“One Ring to Bind Them All”—Part I: The Efficiency of the Macrocyclic Scaffold for G-Quadruplex DNA Recognition

Macrocyclic scaffolds are particularly attractive for designing selective G-quadruplex ligands essentially because, on one hand, they show a poor affinity for the “standard” B-DNA conformation and, on the other hand, they fit nicely with the external G-quartets of quadruplexes. Stimulated by the pioneering studies on the cationic porphyrin TMPyP4 and the natural product telomestatin, follow-up studies have developed, rapidly leading to a large diversity of macrocyclic structures with remarkable-quadruplex binding properties and biological activities. In this review we summarize the current state of the art in detailing the three main categories of quadruplex-binding macrocycles described so far (telomestatin-like polyheteroarenes, porphyrins and derivatives, polyammonium cyclophanes), and in addressing both synthetic issues and biological aspects

“One Ring to Bind Them All”—Part II: Identification of Promising G-Quadruplex Ligands by Screening of Cyclophane-Type Macrocycles

A collection of 26 polyammonium cyclophane-type macrocycles with a large structural diversity has been screened for G-quadruplex recognition. A two-step selection procedure based on the FRET-melting assay was carried out enabling identification of macrocycles of high affinity (ΔT1/2 up to 30°C) and high selectivity for the human telomeric G-quadruplex. The four selected hits possess sophisticated architectures, more particularly the presence of a pendant side-arm as well as the existence of a particular topological arrangement appear to be strong determinants of quadruplex binding. These compounds are thus likely to create multiple contacts with the target that may be at the origin of their high selectivity, thereby suggesting that this class of macrocycles offers unique advantages for targeting G-quadruplex-DNA

Challenges for Biomarker Discovery in Body Fluids Using SELDI-TOF-MS

Protein profiling using SELDI-TOF-MS has gained over the past few years an increasing interest in the field of biomarker discovery. The technology presents great potential if some parameters, such as sample handling, SELDI settings, and data analysis, are strictly controlled. Practical considerations to set up a robust and sensitive strategy for biomarker discovery are presented. This paper also reviews biological fluids generally available including a description of their peculiar properties and the preanalytical challenges inherent to sample collection and storage. Finally, some new insights for biomarker identification and validation challenges are provided

Appréhender le stock de métal monnayé au ive siècle après J.-C.

Parmi les thématiques abordées en numismatique à l’aide de l’archéométrie, les recherches spécifiques sur le stock de métal monnayé au ive siècle s’avèrent fascinantes mais délicates. Afflux de nouvelles ressources métalliques, refontes volontaires de monnaies : tous ces phénomènes sont de grande conséquence pour les monnayages produits dans les différents ateliers de l’Empire romain tardif. Certaines données de ce type sont mesurables. Les alliages métalliques (donc ici monétaires) contiennent en effet des éléments mineurs ou en traces, constituant la « carte d’identité » de l’approvisionnement des ateliers monétaires. Il y a 10 ans Isabelle Bollard et Jean-Noël Barrandon réunissaient plusieurs analyses de monnaies en alliage cuivreux émises au cours du ive siècle. Nos propres observations, obtenues dans le cadre d’un projet de recherche financé par la région Normandie (intitulé FANUM), sur 14 528 monnaies (nummi) de la période constantinienne contenues dans un trésor découvert fortuitement à Saint-Germain-de-Varreville (Manche), nous ont incités à reprendre, de façon plus approfondie, l’étude des alliages utilisés au cours de cette période, en portant notamment notre attention sur les teneurs en or et en argent. En effet, la grande variété des ateliers représentés au sein du trésor favorisait idéalement une telle enquête. Le choix de la fluorescence X portable a permis d’élaborer un protocole d’analyse précis et adapté à un grand nombre de monnaies. L’exploitation de 774 analyses réalisées sur des nummi frappés dans les ateliers occidentaux a permis de compléter les travaux antérieurs, mais également de mettre en évidence l’utilisation de stocks métalliques différents à partir des années 320.Among the thematics studied in numismatics using archeometry, specific research on the stock of metal struck in the 4th century turns out to be fascinating but complex. Influx of new metallic resources, voluntary recasts of coins: these phenomena have great consequences for the coinage produced in all the workshops throughout the late Roman Empire. Some of these data can be measurable. Metallic alloys (coins in this case) contain minor or trace elements that constitute a kind of “identity card” for the supply of monetary workshops. Ten years ago, Isabelle Bollard and Jean-Noël Barrandon collected several analyzes of bronzes struck during the 4th century. Our own observations, obtained within the FANUM project, funded by the Normandy region, on 14 528 coins (nummi) of the Constantinian period, contained in a fortuitously discovered treasure in Saint-Germain-de-Varreville (Manche) have motivated a thorough new study of the alloys used during this period, focusing on the traces of gold and silver. Indeed, the variety of workshops represented in the treasure has encouraged such an inquiry. The choice of portable X-ray fluorescence has enabled to develop a protocol of analysis adapted to a large number of coins. The use of 774 analyzes carried out on nummi struck in the Western workshops has completed previous works, but also enabled to highlight the use of different metal stocks from the 320s

Involvement of G-quadruplex regions in mammalian replication origin activity.

