal conditions for the sulphonylation of DNA using O-methylhydroxylamine and metabisulphite as reactants. The new DNA stain labels all nuclei in either normal human tissue or in tumor cells. For nuclear DNA the staining signal is higher for the sulpho-DNA staining than for the Feulgen staining for nuclear DNA. This new DNA staining technique is suitable for use on tissue sections as well as on cytosmears.

Animals↗
PubMed1989

Quantitative immunohistochemistry: standardization and possible application in research and surgical pathology.

We describe three different methods of calibration of quantitative immunohistochemistry, two of them based on self-made dye-agarose or antigen-agarose substrates. Quantification of immunostained objects (immunosignals) can be done by (1) microscope photometry (plug method) or scanning-photometry; (2) and densitometry of a television transformed microscope image. Both approaches are subjected to the different methods of calibration and three examples of application.

Arginase↗
PubMed1989

Histological classification of synovitis.

A simple system of synovitis classification is proposed based on histomorphological evaluation of 400 random specimens of synovial tissue. It discriminates between two types of diagnosis: 1) one with a high degree of information, being called C-synovitis (synovitis with a characteristic or diagnostic histomorphological pattern), where the clinical diagnosis can be predicted on the basis of the histomorphological pattern, 2) a second type of diagnosis with a lower degree of information, being called NC-synovitis (synovitis with non-characteristic or non-diagnostic morphological findings in the synovial membrane). We subdivided NC-synovitis into 4 subgroups and determined the RA-probability of each subgroup in order to permit clinical use of the proposed histomorphological classification. In addition, by using immunoperoxidase staining methods for immunoglobulins the subgroups were shown to have different quantities of extravasally deposited immunoglobulins in the synovial membrane. The subgroup with the highest RA-probability also turned out to have the highest amount of extravasally deposited immunoglobulins.

Fibroblasts↗
PubMed1988

Immunophotometric quantification of extravascular immunoglobulin deposits in the synovial membrane of patients with osteoarthritis and rheumatoid arthritis.

The amount of extravascular immunoglobulin deposits in the synovial membrane of patients with osteoarthritis and rheumatoid arthritis was studied in comparison with that of patients suffering from non-joint diseases. Immunoglobulin deposits were immunostained using the three-layer immunoperoxidase method. The staining results were quantified with the help of a microscope photometer connected with a scanning stage. Several experiments involving artificial test substrates, diseases with allegedly increased extravascular deposits or diseases not exhibiting extravascular deposits of immunoglobulins are used to validate the described microspectrophotometrical approach for measuring extravascular immunoglobulin deposits. The scanning photometry demonstrates significantly higher amount of extravascular immunoglobulin deposits in rheumatoid arthritis as compared with osteoarthritis and non-joint diseases.

Arthritis, Rheumatoid↗
PubMed1988

Use of lectin-immunohistochemistry in joint diseases.

The lectin binding pattern of the inflamed synovial tissue of patients with OA (osteoarthritis) and RA (rheumatoid arthritis) were examined with 18 different lectins. The synovial lining cells are heavily stained by PHA-L, WGA, PNA, Con A, MPA and RCA. In the subsynovial layer, WGA, PHA-L and RCA were found to be fairly good monocytic markers. The lectin binding pattern of the lectins under study, however, failed to discriminate between patients with OA and RA.

Arthritis↗
PubMed1987

Immunophotometric analysis of immunoglobulin synthesis and deposition in synovial membranes of patients with rheumatoid arthritis.

Immunophotometrical measurements of the Ig synthesis of plasma cells in joint capsules of patients with rheumatoid arthritis and osteoarthritis were compared. Patients with rheumatoid arthritis showed higher mean values in the single plasma cell than patients with osteoarthritis for all Ig isotypes except IgG. These results are indicative for a B cell activation with increased synthesis of Ig in RA.

Arthritis, Rheumatoid↗
PubMed1986

Colour development of immunogold-labelled antibodies for light microscopy.

We describe an improvement of the immunogold technique, which is based on the colour development of silver-intensified immunogold signals. This method (referred to as the coloured-SIG technique) was found to be as sensitive as the silver-intensified immunogold method and more sensitive (in two of the three tested systems) than immunoenzymatic procedures, such as the peroxidase/antiperoxidase technique or the avidin-biotin system. The coloured SIG-method results in either a magenta-red or a cyan-blue final reaction product. Therefore, this new improvement of the immunogold technique should be useful in double-staining methods, immunoblot methods and conventional immunostaining methods.

Antibodies↗
PubMed1986

In situ evaluation of immunoglobulin synthesis of synovial plasma cells in rheumatoid arthritis and osteoarthritis by plug photometry.

Synovial membranes of patients with osteoarthritis--a disease with unaltered immunoregulation--and rheumatoid arthritis were immunostained for the different immunoglobulin isotypes using the PAP-method and a two-layer alkaline phosphatase technique. Randomly selected plasma cells with positive immunostaining were measured by plug photometry. The average extinction of the single plasma cell is considered to be indicative of the immunoglobulin synthesis of the measured plasma cell and the isotype under study. When we compared both disease entities, patients with rheumatoid arthritis were characterized (i) by higher average extinction values for all Ig-isotypes except IgG (ii) by a higher rate of high producer plasma cells. From this data, we conclude that immunophotometry is a new in vivo method for discriminating two types of synovitis (i) one without increased Ig-synthesis at the single cell level and (ii) a second one with increased Ig-synthesis.

