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Supplementary Table 1 from Expression Profiling of t(12;22) Positive Clear Cell Sarcoma of Soft Tissue Cell Lines Reveals Characteristic Up-Regulation of Potential New Marker Genes Including ERBB3

Supplementary Table 2 from Expression Profiling of t(12;22) Positive Clear Cell Sarcoma of Soft Tissue Cell Lines Reveals Characteristic Up-Regulation of Potential New Marker Genes Including ERBB3

Clear cell sarcoma of soft tissue (CCSST), also known as malignant melanoma of soft parts, represents a rare lesion of the musculoskeletal system usually affecting adolescents and young adults. CCSST is typified by a chromosomal t(12;22)(q13;q12) translocation resulting in a fusion between the Ewing sarcoma gene (EWSR1) and activating transcription factor 1 (ATF1), of which the activity in nontransformed cells is regulated by cyclic AMP. Our aim was to identify critical differentially expressed genes in CCSST tumor cells in comparison with other solid tumors affecting children and young adults to better understand signaling pathways regulating specific features of the development and progression of this tumor entity. We applied Affymetrix Human Genome U95Av2 oligonucleotide microarrays representing ∼12,000 genes to generate the expression profiles of the CCSST cell lines GG-62, DTC-1, KAO, MST2, MST3, and Su-CC-S1 in comparison with 8 neuroblastoma, 7 Ewing tumor, and 6 osteosarcoma cell lines. Subsequent hierarchical clustering of microarray data clearly separated all four of the tumor types from each other and identified differentially expressed transcripts, which are characteristically up-regulated in CCSST. Statistical analysis revealed a group of 331 probe sets, representing ∼300 significant (P < 0.001) differentially regulated genes, which clearly discriminated between the CCSST and other tumor samples. Besides genes that were already known to be highly expressed in CCSST, like S100A11 (S100 protein) or MITF (microphthalmia-associated transcription factor), this group shows an obvious portion of genes that are involved in cyclic AMP response or regulation, in pigmentation processes, or in neuronal structure and signaling. Comparison with other expression profile analyses on neuroectodermal childhood tumors confirms the high robustness of this strategy to characterize tumor entities based on their gene expression. We found the avian erythroblastic leukemia viral oncogene homologue 3 (ERBB3) to be one of the most dramatically up-regulated genes in CCSST. Quantitative real-time PCR and Northern blot analysis verified the mRNA abundance and confirmed the absence of the inhibitory transcript variant of this gene. The protein product of the member of the epidermal growth factor receptor family ERBB3 could be shown to be highly present in all of the CCSST cell lines investigated, as well as in 18 of 20 primary tumor biopsies. In conclusion, our data demonstrate new aspects of the phenotype and the biological behavior of CCSST and reveal ERBB3 to be a useful diagnostic marker.

Background Nibrin, as part of the NBN/MRE11/RAD50 complex, is mutated in Nijmegen breakage syndrome (NBS), which leads to impaired DNA damage response and lymphoid malignancy. Results Telomere length (TL) was markedly reduced in homozygous patients (and comparably so in all chromosomes) by ~40% (qPCR) and was slightly reduced in NBS heterozygotes older than 30 years (~25% in qPCR), in accordance with the respective cancer rates. Humanized cancer-free NBS mice had normal TL. Telomere elongation was inducible by telomerase and/or alternative telomere lengthening but was associated with abnormal expression of telomeric genes involved in aging and/or cell growth. Lymphoblastoid cells from NBS patients with long survival times (>12 years) displayed the shortest telomeres and low caspase 7 activity. Conclusions NBS is a secondary telomeropathy. The two-edged sword of telomere attrition enhances the cancer-prone situation in NBS but can also lead to a relatively stable cellular phenotype in tumor survivors. Results suggest a modular model for progeroid syndromes with abnormal expression of telomeric genes as a molecular basis. Methods We studied TL and function in 38 homozygous individuals, 27 heterozygotes, one homozygous fetus, six NBS lymphoblastoid cell lines, and humanized NBS mice, all with the same founder NBN mutation: c.657_661del5.

Serotonin 5-HT2C receptor is a G-protein coupled excitatory receptor that regulates several biochemical pathways and has been implicated in obesity, mental state, sleep cycles, autism, neuropsychiatric disorders and neurodegenerative diseases. The activity of 5-HT2CR is regulated via alternative splicing and A to I editing of exon Vb of its pre-mRNA. Snord115 is a small nucleolar RNA that is expressed in mouse neurons and displays an 18-nucleotide base complementary to exon Vb of 5-HT2CR pre-mRNA. For almost two decades this putative guide element of Snord115 has wandered like a ghost through the literature in attempts to elucidate the biological significance of this complementarity. In mice, Snord115 is expressed in neurons and absent in the choroid plexus where, in contrast, 5-Ht2cr mRNA is highly abundant. Here we report the analysis of 5-Ht2cr pre-mRNA posttranscriptional processing via RNA deep sequencing in a mouse model that ectopically expresses Snord115 in the choroid plexus. In contrast to previous reports, our analysis demonstrated that Snord115 does not control alternative splicing of 5-Ht2cr pre-mRNA in vivo. We identified a modest, yet statistically significant reduction of 5-Ht2cr pre-mRNA A to I editing at the major A, B, C and D sites. We suggest that Snord115 and exon Vb of 5Ht2cr pre-mRNA form a double-stranded structure that is subject to ADAR-mediated A to I editing. To the best of our knowledge, this is the first comprehensive Snord115 gain-of-function analysis based on in vivo mouse models.

Prostate cancer is widely observed to be biologically heterogeneous. Its heterogeneity is manifested histologically as multifocal prostate cancer, which is observed more frequently than unifocal prostate cancer. The clinical and prognostic significance of either focal cancer type is not fully established. To investigate prostate cancer heterogeneity, the genetic profiles of multifocal and unifocal prostate cancers were compared. Here, we report observations deduced from tumor-tumor comparison of copy number alteration data of both focal categories. Forty-one fresh frozen prostate cancer foci from 14 multifocal prostate cancers and eight unifocal prostate cancers were subjected to copy number variation analysis with the Affymetrix SNP 6.0 microarray tool. With the investigated cases, tumors obtained from a single prostate exhibited different genetic profiles of variable degrees. Further comparison identified no distinct genetic pattern or signatures specific to multifocal or unifocal prostate cancer. Our findings suggest that samples obtained from multiple sites of a single unifocal prostate cancer show as much genetic heterogeneity and variability as separate tumors obtained from a single multifocal prostate cancer.

