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Neoadjuvant therapy with abiraterone acetate + prednisone (AAP) + luteinising hormone-releasing hormone agonist (LHRHa) induced significant cytoreduction associated with a lack of recurrence versus LHRHa, though staging did not differ significantly. This first randomised AAP study in localised high-risk prostate cancer explores response heterogeneity and associated molecular effects to guide further research. © 2019 Optimal therapeutic strategy remains an unmet need in localised high-risk prostate cancer (LHRPC). Androgen biosynthesis inhibition in the preoperative setting may improve outcomes. In this single-centre randomised trial, we looked at therapy outcomes of preoperative treatment with abiraterone acetate + prednisone (AAP) + luteinising hormone-releasing hormone agonist (LHRHa) or LHRHa alone followed by radical prostatectomy in 65 men. We did not see a significant difference of organ-confined carcinoma (p = 0.27). However, tumour volume measures were significantly lower for AAP + LHRHa treatment (p ≤ 0.001). Of note, lower tumour epithelium volume correlated with improved biochemical recurrence-free survival at ≥4-yr follow-up (p = 0.0014). Tumours pretreated with AAP + LHRHa had lower proliferation and androgen signalling expression than LHRHa. On multivariate analysis, glucocorticoid receptor (GR) overexpression correlated with persistent tumours in AAP + LHRHa (p = 0.018). The presence of nuclear androgen receptor splice variant (nARV7) correlated with persistent tumours in both arms. No new safety signals were observed. This is the first study investigating the role of preoperative AAP + LHRHa versus LHRHa alone in LHRPC. We report significant cytoreduction by tumour volume measures inversely correlating with biochemical relapse. Validation of these proposed tumour volume measures is planned. A potential role of GR in resistance to androgen biosynthesis inhibition warrants further study. Patient summary: This is the first study of abiraterone acetate plus leuprolide versus leuprolide alone in high-risk localised prostate cancer followed by prostatectomy. Patients in the combination arm had a significantly smaller tumour size. © 2019

This Chapter introduces the need for general Digital Rights Management (DRM) requirements. Further, it intertwines DRM with its spatial counterpart, Geospatial DRM (GeoDRM). However, unlike DRM, GeoDRM is far much complicated due to issues such as the development of Web Mapping technology among other issues. The Chapter discusses the ability of GeoDRM to mitigate transgression of Intellectual Property Rights (IPR). Highlighting economical and environmental wellbeing and other benefits of Spatial Data Infrastructure (SDI) geared towards global sustainable developments, the Chapter focuses on challenges of National Spatial Data Infrastructures (NSDIs) and Regional SDIs and the need to harmonize their standards for the upward mobility of global SDI (GSDI). Emphasizing the undisputed need for Local, Regional and Global Spatial Data Infrastructures (SDIs), in the presence of various Geo-communities and different GeoDRM models, the Chapter concludes that capacity building need to be urgently but carefully harnessed across all levels in order to develop cohesive GeoDRM policies.

Nuclear protein aggregation has been linked to genome instability and disease. The main source of aggregation‐prone proteins in cells is defective ribosomal products (DRiPs), which are generated by translating ribosomes in the cytoplasm. Here, we report that DRiPs rapidly diffuse into the nucleus and accumulate in nucleoli and PML bodies, two membraneless organelles formed by liquid–liquid phase separation. We show that nucleoli and PML bodies act as dynamic overflow compartments that recruit protein quality control factors and store DRiPs for later clearance. Whereas nucleoli serve as constitutive overflow compartments, PML bodies are stress‐inducible overflow compartments for DRiPs. If DRiPs are not properly cleared by chaperones and proteasomes due to proteostasis impairment, nucleoli undergo amyloidogenesis and PML bodies solidify. Solid PML bodies immobilize 20S proteasomes and limit the recycling of free ubiquitin. Ubiquitin depletion, in turn, compromises the formation of DNA repair compartments at fragile chromosomal sites, ultimately threatening cell survival.

