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Radiation-induced fibrosis (RIF) is the main late radiation toxicity in breast cancer patients. Most of the current 3D in vitro breast cancer models are composed by cancer cells only and are unable to reproduce the complex cellular homeostasis within the tumor microenvironment to study RIF mechanisms. In order to account complex cellular interactions within the tumor microenvironment, an advanced 3D spheroid model, consisting of the luminal breast cancer MCF-7 cells and MRC-5 fibroblasts, was developed. The spheroids were generated using the liquid overlay technique in culture media into 96-well plates previously coated with 1% agarose (m/v, in water). In total, 21 experimental setups were tested during the optimization of the model. The generated spheroids were characterized using fluorescence imaging, immunohistology and immunohistochemistry. The expression of ECM components was confirmed in co-culture spheroids. Using α-SMA staining, we confirmed the differentiation of healthy fibroblasts into myofibroblasts upon the co-culturing with cancer cells. The induction of fibrosis was studied in spheroids treated 24 h with 10 ng/mL TGF-β and/or 2 Gy irradiation. Overall, the developed advanced 3D stroma-rich in vitro model of breast cancer provides a possibility to study fibrosis mechanisms taking into account 3D arrangement of the complex tumor microenvironment.

Hypoxia is a key factor responsible for the failure of therapeutic response in most solid tumors and promotes the acquisition of tumor resistance to various antitumor immune effectors. Reshaping the hypoxic immune suppressive tumor microenvironment to improve cancer immunotherapy is still a relevant challenge. We investigated the impact of inhibiting HIF-1α transcriptional activity on cytotoxic immune cell infiltration into B16-F10 melanoma. We showed that tumors expressing a deleted form of HIF-1α displayed increased levels of NK and CD8+effector T cells in the tumor microenvironment, which was associated with high levels of CCL2 and CCL5 chemokines. We showed that combining acriflavine, reported as a pharmacological agent preventing HIF-1α/HIF-1β dimerization, dramatically improved the benefit of cancer immunotherapy based on TRP-2 peptide vaccination and anti-PD-1 blocking antibody. In melanoma patients, we revealed that tumors exhibiting high CCL5 are less hypoxic, and displayed high NK, CD3+, CD4+and CD8+T cell markers than those having low CCL5. In addition, melanoma patients with high CCL5 in their tumors survive better than those having low CCL5. This study provides the pre-clinical proof of concept for a novel triple combination strategy including blocking HIF-1α transcription activity along vaccination and PD-1 blocking immunotherapy.

Publisher's version (útgefin grein), This paper presents the SURATLANT data set (SURveillance ATLANTique). It consists of individual data of temperature, salinity, parameters of the carbonate system, nutrients, and water stable isotopes (δ18O and δD) collected mostly from ships of opportunity since 1993 along transects between Iceland and Newfoundland (https://doi.org/10.17882/54517). We discuss how the data are validated and qualified, their accuracy, and the overall characteristics of the data set. The data are used to reconstruct seasonal cycles and interannual anomalies, in particular of sea surface salinity (SSS); inorganic nutrients; dissolved inorganic carbon (DIC); and its isotopic composition δ13CDIC, total alkalinity (At), and water isotope concentrations. Derived parameters such as fCO2 and pH are also estimated. The relation between salinity and At is estimated from these data to investigate the possibility to replace missing At when estimating other parameters of the carbonate system. When examining the average seasonal cycle in the deep ocean, in both these data with other climatologies, we find a period of small seasonal change between January and late April. On the Newfoundland shelf and continental slope, changes related with spring stratification and blooms occur earlier. The data were collected in a period of multi-decennial variability associated with the Atlantic multi-decadal variability with warming between 1994 and 2004–2007, and with the recent cooling having peaked in 2014–2016. We also observe strong salinification in 2004–2009 and fresher waters in 1994–1995 as well as since 2010 south of 54° N and in 2016–2017 north of 54° N. Indication of multi-decadal variability is also suggested by other variables, such as phosphate or DIC, but cannot be well resolved seasonally with the discrete sampling and in the presence of interannual variability. As a whole, over the 24 years, the ocean fCO2 trend (+1.9 µatm yr−1) is close to the atmospheric trend and associated with an increase in DIC (+0.77 µmol kg−1 yr−1). The data also revealed a canonical pH decrease of −0.0021 yr−1. There is also a decrease in δ13CDIC between 2005 and 2017 (in winter, −0.014 ‰ yr−1, but larger in summer, −0.042 ‰ yr−1), suggesting a significant anthropogenic carbon signal at play together with other processes (mixing, biological activity)., The authors declare that they have no conflict of interest. Easy access on the merchant vessels run or leased by EIMSKIP has been the core of this long effort to maintain surface sampling between Iceland and Newfoundland. The nearly 100 volunteer ship riders and their enthusiasm have been key to the success of this monitoring. The project was initiated when one of the authors, GR, was at LDEO, with initial support from this institution. NOAA/AOML and NOAA/CPO Ocean Observing and Monitoring Division have contributed by maintaining the thermosalinographs and providing XBTs on the different ships that have operated between Iceland and Newfoundland. The French effort was supported by various agencies throughout the years and in particular INSU (direct support to SNO SSS and by the LEFE/CYBER grant CO2SINK and LEFE/GMMC grant GREENGROG since 2016) and IPEV. Support by National Power Company of Iceland Landsvirkjun is acknowledged. The δ 13CDIC sampling was initiated by Paul Quay (University of Washington, Seattle) in 2005–2006. SNAPO-CO2 is acknowledged for analyzing DIC/ At at LODYC/LOCEAN since 2001 and the isotopic platform of OSU Ecce Terra for analyzing δ 13CDIC as well as water stable isotopes at LOCEAN since 2010. Collaboration between French and Icelandic investigators has been supported by PHC Jules Verne 2016 grant 36187YF. Support for Virginie Racapé was most recently provided by IFREMER and for Marion Benetti by a grant from the University of Iceland in Reykjavik. The very supportive help of Christian Brunet for the analysis of DIC/ At samples in 2001–2014 is warmly acknowledged. Support from the European Integrated Project CARBOOCEAN (511176) is also acknowledged. Some figures were plotted using Ocean Data View (ODV) (Schlitzer, 2013). Comments and suggestions by two reviewers have contributed to improve the manuscript.

