15.Radiation survival and colony size of human epidermal keratinocytes in the presence of keratinocyte growth factor (rhKGF)

The capacity of recombinant human keratinocyte growth factor (rhKGF) to ameliorate the radiation response of mouse oral mucosa and other epithelial tissues was recently reported. However, the exact mechanisms of action of KGF remain unclear. The aim of the present study was to investigate the effect of rhKGF on survival and colony size of normal human epidermal keratinocytes in vitro. Primary human neonatal keratinocytes (HEKn) were irradiated with doses of 0 Gy and 2 Gy (200 kV X-rays) and incubated in the presence or absence of 100ng/ml rhKGF. Plating efficiency (PE) and surviving fraction (SF2) were determined in a clonogenic assay. In cell cultures without rhKGF the mean PE was 4.6%. Irradiation with 2 Gy resulted in a SF2 of 51%. In cell cultures with rhKGF, the mean PE was identical (4.6%). After irradiation with 2 Gy, a similar SF2 of 54% was observed, indicating that KGF did not change the survival characteristics of HEKn keratinocytes. Individual colony size, however, in all cultures incubated with rhKGF was significantly increased compared to incubation without rhKGF. The number of extremely large colonies (□2 mm) was clearly higher (p=0.0000) with rhKGF-containing culture medium. In conclusion, rhKGF does not affect keratinocyte survival after irradiation, but stimulates proliferation of surviving cells

26. Chromosomal damage and survival of keratinocytes and fibroblasts after irradiation with 200 kV or 25 kV X-rays

A relative biological effectiveness of 1 is accepted for soft X-rays (25–30 kV), which are applied in diagnostic radiology (mammography). However, it has been shown that soft X-rays can be more effective in cell killing and chromosomal damage. The present study was initiated to define biological effects of low-energy X-rays in vitro. Experiments were performed with 25 kV X-rays and 200 kV reference X-rays on neonatal human keratinocytes (HEKn), and NIH/3T3 mouse fibroblasts. Cell survival was studied with graded doses in a clonogenic assay, chromosomal damage in a micronucleus (MN) assay. The surviving fraction at 2 Gy for keratinocytes was 46±5% after 200 kV and 33±11% after 25 kV X-rays. Linear-quadratic cell survival analysis yielded α=0.305±0.033 Gy-1 and β=0.048±0.011 Gy-2 for 200 kV and α=0.399±0.103 Gy-1 and β=0.048±0.054 Gy-2 for 25 kV. For 3T3 fibroblasts an SF2 of 53±3% after 200 kV and 61±18% after 25 kV was observed. Values of α=0.24±0.02 Gy-1 and β=0.022±0.002 Gy-2 for 200 kV and α=0.10±0.05 Gy-1 and β=0.070±0.010 Gy-2 for 25 kV were derived. In conclusion, keratinocyte survival was similar for both radiation qualities. For fibroblasts, a reduction in survival at higher doses was observed. Results from MN studies will be presented

26. Chromosomal damage and survival of keratinocytes and fibroblasts after irradiation with 200 kV or 25 kV X-rays

A relative biological effectiveness of 1 is accepted for soft X-rays (25–30 kV), which are applied in diagnostic radiology (mammography). However, it has been shown that soft X-rays can be more effective in cell killing and chromosomal damage. The present study was initiated to define biological effects of low-energy X-rays in vitro. Experiments were performed with 25 kV X-rays and 200 kV reference X-rays on neonatal human keratinocytes (HEKn), and NIH/3T3 mouse fibroblasts. Cell survival was studied with graded doses in a clonogenic assay, chromosomal damage in a micronucleus (MN) assay. The surviving fraction at 2 Gy for keratinocytes was 46±5% after 200 kV and 33±11% after 25 kV X-rays. Linear-quadratic cell survival analysis yielded α=0.305±0.033 Gy-1 and β=0.048±0.011 Gy-2 for 200 kV and α=0.399±0.103 Gy-1 and β=0.048±0.054 Gy-2 for 25 kV. For 3T3 fibroblasts an SF2 of 53±3% after 200 kV and 61±18% after 25 kV was observed. Values of α=0.24±0.02 Gy-1 and β=0.022±0.002 Gy-2 for 200 kV and α=0.10±0.05 Gy-1 and β=0.070±0.010 Gy-2 for 25 kV were derived. In conclusion, keratinocyte survival was similar for both radiation qualities. For fibroblasts, a reduction in survival at higher doses was observed. Results from MN studies will be presented

On the HU Aquarii planetary system hypothesis

In this work, we investigate the eclipse timing of the polar binary HU Aquarii that has been observed for almost two decades. Recently, Qian et al. attributed large (O-C) deviations between the eclipse ephemeris and observations to a compact system of two massive jovian companions. We improve the Keplerian, kinematic model of the Light Travel Time (LTT) effect and re-analyse the whole currently available data set. We add almost 60 new, yet unpublished, mostly precision light curves obtained using the time high-resolution photo-polarimeter OPTIMA, as well as photometric observations performed at the MONET/N, PIRATE and TCS telescopes. We determine new mid--egress times with a mean uncertainty at the level of 1 second or better. We claim that because the observations that currently exist in the literature are non-homogeneous with respect to spectral windows (ultraviolet, X-ray, visual, polarimetric mode) and the reported mid--egress measurements errors, they may introduce systematics that affect orbital fits. Indeed, we find that the published data, when taken literally, cannot be explained by any unique solution. Many qualitatively different and best-fit 2-planet configurations, including self-consistent, Newtonian N-body solutions may be able to explain the data. However, using high resolution, precision OPTIMA light curves, we find that the (O-C) deviations are best explained by the presence of a single circumbinary companion orbiting at a distance of ~4.5 AU with a small eccentricity and having ~7 Jupiter-masses. This object could be the next circumbinary planet detected from the ground, similar to the announced companions around close binaries HW Vir, NN Ser, UZ For, DP Leo or SZ Her, and planets of this type around Kepler-16, Kepler-34 and Kepler-35.Comment: 20 pages, 18 figures, accepted to Monthly Notices of the Royal Astronomical Society (MNRAS

