Unspecific redness of the eyelid--two malignant tumours.

BACKGROUND: Chronic, non- or little pigmented reddish or brownish lesions on the lid margin and the surrounding skin are frequently underestimated and thus carry a clear risk of malignancy. HISTORY AND SIGNS: A 61-year-old lady was referred with a chronic, reddish lesion in the medial third of the left lower lid after a topical therapy over a period of 4 months had not improved the situation. She noticed the lesion for the first time 10 previously. Recurrent trichiasis and a slight ulceration had developed during the last year. THERAPY AND OUTCOME: After an unclear result of the biopsy (2)/ (3) of the left lower lid were removed because of atypical cells ascending into the epidermis. A superficial spreading melanoma 0.4 mm depth in the Breslow classification was detected and right next to it a superficial basal cell carcinoma. The lid defect was reconstructed with a tarsoconjunctival flap and a free full thickness skin graft as described by Hughes. CONCLUSION: Chronic redness at or around the lid margin which does not respond to therapy should be biopsied as malignant tumours may well be hidden behind it.

Contribution to the biological assessment of orthodontic acrylic materials. Measurement of their residual monomer output and cytotoxicity.

The acrylic materials used in orthodontics for the fabrication of removable appliances are subjected in the oral cavity to processes of change which influence their physical, mechanical and biological properties. It is therefore essential that every newly developed material must be judged in terms of its clinical value. In the present study, 2 orthodontic cold-cure acrylics, Orthocryl and Forestacryl, and 4 orthodontic photocure acrylics, Triad, Wil-O-Dont, Odontolux and Lux-A-Tech, were investigated and compared with 2 prosthetic acrylic materials: the cold-cure acrylic Palapress and the hot-cure acrylic Paladon. The quantity of residual monomers from methyl methacrylate (MAA) or urethane dimethacrylate (UDMA) eluted from the sample in a given time after the processing was estimated by high pressure liquid chromatography (HPLC). The cytotoxic properties of the materials were examined by Mosmann's proliferation-inhibition test with an established culture of fibroblasts (= MTT test). The hot-cure acrylic Paladon produced by far the smallest amount of eluted residual monomer and the least growth inhibition in the MTT test. The prosthetic cold-cure acrylic Palapress achieved significantly better results than the orthodontic cold-cure materials Orthocryl and Forestacryl. The photocure acrylics released less UDMA than did the cold-cure acrylics MMA. In the cell culture test, all the orthodontic materials examined were assessed as "slightly cytotoxic"; the prosthetic acrylics were graded under ISO-standard 10993-5 as "noncytotoxic". After soaking the plastic material in water for 3 days its cytotoxic properties, as exemplified by the cold-cure acrylic Forestacryl and the photocure acrylic Triad, were reduced, and during the following investigation no more inhibition of growth was observed. It was possible to confirm with the tests used that, for Triad, it is necessary to carefully remove the oxygen-inhibition layer of the photocure acrylic in order to improve the biological properties. The influence of the plastic material on fibroblast cultures was assessed, among other methods, by the quantity of residual monomers liberated. These were significantly reduced after soaking the manufactured substance in water for 3 days. Careful laboratory treatment of the photocure acrylics is necessary in order to improve their biological properties.

Radiation-induced alterations of the proliferation dynamics of human skin fibroblasts after repeated irradiation in the subtherapeutic dose range.

PURPOSE: The aim of this study was to determine the effect of single and multiple irradiations on the differentiation and proliferation pattern of the stem cell system of human fibroblasts. METHODS: The pattern of differentiation of fibroblast cultures was analyzed by morphological criteria using colony forming assays. Proliferation rates were assessed by cell counting and measuring the incorporation of BrdU. RESULTS: Ionizing radiation both in low and high dose ranges exerts differential effects on the cellular processes of differentiation and proliferation in human fibroblasts. Single irradiations of fibroblasts in the dose range of 1 to 8 Gy induced terminal differentiation into postmitotic fibrocytes at high percentage level. Irradiation of long-term cultures of fibroblasts with repeated doses of 0.2, 0.6 and 1.0 Gy revealed that only in cultures, which were irradiated repeatedly (x 10) with 0.6 and 1.0 Gy a marked reduction of the proliferation capacity was apparent. Inhibition of proliferation by repeated irradiations with cumulative doses up to 10 Gy was not more pronounced as compared to single irradiations. CONCLUSION: 1. These results of radiation-induced changes in the proliferation and differentiation pattern of cells may be a basis for the understanding of the cellular processes leading to radiation-induced fibrosis and tissue aging. 2. Multiple irradiations with single doses up to 1 Gy and cumulative doses up to 10 Gy did not change the radiosensitivity of fibroblast cultures regarding effects on cell proliferation.

Mutant analysis suggests that cyclic GMP mediates the cyclic AMP-induced Ca2+ uptake in Dictyostelium.

