Differential effects of overexpression of ERalpha and ERbeta in MCF10A immortalised, non-transformed human breast epithelail cells

Background: Cellular effects of oestrogen are mediated by two intracellular receptors ERα and ERβ. However, to compare responses mediated through these two receptors, experimental models are needed where ERα and ERβ are individually stably overexpressed in the same cell type. Methods: We compared the effects of stable overexpression of ERα and ERβ in the MCF10A cell line, which is an immortalised but non-transformed breast epithelial cell line without high endogenous ER expression. Results: Clones of MCF10A cells were characterised which stably overexpressed ERα (10A-ERα2, 10A-ERα13) or which stably overexpressed ERβ (10A-ERβ12, 10A-ERβ15). Overexpression of either ERα or ERβ allowed induction of an oestrogen-regulated ERE-LUC reporter gene by oestradiol which was not found in the untransfected cells. Oestradiol also increased proliferation of 10A-ERα13 and 10A-ERβ12 cells, but not untransfected cells, by 1.3-fold over 7 days. The phytoestrogen, genistein, which is reported to bind more strongly to ERβ than to ERα, could induce luciferase gene expression from an ERE-LUC reporter gene at concentrations of 10−6 M and 10−5 M but only in the clones overexpressing ERβ and not in those overexpressing ERα. Clone 10A-ERβ12 also yielded growth stimulation with 10-6 M genistein. Finally, the overexpression of ERα, but not ERβ, gave rise to increased growth in semi-solid methocel suspension culture in the presence of 70 nM oestradiol, suggesting that overexpression of ERα, but not ERβ, produces characteristics of a transformed phenotype. Conclusions: This provides a model system to compare effects of oestradiol with other oestrogenic ligands in cells stably overexpressing individually ERα or ERβ.

Crosstalk with insulin and dependence on PI3K/Akt/mTOR rather than MAPK pathways in upregulation of basal growth following long-term oestrogen deprivation in three human breast cancer cell lines

Background: MCF-7, T-47-D, ZR-75-1 human breast cancer cell lines are dependent on oestrogen for growth but can adapt to grow during long-term oestrogen deprivation. This serves as a model for identification of therapeutic targets in endocrine-resistant breast cancer. Methods: An overlooked complication of this model is that it involves more than non-addition of oestrogen, and inadequate attention has been given to separating molecular events associated with each of the culture manipulations. Results: Insulin and oestradiol were shown to protect MCF-7 cells against upregulation of basal growth, demonstrating a crosstalk in the growth adaptation process. Increased phosphorylation of p44/42MAPK and c-Raf reflected removal of insulin from the medium and proliferation of all three cell lines was inhibited to a lesser extent by PD98059 and U0126 following long-term oestrogen/insulin withdrawal, demonstrating a reduced dependence on the MAPK pathway. By contrast, long-term oestrogen/insulin deprivation did not alter levels of phosphorylated Akt and did not alter the dose-response of growth inhibition with LY294002 in any of the three cell lines. The IGF1R inhibitor picropodophyllin inhibited growth of all MCF-7 cells but only in the long-term oestrogen/insulin-deprived cells was this paralleled by reduction in phosphorylated p70S6K, a downstream target of mTOR. Long-term oestrogen/insulin-deprived MCF-7 cells had higher levels of phosphorylated p70S6K and developed increased sensitivity to growth inhibition by rapamycin. Conclusions: The greater sensitivity to growth inhibition by rapamycin in all three cell lines following long-term oestrogen/insulin deprivation suggests rapamycin-based therapies might be more effective in breast cancers with acquired oestrogen resistance. Keywords Akt, breast cancer cells, endocrine resistance, insulin, MAPK, MCF-7 cells, mTOR, oestrogen, oestrogen-deprived, PI3K, picropodophyllin, rapamycin, T-47-D cells, ZR-75-1 cells

Unusual censor readings: G.D.R. science fiction and the Ministry of Culture

Covers the censorship by the Hauptverwaltung Verlage und Buchhandel in the Ministry of Culture of science fiction works in East Germany. These ranged from propaganda pieces supporting the scientific-technological revolution and the progress of socialism, to veiled criticisms of the system. The chapter argues that publishers became dependent on the high sales generated by popular culture, creating a relatively safe space for this genre.

