Establishment of a normal-derived estrogen receptor-positive cell line comparable to the prevailing human breast cancer subtype

Understanding human cancer increasingly relies on insight gained from subtype specific comparisons between malignant and non-malignant cells. The most frequent subtype in breast cancer is the luminal. By far the most frequently used model for luminal breast cancer is the iconic estrogen receptor-positive (ER(pos)) MCF7 cell line. However, luminal specific comparisons have suffered from the lack of a relevant non-malignant counterpart. Our previous work has shown that transforming growth factor-β receptor (TGFβR) inhibition suffices to propagate prospectively isolated ER(pos) human breast luminal cells from reduction mammoplasties (HBEC). Here we demonstrate that transduction of these cells with hTERT/shp16 renders them immortal while remaining true to the luminal lineage including expression of functional ER (iHBEC(ERpos)). Under identical culture conditions a major difference between MCF7 and normal-derived cells is the dependence of the latter on TGFβR inhibition for ER expression. In a breast fibroblast co-culture model we further show that whereas MCF7 proliferate concurrently with ER expression, iHBEC(ERpos) form correctly polarized acini, and segregate into proliferating and ER expressing cells. We propose that iHBEC(ERpos) may serve to shed light on hitherto unappreciated differences in ER regulation and function between normal breast and breast cancer

Propagation of oestrogen receptor-positive and oestrogen-responsive normal human breast cells in culture

Investigating the susceptibility of oestrogen receptor-positive (ER(pos)) normal human breast epithelial cells (HBECs) for clinical purposes or basic research awaits a proficient cell-based assay. Here we set out to identify markers for isolating ER(pos) cells and to expand what appear to be post-mitotic primary cells into exponentially growing cultures. We report a robust technique for isolating ER(pos) HBECs from reduction mammoplasties by FACS using two cell surface markers, CD166 and CD117, and an intracellular cytokeratin marker, Ks20.8, for further tracking single cells in culture. We show that ER(pos) HBECs are released from growth restraint by small molecule inhibitors of TGFβ signalling, and that growth is augmented further in response to oestrogen. Importantly, ER signalling is functionally active in ER(pos) cells in extended culture. These findings open a new avenue of experimentation with normal ER(pos) HBECs and provide a basis for understanding the evolution of human breast cancer

Medicin, narkotika og færdselsuheld

Projektet IMMORTAL - “Impaired Motorists, Methods of Roadside Testing and Assessment for Licensing” har indhentet ny viden om forskellige forhold, der kan påvirke køreegenskaberne. Det drejer sig bla. om påvirkning af medicin, narkotika og alkoholI ét af delprojekterne blev der taget blod- eller spytprøver fra førere af motorkøretøjer, der blev behandlet efter færdselsuheld på Odense Universitetshospital eller Glostrup sygehus. Prøverne blev analyseret for forskellige typer af medicin, narkotika samt alkohol. Patienter, der blev fundet positive eller selv havde oplyst brug af medicin eller narkotika blev efterfølgende kontaktet med henblik på at gennemføre et kvalitativt interview om uheldet og dets konsekvenser. Baseret på interviewene kan det sluttes, at i de fleste uheld var der tale om forskellige uheldsfaktorer, der kunne relateres til interviewpersonen, såsom for høj hastighed, ulovlig overhaling, manglende opmærksomhed, misopfattelse af situationen samt psykisk uligevægt. For halvdelen af de førere, der var positive for et eller flere stoffer, da uheldet skete, kan det ud fra koncentrationen antages, at påvirkningen af dette eller disse stoffer også havde medvirket til uheldets opståen. Interviewene viste, at de påvirkede førere kunne inddeles i 3 grupper: 1. Unge, velfungerende mænd, som tog illegale stoffer (overvejende hash eller amfetamin), enten om aftenen eller i weekenden. 2. Midaldrende mænd og kvinder (35-54 år), der var gået på førtidspension, givet på grund af deres afhængighed af alkohol og/eller lægeordineret medicin. 3. Førere over 55 år, som stadig var aktive på arbejdsmarkedet eller pensionerede, og som anvendte ordineret medicin eller håndkøbsmedicin

Evidence of two distinct functionally specialized fibroblast lineages in breast stroma

BACKGROUND: The terminal duct lobular unit (TDLU) is the most dynamic structure in the human breast and the putative site of origin of human breast cancer. Although stromal cells contribute to a specialized microenvironment in many organs, this component remains largely understudied in the human breast. We here demonstrate the impact on epithelium of two lineages of breast stromal fibroblasts, one of which accumulates in the TDLU while the other resides outside the TDLU in the interlobular stroma. METHODS: The two lineages are prospectively isolated by fluorescence activated cell sorting (FACS) based on different expression levels of CD105 and CD26. The characteristics of the two fibroblast lineages are assessed by immunocytochemical staining and gene expression analysis. The differentiation capacity of the two fibroblast populations is determined by exposure to specific differentiating conditions followed by analysis of adipogenic and osteogenic differentiation. To test whether the two fibroblast lineages are functionally imprinted by their site of origin, single cell sorted CD271(low)/MUC1(high) normal breast luminal epithelial cells are plated on fibroblast feeders for the observation of morphological development. Epithelial structure formation and polarization is shown by immunofluorescence and digitalized quantification of immunoperoxidase-stained cultures. RESULTS: Lobular fibroblasts are CD105(high)/CD26(low) while interlobular fibroblasts are CD105(low)/CD26(high). Once isolated the two lineages remain phenotypically stable and functionally distinct in culture. Lobular fibroblasts have properties in common with bone marrow derived mesenchymal stem cells and they specifically convey growth and branching morphogenesis of epithelial progenitors. CONCLUSIONS: Two distinct functionally specialized fibroblast lineages exist in the normal human breast, of which the lobular fibroblasts have properties in common with mesenchymal stem cells and support epithelial growth and morphogenesis. We propose that lobular fibroblasts constitute a specialized microenvironment for human breast luminal epithelial progenitors, i.e. the putative precursors of breast cancer. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13058-016-0769-2) contains supplementary material, which is available to authorized users

