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

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

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

Laboratory-Generated Autologous Skin Substitutes for Burn Treatment in Europe: Narrative Review, Experts' Opinion, and Legal Considerations.

Autologous skin substitutes represent a promising advancement in the treatment of burn injuries, offering personalized solutions for patients with extensive skin loss. This white paper synthesizes the current knowledge on laboratory-generated autologous skin substitutes in Europe, incorporating expert opinions and legal considerations. The white paper examines the scientific principles underlying autologous skin substitute development, including cell sourcing, bioengineering techniques, and clinical applications. The regulatory framework governing the production and use of these advanced therapies in Europe is also examined, highlighting challenges in standardization, safety, and approval pathways. The text features expert insights that offer a real-world perspective on the clinical viability and translational hurdles of autologous skin substitutes. The findings highlight the potential of autologous skin substitutes to improve burn treatment outcomes while emphasizing the need for harmonized regulations to facilitate clinical implementation. Despite technological advancements, significant challenges persist, including production costs, scalability, and long-term efficacy. Another focus of this white paper are the legal changes, which have significantly impacted the production and availability of these technologies. The review concludes that while autologous skin substitutes hold great promise, further research, regulatory refinement, and interdisciplinary collaboration are essential to optimize their integration into clinical practice

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