Contemporary biological cells are sophisticated and highly compartmentalized. Compartmentalization is an essential principle of prebiotic life as well as a key feature in bottom-up synthetic biology research. In this review, the dynamic growth of compartments as an essential prerequisite for enabling self-reproduction as a fundamental life process is discussed. The micrometer-sized compartments are focused on due to their cellular dimensions. Two types of compartments are considered, membraneless droplets and membrane-bound microcompartments. Growth mechanisms of aqueous droplets such as protein (condensates) or macromolecule-rich droplets (aqueous two phase systems) and coacervates are discussed, for which growth occurs via Ostwald ripening or coalescence. For membrane-bound compartments, vesicles are considered, which are composed of fatty acids, lipids, or polymers, where directed growth can occur via fusion or uptake of material from the surrounding. The development of novel approaches for growth of biomimetic microcompartments can eventually be utilized to construct new synthetic cells.

Many industrial activities impose a threat on biodiversity, and it is unclear to what extent environmental regulations can reduce the threat of such activities. Bycatch in industrial fisheries is one of the greatest sources of mortality for seabirds, but a threat for which effective mitigation exists. Here we quantify whether the introduction of a new regulation that required the use of bird-scaring lines reduced seabird mortality in two of the most hazardous fisheries in the South Atlantic. The Namibian hake demersal trawl and longline fisheries, estimated to be killing 20,000–30,000 birds/year, have been required to use bird-scaring lines since 2015. We used data from BirdLife International's Albatross Task Force and the Namibian Fisheries Observer Agency to quantify changes in seabird mortality in these fisheries before and after the introduction of these regulations. Our estimated bycatch rates in the longline fleet were 0.468 birds/1000 hooks (95% confidence interval 0.067–1.450) before regulations and 0.004 birds/1000 hooks (0.001–0.013) following their introduction, a 98.4% reduction. Our estimate suggests that 215 (1–751) seabirds were killed across this fleet in 2018 compared to 22,222 (3187–68,786) in 2009. In the trawl fleet, observers recorded seabird mortality resulting from interactions with trawl cables. The average rate of heavy interactions was 1.09 interactions/h (0.81–1.39) before the regulation came into effect, and 0.49 interactions/h (0.23–0.84) since then. Extrapolations based on the number of observed fatal interactions suggest 1452 (0–3865) birds were killed by this fleet in 2017 compared to 7030 (0–16,374) in 2009. The lower mortality reduction in the trawl fleet is likely due to incomplete implementation of regulations and highlights the importance of adequate enforcement for effective bycatch mitigation. Overall, we demonstrate that regulations that mandate that well-tested safeguards are used during industrial operations can have enormous benefits for the conservation of threatened species.