International audience; Hirsutism induced by hyperandrogenism can be associated with polycystic ovary syndrome, 21-hydroxylase (OH) deficiency or androgen-secreting tumors, including ovarian and adrenal tumors. Adrenal androgen-secreting tumors are frequently malignant. Adrenal oncocytomas represent rare causes of hyperandrogenism. The aim of the study was to investigate steroidogenic enzyme expression and steroid secretion in an androgen-secreting adrenal oncocytoma in a young woman presenting with hirsutism. Hyperandrogenism was diagnosed on the basis of elevated plasma Δ4-androstenedione and testosterone levels. Pelvic ultrasound was normal, CT scanning revealed a right adrenal mass. Androgens were assessed in adrenal and ovarian vein samples and proved a right adrenal origin. Adrenalectomy normalized androgen levels and the adrenal tumor was diagnosed as an oncocytoma. Real time-PCR, immunohistochemistry and cell culture studies were performed on tumor explants to investigate the steroid secretion profile. Among enzymes required for cortisol synthesis, 17α-OH and 3β-hydroxysteroid dehydrogenase 2 (3β-HSD2) were highly expressed whereas 21-OH and 11β-OH were weakly produced at the mRNA and/or protein levels. Enzymes involved in testosterone production, 17β-HSD5 and 17β-HSD3, were also detected. ACTH receptor was present in the tissue. Cortisol, Δ4-androstenedione and testosterone secretions by cultured cells were increased by ACTH. These results provide the first demonstration, to our knowledge, of abnormal expression profile of steroidogenic enzymes in an adrenocortical oncocytoma. Our results also indicate that Δ4-androstenedione hypersecretion resulted from high 17α-OH and 3β-HSD2 expression in combination with low expression of 21-OH and 11β-OH. Testosterone production was ascribed to occurrence of 17β-HSD5 and 17β-HSD3. Finally, our results indicate that androgen secretion was stimulated by ACTH.

This paper describes the ways and means of assembling and quality controling the Irminger Sea and Iceland Sea time-series biogeochemical data which are included in the CARINA data set. The Irminger Sea and the Iceland Sea are hydrographically different regions where measurements of sea water carbon and nutrient chemistry were started in 1983. The sampling is seasonal, four times a year. The carbon chemistry is studied with measurements of the partial pressure of carbon dioxide in seawater, pCO2, and total dissolved inorganic carbon, TCO2. The carbon chemistry data are for surface waters only until 1991 when water column sampling was initiated. Other measured parameters are salinity, dissolved oxygen and the inorganic nutrients nitrate, phosphate and silicate. Because of the CARINA criteria for secondary quality control, depth >1500 m, the IRM-TS could not be included in the routine QC and the IS-TS only in a limited way. However, with the information provided here, the quality of the data can be assessed, e.g. on the basis of the results obtained with the use of reference materials.

This paper describes the ways and means of assembling and quality controling the Irminger Sea and Iceland Sea time-series biogeochemical data which are included in the CARINA data set. The Irminger Sea and the Iceland Sea are hydrographically different regions where measurements of sea water carbon and nutrient chemistry were started in 1983. The sampling is seasonal, four times a year. The carbon chemistry is studied with measurements of the partial pressure of carbon dioxide in seawater, pCO2, and total dissolved inorganic carbon, TCO2. The carbon chemistry data are for surface waters only until 1994 when water column sampling was initiated. Other measured parameters are salinity, dissolved oxygen and the inorganic nutrients nitrate, phosphate and silicate. Because of the CARINA criteria for secondary quality control, depth >1500 m, the IRM-TS could not be included in the routine QC and the IS-TS only in a limited way. However, with the information provided here, the quality of the data can be assessed e.g. on the basis of the results obtained with the use of reference materials.

The Iceland Sea is one part of the Nordic Seas. Cold Arctic Water prevails there and the deep-water is an important source of North Atlantic Deep Water. We have evaluated time series observations of measured pCO2 and total CO2 concentration from discrete seawater samples during 1985–2008 for the surface and 1994–2008 for deep-water, and following changes in response to increasing atmospheric carbon dioxide. The surface pH in winter decreases at a rate of 0.0024 yr−1, which is 50% faster than average yearly rates at two subtropical time series stations, BATS and ESTOC. In the deep-water regime (>1500 m), the rate of pH decline is a quarter of that observed in surface waters. The surface seawater carbonate saturation states (Ω) are about 1.5 for aragonite and 2.5 for calcite, about half of levels found in subtropical surface waters. During 1985–2008, the degree of saturation (Ω) decreased at an average rate of 0.0072 yr−1 for aragonite and 0.012 yr−1 for calcite. The aragonite saturation horizon is currently at 1710 m and shoaling at 4 m yr−1. Based on this rate of shoaling and on the local hypsography, each year another 800 km2 of seafloor becomes exposed to waters that have become undersaturated with respect to aragonite.