Genome-wide studies of DNA replication origins revealed that origins preferentially associate with an Origin G-rich Repeated Element (OGRE), potentially forming G-quadruplexes (G4). Here, we functionally address their requirements for DNA replication initiation in a series of independent approaches. Deletion of the OGRE/G4 sequence strongly decreased the corresponding origin activity. Conversely, the insertion of an OGRE/G4 element created a new replication origin. This element also promoted replication of episomal EBV vectors lacking the viral origin, but not if the OGRE/G4 sequence was deleted. A potent G4 ligand, PhenDC3, stabilized G4s but did not alter the global origin activity. However, a set of new, G4-associated origins was created, whereas suppressed origins were largely G4-free. In vitro Xenopus laevis replication systems showed that OGRE/G4 sequences are involved in the activation of DNA replication, but not in the pre-replication complex formation. Altogether, these results converge to the functional importance of OGRE/G4 elements in DNA replication initiation

Biomarker discovery for inflammatory bowel disease, using proteomic serum profiling

peer reviewedCrohn's disease and ulcerative colitis known as inflammatory bowel diseases (IBD) are chronic immuno-inflammatory pathologies of the gastrointestinal tract. These diseases are multifactorial, polygenic and of unknown etiology. Clinical presentation is non-specific and diagnosis is based on clinical, endoscopic, radiological and histological criteria. Novel markers are needed to improve early diagnosis and classification of these pathologies. We performed a study with 120 serum samples collected from patients classified in 4 groups (30 Crohn, 30 ulcerative colitis, 30 inflammatory controls and 30 healthy controls) according to accredited criteria. We compared protein sera profiles obtained with a Surface Enhanced Laser Desorption Ionization-Time of Flight-Mass Spectrometer (SELDI-TOF-MS). Data analysis with univariate process and a multivariate statistical method based on multiple decision trees algorithms allowed us to select some potential biomarkers. Four of them were identified by mass spectrometry and antibody based methods. Multivariate analysis generated models that could classify samples with good sensitivity and specificity (minimum 80%) discriminating groups of patients. This analysis was used as a tool to classify peaks according to differences in level on spectra through the four categories of patients. Four biomarkers showing important diagnostic value were purified, identified (PF4, MRP8, FIBA and Hpalpha2) and two of these: PF4 and Hpalpha2 were detected in sera by classical methods. SELDI-TOF-MS technology and use of the multiple decision trees method led to protein biomarker patterns analysis and allowed the selection of potential individual biomarkers. Their downstream identification may reveal to be helpful for IBD classification and etiology understanding

Monitoring DNA–Ligand Interactions in Living Human Cells Using NMR Spectroscopy

International audienceStudies on DNA−ligand interactions in the cellular environment are problematic due to the lack of suitable biophysical tools. To address this need, we developed an in-cell NMR-based approach for monitoring DNA−ligand interactions inside the nuclei of living human cells. Our method relies on the acquisition of NMR data from cells electroporated with preformed DNA−ligand complexes. The impact of the intracellular environment on the integrity of the complexes is assessed based on in-cell NMR signals from unbound and ligand-bound forms of a given DNA target. This technique was tested on complexes of two model DNA fragments and four ligands, namely, a representative DNA minor-groove binder (netropsin) and ligands bindin

Author Correction: Involvement of G-quadruplex regions in mammalian replication origin activity

An amendment to this paper has been published and can be accessed via a link at the top of the paper

Innovations in physical and rehabilitation medicine

peer reviewedAu cours de la dernière décennie, la Médecine Physique et Réadaptation (MPR) est une spécialité médicale qui a fortement évolué dans les différents domaines qui la concernent : de la prise en charge des lombalgies et lombosciatalgies ou encore de l’ostéoporose de manière pluridisciplinaire, en passant par l’utilisation des nouvelles technologies en rééducation neuro-locomotrice et de la robotisation en rééducation, chez les patients amputés par exemple, le développement de la médecine à vocation régénérative et la prévention en traumatologie du sport et, enfin, les progrès des techniques d’électrophysiologie pour le diagnostic des neuropathies à petites fibres. Ces différentes avancées seront abordées dans cet article.Over the last decade, Physical and Rehabilitation Medicine (PRM) is a medical specialty that has evolved considerably in the various fields that concern it : from the management of low back pain and lumbosciatalgia or osteoporosis in a multidisciplinary manner, through the use of new technologies in neuro-locomotor rehabilitation and robotisation in amputee patients for example, the development of regenerative medicine and prevention in sports traumatology and, finally, the progress of electrophysiology techniques for the diagnosis of small-fibre neuropathies. These various advances will be discussed in this article