Cost733cat: a database of weather and circulation type classifications

A new database of weather and circulation type catalogs is presented comprising 17 automated classification methods and five subjective classifications. It was compiled within COST Action 733 "Harmonisation and Applications of Weather Type Classifications for European regions" in order to evaluate different methods for weather and circulation type classification. This paper gives a technical description of the included methods using a new conceptual categorization for classification methods reflecting the strategy for the definition of types. Methods using predefined types include manual and threshold based classifications while methods producing types derived from the input data include those based on eigenvector techniques, leader algorithms and optimization algorithms. In order to allow direct comparisons between the methods, the circulation input data and the methods' configuration were harmonized for producing a subset of standard catalogs of the automated methods. The harmonization includes the data source, the climatic parameters used, the classification period as well as the spatial domain and the number of types. Frequency based characteristics of the resulting catalogs are presented, including variation of class sizes, persistence, seasonal and inter-annual variability as well as trends of the annual frequency time series. The methodological concept of the classifications is partly reflected by these properties of the resulting catalogs. It is shown that the types of subjective classifications compared to automated methods show higher persistence, inter-annual variation and long-term trends. Among the automated classifications optimization methods show a tendency for longer persistence and higher seasonal variation. However, it is also concluded that the distance metric used and the data preprocessing play at least an equally important role for the properties of the resulting classification compared to the algorithm used for type definition and assignment.

How to quantify the resolution of surface climate by circulation types: an example for Alpine precipitation

A novel approach is presented for the evaluation of circulation type classifications (CTCs) in terms of their capability to predict surface climate variations. The approach is analogous to that for probabilistic meteorological forecasts and is based on the Brier skill score. This score is shown to take a particularly simple form in the context of CTCs and to quantify the resolution of a climate variable by the classifications. The sampling uncertainty of the skill can be estimated by means of nonparametric bootstrap resampling. The evaluation approach is applied for a systematic intercomparison of 71 CTCs (objective and manual, from COST Action 733) with respect to their ability to resolve daily precipitation in the Alpine region. For essentially all CTCs, the Brier skill score is found to be higher for weak and moderate compared to intense precipitation, for winter compared to summer, and over the north and west of the Alps compared to the south and east. Moreover, CTCs with a higher number of types exhibit better skill than CTCs with few types. Among CTCs with comparable type number, the best automatic classifications are found to outperform the best manual classifications. It is not possible to single out one ‘best’ classification for Alpine precipitation, but there is a small group showing particularly high skill.

Seasonality and interannual variability of the westerly jet in the Tibetan Plateau region

In this study, 40-yr ECMWF Re-Analysis (ERA-40) data are used for the description of the seasonal cycle and the interannual variability of the westerly jet in the Tibetan Plateau region. To complement results based on the analysis of monthly mean horizontal wind speeds, an occurrence-based jet climatology is constructed by identifying the locations of the jet axes at 6-hourly intervals throughout 1958–2001. Thus, a dataset describing the highly transient and localized features of jet variability is obtained. During winter and summer the westerly jet is located, respectively, to the south and north of the Tibetan Plateau. During the spring and autumn seasons there are jet transitions from south to north and vice versa. The median dates for these transitions are 28 April and 12 October. The spring transition is associated with large interannual variations, while the fall transition occurs more reliably within a 3-week period. The strength of the jet exhibits a peculiar seasonal cycle. During northward migration in April/May, the jet intensity weakens and its latitudinal position varies largely. In some springs, there are several transitions and split configurations occur before the jet settles in its northern summer position. In June, a well-defined and unusually strong jet reappears at the northern flanks of the Tibetan Plateau. In autumn, the jet gradually but reliably recedes to the south and is typically more intense than in spring. The jet transitions between the two preferred locations follow the seasonal latitudinal migration of the jet in the Northern Hemisphere. An analysis of interannual variations shows the statistical relationship between the strength of the summer jet, the tropospheric meridional temperature gradient, and the all-India rainfall series. Both this analysis and results from previous studies point to the particular dynamical relevance of the onsetting Indian summer monsoon precipitation and the associated diabatic heating for the formation of the strong summer jet. Finally, an example is provided that illustrates the climatological significance of the jet in terms of the covariation between the jet location and the spatial precipitation distribution in central Asia.

The precipitation climate of Central Asia: intercomparison of observational and numerical data sources in a remote semiarid region

In this study, we systematically compare a wide range of observational and numerical precipitation datasets for Central Asia. Data considered include two re-analyses, three datasets based on direct observations, and the output of a regional climate model simulation driven by a global re-analysis. These are validated and intercompared with respect to their ability to represent the Central Asian precipitation climate. In each of the datasets, we consider the mean spatial distribution and the seasonal cycle of precipitation, the amplitude of interannual variability, the representation of individual yearly anomalies, the precipitation sensitivity (i.e. the response to wet and dry conditions), and the temporal homogeneity of precipitation. Additionally, we carried out part of these analyses for datasets available in real time. The mutual agreement between the observations is used as an indication of how far these data can be used for validating precipitation data from other sources. In particular, we show that the observations usually agree qualitatively on anomalies in individual years while it is not always possible to use them for the quantitative validation of the amplitude of interannual variability. The regional climate model is capable of improving the spatial distribution of precipitation. At the same time, it strongly underestimates summer precipitation and its variability, while interannual variations are well represented during the other seasons, in particular in the Central Asian mountains during winter and spring