The effect of chronic otitis media on the immunoreactivity of human inner ear.

Twenty temporal bones (TBs) were removed from autopsy cases and prepared for immunohistochemical examination. Ten TBs were free of ear disease whereas the other ten TBs showed the signs of chronic otitis media. Expression of markers for monocyte-macrophages (25F9, 27E10) and natural killer cells (anti-Leu-11) was examined immunohistochemically. There were no specific positive stainings with 25F9 or anti-Leu-11 antibodies in any of the specimens. Staining for 27E10 was found to be negative in each section obtained from normal cochlea. However, 27E10 positivity was detected in three of ten TBs with signs of chronic ear inflammation. This positivity can be explained by two theories: (1) activated monocytes can enter the inner ear from the systemic circulation as a consequence of chronic antigen challenge; (2) mesothelial cells could become activated as a result of a cross-reaction, with resultant positivity. Development of sensorineural hearing loss in some cases of chronic otitis media may be due to these immunological reactions.

[Virologic aspects of juvenile laryngeal papillomatosis]. / A juvenilis gégepapillomatosis virológiai vonatkozásai.

Juvenile laryngeal papillomatosis is the most common benign tumor of the larynx in childhood. The specific etiological factors are non-oncogenic human papillomavirus types 6 and 11. In the present study two cases (a 6-year-old male and a 5 and a half-year-old female) operated five times each and harbouring type 11 DNA in papillomas excised in the first operations are analysed from the following virological aspects: 1. the examination of vertical transmission by general primer-polymerase chain reaction of maternal cervical exfoliation; 2. sites of papilloma predilections in the larynx; 3. histopathology; 4. viral DNA detection from the formalin-fixed and paraffin-embedded archive tissues and from a fresh papilloma tissue in one case by polymerase chain reaction applying type-specific primers. We did not find any signs of maternofoetal transmission in the anamnesis and the maternal cervix proved to be negative for viral DNA. However, the vertical route of transmission can not be excluded due to the special natural history of papillomavirus infections. Papillomas usually localised in normal squamociliary junctions of the larynx. The histopathologic review did not reveal any signs of malignancy. Koilocytosis referring to productive viral infection and the signs of abnormal keratinisation were present in each tissue. All tissues of the patients proved to be positive for the short amplimer deriving from the genome of human papillomavirus type 11.

Apoptosis in the human inner ear. Detection by in situ end-labeling of fragmented DNA and correlation with other markers.

The aim of this study was to obtain baseline data on the recently described special form of single cell death, apoptosis, in normal human inner ears. For this purpose, in situ end-labeling of the fragmented DNA was applied, in conjunction with apoptosis-related markers, to detect cellular elements showing programmed cell death in decalcified and paraffin-embedded tissues. Over 20 specimens were analyzed which were obtained from autopsy cases with no history of acoustic lesions confirmed by histopathology. Based on staining results, we saw no apoptotic signs in the majority of normal adult inner ears. An apoptotic cell captured in the Reissner's membrane of the cochlea from an old patient may, however, indicate an age-related subtle cell loss with the process of apoptosis. Nevertheless, the fact that more apoptosis was not found in our cases suggests that this phenomenon does not contribute significantly to the tissue homeostasis in the adult inner ear under normal conditions. These data are in accordance with our immunohistochemical findings on the p53 nucleoprotein, and proliferating cell nuclear antigen expression since there was no staining in any of the cellular elements, including the mesenchymal cells. This reflects a stationary and stable condition of cells of the vestibular and the cochlear structures, probably to maintain their integrity and the fine sensory functions. As opposed to the above findings, during inner ear development, the epithelial cells lining the cochlear lumen, the ossifying cartilage of the temporal bone, and the mesenchymal cells show different degrees of proliferation in combination with single cell death as signs of maturation of the vestibular and the cochlear apparatus. In addition, apoptosis has been demonstrated in cells of the cochlear stria vascularis from an adult patient treated with high doses of cisplatin, vinblastine and bleomycin prior to death. Furthermore, a wide range of apoptosis could be induced experimentally in a normal ear by an external perfusion of actinomycin D (ActD), which is known to produce programmed cell death in many cell types of different origins. The potential role of cytostatic agents in the apoptotic process of the inner ear needs, however, to be confirmed in large-scale specimens from patients treated with genotoxins. The fact, however, that apoptotic cells are also seen in association with ActD indicates that the fine sensory structure of the cochlea may also be a target for certain chemotherapeutic agents when administered in high doses.