Study question Is the molecular profile of human spermatogonia homogeneous or heterogeneous when analysed at the single-cell level? Summary answer Heterogeneous expression profiles may be a key characteristic of human spermatogonia, supporting the existence of a heterogeneous stem cell population. What is known already Despite the fact that many studies have sought to identify specific markers for human spermatogonia, the molecular fingerprint of these cells remains hitherto unknown. Study design, size, duration Testicular tissues from patients with spermatogonial arrest (arrest, n = 1) and with qualitatively normal spermatogenesis (normal, n = 7) were selected from a pool of 179 consecutively obtained biopsies. Gene expression analyses of cell populations and single-cells (n = 105) were performed. Two OCT4-positive individual cells were selected for global transcriptional capture using shallow RNA-seq. Finally, expression of four candidate markers was assessed by immunohistochemistry. Participants/materials, setting, methods Histological analysis and blood hormone measurements for LH, FSH and testosterone were performed prior to testicular sample selection. Following enzymatic digestion of testicular tissues, differential plating and subsequent micromanipulation of individual cells was employed to enrich and isolate human spermatogonia, respectively. Endpoint analyses were qPCR analysis of cell populations and individual cells, shallow RNA-seq and immunohistochemical analyses. Main results and the role of chance Unexpectedly, single-cell expression data from the arrest patient (20 cells) showed heterogeneous expression profiles. Also, from patients with normal spermatogenesis, heterogeneous expression patterns of undifferentiated (OCT4, UTF1 and MAGE A4) and differentiated marker genes (BOLL and PRM2) were obtained within each spermatogonia cluster (13 clusters with 85 cells). Shallow RNA-seq analysis of individual human spermatogonia was validated, and a spermatogonia-specific heterogeneous protein expression of selected candidate markers (DDX5, TSPY1, EEF1A1 and NGN3) was demonstrated. Limitations, reasons for caution The heterogeneity of human spermatogonia at the RNA and protein levels is a snapshot. To further assess the functional meaning of this heterogeneity and the dynamics of stem cell populations, approaches need to be developed to facilitate the repeated analysis of individual cells. Wider implications of the findings Our data suggest that heterogeneous expression profiles may be a key characteristic of human spermatogonia, supporting the model of a heterogeneous stem cell population. Future studies will assess the dynamics of spermatogonial populations in fertile and infertile patients. Large scale data RNA-seq data is published in the GEO database: GSE91063. Study funding/competing interest(s) This work was supported by the Max Planck Society and the Deutsche Forschungsgemeinschaft DFG-Research Unit FOR 1041 Germ Cell Potential (grant numbers SCHO 340/7-1, SCHL394/11-2). The authors declare that there is no conflict of interest.

Activation of the transcription factor nuclear factor-kappaB (NFkappaB) by inflammatory cytokines like tumor necrosis (TNF) factor and interleukin-1 (IL-1) is generally associated with the induction of antiapoptotic pathways. Therefore, NFkappaB inhibits both intrinsically and extrinsically induced apoptosis and thus is regarded to act universally in an antiapoptotic fashion. Accordingly, activation of NFkappaB by IL-1 was shown to result in reduction of death ligand-induced apoptosis via up-regulation of antiapoptotic inhibitor of apoptosis proteins (IAPs). In contrast, apoptosis induced by ultraviolet-B radiation (UVB) was shown to be enhanced in an NFkappaB-dependent manner, indicating that NFkappaB can also act in a proapoptotic fashion. This study investigates the molecular mechanisms underlying IL-1-mediated enhancement of UVB-induced apoptosis. We show that NFkappaB activation in costimulation with UVB treatment results in repression of antiapoptotic genes and consequently in down-regulation of the respective proteins, like c-IAP, FLICE-inhibitory protein (FLIP), and some members of the TNF receptor-associated (TRAF)2 protein family. In parallel, TNFalpha is released, leading to activation of signaling pathways mediated by TNF receptor-1 (TNF-R1). Although TNF is well known to induce both proapoptotic and antiapoptotic effects, the down-regulated levels of TRAF-1, -2, and -6 proteins by IL-1 plus UVB action leads to a shift toward promotion of the proapoptotic pathway. In concert with the down-regulation of IAPs and FLIP, TNF-R1 activation as an additional proapoptotic stimulus now results in significant enhancement of UVB-induced apoptosis. Taken together, elucidation of the molecular mechanisms underlying IL-1-mediated enhancement of UVB-induced apoptosis revealed that NFkappaB does not exclusively act in an antiapoptotic fashion but may also mediate proapoptotic effects.

Distinct parallel cytogenetic pathways in breast carcinogenesis could be identified in recent years. Nevertheless, it remained unclear as to which tumours may have progressed in grade or which patterns of cytogenetic alteration may define the switch from an in situ towards an invasive lesion. In order to gain more detailed insights into cytogenetic mechanisms of the pathogenesis of breast cancer, the chromosomal imbalances of 206 invasive breast cancer cases were characterised by means of comparative genomic hybridisation (CGH). CGH data were subjected to hierarchical cluster analysis and the results were further compared with immunohistochemical findings on tissue arrays from the same breast cancer cases. The combined analysis of immunohistochemical and cytogenetic data provided evidence that carcinomas with gains of 7p, and to a lesser extent losses of 9q and gains of 5p, are a distinct subgroup within the spectrum of ductal invasive grade 3 breast carcinomas. These aberrations were associated with a high degree of cytogenetic instability (16.6 alterations per case on average), 16q-losses in over 70% of these cases, strong oestrogen receptor expression and absence of strong expression of p53, c-erbB2 and Ck 5. These characteristics provide strong support for the hypothesis that these tumours may develop through stages of well- and perhaps intermediately differentiated breast cancers. Our results therefore underline the existence of several parallel and also stepwise progression pathways towards breast cancer.

Background Logistic regression (LR) is commonly used to estimate risk of coronary heart disease. We investigated if neural networks improved on the risk estimate of LR by analysing data from the Prospective Cardiovascular Munster Study (PROCAM), a large prospective epidemiological study of risk factors for coronary heart disease among men and women at work in northern Germany. Methods We used a multi-layer perceptron (MLP) and probabilistic neural networks (PNN) to estimate the risk of myocardial infarction or acute coronary death (coronary events) during 10 years' follow-up among 5159 men aged 35-65 years at recruitment into PROCAM. In all, 325 coronary events occurred in this group. We assessed the performance of each procedure by measuring the area under the receiver-operating characteristics curve (AUROC). Results The AUROC of the MLP was greater than that of the PNN (0.897 versus 0.872), and both exceeded the AUROC for LR of 0.840. If 'high risk' is defined as an event risk >20% in 10 years, LR classified 8.4% of men as high risk, 36.7% of whom suffered an event in 10 years (45.8% of all events). The MLP classified 7.9% as high risk, 64.0% of whom suffered an event (74.5% of all events), while with the PNN, only 3.9% were at high risk, 58.6% of whom suffered an event (33.5% of all events). Conclusion Intervention trials indicate that about one in three coronary events can be prevented by 5 years of lipid-lowering treatment. Our analysis suggests that use of the MLP to identify high-risk individuals as candidates for drug treatment would allow prevention of 25% of coronary events in middle-aged men, compared to 15% and 11% with LR and the PNN, respectively.