The rapid growth of Botswana's economy since independence in 1966 has brought more tarred roads and vehicles, accompanied by an escalating road crash fatality rate. We tested the hypothesis that motor vehicle crash fatality increases resulted from, rather than just corresponded with, annual gross domestic product (GDP) increases. Data from Zambia, adjacent to Botswana, were used for comparison.Annual social and economic indicators and motor vehicle crash fatality rates in Botswana and Zambia were accessed from 1960 to 2012 and analysed using vector autoregressive analysis and Granger causality tests.In Botswana, annual changes in per capita GDP predicted annual changes in motor vehicle crash fatality rates (Road crash fatalities increased in recent decades in both Zambia and Botswana. But the rapid economic development in Botswana over this time period appears to have driven proportionate road traffic fatality increases. There are opportunities for newly emerging economies such as Zambia, Angola, and others to learn from the Botswana experience. Evidence-based investments in road safety interventions should be concomitant with economic development.La croissance rapide de l’économie du Botswana depuis l’indépendance en 1966 s’est traduite par le développement du nombre de routes goudronnées et de véhicules, accompagnés d’un taux de mortalité due aux accidents de la route qui va s’accélérant. Nous avons testé l’hypothèse selon laquelle les hausses de la mortalité due aux accidents de véhicules motorisés seraient attribuables aux augmentations du produit intérieur brut (PIB), plutôt que d’en être un simple reflet. Des données provenant de Zambie, pays adjacent au Botswana, ont été utilisées pour établir une comparaison.Des indicateurs économiques et sociaux annuels et les taux de mortalité due aux accidents de la route au Botswana et en Zambie ont été examinés sur la période 1960–2012 et analysés en utilisant une analyse vectorielle autorégressive et des tests de causalité au sens de Granger.Au Botswana, les variations annuelles de PIB par habitant ont prédit les variations annuelles des taux de mortalité due aux accidents de véhicule motorisés (p = 0,042). L’inverse n’a pas été observé; les variations annuelles de taux de mortalité due aux accidents de véhicules motorisés ne permettent pas de prédire les variations annuelles de PIB. Ces résultats suggèrent que la croissance du PIB pour une année donnée a causé des décès occasionnés par des accidents de la route au Botswana et qu’en moyenne, chaque augmentation d’un milliard de dollars du PIB a produit une augmentation du taux de décès occasionnés par des accidents de la route. En Zambie, les variations annuelles de PIB ont prédit les variations annuelles du taux de mortalité trois ans plus tard (p = 0,029), mais les variations annuelles des taux de mortalité des accidents de la route ont également prédit les augmentations annuelles de PIB par habitant (p = 0,026) trois ans plus tard, ce qui suggère une corrélation entre les tendances mais pas un effet de causalité du PIB.Les décès occasionnés par les accidents de la route ont augmenté au cours des dernières décennies en Zambie comme au Botswana. Mais le développement économique rapide au Botswana au cours de cette période semble avoir entraîné des augmentations proportionnelles des décès dus aux accidents de la route. Il est possible, pour les nouvelles économies émergentes comme la Zambie, l’Angola, et d’autres, de tirer des leçons de l’expérience du Botswana. Des investissements dans des interventions en matière de sécurité routière, fondés sur des données concrètes, doivent être concomitants au développement économique.

Summary Cells sense elevated temperatures and mount an adaptive heat shock response that involves changes in gene expression, but the underlying mechanisms, particularly on the level of translation, remain unknown. Here we report that, in budding yeast, the essential translation initiation factor Ded1p undergoes heat-induced phase separation into gel-like condensates. Using ribosome profiling and an in vitro translation assay, we reveal that condensate formation inactivates Ded1p and represses translation of housekeeping mRNAs while promoting translation of stress mRNAs. Testing a variant of Ded1p with altered phase behavior as well as Ded1p homologs from diverse species, we demonstrate that Ded1p condensation is adaptive and fine-tuned to the maximum growth temperature of the respective organism. We conclude that Ded1p condensation is an integral part of an extended heat shock response that selectively represses translation of housekeeping mRNAs to promote survival under conditions of severe heat stress.

Stressed cells shut down translation, release mRNA molecules from polysomes, and form stress granules (SGs) via a network of interactions that involve G3BP. Here we focus on the mechanistic underpinnings of SG assembly. We show that, under non-stress conditions, G3BP adopts a compact auto-inhibited state stabilized by electrostatic intramolecular interactions between the intrinsically disordered acidic tracts and the positively charged arginine-rich region. Upon release from polysomes, unfolded mRNAs outcompete G3BP auto-inhibitory interactions, engendering a conformational transition that facilitates clustering of G3BP through protein-RNA interactions. Subsequent physical crosslinking of G3BP clusters drives RNA molecules into networked RNA/protein condensates. We show that G3BP condensates impede RNA entanglement and recruit additional client proteins that promote SG maturation or induce a liquid-to-solid transition that may underlie disease. We propose that condensation coupled to conformational rearrangements and heterotypic multivalent interactions may be a general principle underlying RNP granule assembly.

© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim Among the best-performing homogeneous catalysts for the direct amination of activated secondary alcohols with electron-poor amine derivatives, metal triflates, such as aluminum triflate, Al(OTf)3, stand out. Herein we report the extension of this reaction to electron-rich amines and activated primary alcohols. We provide detailed insight into the structure and reactivity of the catalyst under working conditions in both nitromethane and toluene solvent, through experiment (cyclic voltammetry, conductimetry, NMR spectroscopy), and density functional theory (DFT) simulations. Competition between aniline and benzyl alcohol for Al in the two solvents explains the different reactivities. The catalyst structures predicted from the DFT calculations were validated by the experiments. Whereas a SN1-type mechanism was found to be active in nitromethane, we propose a SN2 mechanism in toluene to rationalize the much higher selectivity observed when using this solvent. Also, unlike what is commonly assumed in homogeneous catalysis, we show that different active species may be active instead of only one.

RNA and membraneless organelles Membraneless compartments can form in cells through liquidliquid phase separation (see the Perspective by Polymenidou). But what prevents these cellular condensates from randomly fusing together? Using the RNA-binding protein (RBP) Whi3, Langdon et al. demonstrated that the secondary structure of different RNA components determines the distinct biophysical and biological properties of the two types of condensates that Whi3 forms. Several RBPs, such as FUS and TDP43, contain prion-like domains and are linked to neurodegenerative diseases. These RBPs are usually soluble in the nucleus but can form pathological aggregates in the cytoplasm. Maharana et al. showed that local RNA concentrations determine distinct phase separation behaviors in different subcellular locations. The higher RNA concentrations in the nucleus act as a buffer to prevent phase separation of RBPs; when mislocalized to the cytoplasm, lower RNA concentrations trigger aggregation. Science , this issue p. 922 , p. 918 ; see also p. 859

The formation of membrane-less organelles and compartments by protein phase separation is an important way in which cells organize their cytoplasm and nucleoplasm. In vitro phase separation assays with purified proteins have become the standard way to investigate proteins that form membrane-less compartments. By now, various proteins have been purified and tested for their ability to phase separate and form liquid condensates in vitro. However, phase-separating proteins are often aggregation-prone and difficult to purify and handle. As a consequence, the results from phase separation assays often differ between labs and are not easily reproduced. Thus, there is an urgent need for high-quality proteins, standardized procedures, and generally agreed-upon practices for protein purification and conducting phase separation assays. This paper provides protocols for protein purification and guides the user through the practicalities of in vitro protein phase separation assays, including best-practice approaches and pitfalls to avoid. We believe that this compendium of protocols and practices will provide a useful resource for scientists studying the phase behavior of proteins., Graphical Abstract Unlabelled Image, Highlights • Membrane-less organelles form by phase separation. • Membrane-less organelles can be reconstituted from minimal components. • Phase-separating proteins are difficult to purify and handle. • We provide guidelines and protocols for working with phase-separating proteins.

Provided is a method for producing an alkene. The method comprises a step of converting a compound (I) comprising at least two hydroxyl functions into a compound (II) comprising at least two alkenyl functions with cerium oxide particles as catalyst. It notably permits the conversion of 1,3-butanediol,1,4-butanediol into butadiene.; L'invention concerne un procédé de production d'un alcène. Le procédé comprend une étape de conversion d'un composé (I) comprenant au moins deux fonctions hydroxyle en un composé (II) comprenant au moins deux fonctions alcényle avec des particules d'oxyde de cérium comme catalyseur. Elle permet notamment la conversion de 1,3-butanediol,1,4-butanediol en butadiène.

