Complete androgen insensitivity syndrome due to a new frameshift deletion in exon 4 of the androgen receptor gene: Functional analysis of the mutant receptor

We studied the androgen receptor gene in a large kindred with complete androgen insensitivity syndrome and negative receptor-binding activity, single-strand conformation polymorphism (SSCP) analysis and sequencing identified a 13 base pair deletion within exon 4. This was responsible for a predictive frameshift in the open reading frame and introduction of a premature stop codon at position 783 instead of 919. The deletion was reproduced in androgen receptor wildtype cDNA and transfected into mammalian cells. Western blot showed a smaller androgen receptor of 94 kDa for the transfected mutated cDNA instead of 110 kDa. Androgen-binding assay of the mutated transfected cells assessed the lack of androgen-binding. Gel retardation assay demonstrated the ability of the mutant to bind target DNA; however, the mutant was unable to transactivate a reporter gene. Although the role of the partial deletion in the lack of androgen action was expected, in vitro analyses highlight the role of the abnormal C-terminal portion in the inhibition of the receptor transregulatory activity of the protein causing androgen resistance in this family

Modern relationships between microscopic charcoal in marine sediments and fire regimes on adjacent landmasses to refine the interpretation of marine paleofire records: An Iberian case study

Marine microcharcoal records provide invaluable information to understand changes in biomass burning and its drivers over multiple glacial and interglacial cycles and to evaluate fire models under warmer climates than today. However, quantitative reconstructions of burnt area, fire intensity and frequency from these records need calibration studies of the current fire-microcharcoal relationship. Here, we present the analysis of microcharcoal concentration and morphology in 102 core-top sediment samples collected in the Iberian margin and the Gulf of Cádiz. We show that microcharcoal concentrations are influenced by the water depth or the distance from the river mouth. At regional scale, the mean microcharcoal concentrations and microcharcoal elongation (length to width ratio) show a marked latitudinal variation in their distribution, primarily controlled by the type of burnt vegetation in the adjacent continent. High microcharcoal concentrations in marine sediments represent rare, large and intense fires in open Mediterranean woodlands. Based on these results, the increasing trend of microcharcoal concentrations recorded since 8 ka in the well-known marine sedimentary core MD95-2042 off the Iberian margin indicates the occurrence of large and infrequent fires of high intensity due to the progressive degradation of the Mediterranean forest and the expansion of shrublands

Androgen receptor signaling regulates the transcriptome of prostate cancer cells by modulating global alternative splicing

Androgen receptor (AR), is a transcription factor and a member of a hormone receptor superfamily. AR plays a vital role in the progression of prostate cancer and is a crucial target for therapeutic interventions. While the majority of advanced-stage prostate cancer patients will initially respond to the androgen deprivation, the disease often progresses to castrate-resistant prostate cancer (CRPC). Interestingly, CRPC tumors continue to depend on hyperactive AR signaling and will respond to potent second-line antiandrogen therapies, including bicalutamide (CASODEX®) and enzalutamide (XTANDI®). However, the progression-free survival rate for the CRPC patients on antiandrogen therapies is only 8–19 months. Hence, there is a need to understand the mechanisms underlying CRPC progression and eventual treatment resistance. Here, we have leveraged next-generation sequencing and newly developed analytical methodologies to evaluate the role of AR signaling in regulating the transcriptome of prostate cancer cells. The genomic and pharmacologic stimulation and inhibition of AR activity demonstrates that AR regulates alternative splicing within cancer-relevant genes. Furthermore, by integrating transcriptomic data from in vitro experiments and in prostate cancer patients, we found that a significant number of AR-regulated splicing events are associated with tumor progression. For example, we found evidence for an inadvertent AR-antagonist-mediated switch in IDH1 and PL2G2A isoform expression, which is associated with a decrease in overall survival of patients. Mechanistically, we discovered that the epithelial-specific splicing regulators (ESRP1 and ESRP2), flank many AR-regulated alternatively spliced exons. And, using 2D invasion assays, we show that the inhibition of ESRPs can suppress AR-antagonist-driven tumor invasion. Our work provides evidence for a new mechanism by which AR alters the transcriptome of prostate cancer cells by modulating alternative splicing. As such, our work has important implications for CRPC progression and development of resistance to treatment with bicalutamide and enzalutamide

Constraint solving in uncertain and dynamic environments - a survey

International audienceThis article follows a tutorial, given by the authors on dynamic constraint solving at CP 2003 (Ninth International Conference on Principles and Practice of Constraint Programming) in Kinsale, Ireland. It aims at offering an overview of the main approaches and techniques that have been proposed in the domain of constraint satisfaction to deal with uncertain and dynamic environments

Sulfur K-Edge XAS Study of Sulfidic Crosslinks in Vulcanised Rubbers

This feasibility study allowed us to establish three main results : (i) With high quality XANES spectra, it becomes quite possible to resolve the characteristic σ*(S-S), π*(S-S) and σ*(S-C) resonances; (ii) The "undulator gap scan" technique can yield excellent EXAFS spectra at the sulfur K-edge over more than 1300 eV; (iii) S K-edge EXAFS spectra made it possible to discriminate between vulcanizates with variable amounts of sulfur crosslinks and zinc sulfide

Involvement of HP1 alpha protein in irreversible transcriptional inactivation by antiestrogens in breast cancer cells

AbstractResistance to 4-hydroxy-tamoxifen (OHT), which appears in breast cancer cells after long-term antiestrogen treatment, may involve irreversible changes of gene expression. We previously developed a MCF-7 derived cell line (MVLN), in which OHT rapidly and irreversibly inactivates the expression of an estrogen-regulated luciferase transgene (Vit-tk-luciferase). In chromatin immunoprecipitation experiments, heterochromatin protein 1 (HP1α) was found to be associated with the Vit-tk-luciferase transgene, only when it was inactivated by OHT treatment. Chimeras composed of either HP1α or the Krupple-associated box (KRAB) module of KOX-1 protein (known to repress gene expression by recruitment of HP1 proteins), fused to the estrogen receptor (ER)–DNA binding domain (DBD) and the androgen receptor (AR)–ligand binding domain (LBD) were generated and appeared as potent transcriptional repressors. In stably transfected MVLN cells, irreversible inactivation of the luciferase transgene expression obtained with HP1α–ER(DBD)–AR(LBD) was partial, whereas inactivation obtained with KRAB–ER(DBD)–AR(LBD) was comparable to that obtained with OHT, although with a slower kinetics. Altogether, these data suggest that HP1α is involved in the silencing effects associated with long-term OHT treatments