Changes in variability and extremes of the hydrological cycle are studied in two 30 year simulations using a general circulation model at high horizontal resolution. The simulations represent the present-day climate and a period in which the radiative forcing corresponds to a doubling of the present-day concentrations of atmospheric greenhouse gases. In most regions and seasons the probability density function of daily precipitation experiences a stretching associated with a higher probability of heavy precipitation events in the warmer climate. Whereas extremely long wet spells show only moderate changes, the extremely long dry spells are extended at middle latitudes over most land areas. At high latitudes the changes in annual maximum river runoff are mainly controlled by changes in snow budget. Eight out of 14 selected major rivers show a statistically significant change in 10 year return values of the annual maximum discharge. In two cases a significant decrease is found and in six cases there is a significant increase. Copyright © 2002 Royal Meteorological Society

Our earlier studies demonstrated that high-density lipoproteins (HDLs) stimulate multiple signaling pathways, including activation of phosphatidylcholine-specific phospholipases C and D (PC-PLs) and phosphatidylinositol-specific phospholipase C (PI-PLC). However, only activation of PC-PLs was linked to the HDL-induced cholesterol efflux. In the study presented here, the role of HDL-induced PI-PLC activation was studied. In human skin fibroblasts, HDL potently induced PI-PLC as inferred from enhanced phosphatidylinositol bisphosphate (PtdInsP(2)) turnover and Ca(2+) mobilization. The major protein component of HDL, apo A-I, did not induce PtdInsP(2) turnover or Ca(2+) mobilization in these cells. Both HDL and apo A-I promoted cellular cholesterol efflux, whereas only HDL induced fibroblast proliferation. Inhibition of PI-PLC with U73122 or blocking intracellular Ca(2+) elevation with Ni(2+) or EGTA markedly reduced the extent of HDL-induced cell proliferation but had no effect on cholesterol efflux. In fibroblasts from patients with Tangier disease which are characterized by defective cholesterol efflux, neither HDL-induced PtdInsP(2) breakdown and Ca(2+) mobilization nor cell proliferation was impaired. HDL-induced fibroblast proliferation, PtdInsP(2) turnover, and Ca(2+) mobilization were fully mimicked by the lipid fraction isolated from HDL. Analysis of this fraction with high-performance liquid chromatography (HPLC) and time-of-flight secondary ion mass spectroscopy (TOF-SIMS) revealed that the PI-PLC-inducing activity is identical with two bioactive lysosphingolipids, namely, lysosulfatide (LSF) and sphingosylphosphorylcholine (SPC). Like native HDL, LSF and SPC induced PtdInsP(2) turnover, Ca(2+) mobilization, and fibroblast proliferation. However, both compounds did not promote cholesterol efflux. In conclusion, two agonist activities are carried by HDL. Apo A-I stimulates phosphatidylcholine breakdown and thereby facilitates cholesterol efflux, whereas LSF and SPC trigger PI-PLC activation and thereby stimulate cell proliferation.

In this study we investigate the role of heat, freshwater and momentum fluxes in changing the oceanic climate and thermohaline circulation as a consequence of increasing atmospheric CO2 concentration. Two baseline integrations with a fully coupled ocean atmosphere general circulation model with either fixed or increasing atmospheric CO2 concentrations have been performed. In a set of sensitivity experiments either freshwater (precipitation, evaporation and runoff from the continents) and/or momentum fluxes were no longer simulated, but prescribed according to one of the fully coupled baseline experiments. This approach gives a direct estimate of the contribution from the individual flux components. The direct effect of surface warming and the associated feedbacks in ocean circulation are the dominant processes in weakening the Atlantic thermohaline circulation in our model. The relative contribution of momentum and freshwater fluxes to the total response turned out to be less than 25%, each. Changes in atmospheric water vapour transport lead to enhanced freshwater input into middle and high latitudes, which weakens the overturning. A stronger export of freshwater from the Atlantic drainage basin to the Indian and Pacific ocean, on the other hand, intensifies the Atlantic overturning circulation. In total the modified freshwater fluxes slightly weaken the Atlantic thermohaline circulation. The contribution of the modified momentum fluxes has a similar magnitude, but enhances the formation of North Atlantic deep water. Salinity anomalies in the Atlantic as a consequence of greenhouse warming stem in almost equal parts from changes in net freshwater fluxes and from changes in ocean circulation caused by the surface warming due to atmospheric heat fluxes. Important effects of the momentum fluxes are a poleward shift of the front between Northern Hemisphere subtropical and subpolar gyres and a southward shift in the position of the Antarctic circumpolar current, with a clear signal in sea level.

This paper studies the variability of deep-ocean mass transport using four 1000-yr integrations performed with coupled general circulation models. Statistics describing the spectral and spatial features are considered. It is shown that these features depend crucially on the time-mean state. For the transport of tropical and subtropical water masses in three of the integrations, the spectral levels continually increase with decreasing frequency and do not show isolated peaks at low frequencies. The slope of the low-frequency spectrum (in a log–log plot) changes with increasing depth. It has values of about 0 near the surface, about −1 at intermediate depth, and about −2 at or near the bottom. The result indicates that the maximal memory timescale for deep-ocean mass transport is longer than a few centuries. The situation is different in the fourth integration, which has a different mean circulation pattern. In this case, the low-frequency spectrum is more or less flat in the tropical and subtropic...

The leading variability mode of the coupled troposphere-stratosphere winter circulation in the Northern Hemisphere (NH) describes a close relationship between the strength of the stratospheric cyclonic vortex and the index of a tropospheric wave-like pattern covering the North Atlantic-Eurasian region. This mode can be determined by applying singular value decomposition analysis between the time series of winter mean NH 50- and 500-hPa geopotential heights. We compared the features of the leading coupled variability mode between two climate regimes, determined from a 1900-year integration with the coupled atmosphere-ocean climate model ECHAM3-LSG. The two regimes differ on the interdecadal timescale in the strength of the stratospheric polar vortex and therefore in the transmission-refraction properties of vertically propagating tropospheric waves. The spatial structures of the leading coupled variability mode of observational data better match the corresponding structures of the model's weak polar vortex regime (PVR) than those of the strong one. Because of the more effective tropospheric trapping of stationary wave energy of zonal wave number (ZWN) 2 at midlatitudes, the zonal variability structure of this wave is changed by barotropic effects in the troposphere as well as in the stratosphere. The coupled troposphere-stratosphere mode and the response of winter circulation were studied in a climate-change experiment carried out with the same model. We could show that under increased greenhouse gas forcing, both the response and the coupled variability mode between tropospheric and stratospheric circulation itself has a high similarity to the leading coupled mode in the strong PVR.

This paper discusses two aspects of climate modeling, the deep water formation in the North Atlantic and precipitation changes due to climate change caused by anthropogenic emissions of greenhouse gases. The deep water formation is strongly influenced by the precipitation, and the precipitation is affected by the concentration of the greenhouse gases in the atmosphere and by the atmospheric and oceanic circulation. The experiments discussed here have been performed independently to test the stability of the thermohaline circulation of the North Atlantic and to investigate changes in precipitation due to anthropogenic greenhouse gas emissions. The precipitation changes in a climate change environment are sufficient in some simulations to decrease the thermohaline circulation noticeably. However, it appears that the amount of freshwater needed to bring the circulation to a collapse is magnitudes larger than the anticipated change in precipitation due to anthropogenic activities within the next 100 years. The precipitation changes, on the other hand, might change regionally quite drastically towards more extreme situations, thereby putting additional stress on vegetation and enhancing soil erosion.

To estimate the effect of the solar variability on the climate, two estimates of the solar intensity variations during the last three centuries have been used as forcing in numerical simulations. The model employed to carry out the experiments was the same coupled global ocean-atmosphere model used in a number of studies to assess the effect of the anthropogenic greenhouse gases on climate. The near surface temperature and the tropospheric temperature distribution shows a clear response to the variability of the solar input. Even the thermohaline circulation reacts on the large amplitudes in the forcing. In the stratosphere, the response pattern is similar as in the observations, however, the 11-year cycle found in the forcing data does not excite an appreciable response. This might be due to the missing parameterisation of the increase in the UV-radiation at the solar cycle maximum and the connected increase of the stratospheric ozone concentration.