INTRODUCTION The formation of dynamic, membraneless compartments using intracellular phase transitions such as phase separation and gelation provides an efficient way for cells to respond to environmental changes. Recent work has identified a special class of intrinsically disordered domains enriched for polar amino acids such as glycine, glutamine, serine, or tyrosine as potential drivers of phase separation in cells. However, more traditional work has highlighted the ability of these domains to drive the formation of fibrillar aggregates. Such domains are also known as prion domains. They have first been identified in budding yeast proteins that form amyloid-like aggregates. Because these aggregates are heritable and change the activity of the prion-domain–containing protein, they are thought to be a common mechanism for phenotypic inheritance in fungi and other organisms. However, the aggregation of prion domains has also been associated with neurodegenerative diseases in mammals. Therefore, the relationship between the role of these domains as drivers of phase separation and their ability to form prion-like aggregates is unknown. RATIONALE The budding yeast translation termination factor Sup35 is an archetypal prion-domain–containing protein. Sup35 forms irreversible heritable aggregates, and these aggregates have been proposed to be either a disease or an adaptation that generates heritable phenotypic variation in populations of budding yeast. Despite having been described almost 25 years ago, the physiological functions of the Sup35 prion domain and other prion-like domains remain unclear. Uncovering these functions is a prerequisite for understanding the evolutionary pressures shaping prion-like sequences and how their physiological and pathological transitions affect cellular fitness. RESULTS Here, we show that the prion domain of Sup35 drives the reversible phase separation of the translation termination factor into biomolecular condensates. These condensates are distinct and different from fibrillar amyloid-like prion particles. Combining genetic analysis in cells with in vitro reconstitution protein biochemistry and quantitative biophysical methods, we demonstrate that Sup35 condensates form by pH-induced liquid-like phase separation as a response to sudden stress. The condensates are liquid-like initially but subsequently solidify to form protective protein gels. Cryo–electron tomography demonstrates that these gel-like condensates consist of cross-linked Sup35 molecules forming a porous meshwork. A cluster of negatively charged amino acids functions as a pH sensor and regulates condensate formation. The ability to form biomolecular condensates is shared among distantly related budding yeast and fission yeast. This suggests that condensate formation is a conserved and ancestral function of the prion domain of Sup35. In agreement with an important physiological function of the prion domain, the catalytic guanosine triphosphatase (GTPase) domain of the translation termination factor Sup35 readily forms irreversible aggregates in the absence of the prion domain. Consequently, cells lacking the prion domain exhibit impaired translational activity and a growth defect when recovering from stress. These data demonstrate that the prion domain rescues the essential GTPase domain of Sup35 from irreversible aggregation, thus ensuring that the translation termination factor remains functional during harsh environmental conditions. CONCLUSION The prion domain of Sup35 is a highly regulated molecular device that has the ability to sense and respond to physiochemical changes within cells. The N-terminal prion domain provides the interactions that drive liquid phase separation. Phase separation is regulated by the adjacent stress sensor. The synergy of these two modules enables the essential translation termination factor to rapidly form protective condensates during stress. This suggests that prion domains are protein-specific stress sensors and modifiers of protein phase transitions that allow cells to respond to specific environmental conditions.

SummaryMany proteins contain disordered regions of low-sequence complexity, which cause aging-associated diseases because they are prone to aggregate. Here, we study FUS, a prion-like protein containing intrinsically disordered domains associated with the neurodegenerative disease ALS. We show that, in cells, FUS forms liquid compartments at sites of DNA damage and in the cytoplasm upon stress. We confirm this by reconstituting liquid FUS compartments in vitro. Using an in vitro “aging” experiment, we demonstrate that liquid droplets of FUS protein convert with time from a liquid to an aggregated state, and this conversion is accelerated by patient-derived mutations. We conclude that the physiological role of FUS requires forming dynamic liquid-like compartments. We propose that liquid-like compartments carry the trade-off between functionality and risk of aggregation and that aberrant phase transitions within liquid-like compartments lie at the heart of ALS and, presumably, other age-related diseases.Video Abstract

International audience; no abstract

This Chapter introduces the need for general Digital Rights Management (DRM) requirements. Further, it intertwines DRM with its spatial counterpart, Geospatial DRM (GeoDRM). However, unlike DRM, GeoDRM is far much complicated due to issues such as the development of Web Mapping technology among other issues. The Chapter discusses the ability of GeoDRM to mitigate transgression of Intellectual Property Rights (IPR). Highlighting economical and environmental wellbeing and other benefits of Spatial Data Infrastructure (SDI) geared towards global sustainable developments, the Chapter focuses on challenges of National Spatial Data Infrastructures (NSDIs) and Regional SDIs and the need to harmonize their standards for the upward mobility of global SDI (GSDI). Emphasizing the undisputed need for Local, Regional and Global Spatial Data Infrastructures (SDIs), in the presence of various Geo-communities and different GeoDRM models, the Chapter concludes that capacity building need to be urgently but carefully harnessed across all levels in order to develop cohesive GeoDRM policies.