Antibody-Producing Cells↗
PubMed1985

Distribution of albumin in normal and regenerating livers of mice. A light microscopic immunohistochemical and autoradiographic study.

The conditions affecting the immunohistochemical identification of albumin in livers of male NMRI-mice were investigated by light microscopy. In normal livers albumin is randomly distributed, revealing a pancytoplasmic nearly homogen reaction in groups of hepatocytes or single parenchymal cells. However, combined autoradiographic studies after pulse labelling with 3H-valine and perfusion experiments with human albumin indicate that this distribution is caused by albumin from blood plasma and does not reflect true protein synthesis. After perfusion of the livers followed by immunohistochemical amplification techniques which allowed to dilute the primary antibody up to 1:30,000, albumin could be detected nearly in all liver parenchymal cells as granular deposits decreasing in its density from periportal fields towards the terminal hepatic venules. In regenerating livers due to partial hepatectomy no remarkable differences in granular albumin deposits between G1- and S-phase of the cell cycle could be detected as was demonstrated by combined immunohistochemistry and 3H-dThd-autoradiography. However, during mitosis the content of albumin was often considerably reduced.

Albumins↗
PubMed1985

[Immunohistochemistry: theoretical potentials and practical application].

Immunhistochemical methods are increasingly used and their application in surgical pathology is obvious. Nine different immunohistochemical techniques are compared in this review article. The peroxidase anti-peroxidase method of Sternberger et al. (1970), the avidin biotin complex method of Hsu et al. (1981) and the labeled avidin biotin technique of Guesdon et al. (1979) are to be preferred. Two step methods using labeled antibodies such as alkaline phosphatase or peroxidase-labeled second antibodies are less sensitive. However, two-step methods using alkaline phosphatase-labeled antibodies offer the advantage of double staining procedures since alkaline phosphatase reacts with different azo dyes and results in a wide spectrum of colours in the final reaction product. At present, the major problems of immunohistochemistry in daily work are the selection of the fixative and of appropriate controls. Though both problems remain unresolved, immunohistochemical methods should be used for special problems in surgical pathology.

Alkaline Phosphatase↗
PubMed1977

Synchronization of hepatocellular DNA synthesis in regenerating rat liver by continuous infusion of hydroxyurea.

Hydroxyurea (HU) inhibited DNA synthesis in the livers of partially hepatectomized rats. After an i.v. infusion of HU begun in the late G1 phase and continued for periods of up to 30 hr, all hepatocytes scheduled to embark on DNA synthesis in a characteristic intralobular sequence during this time interval accumulated at the G1-S boundary. The effective dose was 1.25 mmoles/kg/hr, preceded by a single injection of 1.69 mmoles/kg. Serum levels of HU rose slightly during continuous infusion and decreased after termination, with a half-life of about 80 min. Liver weight increased during HU infusion. The G1-S blockade was rapidly reversed in the liver after the end of HU infusion. The specific activity of DNA increased to a maximum between 3 and 5 hr. [3H]Thymidine labeling indices reached about 80%. Intralobular distribution of labeled hepatocytes was congruent to the pattern seen in partially hepatectomized rats after a continuous [3H]thymidine infusion of equal duration. The beginning of DNA synthesis in nonparenchymal cells was delayed, as compared with hepatocytes. Vincristine infusion for 12 hr after release from the HU block arrested about 40% of the hepatocytes in mitosis, indicating that a large fraction of cells progressed through the cycle after the prolonged HU block. Partially resected rat liver appeared to be rather resistant to the unfavorable consequences of "unbalanced growth" during the protracted inhibition of DNA synthesis, providing a useful model for synchronization of DNA synthesis in a differentiated resting organ triggered into active growth.

Animals↗
PubMed1976

Analysis of cell cycle compartments of hepatocytes after partial hepatecomy.

An analysis of cell kinetic parameters as a function of intralobular localization of hepatocytes was performed using autoradiographic methods to obtain a basis for a quantitative description of cell cycle compartments after partial hepatectomy. The influx into the S compartment revealed a maximum in a lobular zone not immediately adjacent to the portal tract, with a decrease towards the perivenous and periportal area of the lobule. The maximum influx was found in the intermediate zone at 34 hr and a lower one in the perivenous parts of the lobule at 40 hr. The influx pattern at 56 hr was similar to the situation at 18 hr. The fraction of labelled mitoses in the whole liver and in subunits of the lobule disclosed a constant duration of S whereas the duration of G2 + M was prolonged from the periportal to the perivenous zone. A graphical model of kinetic events after partial hepatectomy is proposed, which describes the sizes of the various cell cycle compartments; the model successfully predicted the results of a continuous labelling experiment.

Animals↗