The El Nino-Southern Oscillation (ENSO) is investigated in a multicentury integration conducted with the coupled general circulation model (CGCM) ECHAM3/LSG. The quasiperiodic interannual oscillations of the simulated equatorial Pacific climate system are due to subsurface temperature anomaly propagation and a positive atmosphere-ocean feedback. The gravest internal wave modes contribute to the generation of these anomalies. The simulated ENSO has a characteristic period of 5–8 years. Due to the coarse resolution of the ocean model the ENSO amplitude is underestimated by a factor of three as compared to observations. The model ENSO is associated with the typical atmospheric teleconnection patterns. Using wavelet statistics two characteristic interdecadal modulations of the ENSO variance are identified. The origins of a 22 and 35 y ENSO modulation as well as the characteristic ENSO response to greenhouse warming simulated by our model are discussed.

The predictability of the coupled ocean–atmosphere climate system on interannual to decadal timescales has been studied by means of ensemble forecast experiments with a global coupled ocean–atmosphere general circulation model. Over most parts of the globe the model’s predictability can be sufficiently explained by damped persistence as expected from the stochastic climate model concept with damping times of considerably less than a year. Nevertheless, the tropical Pacific and the North Atlantic Ocean exhibit oscillatory coupled ocean–atmosphere modes, which lead to longer predictability timescales. While the tropical mode shares many similarities with the observed ENSO phenomenon, the coupled mode within the North Atlantic region exhibits a typical period of about 30 yr and relies on an interaction of the oceanic thermohaline circulation and the atmospheric North Atlantic oscillation. The model’s ENSO-like oscillation is predictable up to one-third to one-half (2–3 yr) of the oscillation period both in the ocean and the atmosphere. The North Atlantic yields considerably longer predictability timescales (of the order of a decade) only for quantities describing the model’s thermohaline circulation. For surface quantities and atmospheric variables only marginal predictability (of the order of a year) was obtained. The predictability of the coupled signal at the surface is destroyed by the large amount of internally generated (weather) noise. This is illustrated by means of a simple conceptual model for coupled ocean–atmosphere variability and predictability.

A new periodically synchronous coupling scheme has been applied to an atmosphere-ocean general circulation model. Due to a temporary switching off of the atmospheric model this scheme can considerably reduce computer requirements of coupled model experiments. In order to evaluate the new coupling scheme the model results are compared to corresponding synchronously coupled integrations. Experiments with fixed present-day CO2 concentration and a gradual increase of CO2 show a good reproduction of the mean state and the climate-change pattern, respectively. The deviations from the synchronously coupled experiments are in the range of the variability of the corresponding synchronously coupled runs. Due to the forcing during the ocean-only periods the short-term fluctuations are underestimated and the long-term variability is overestimated.

IntroductionApoptosis is a crucial pathway in tumor growth and metastatic development. Apoptotic proteins regulate the underlying molecular cascades and are thought to modulate the tumor response to chemotherapy and radiation. However, the prognostic value of the expression of apoptosis regulators in localized non–small-cell lung cancer (NSCLC) is still unclear.MethodsWe investigated the protein expression of apoptosis regulators Bcl-2, Bcl-xl, Mcl-1, and pp32/PHAPI, and the expression of the lncRNA MALAT-1 in tumor samples from 383 NSCLC patients (median age: 65.6 years; 77.5% male; paraffin-embedded tissue microarrays). For statistical analysis correlation tests, Log rank tests and Cox proportional hazard models were applied.ResultsTumor histology was significantly associated with the expression of Bcl-2, Bcl-xl and Mcl-1 (all p < 0.001). Among the tested apoptotic markers only Bcl-2 demonstrated prognostic impact (hazard ratio = 0.64, p = 0.012). For NSCLC patients with non-adenocarcinoma histology, Bcl-2 expression was associated with increased overall survival (p = 0.036). Besides tumor histology, prognostic impact of Bcl-2 was also found to depend on MALAT-1 lncRNA expression. Gene expression analysis of A549 adenocarcinoma cells with differential MALAT-1 lncRNA expression demonstrated an influence on the expression of Bcl-2 and its interacting proteins.ConclusionsBcl-2 expression was specifically associated with superior prognosis in localized NSCLC. An interaction of Bcl-2 with MALAT-1 lncRNA expression was revealed, which merits further investigation for risk prediction in resectable NSCLC patients.

A multi-fingerprint analysis is applied to the detection and attribution of anthropogenic climate change. While a single fingerprint is optimal for the detection of climate change, further tests of the statistical consistency of the detected climate change signal with model predictions for different candidate forcing mechanisms require the simultaneous application of several fingerprints. Model-predicted climate change signals are derived from three anthropogenic global warming simulations for the period 1880 to 2049 and two simulations forced by estimated changes in solar radiation from 1700 to 1992. In the first global warming simulation, the forcing is by greenhouse gas only, while in the remaining two simulations the direct influence of sulfate aerosols is also included. From the climate change signals of the greenhouse gas only and the average of the two greenhouse gas-plus-aerosol simulations, two optimized fingerprint patterns are derived by weighting the model-predicted climate change patterns towards low-noise directions. The optimized fingerprint patterns are then applied as a filter to the observed near-surface temperature trend patterns, yielding several detection variables. The space-time structure of natural climate variability needed to determine the optimal fingerprint pattern and the resultant signal-to-noise ratio of the detection variable is estimated from several multi-century control simulations with different CGCMs and from instrumental data over the last 136 y. Applying the combined greenhouse gas-plus-aerosol fingerprint in the same way as the greenhouse gas only fingerprint in a previous work, the recent 30-y trends (1966–1995) of annual mean near surface temperature are again found to represent a significant climate change at the 97.5% confidence level. However, using both the greenhouse gas and the combined forcing fingerprints in a two-pattern analysis, a substantially better agreement between observations and the climate model prediction is found for the combined forcing simulation. Anticipating that the influence of the aerosol forcing is strongest for longer term temperature trends in summer, application of the detection and attribution test to the latest observed 50-y trend pattern of summer temperature yielded statistical consistency with the greenhouse gas-plus-aerosol simulation with respect to both the pattern and amplitude of the signal. In contrast, the observations are inconsistent with the greenhouse-gas only climate change signal at a 95% confidence level for all estimates of climate variability. The observed trend 1943–1992 is furthermore inconsistent with a hypothesized solar radiation change alone at an estimated 90% confidence level. Thus, in contrast to the single pattern analysis, the two pattern analysis is able to discriminate between different forcing hypotheses in the observed climate change signal. The results are subject to uncertainties associated with the forcing history, which is poorly known for the solar and aerosol forcing, the possible omission of other important forcings, and inevitable model errors in the computation of the response to the forcing. Further uncertainties in the estimated significance levels arise from the use of model internal variability simulations and relatively short instrumental observations (after subtraction of an estimated greenhouse gas signal) to estimate the natural climate variability. The resulting confidence limits accordingly vary for different estimates using different variability data. Despite these uncertainties, however, we consider our results sufficiently robust to have some confidence in our finding that the observed climate change is consistent with a combined greenhouse gas and aerosol forcing, but inconsistent with greenhouse gas or solar forcing alone.