National audience

Objective The present study reviews a series of patients who underwent thyroid surgery in Eldoret, Kenya, to demonstrate the feasibility of conducting long-term (1 year) outcomes research in a resource-limited setting, impact on the quality of life of the recipient population, and inform future humanitarian collaborations. Study Design Case series with chart review. Setting Tertiary public referral hospital in Eldoret, Kenya. Subjects and Methods Twenty-one patients were enrolled during the study period. A retrospective chart review was performed for all adult patients who underwent thyroid surgery during humanitarian trips (2010-2015). Patients were contacted by mobile telephone. Medical history and physical examination, including laryngoscopy, were performed, and the SF-36 was administered (a quality-of-life questionnaire). Laboratory measurements of thyroid function and neck ultrasound were obtained. Results The mean follow-up was 33.6 ± 20.2 months after surgery: 37.5% of subtotal thyroidectomy patients and 15.4% of lobectomy patients were hypothyroid postoperatively according to serologic studies. There were no cases of goiter recurrence or malignancy. All patients reported postoperative symptomatic improvement and collectively showed positive pre- and postoperative score differences on the SF-36. Conclusion Although limited by a small sample size and the retrospective nature, our study demonstrates the feasibility of long-term surgical and quality-of-life outcomes research in a resource-limited setting. The low complication rates suggest minimal adverse effects of performing surgery in this context. Despite a considerable rate of postoperative hypothyroidism, it is in accordance with prior studies and emphasizes the need for individualized, longitudinal, and multidisciplinary care. Quality-of-life score improvements suggest benefit to the recipient population.

Cells under stress must adjust their physiology, metabolism, and architecture to adapt to the new conditions. Most importantly, they must down-regulate general gene expression, but at the same time induce synthesis of stress-protective factors, such as molecular chaperones. Here, we investigate how the process of phase separation is used by cells to ensure adaptation to stress. We summarize recent findings and propose that the solubility of important translation factors is specifically affected by changes in physical-chemical parameters such temperature or pH and modulated by intrinsically disordered prion-like domains. These stress-triggered changes in protein solubility induce phase separation into condensates that regulate the activity of the translation factors and promote cellular fitness. Prion-like domains play important roles in this process as environmentally regulated stress sensors and modifier sequences that determine protein solubility and phase behavior. We propose that protein phase separation is an evolutionary conserved feature of proteins that cells harness to regulate adaptive stress responses and ensure survival in extreme environmental conditions.

The design of proteins that self-assemble into well-defined, higher order structures is an important goal that has potential applications in synthetic biology, materials science, and medicine. We previously designed a two-component protein system, designated A-(+) and A-(−), in which self-assembly is mediated by complementary electrostatic interactions between two coiled-coil sequences appended to the C-terminus of a homotrimeric enzyme with C3 symmetry. The coiled-coil sequences are attached through a short, flexible spacer sequence providing the system with a high degree of conformational flexibility. Thus, the primary constraint guiding which structures the system may assemble into is the symmetry of the protein building block. We have now characterized the properties of the self-assembling system as a whole using native gel electrophoresis and analytical ultracentrifugation (AUC) and the properties of individual assemblies using cryo-electron microscopy (EM). We show that upon mixing, A-(+) and A-(−) form only six different complexes in significant concentrations. The three predominant complexes have hydrodynamic properties consistent with the formation of heterodimeric, tetrahedral, and octahedral protein cages. Cryo-EM of size-fractionated material shows that A-(+) and A-(−) form spherical particles with diameters appropriate for tetrahedral or octahedral protein cages. The particles varied in diameter in an almost continuous manner suggesting that their structures are extremely flexible.