Constraints on Civil Society's Capacity to Curb Corruption

Summaries This article suggests reasons why it is particularly difficult for civil society to organise effectively to curb forms of corruption which disproportionately afflict the poor. We take issue with currently fashionable views on the state's ability to ‘foster’ the emergence of civil?society organisations which effectively articulate the interests of socially excluded people. The Indian case demonstrates the potential for state?fostering to produce precisely the kinds of ‘compromised’ civil?society organisations which inhibit the emergence of the most effective form of anti?corruption movement: one based upon the premise that citizens have a right to audit government finances in minute detail. Indeed, the organisation in India that has pioneered this radical means for increasing accountability to the poor consciously distances itself from state?sponsored efforts to influence the development of civil society. In Latin America — whence most of the evidence for the optimistic view of state?fostered civil society has been drawn — efforts to promote popular auditing (as opposed to participatory planning) are conspicuous by their absence. This raises suspicions that state?fostering of civil society, whatever its merits, comes at a heavy price: the proliferation of associations unwilling to migrate from ‘safe’ forms of participation to those which involve confronting powerful elites and challenging the state's prerogative of auditing its own financial conduct. Such groups appear disinclined to demand and obtain sensitive government records, let alone organise ordinary people to analyse such information to determine where funds allocated for their benefit have actually gone

Analysis of mortality metrics associated with a comprehensive range of disorders in Denmark, 2000 to 2018: A population-based cohort study

Background: The provision of different types of mortality metrics (e.g., mortality rate ratios [MRRs] and life expectancy) allows the research community to access a more informative set of health metrics. The aim of this study was to provide a panel of mortality metrics associated with a comprehensive range of disorders and to design a web page to visualize all results. Methods and findings: In a population-based cohort of all 7,378,598 persons living in Denmark at some point between 2000 and 2018, we identified individuals diagnosed at hospitals with 1,803 specific categories of disorders through the International Classification of Diseases-10th Revision (ICD-10) in the National Patient Register. Information on date and cause of death was obtained from the Registry of Causes of Death. For each of the disorders, a panel of epidemiological and mortality metrics was estimated, including incidence rates, age-of-onset distributions, MRRs, and differences in life expectancy (estimated as life years lost [LYLs]). Additionally, we examined models that adjusted for measures of air pollution to explore potential associations with MRRs. We focus on 39 general medical conditions to simplify the presentation of results, which cover 10 broad categories: circulatory, endocrine, pulmonary, gastrointestinal, urogenital, musculoskeletal, hematologic, mental, and neurologic conditions and cancer. A total of 3,676,694 males and 3,701,904 females were followed up for 101.7 million person-years. During the 19-year follow-up period, 1,034,273 persons (14.0%) died. For 37 of the 39 selected medical conditions, mortality rates were larger and life expectancy shorter compared to the Danish general population. For these 37 disorders, MRRs ranged from 1.09 (95% confidence interval [CI]: 1.09 to 1.10) for vision problems to 7.85 (7.77 to 7.93) for chronic liver disease, while LYLs ranged from 0.31 (0.14 to 0.47) years (approximately 16 weeks) for allergy to 17.05 (16.95 to 17.15) years for chronic liver disease. Adjustment for air pollution had very little impact on the estimates; however, a limitation of the study is the possibility that the association between the different disorders and mortality could be explained by other underlying factors associated with both the disorder and mortality. Conclusions: In this study, we show estimates of incidence, age of onset, age of death, and mortality metrics (both MRRs and LYLs) for a comprehensive range of disorders. The interactive data visualization site (https://nbepi.com/atlas) allows more fine-grained analysis of the link between a range of disorders and key mortality estimates.publishedVersio

Long-term thermal sensitivity of Earth’s tropical forests

The sensitivity of tropical forest carbon to climate is a key uncertainty in predicting global climate change. Although short-term drying and warming are known to affect forests, it is unknown if such effects translate into long-term responses. Here, we analyze 590 permanent plots measured across the tropics to derive the equilibrium climate controls on forest carbon. Maximum temperature is the most important predictor of aboveground biomass (−9.1 megagrams of carbon per hectare per degree Celsius), primarily by reducing woody productivity, and has a greater impact per °C in the hottest forests (>32.2°C). Our results nevertheless reveal greater thermal resilience than observations of short-term variation imply. To realize the long-term climate adaptation potential of tropical forests requires both protecting them and stabilizing Earth’s climate