The stability of the Atlantic thermohaline circulation against meltwater input is investigated in a coupled ocean-atmosphere general circulation model. The meltwater input to the Labrador Sea is increased linearly for 250 years to a maximum input of 0.625 Sv and then reduced again to 0 (both instantaneously and linearly decreasing over 250 years). The resulting freshening forces a shutdown of the formation of North Atlantic deepwater and a subsequent reversal of the thermohaline circulation of the Atlantic, filling the deep Atlantic with Antarctic bottom water. The change in the overturning pattern causes a drastic reduction of the Atlantic northward heat transport, resulting in a strong cooling with maximum amplitude over the northern North Atlantic and a south-ward shift of the sea-ice margin in the Atlantic. Due to the increased meridional temperature gradient, the intertropical convergence zone over the Atlantic is displaced south-ward and the westerlies in the Northern Hemisphere gain strength. We identify four main feedbacks affecting the stability of the thermohaline circulation: the change in the overturning circulation of the Atlantic leads to longer residence times of the surface water in high-northern latitudes, which allows them to accumulate more precipitation and runoff from the continents. As a consequence the stratification in the North Atlantic becomes more stable. This effect is further amplified by an enhanced northward atmospheric water vapour transport, which increases the freshwater input into the North Atlantic. The reduced northward oceanic heat transport leads to colder seasurface temperatures and an intensification of the atmospheric cyclonic circulation over the Norwegian Sea. The associated Ekman transports cause increased upwelling and increased freshwater export with the East Greenland Current. Both the cooling and the wind-driven circulation changes largely compensate for the effects of the first two feedbacks. The wind-stress feedback destabilizes modes without deep water formation in the North Atlantic, but has been neglected in almost all studies so far. After the meltwater input stops, the North Atlantic deepwater formation resumed in all experiments and the meridional overturning returned within 200 years to a conveyor belt pattern. This happened although the formation of North Atlantic deep water was suppressed in one experiment for more than 300 years and the Atlantic overturning had settled into a circulation pattern with Antarctic bottom water as the only source of deep water. It is a clear indication that cooling and wind-stress feedback are more effective, at least in our model, than advection feedback and increased atmospheric water vapour transport. We conclude that the conveyor belt-type thermohaline circulation seems to be much more stable than hitherto assumed from experiments with simpler models.

Asynchronous and periodically-synchronous schemes for coupling atmosphere and ocean models are presented. The performance of the schemes is tested by simulating the climatic response to a step function forcing and to a gradually increasing forcing with a simple zero-dimensional non-linear energy balance model. Both the initial transient response and the asymptotic approach of the equilibrium state are studied. If no annual cycle is allowed the asynchronous coupling technique proves to be a suitable tool. However, if the annual cycle is retained, the periodically synchronous coupling technique reproduces the results of the synchronously coupled runs with smaller bias. In this case it is important that the total length of one synchronous period and one ocean only period is not a multiple of 6 months.

We describe a rapid and sensitive method involving time-of-flight secondary-ion mass spectrometry (TOF-SIMS) for specific laboratory diagnosis of the Smith-Lemli-Opitz syndrome, which is characterized by massive (approximately 1000-fold) accumulation of the biosynthetic cholesterol precursor 7-dehydrocholesterol. Minute amounts of blood (1-50 microL) were extracted with n-hexane, and aliquots were analyzed by TOF-SIMS. 7-Dehydrocholesterol and its isomers were detected at 491.3 mass units ([M + 107Ag]+) and cholesterol at 495.3 mass units ([M + 109Ag]+). Quantitation of 7-dehydrocholesterol and cholesterol was achieved after saponification and addition of stigmasterol as internal standard. Whereas 7-dehydrocholesterol and isomeric dehydrocholesterol were not detectable in controls, the patients revealed concentrations ranging between 0.84 and 1.25 mmol/L. Comparison with results obtained by gas chromatography indicated that quantitation by TOF-SIMS yielded the sum of 7-dehydrocholesterol, isomeric dehydrocholesterol II, and sterol III, the latter two also being increased in the patients. Consistent with quantitation by gas chromatography, the cholesterol concentrations in the patients ranged between 1.54 and 2.12 mmol/L (controls: 6.10 +/- 1.37 mmol/L).

Finite computer resources force compromises in the design of transient numerical experiments with coupled atmosphere-ocean general circulation models which, in the case of global warming simulations, normally preclude a full integration from the undisturbed pre-industrial state. The start of the integration at a later time from a climate state which, in contrast to the true climate, is initially in equilibrium then induces a cold start error. Using linear response theory a general expression for the cold start error is derived. The theory is applied to the Hamburg CO2 scenario simulations. An attempt to estimate the global-mean-temperature response function of the coupled model from the response of the model to a CO2 doubling was unsuccessful because of the non-linearity of the system. However, an alternative derivation, based on the transient simulation itself, yielded a cold start error which explained the initial retardation of the Hamburg global warming curve relative to the IPCC results obtained with a simple box-diffusion-upwelling model. In the case of the sea level the behaviour of the model is apparently more linear. The cold start error estimations based on a CO2 doubling experiment and on an experiment with gradually increasing CO2 (scenario A) are very similar and explain about two thirds of the coupled model retardation relative to the IPCC results.

Background We have previously shown that genetically induced smooth muscle cell (SMC) cycle reentry in transgenic mouse models expressing the SV40 T antigen (TAg) resulted in adaptive arterial remodeling. The present investigation targeted the in vitro expression pattern of the collageneous matrix associated with TAg-induced SMC cycle modulation. Methods SMC cultures were established from the transgenic model expressing temperature-sensitive TAg. This allowed inducible transgene expression at the permissive temperature of 33°C compared with the restrictive temperature of 39.5°C. To distinguish a transgene effect from a temperature effect, SMCs with constitutively expressed TAg were used as controls. Data were obtained using array technology, Northern blotting, reverse transcription polymerase chain reaction, and zymography. Results TAg-induced SMC cycle reentry resulted in significant down-regulation of matrix metalloproteinase (MMP)-3, whereas MMP-2, -9, and -11 were not influenced. In addition, SMC cycle reentry resulted in significantly increased RNA levels of procollagen α2(IV), procollagen α2(V), and procollagen α1(XI), whereas procollagen α1(III) and procollagen α1(VIII) were down-regulated. Studies of the RNA expression levels of granulocyte–macrophage colony-stimulating factor revealed an up-regulation of this proinflammatory and matrix-modulating cytokine. Conclusions This transgenic model provides evidence that TAg-induced cell cycle reentry is associated with a complex modulation of the collageneous matrix. Factors identified in this in vitro study reveal a comprehensive expression pattern of candidates, which might allow the vessel to undergo adaptive arterial remodeling under in vivo conditions. Our results will give rise to further investigations to elaborate on this hypothesis and to improve understanding of the role of such factors in vascular diseases.

