Nucleosomes, which are typical cell death products, are elevated in the serum of cancer patients and are known to rapidly increase during radiotherapy. As both normal and malignant cells are damaged by irradiation, we investigated to which extent both cell types contribute to the release of nucleosomes. We cultured monolayers of normal bronchoepithelial lung cells (BEAS-2B, n = 18) and epithelial lung cancer cells (EPLC, n = 18), exposed them to various radiation doses (0, 10 and 30 Gy) and observed them for 5 days. Culture medium was changed every 24 h. Subsequently, nucleosomes were determined in the supernatant by the Cell Death Detection-ELISA^plus (Roche Diagnostics). Additionally, the cell number was estimated after harvesting the cells in a second preparation. After 5 days, the cell number of BEAS-2B cultures in the irradiated groups (10 Gy: median 0.03 × 10⁶ cells/culture, range 0.02–0.08 × 10⁶ cells/culture; 30 Gy: median 0.08 × 10⁶ cells/culture, range 0.02–0.14 × 10⁶ cells/culture) decreased significantly (10 Gy: p = 0.005; 30 Gy p = 0.005; Wilcoxon test) compared to the non-irradiated control group (median 4.81 × 10⁶ cells/culture, range 1.50–9.54 × 10⁶ cells/culture). Consistently, nucleosomes remained low in the supernatant of non-irradiated BEAS-2B. However, at 10 Gy, BEAS-2B showed a considerably increasing release of nucleosomes, with a maximum at 72 h (before irradiation: 0.24 × 10³ arbitrary units, AU, range 0.13–4.09 × 10³ AU, and after 72 h: 1.94 × 10³ AU, range 0.11–5.70 × 10³ AU). At 30 Gy, the release was even stronger, reaching the maximum earlier (at 48 h, 11.09 × 10³ AU, range 6.89–18.28 × 10³ AU). In non-irradiated EPLC, nucleosomes constantly increased slightly. At 10 Gy, we observed a considerably higher release of nucleosomes in EPLC, with a maximum at 72 h (before irradiation: 2.79 × 10³ AU, range 2.42–3.80 × 10³ AU, and after 72 h: 7.16 × 10³ AU, range 4.30–16.20 × 10³ AU), which was more than 3.5 times higher than in BEAS-2B. At 30 Gy, the maximum (6.22 × 10³ AU, range 5.13–9.71 × 10³ AU) was observed already after 24 h. These results indicate that normal bronchoepithelial and malignant lung cancer cells contribute to the release of nucleosomes during irradiation in a dose- and time-dependent manner with cancer cells having a stronger impact at low doses.

No abstract

A cyclosporine (CsA)-based immunosuppression is associated with an increased incidence of cholelithiasis after heart transplantation. It is not known if tacrolimus (Tac) has comparable biliary side effects in humans.We evaluated the incidence of gallbladder sludge and cholelithiasis under Tac-based immunosuppression by ultrasound examinations in 31 cardiac transplants (25 male, 6 female, mean age: 59 ± 11 years). Data were compared to 57 patients (47 male, 10 female, mean age: 58 ± 11 years) who received CsA-based immunosuppression.6 patients receiving Tac and 6 patients receiving CsA had already gallstones prior to transplantation so that finally 25 patients of the Tac group and 51 patients of the CsA group could be evaluated.In the Tac group the incidence of biliary sludge was 4% (1 of 25), of gallstones 28% (7 of 25). In comparison, patients receiving CsA developed biliary sludge in also 4% (2 of 51) and gallstones in 25% (13 of 51). Nine of 42 males in the CsA group (21%) and eight of 20 males in the Tac group (40%) developed either gallstones or sludge (n.s). Six of nine females in the CsA group (67%), but none of five females in the Tac group (0%) developed either gallstones or sludge (p = 0.01).In summary, the incidence of biliary disease in patients with Tac is comparable with CsA-based immunosuppression. We recommend regular sonographical examinations to detect biliary diseases as early as possible. In cases of clinically, laboratory and sonographical signs of cholecystitis cholecystectomy is indicated. It seems that towards lithogenicity female patients benefit more from a Tac-based treatment because the occurrence of gallstones is rare.