S-methylthio-cysteine and cystamine are potent stimulators of thiol production and glutathione synthesis.

The effects of methylthio-cysteine disulfide (MT-Cy) and cystamine (CAM) on the thiol production and glutathione content of a human T cell line (CEM-SS) have been investigated. MT-Cy per se and CAM in the presence of cystine greatly enhanced thiol production and glutathione content of cells while cystine alone exerted no or slight influence in the first hours. The MT-Cy- or CAM-induced extracellular SH-generation was observed both in a complete nutrient medium and even more in SH-free D-PBS. The acid-soluble thiol level and glutathione content of cells elevated markedly (up to 5-6 fold in two hours) when incubating cells in complete medium. Inhibition of glutathione synthesis by DL-buthionine (S,R)-sulfoximine did not alter the MT-Cy- or CAM-induced extracellular thiol production indicating that glutathione synthesis is not involved in this effect. The results suggest that MT-Cy easily enters the cells thus accelerating the thiol cycle in SH-poor medium while CAM promotes cystine uptake into the cells. Phenylalanine and leucine inhibited both MT-Cy- and CAM-dependent thiol production in D-PBS most effectively suggesting the involvement of the L membrane transport system in these effects.

Sulfhydryl groups generated by macrophages into the culture medium.

Previous data showing that thiols can functionally replace macrophages in certain in vitro lymphocyte reactions have raised the possibility that macrophages generate SH groups in the medium. The SH activity of cell-free medium decreases considerably due to auto-oxidation. The presence of macrophages inhibited the spontaneous decrease of SH activity and even increased the number of free SH groups in the culture media. This was designated as SH generation. Resident as well as in vivo stimulated (NaIO4, thioglycollate medium, paraffin oil or BCG) macrophages have nearly the same capacity to generate SH groups when cultured in vitro. The SH-generating ability of macrophages depends on the viability and density of cells, and is greatly influenced by the serum concentration of the media. The majority of SH groups produced by macrophages could be demonstrated as albumin SH groups and proved to be more resistant to autooxidation than non-protein thiols. Moreover albumin could replace whole serum in respect of SH generation. It is suggested that macrophages by generating SH activity, may exert a non-specific helper function to lymphocytes or other cells in their environment.

Effect of heparin on the reactivity of SH-groups in the macrophage membrane.

The effect of heparin on macrophage (M phi) adherence and on the reactivity of membrane SH-groups to the specific SH-oxidizing agent 4,4'-dithiodipyridine (PDS) was studied. Various types of SH-reactive agents, except 5,5'-dithio-bis (2-nitrobenzoate) (DTNB), were found to inhibit adherence of mouse peritoneal M phi to serum-coated Falcon surfaces. Heparin inhibited M phi-adherence in serum containing medium and in higher concentrations stimulated the adherence inhibitory effect of PDS, especially in Ca-depleted medium. This effect of heparin may be due to its polyanionic character, as dextran sulphate but not dextran induced similar changes. The effect of heparin to increase the sensitivity of membrane SH-groups against SH-reactive agents was demonstrated also by cytotoxicity experiments. It is concluded that heparin makes the M phi-membrane unstable, by exposing some hidden SH-groups playing a role in membrane function.

Immunodiffusion analysis of the antigenic structure of the transplantable MC 29 tumor.

By the use of specific anti-chicken liver sera it could be proved with immunodiffusion techniques that the transplantable MC 29 tumor line contains liver-specific antigens. Production of chicken serum-proteins by the tumor could also be demonstrated. Thus, the transplantable MC 29 tumor line can be regarded as a hepatoma.