Clear cell sarcoma of soft tissue (CCSST), also known as malignant melanoma of soft parts, represents a rare lesion of the musculoskeletal system usually affecting adolescents and young adults. CCSST is typified by a chromosomal t(12;22)(q13;q12) translocation resulting in a fusion between the Ewing sarcoma gene (EWSR1) and activating transcription factor 1 (ATF1), of which the activity in nontransformed cells is regulated by cyclic AMP. Our aim was to identify critical differentially expressed genes in CCSST tumor cells in comparison with other solid tumors affecting children and young adults to better understand signaling pathways regulating specific features of the development and progression of this tumor entity. We applied Affymetrix Human Genome U95Av2 oligonucleotide microarrays representing ∼12,000 genes to generate the expression profiles of the CCSST cell lines GG-62, DTC-1, KAO, MST2, MST3, and Su-CC-S1 in comparison with 8 neuroblastoma, 7 Ewing tumor, and 6 osteosarcoma cell lines. Subsequent hierarchical clustering of microarray data clearly separated all four of the tumor types from each other and identified differentially expressed transcripts, which are characteristically up-regulated in CCSST. Statistical analysis revealed a group of 331 probe sets, representing ∼300 significant (P < 0.001) differentially regulated genes, which clearly discriminated between the CCSST and other tumor samples. Besides genes that were already known to be highly expressed in CCSST, like S100A11 (S100 protein) or MITF (microphthalmia-associated transcription factor), this group shows an obvious portion of genes that are involved in cyclic AMP response or regulation, in pigmentation processes, or in neuronal structure and signaling. Comparison with other expression profile analyses on neuroectodermal childhood tumors confirms the high robustness of this strategy to characterize tumor entities based on their gene expression. We found the avian erythroblastic leukemia viral oncogene homologue 3 (ERBB3) to be one of the most dramatically up-regulated genes in CCSST. Quantitative real-time PCR and Northern blot analysis verified the mRNA abundance and confirmed the absence of the inhibitory transcript variant of this gene. The protein product of the member of the epidermal growth factor receptor family ERBB3 could be shown to be highly present in all of the CCSST cell lines investigated, as well as in 18 of 20 primary tumor biopsies. In conclusion, our data demonstrate new aspects of the phenotype and the biological behavior of CCSST and reveal ERBB3 to be a useful diagnostic marker.

Background: Bilaterality in breast cancer is a rare event and together with an early onset of disease points towards inheritance of the disease. However, most cases seem to occur sporadically, either in a synchronous or metachronous manner. Methods: Thirty two invasive carcinomas and one in situ carcinoma from 16 patients with synchronous, bilateral breast cancer were investigated by means of comparative genomic hybridisation (CGH) and polymerase chain reaction based multiplex microsatellite analysis. The results were analysed conventionally and were also subjected to a biomathematical cluster analysis. Results: On average, bilateral breast cancer cases showed a low number of genetic alterations, a low frequency of genetic amplifications, and a high rate of chromosomal 16q losses. A distinct, characteristic genetic alteration associated with bilateral breast disease could not be found. Although two tumour pairs appeared to be related using biomathematical processing for microsatellite analysis, this result was reproduced by CGH data processing in one patient only. Conclusions: Most synchronous, bilateral breast cancer cases seem to represent independent tumours rather than metastatic events. Nevertheless, the possibility of a specific susceptibility remains.

We describe the behaviour of 23 dynamical ocean-atmosphere models, in the context of comparison with observations in a common framework. Fields of tropical sea surface temperature (SST), surface wind stress and upper ocean vertically averaged temperature (VAT) are assessed with regard to annual mean, seasonal cycle, and interannual variability characteristics. Of the participating models, 21 are coupled GCMs, of which 13 use no form of flux adjustment in the tropics. The models vary widely in design, components and purpose: nevertheless several common features are apparent. In most models without flux adjustment, the annual mean equatorial SST in the central Pacific is too cool and the Atlantic zonal SST gradient has the wrong sign. Annual mean wind stress is often too weak in the central Pacific and in the Atlantic, but too strong in the west Pacific. Few models have an upper ocean VAT seasonal cycle like that observed in the equatorial Pacific. Interannual variability is commonly too weak in the models: in particular, wind stress variability is low in the equatorial Pacific. Most models have difficulty in reproducing the observed Pacific 'horseshoe' pattern of negative SST correlations with interannual Nino3 SST anomalies, or the observed Indian-Pacific lag correlations. The results for the fields examined indicate that several substantial model improvements are needed, particularly with regard to surface wind stress.

The response of the hydrological cycle in Europe to the increasing atmospheric concentrations of the important greenhouse gases is investigated on the basis of a time-slice experiment. The time-slice experiment has been performed with the ECHAM4 atmospheric general circulation model at a high horizontal resolution of T106 for two time-slices representing the present-day climate (1970–1999) and the future climate after an effective doubling of the carbon dioxide concentration in the atmosphere (2060–2089). The changes in the greenhouse gas concentrations have been prescribed according to the Intergovernmental Panel on Climate Change scenario IS92a. Both changes in the mean state and changes in the extremes of the hydrological cycle are considered.

The North Atlantic climate system is characterized by considerable interdecadal variability. We show examples of interdecadal variability in Figs. 9.1 and 9.2. One of the main modes of the atmosphere over the North Atlantic is the North Atlantic Oscillation (NAO) (e.g. van Loon and Rogers (1978), Hurrell (1995)). The NAO is a dipole in sea level pressure (SLP), with centers of action near Island and the Azores (Fig. 9.Ib), originally described by Walker (1924) and Walker and Bliss (1932). Hurrell (1995) defined an index of the NAO by the difference of the SLPs measured at Lisbon (Portugal) and Stykkisholmur (Iceland). Its time evolution (Fig. 9.1a) exhibits considerable interdecadal variability, with a maximum during the beginning of this century, a minimum during the 1960s, and strongly increasing values thereafter up to present. Moreover, there are observed fairly regular quasidecadal [O(10 years)] variations during the most recent decades. The relatively strong upward trend observed during the last 20 years which contributed strongly to the rise in mean Northern Hemisphere surface temperature (Hurrell (1996)) has been the matter of intense scientific debate, since it is not clear as to whether this trend reflects greenhouse warming or is simply an expression of interdecadal variability.