To assess the effects of radiation on bronchial epithelium, BEAS 2B cells cultured as monolayers and human bronchial epithelium cultured as organ cultures were exposed to single doses of 0, 10 and 30 Gy. The lactate dehydrogenase in the supernatant of the BEAS 2B cells increased markedly 24 h after irradiation, whereas in the organ cultures only a minor increase was found after 48 h. The nucleosomes in the supernatant of the BEAS 2B cells showed a massive increase in response to irradiation, whereas in the organ cultures no change could be seen. The number of BEAS 2B cells was dramatically diminished after 96 h, whereas in the organ cultures a smaller decrease was observed no earlier than 21 days after irradiation. To assess the effects of brachytherapy in bronchial epithelium in vivo, brachytherapy with 30 Gy was performed in Goettinger minipigs, and histological sections of the bronchi were analyzed for morphological alterations and cell numbers. After 2 weeks, only slight cell damage was observable, and after 3 weeks, moderate morphological changes and decreased cell numbers were found. However, after 8 weeks, the epithelium had nearly regained its normal structure. We conclude that the bronchial epithelium has a remarkably high radioresistance and that organ cultures, but not monolayers of BEAS 2B cells, reflect the effects of radiation in vivo.

OriginalienHintergrund und Fragestellung: Nach Herztransplantation (HTx) kommt es gehäuft zu einer Osteoporose. In wie weit das Auftreten eines Cyclosporin-induzierten Hypogonadismus die Posttransplantations-Osteoporose fördert und welche Therapiemaßnahmen geeignet sind, ist bislang unklar. Ziel dieser Studie war es zu untersuchen, wie häufig bei Männern nach HTx ein Hypogonadismus auftritt und inwiefern dieser sich auf die Knochendichte auswirkt. Des weiteren sollte der Effekt einer Testosteronsubstitution auf die Knochendichte hypogonadaler Patienten überprüft werden. Patienten und Methode: Anhand des Hormonstatus wurden 88Herztransplantierte einer normogonadalen bzw. hypogonadalen Gruppe zugeteilt. Bei Einschluss sowie nach 1 (n = 76) und 2 (n = 38) Jahren wurde die Knochendichte der Lendenwirbelsäule mittels Dual Energy X-Ray Absorptiometrie (DEXA) (g/cm 2 , TScore) untersucht. Alle Patienten erhielten als Basismedikation Kalzium und Vitamin D, hypogonadale Patienten wurden zusätzlich mit Testosteron substituiert.Ergebnisse: 21 Patienten (24%) zeigten unabhängig vom Alter einen Hypogonadismus. Die Knochendichte hypogonadaler Patienten (BMD = 0,8070 g/cm 2 , T-Wert = -2,6514) war gegenüber der normogonadaler (BMD = 0,9882 g/cm 2 , T-Wert = -1,0568) zusätzlich signifikant vermindert (p<0,001). Obwohl hypogonadale Patienten mit Testosteron substituiert wurden, zeigte sich bei den im Verlauf nach 1 bzw. 2 Jahren untersuchten Patienten keine signifikant stärkere Zunahme der Knochendichte gegenü-ber normogonadalen Patienten.Folgerung: Bei männlichen Herztransplantierten fand sich gehäuft ein Hypogonadismus, welcher mit einer signifikant verminderten Knochendichte einherging. Unter einer additiven Testosteronsubstitution war keine signifikante Steigerung der Knochendichte zu beobachten.Background and objective: Accelerated bone loss is a well recognized complication after cardiac transplantation (HTx). The role of an immunosuppressive-induced hypogonadism, a wellknown cause of osteoporosis in men and its prevention are less defined after HTx. The aim of this study was first, to evaluate the incidence of hypogonadism after HTx and its influence on bone mineral metabolism and second, to assess the effect of a testosterone replacement therapy in hypogonadal transplants. Patients and methods:Due to hormonal status, 88 male cardiac transplants were randomised to a normogonadal or hypogonadal group. At baseline as well as after 1 and 2 years bone mineral density (BMD g/cm 2 , T-score) was measured at the lumbar spine with DEXA. All patients received a basic therapy of calcium and vitamin D. The hypogonadal patients received additional testosterone.Results: 21 patients (24%) showed an age-independent hypogonadism. Hypogonadal transplants showed a significant lower BMD (p<0,001) (BMD = 0,8070 g/cm 2 , T-value = -2,6514) than normogonadal patients (BMD = 0,9882 g/cm 2 , T-value = -1,0568). Despite testosterone replacement hypogonadal patients showed no significant additional increase in BMD over 1-2 years compared with the n...