Multidisciplinary approach of early breast cancer: The biology applied to radiation oncology

Early breast cancer treatment is based on a multimodality approach with the application of clinical and histological prognostic factors to determine locoregional and systemic treatments. The entire scientific community is strongly involved in the management of this disease: radiologists for screening and early diagnosis, gynecologists, surgical oncologists and radiation oncologists for locoregional treatment, pathologists and biologists for personalized characterization, genetic counselors for BRCA mutation history and medical oncologists for systemic therapies

Chromosomal radiosensitivity and acute radiation side effects after radiotherapy in tumour patients - a follow-up study

Radiotherapists are highly interested in optimizing doses especially for patients who tend to suffer from side effects of radiotherapy (RT). It seems to be helpful to identify radiosensitive individuals before RT. Thus we examined aberrations in FISH painted chromosomes in in vitro irradiated blood samples of a group of patients suffering from breast cancer. In parallel, a follow-up of side effects in these patients was registered and compared to detected chromosome aberrations. METHODS: Blood samples (taken before radiotherapy) were irradiated in vitro with 3 Gy X-rays and analysed by FISH-painting to obtain aberration frequencies of first cycle metaphases for each patient. Aberration frequencies were analysed statistically to identify individuals with an elevated or reduced radiation response. Clinical data of patients have been recorded in parallel to gain knowledge on acute side effects of radiotherapy. RESULTS: Eight patients with a significantly elevated or reduced aberration yield were identified by use of a t-test criterion. A comparison with clinical side effects revealed that among patients with elevated aberration yields one exhibited a higher degree of acute toxicity and two patients a premature onset of skin reaction already after a cumulative dose of only 10 Gy. A significant relationship existed between translocations in vitro and the time dependent occurrence of side effects of the skin during the therapy period. CONCLUSIONS: The results suggest that translocations can be used as a test to identify individuals with a potentially elevated radiosensitivity

Radiosensitivity in breast cancer assessed by the Comet and micronucleus assays

Spontaneous and radiation-induced genetic instability of peripheral blood mononuclear cells derived from unselected breast cancer (BC) patients (n=50) was examined using the single-cell gel electrophoresis (Comet) assay and a modified G2 micronucleus (MN) test. Cells from apparently healthy donors (n=16) and from cancer patients (n=9) with an adverse early skin reaction to radiotherapy (RT) served as references. Nonirradiated cells from the three tested groups exhibited similar baseline levels of DNA fragmentation assessed by the Comet assay. Likewise, the Comet analysis of in vitro irradiated (5 Gy) cells did not reveal any significant differences among the three groups with respect to the initial and residual DNA fragmentation, as well as the DNA repair kinetics. The G2 MN test showed that cells from cancer patients with an adverse skin reaction to RT displayed increased frequencies of both spontaneous and radiation-induced MN compared to healthy control or the group of unselected BC patients. Two patients from the latter group developed an increased early skin reaction to RT, which was associated with an increased initial DNA fragmentation in vitro only in one of them. Cells from the other BC patient exhibited a striking slope in the dose–response curve detected by the G2 MN test. We also found that previous RT strongly increased both spontaneous and in vitro radiation-induced MN levels, and to a lesser extent, the radiation-induced DNA damage assessed by the Comet assay. These data suggest that clinical radiation may provoke genetic instability and/or induce persistent DNA damage in normal cells of cancer patients, thus leading to increased levels of MN induction and DNA fragmentation after irradiation in vitro. Therefore, care has to be taken when blood samples collected postradiotherapeutically are used to assess the radiosensitivity of cancer patients

Drosophila Eggshell Production: Identification of New Genes and Coordination by Pxt

Drosophila ovarian follicles complete development using a spatially and temporally controlled maturation process in which they resume meiosis and secrete a multi-layered, protective eggshell before undergoing arrest and/or ovulation. Microarray analysis revealed more than 150 genes that are expressed in a stage-specific manner during the last 24 hours of follicle development. These include all 30 previously known eggshell genes, as well as 19 new candidate chorion genes and 100 other genes likely to participate in maturation. Mutations in pxt, encoding a putative Drosophila cyclooxygenase, cause many transcripts to begin expression prematurely, and are associated with eggshell defects. Somatic activity of Pxt is required, as RNAi knockdown of pxt in the follicle cells recapitulates both the temporal expression and eggshell defects. One of the temporally regulated genes, cyp18a1, which encodes a cytochromome P450 protein mediating ecdysone turnover, is downregulated in pxt mutant follicles, and cyp18a1 mutation itself alters eggshell gene expression. These studies further define the molecular program of Drosophila follicle maturation and support the idea that it is coordinated by lipid and steroid hormonal signals