Prognosis of T-cell lymphoblastic lymphoma (T-LBL) in children and adolescents is poor in case of relapse, but pathogenetic knowledge of the disease which may lead to the establishment of prognostic parameters is still extremely limited. Recently, it was reported that molecular aberrations like NOTCH1 mutations or loss of heterozygosity at chromosome 6q (LOH6q) significantly correlate with relapse risk and survival in patients with T-LBL treated according to NHL-BFM protocols. PI3-kinase (PI3K) consists of two subunits (PIK3R1 and PIK3CA); its activity is repressed by the tumor suppressor PTEN. Mutations in PIK3R1, PIK3CA and PTEN leading to increased cell proliferation have been reported for different cancer types. Next generation sequencing (NGS) technologies facilitate in-depth genome-wide insight into different types of cancer. We performed NGS of pediatric T-LBL patients and validated results regarding the PI3K pathway in a large cohort of patients with respect to mutational frequencies and prognostic relevance. We analyzed five pediatric T-LBL cases by whole exome sequencing. A total of 107 cases were available for validation of candidate genes. All patients were treated according to NHL-BFM regimen for LBL. A lymphoma-specific aberration in PIK3R1 was identified via whole exome sequencing and verified by Sanger sequencing. Mutation frequencies in selected exons of PIK3R1, PIK3CA and PTEN were investigated in 107 cases. Five patients (5%) showed mutations in PIK3R1, 7 (7%) in PIK3CA, and 16 (15%) in PTEN. In 22% of the cases (24/107), mutations in at least one of the genes could be detected. These aberrations correlated with inferior probability of event-free survival (pEFS) in the analyzed cohort (pEFS at 5 years 0.80±0.05 vs. 0.61±0.10; p Although PTEN is reported to be transcriptionally repressed by NOTCH1, NOTCH1 mutations seem to set off the negative effect of PTEN mutations on outcome instead of acting synergistically in T-LBL. The pEFS of patients with mutations in both genes was 0.88±0.12 (N=10), whereas patients with mutations in PTEN, but wild-type NOTCH1 (N=6), displayed a significantly inferior pEFS compared to all other patients (0.17±0.15 vs. 0.80±0.04; p Thus, we identified mutations in PTEN as another molecular parameter which is significantly associated with prognosis in pediatric T-LBL patients treated according to NHL-BFM protocols. Validation studies on larger series are needed to evaluate the genetic interplay of these alterations and their impact on pediatric T-LBL pathogenesis. Citation Format: Bettina R. Bonn, Andreas Huge, Marius Rohde, Martin Zimmermann, Reinhard Voss, Wilhelm Woessmann, Lorenz Trümper, Claudia Rossig, Heribert Juergens, Jochen Seggewiss, Birgit Burkhardt. PTEN mutations correlate with relapse risk in pediatric T-cell lymphoblastic lymphoma patients: Validation of whole exome sequencing results. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3092. doi:10.1158/1538-7445.AM2014-3092

Impulse-response-function (IRF) models are designed for applications requiring a large number of climate change simulations, such as multi-scenario climate impact studies or cost-benefit integrated-assessment studies. The models apply linear response theory to reproduce the characteristics of the climate response to external forcing computed with sophisticated state-of-the-art climate models like general circulation models of the physical ocean-atmosphere system and three-dimensional oceanic-plus-terrestrial carbon cycle models. Although highly computer efficient, IRF models are nonetheless capable of reproducing the full set of climate-change information generated by the complex models against which they are calibrated. While limited in principle to the linear response regime (less than about 3 degreesC global-mean temperature change), the applicability of the IRF model presented has been extended into the nonlinear domain through explicit treatment of the climate system's dominant nonlinearities: CO2 chemistry in ocean water, CO2 fertilization of land biota, and sublinear radiative forcing. The resultant nonlinear impulse-response model of the coupled carbon cycle- climate system (NICCS) computes the temporal evolution of spatial patterns of climate change for four climate variables of particular relevance for climate impact studies: near- surface temperature, cloud cover, precipitation, and sea level. The space-time response characteristics of the model are derived from an EOF analysis of a transient 850-year greenhouse warming simulation with the Hamburg atmosphere-ocean general circulation model ECHAM3-LSG and a similar response experiment with the Hamburg carbon cycle model HAMOCC. The model is applied to two long-term CO2 emission scenarios, demonstrating that the use of all currently estimated fossil fuel resources would carry the Earth's climate far beyond the range of climate change for which reliable quantitative predictions are possible today, and that even a freezing of emissions to present-day levels would cause a major global warming in the long term.

The response of a coupled atmosphere-ocean general circulation model and an uncoupled atmosphere general circulation model to boundary conditions for 6000 years ago has been studied. The set of quasi-equilibrium simulations enables a separation of the contributions from changes in the ocean circulation, the CO2 concentration, and the insolation to the total response. The results indicate that all three factors have considerable impact on the climate change signal and here especially on the northern African and Indian monsoon. Multi-century adjustment processes associated with the inertia of the deep ocean play only a minor role for the climate change signal of the upper ocean and the atmosphere.

Most global climate models simulate a weakening of the North Atlantic thermohaline circulation (THC) in response to enhanced greenhouse warming. Both surface warming and freshening in high latitudes, the so-called sinking region, contribute to the weakening of the THC. Some models even simulate a complete breakdown of the THC at sufficiently strong forcing. Here results are presented from a state-of-the-art global climate model that does not simulate a weakening of the THC in response to greenhouse warming. Large-scale air–sea interactions in the Tropics, similar to those operating during present-day El Niños, lead to anomalously high salinities in the tropical Atlantic. These are advected into the sinking region, thereby increasing the surface density and compensating the effects of the local warming and freshening.

Strategies are developed to analyze and represent spatially resolved biosphere models for carbon sequestration in response to changes in atmospheric CO2 and climate by reduced-form, substitute models. We explore the High-Resolution Terrestrial Biosphere Model as implemented in the Community Terrestrial Biosphere Model (HRBM/CTBM), the Frankfurt Biosphere Model (FBM), and the box-type biosphere of the Bern model. Storage by CO2 fertilization is described by combining analytical representations of (1) net primary productivity (NPP) as a function of atmospheric CO2 and (2) a decay impulse response function to characterize the timescales of biospheric carbon turnover. Storage in response to global warming is investigated for the HRBM/CTBM. The relation between the evolution of radiative forcing and climate change is expressed by a combination of impulse response functions and empirical orthogonal functions extracted from results of the European Center/Hamburg (ECHAM3) coupled atmosphere-ocean general circulation model. A box-type, differential-analogue substitute model is developed to represent global carbon storage of the HRBM/CTBM in response to regional changes in Temperature, Precepitation and cloud cover. The substitute models represent the spatially resolved models accurately and cost-efficiently for carbon sequestration in response to changes in CO2 or in CO2 and climate and for simulations of the global isotopic signals. Deviations in carbon uptake simulated by the spatially resolved models and their substitutes are less than a few percent.

A coupled air–sea mode in the Northern Hemisphere with a period of about 35 years is described. The mode was derived from a multicentury integration with a coupled ocean–atmosphere general circulation model and involves interactions of the thermohaline circulation with the atmosphere in the North Atlantic and interactions between the ocean and the atmosphere in the North Pacific.The authors focus on the physics of the North Atlantic interdecadal variability. If, for instance, the North Atlantic thermohaline circulation is anomalously strong, the ocean is covered by positive sea surface temperature (SST) anomalies. The atmospheric response to these SST anomalies involves a strengthened North Atlantic Oscillation, which leads to anomalously weak evaporation and Ekman transport off Newfoundland and in the Greenland Sea, and the generation of negative sea surface salinity (SSS) anomalies. These SSS anomalies weaken the deep convection in the oceanic sinking regions and subsequently the strength of the thermohaline circulation. This leads to a reduced poleward heat transport and the formation of negative SST anomalies, which completes the phase reversal.The Atlantic and Pacific Oceans seem to be coupled via an atmospheric teleconnection pattern and the interdecadal Northern Hemispheric climate mode is interpreted as an inherently coupled air–sea mode. Furthermore, the origin of the Northern Hemispheric warming observed recently is investigated. The observed temperatures are compared to a characteristic warming pattern derived from a greenhouse warming simulation with the authors’ coupled general circulation model and also with the Northern Hemispheric temperature pattern associated with the 35-yr climate mode. It is shown that the recent Northern Hemispheric warming projects well onto the temperature pattern of the interdecadal mode under consideration.