Previous work has shown that streamer F (stmF) mutants of Dictyostelium discoideum exhibit prolonged chemotactic elongation in aggregation fields. The mutants carry an altered structural gene for cyclic GMP phosphodiesterase resulting in low activities of this enzyme. Chemotactic stimulation by cyclic AMP causes a rapid transient increase in the cyclic GMP concentration followed by association of myosin heavy chains with the cytoskeleton. Both events persist several times longer in stmF mutants than in the parental strain, indicating that the change in association of myosin with the cytoskeleton is transmitted directly or indirectly by cyclic GMP. We measured the cyclic AMP-induced Ca2+ uptake with a Ca(2+)-sensitive electrode and found that Ca2+ uptake was prolonged in stmF mutants but not in the parental strain. The G alpha 2 mutant strain HC33 (fgdA), devoid of InsP3 release and receptor/guanylate cyclase coupling, lacked Ca2+ uptake. However, the latter response and cyclic GMP formation were normal in the signal-relay mutant strain agip 53 where cyclic AMP-stimulated cyclic AMP synthesis is absent. LiCl, which inhibits InsP3 formation in Dictyostelium, blocked Ca2+ uptake in a dose-dependent manner. The data indicate that the receptor-mediated Ca2+ uptake depends on the InsP3 pathway and is regulated by cyclic GMP. The rate of Ca2+ uptake was correlated in time with the association of myosin with the cytoskeleton, suggesting that Ca2+ uptake is involved in the motility response of the cells.

A high-affinity plasma membrane Ca2+-ATPase in Dictyostelium discoideum: its relation to cAMP-induced Ca2+ fluxes.

Chemotactic stimulation of Dictyostelium discoideum induces an uptake of Ca2+ by the cells followed by a release of Ca2+. In this study we investigated the mechanism of Ca2+ release and found that it was inhibited by La3+, Cd2+ and azide. Ca2+ release occurred in the absence of external Na+, indicating that an Na+/Ca2+ exchange was not involved. Plasma membranes contained high- and low-affinity ATPase activities. Apparent K0.5 values were 8 microM for the major Mg2+-ATPase and 1.1 microM for the high-affinity Ca2+-ATPase, respectively. The Mg2+-ATPase activity was inhibited by elevated concentrations of Ca2+, whereas both Ca2+-ATPases were active in the absence of added Mg2+. The activities of the Ca2+-ATPases were not modified by calmodulin. The high-affinity Ca2+-ATPase was competitively inhibited by La3+ and Cd2+; we suggest that this high-affinity enzyme mediates the release of Ca2+ from D. discoideum cells.

Oscillations of Ca++ concentration during the cell differentiation of Dictyostelium discoideum: their relation to oscillations in cyclic AMP and other components.

Periodic cyclic-AMP pulses control the cell aggregation and differentiation of Dictyostelium discoideum. Another component required for the aggregation and differentiation of these cells appears to be extracellular Ca+ +. Oscillations in extracellular Ca+ + concentration were investigated in suspensions of differentiating cells. We observed spike-shaped and sinusoidal Ca+ + oscillations. In the course of differentiation, spike-shaped Ca+ + oscillations preceded sinusoidal oscillations, and no phase change occurred at the transition from spike-shaped to sinusoidal Ca+ + oscillations. Spike-shaped and sinusoidal Ca+ + oscillations were related to oscillations in (1) the cyclic-AMP and cyclic-GMP content of cells, (2) the light-scattering properties of cells, and (3) the extracellular pH. Spikeshaped Ca+ + oscillations were observed together with cyclic-AMP oscillations. The minima of the extracellular Ca+ + concentration trailed the maxima of the cyclic-AMP concentration by about 30 s. Sinusoidal Ca+ + oscillations were not accompanied by measurable cyclic-AMP oscillations. The amplitudes of the sinusoidal Ca+ + oscillations were smaller than those of the spike-shaped Ca+ + oscillations. A Ca+ + oscillation of small amplitude (instead of a spike-shaped oscillation) was observed when one cyclic-AMP spike was skipped. Our results provide evidence for the existence of a sinusoidal cyclic-AMP-independent Ca+ + oscillation of small amplitude, and they also suggest that spike-shaped Ca+ + oscillations may be superimposed on such small-amplitude oscillations. When D. discoideum cells produce cyclic-AMP spikes, the uptake of additional Ca+ + is induced, resulting in Ca+ + oscillations of a large amplitude.

Attractant-induced changes and oscillations of the extracellular Ca++ concentration in suspensions of differentiating Dictyostelium cells.

We used a Ca++-sensitive electrode to measure changes in extracellular Ca++ concentration in cell suspensions of Dictyostelium discoideum during differentiation and attractant stimulation. The cells maintained an external level of 3-8 microM Ca++ until the beginning of aggregation and then started to take up Ca++. The attractants, folic acid, cyclic AMP, and cyclic GMP, induced a transient uptake of Ca++ by the cells. The response was detectable within 6 s and peaked at 30 s. Half-maximal uptake occurred at 5 nM cyclic AMP or 0.2 microM folic acid, respectively. The apparent rate of uptake amounted to 2 X 10(7) Ca++ per cell per min. Following uptake, Ca++ was released by the cells with a rate of 5 X 10(6) ions per cell per min. Specificity studies indicated that the induced uptake of Ca++ was mediated by cell surface receptors. The amount of accumulated Ca++ remained constant as long as a constant stimulus was provided. No apparent adaptation occurred. The cyclic AMP-induced uptake of Ca++ increased during differentiation and was dependent on the external Ca++ concentration. Saturation was found above 10 microM external Ca++. The time course and magnitude of the attractant-induced uptake of external Ca++ agree with a role of Ca++ during contraction. During development the extracellular Ca++ level oscillated with a period of 6-11 min. The change of the extracellular Ca++ concentration during one cycle would correspond to a 30-fold change of the cellular free Ca++ concentration.