The influence of interleukin-1 and interleukin-6 on expression of proteins involved in steroidogenesis in Leydig cells and on their apoptosis or necrosis in vitro.

This paper presents a study on the expression of proteins involved in steroidogenesis in rat Leydig cells: StAR protein and aromatase, in relation to the age of animals and to the influence of interleukin-1 and interleukin-6. In addition, their effect on apoptosis and damage of Leydig cells was investigated. Leydig cells were isolated from rats at certain age intervals and exposed to different concentrations of the above-mentioned cytokines. Next, total RNA was isolated from Leydig cells and RT-PCR with primers specific for rat StAR protein and aromatase was performed. The products were analysed by agarose gel electrophoresis. Leydig cells were also stained with propidium iodide and Hoechst 33342 to estimate the percentage of apoptotic or necrotic cells. The results seem to confirm the role of interleukins in the control of expression of enzymes and regulatory factors involved in steroidogenesis in rat Leydig cells, and in the regulation of their apoptosis and necrosis. The effect of the studied interleukins on Leydig cells is exerted during development and after adolescence.

An example of application of molecular techniques to the study on the local regulatory system controlling the function of the rat male gonad.

Molecular biology techniques help to study details of testicular cell interactions, which play an important role in the control of spermatogenesis and secretory function of the male gonad. This paper presents an example of application of such techniques, with evaluation of their usability, in the study on expression of some cytokines involved in cellular interactions in the rat testis. Testicular cells were isolated by digestion with collagenase followed by percoll gradient centrifugation or by the adherence technique (macrophages). Then RNA was isolated from cells using Chomczynski-Sacchi method and analysed by electrophoresis and spectrophotometry. RNA was reversely transcribed using oligo-dT15 primer and AMV (Avian Myeloblastosis Virus) transcriptase. cDNA was amplified by PCR with primers for rat IL-1alpha, IL-1beta, IL-6 and Tbr (Thermus brockianus) polymerase. RT-PCR products were analysed by electrophoresis. Chomczynski-Sacchi method proved to be effective for isolation of good quality RNA even from small number of cells. RT-PCR revealed the presence of mRNA for studied cytokines in isolated cell. The results indicate that the applied techniques are useful in studies on the function of male gonad and indicate the need for use of semi-quantitative methods for evaluation of cytokine expression.

Immunocytochemical observations on the organ distribution of exogenous monomeric and dimerized lysozyme.

Using commercial anti-lysozyme antibodies and anti-dimerized lysozyme rabbit serum produced by us we demonstrated by immunohistochemistry, in some organs of rats, the expression of exogenous egg white lysozyme preparations beside the native lysozyme. After oral administration, the egg white lysozyme was detected in intestinal epithelium, proximal and distal tubules of some nephrons, pulmonary alveolar walls and hepatocytes in the 3rd zone of liver acini, whereas native lysozyme was strongly expressed in intestinal and pulmonary macrophages in both the experimental and control animals. However, expression of the dimerized lysozyme released from the intraperitoneally implanted mini-osmotic pumps and detected using specific antisera was evident only on erythrocytes in intestinal blood vessels. It is concluded that the lysozyme preparations administered per os or parenterally are resorbed to blood circulation and distributed among various organs in an active form and maintaining their antigenic specificity. It may speak for their direct anti-inflammatory and immunomodulatory effects in respiratory, urinary, digestive and other systems.

[The treatment of tracheal stenosis in Wegener's granulomatosis by laser surgery]. / Leczenie zwezenia tchawicy w przebiegu choroby Wegenera za pomoca lasera.

A case of Wegener's disease is described. In eight years of remission a patient, treated with cyclophosphamid and steroids demonstrated sterosis of the trachea and upper way obstruction. We have performed carbon dioxide laser photoresection of granulotion mucosis and scars with an excellent results.