Due to restrictions in the available computing resources and a lack of suitable observational data, transient climate change experiments with global coupled ocean-atmosphere models have been started from an initial state at equilibrium with the present day forcing. The historical development of greenhouse gas forcing from the onset of industrialization until the present has therefore been neglected. Studies with simplified models have shown that this "cold start" error leads to a serious underestimation of the anthropogenic global warming. In the present study, a 150-year integration has been carried out with a global coupled ocean-atmosphere model starting from the greenhouse gas concentration observed in 1935, i.e., at an early time of industrialization. The model was forced with observed greenhouse gas concentrations up to 1985, and with the equivalent C02 concentrations stipulated in Scenario A ("Business as Usual") of the Intergovernmental Panel on Climate Change from 1985 to 2085. The early starting date alleviates some of the cold start problems. The global mean near surface temperature change in 2085 is about 0.3 K (ca. 10) higher in the early industrialization experiment than in an integration with the same model and identical Scenario A greenhouse gas forcing, but with a start date in 1985. Comparisons between the experiments with early and late start dates show considerable differences in the amplitude of the regional climate change patterns, particularly for sea level. The early industrialization experiment can be used to obtain a first estimate of the detection time for a greenhouse-gas-induced near-surface temperature signal. Detection time estimates are obtained using globally and zonally averaged data from the experiment and a long control run, as well as principal component time series describing the evolution of the dominant signal and noise modes. The latter approach yields the earliest detection time (in the decade 1990-2000) for the time-evolving near-surface temperature signal. For global-mean temperatures or for temperatures averaged between 45°N and 45°S, the signal detection times are in the decades 2015-2025 and 2005-2015, respectively. The reduction of the "cold start" error in the early industrialization experiment makes it possible to separate the near-surface temperature signal from the noise about one decade earlier than in the experiment starting in 1985. We stress that these detection times are only valid in the context of the coupled model's internally-generated natural variability, which possibly underestimates low frequency fluctuations and does not incorporate the variance associated with changes in external forcing factors, such as anthropogenic sulfate aerosols, solar variability or volcanic dust. © 1995 Springer-Verlag.

Four time-dependent greenhouse warming experiments were performed with the same global coupled atmosphere-ocean model, but with each simulation using initial conditions from different ''snapshots'' of the control run climate. The radiative forcing - the increase in equivalent CO2 concentrations from 19852035 specified in the Intergovernmental Panel on Climate Change (IPCC) scenario A - was identical in all four 50-year integrations. This approach to climate change experiments is called the Monte Carlo technique and is analogous to a similar experimental set-up used in the field of extended range weather forecasting. Despite the limitation of a very small sample size, this approach enables the estimation of both a mean response and the ''between-experiment'' variability, information which is not available from a single integration. The use of multiple realizations provides insights into the stability of the response, both spatially, seasonally and in terms of different climate variables. The results indicate that the time evolution of the global mean warming signal is strongly dependent on the initial state of the climate system. While the individual members of the ensemble show considerable variation in the pattern and amplitude of near-surface temperature change after 50 years, the ensemble mean climate change pattern closely resembles that obtained in a 100-year integration performed with the same model. In global mean terms, the climate change signals for near surface temperature, the hydrological. cycle and sea level significantly exceed the variability among the members of the ensemble. Due to the high internal variability of the modelled climate system, the estimated detection time of the global mean temperature change signal is uncertain by at least one decade. While the ensemble mean surface temperature and sea level fields show regionally significant responses to greenhouse-gas forcing, it is not possible to identify a significant response in the precipitation and soil moisture fields, variables which are spatially noisy and characterized by large variability between the individual integrations.

Evidence for a cooling event synchronous with the Younger Dryas (12,000 calendar years before present) has been found in the North Pacific Ocean north of 30° N in records of surface1–5 and subsurface water properties6,7. These changes may be related to a temporary shut-down of North Atlantic Deep Water formation and associated surface cooling over the North Atlantic. It has remained unclear, however, whether this North Atlantic cooling was communicated to the North Pacific Ocean through the atmosphere or the ocean. Here we report results of a sensitivity experiment with a coupled ocean–atmosphere general circulation model that support a primarily atmospheric forcing of North Pacific climate variations. Changes in wind strongly affect coastal upwelling at the North American west coast, and surface cooling by the atmosphere causes better ventilation of the thermocline waters of the northeast Pacific. This effect is amplified by oceanic progagation to the Pacific of the signal arising from collapse of North Atlantic Deep Water formation. These teleconnections may also explain earlier North Pacific and western North American millenial-scale cooling events of a similar nature8–12.

Tangier disease (TD) is an inherited disorder of lipid metabolism characterized by very low high density lipoprotein (HDL) plasma levels, cellular cholesteryl ester accumulation and reduced cholesterol excretion in response to HDL apolipoproteins. Molecular defects in the ATP binding cassette transporter 1 (ABCA1) have recently been identified as the cause of TD. ABCA1 plays a key role in the translocation of cholesterol across the plasma membrane, and defective ABCA1 causes cholesterol storage in TD cells. Not only cholesterol efflux, but also phospholipid efflux was shown to be impaired in TD cells. By use of thin layer chromatography, high performance liquid chromatography and time-of-flight secondary ion mass spectrometry, we characterized the cellular phospholipid content in fibroblasts from three homozygous TD patients. The cellular content of the major phospholipids was not found to be significantly altered in TD fibroblasts. However, the two phospholipids cardiolipin and lysocardiolipin, which make up minute amounts in normal cells, were at least 3–5-fold enriched in fibroblasts from TD subjects. A structurally closely related phospholipid (lysobisphosphatidic acid) has recently been shown to be enriched in Niemann–Pick type C, another lipid storage disorder. Altogether these data may indicate that the role of these phospholipids is a regulatory one rather than that of a bulk mediator of cholesterol solubilization in sterol trafficking and efflux.

The introduction of a concept proposing multiple cellular subgroups in the normal female breast, including cytokeratin 5/6 (Ck 5/6)-positive progenitor cells, offers a new explanation for the existence of highly aggressive breast cancers with and without Ck 5/6 expression. Using the tissue microarray technique, 166 breast cancer cases, all characterized by comparative genomic hybridization, were evaluated by immunohistochemistry, using 15 different antibodies (estrogen receptor, progesterone receptor, p53, Ki-67, c-erbB2, epidermal growth factor receptor, cyclins A, D1, and E, bcl-2, p21, p27, Ck 5/6, Ck 8/18, and smooth muscle actin) and chromogenic in situ hybridization for c-erbB2. Biomathematical cluster analysis was applied to confirm the conventional interpretation of the results by an independent approach. Ck 5/6-positive breast carcinomas were in general negative for estrogen receptor and progesterone receptor, were highly proliferating (as reflected by Ki67 and cyclin A), and were associated with specific protein expression patterns, such as expression of p53 and epithelial growth factor receptor (all related to more aggressive tumor behavior), which could further be demonstrated by biomathematical cluster analysis. In contrast Ck 5/6-negative breast carcinomas revealed a lower tumor proliferation rate, an increased expression of p21, p27, c-erbB2, and bcl-2, and a significantly lower number of genetic alterations, with losses of chromosomal material of 16q as the most common genetic alteration. Our data give the first hints to the hypothesis that different cellular subgroups in the female breast give rise to subgroups of breast carcinomas with differing protein expression and cytogenetic alteration patterns that may be related to clinical behavior.