Lanthanide-based peptide biosensor to monitor CDK4/cyclin D kinase activity

We describe a lanthanide biosensor that responds to CDK4 kinase activity in melanoma cell extracts through a significant and dose dependent increase in luminescence, thanks to sensitization of a DOTA[Tb3+] complex incorporated into a CDK4 substrate peptide by a unique tryptophan residue in an adjacent phosphoaminoacid binding moietyThis work was funded by the CNRS (Centre National de la Recherche Scientifique) and a Marie-Curie fellowship EC-FP7 Framework (PIEF-GA-2013-623151) supporting JAGV. CB was funded by the INCA (PRTK-2014). Financial support from the Spanish MINECO (CTQ2015-70698-R), the Xunta de Galicia (Centro singular de investigacio´n de Galicia accreditation 2016–2019), and the European Union (European Regional Development Fund – ERDF), are gratefully acknowledgedS

New observations with the gas electron multiplier (GEM)

We describe recent measurements realized with the Gas Electron Multiplier (GEM) mesh added as pre-amplification element to a multiwire and a micro-strip chamber. Large, stable combined gains are obtained, with good uniformity and energy resolution, in a wide range of filling gases including non-flammable mixtures; coupled to a micro-strip plate, the pre-amplification element allows to maintain the high rate capability and resolution at considerably lower operating voltages, completely eliminating discharge problems. Charge gains are large enough to allow detection of signals in the ionization mode on the last element, permitting the use of a simple printed circuit as read-out electrode; two-dimensional read-out can then be easily implemented. The absence of charge multiplication in the last stage avoids charge build-up on the substrate and prevents ageing phenomena. A new generation of simple, reliable and cheap fast position sensitive detectors seems at hand

The gas electron multiplier (GEM)

We describe operating priciples and results obtained with a new detector component: the Gas Electrons Multiplier (GEM). Consisting of a thin composite sheet with two metal layers separated by a thin insulator, and pierced by a regular matrix of open channels, the GEM electrode, inserted on the path of electrons in a gas detector, allows to transfer the charge with an amplification factor approaching ten. Uniform response and high rate capability are demonstrated. Coupled to another device, multiwire or micro-strip chamber, the GEM electrode permit to obtain higher gains or less critical operation; separation of the sensitive (conversion) volume and the detection volume has other advantages, as a built-in delay (useful for triggering purposes) and the possibility of applying high fields on the photo-cathode of ring imaging detectors to improve efficiency. Multiple GEM grids in the same gas volume allow to obtain large amplification factors in a succession of steps, leading to the realization of an effective gas-filled photomultiplier

Optimization of design and beam test of microstrip gas chambers

We describe recent experimental and theoretical work aimed at optimizing the geometry and the operation of micro-strip gas chambers in order to improve their performance and reliability. With the help of a simulation program, we have studied the mechanism of signal propagation and analyzed the effects on signal shape and size of resistivity of strips, grouping of biased strips and presence of a back-plane. Several detectors manufactured according to the results of the study and equipped with fast amplifiers have been installed in a test beam to study general operating characteristics, efficiency and localization accuracy; preliminary results of the data analysis are discussed

Studies of aging and HV break down problems during development and operation of MSGC and GEM detectors for the Inner Tracking System of HERA-B

The results of five years of development of the inner tracking system of the HERA-B experiment and first experience from the data taking period of the year 2000 are reported. The system contains 184 chambers, covering a sensitive area of about 20 * 20 cm2 each. The detector is based on microstrip gas counters (MSGCs) with diamond like coated (DLC) glass wafers and gas electron multipliers (GEMs). The main problems in the development phase were gas discharges in intense hadron beams and aging in a high radiation dose environment. The observation of gas discharges which damage the electrode structure of the MSGC led to the addition of the GEM as a first amplification step. Spurious sparking at the GEM cannot be avoided completely. It does not affect the GEM itself but can produce secondary damage of the MSGC if the electric field between the GEM and the MSGC is above a threshold depending on operation conditions. We observed that aging does not only depend on the dose but also on the spot size of the irradiated area. Ar-DME mixtures had to be abandoned whereas a mixture of 70% Ar and 30% CO2 showed no serious aging effects up to about 40 mC/cm deposited charge on the anodes. X-ray measurements indicate that the DLC of the MSGC is deteriorated by the gas amplification process. As a consequence, long term gain variations are expected. The Inner Tracker has successfully participated in the data taking at HERA-B during summer 2000.Comment: 29 pages, 22 figure

The Cleo Rich Detector

We describe the design, construction and performance of a Ring Imaging Cherenkov Detector (RICH) constructed to identify charged particles in the CLEO experiment. Cherenkov radiation occurs in LiF crystals, both planar and ones with a novel ``sawtooth''-shaped exit surface. Photons in the wavelength interval 135--165 nm are detected using multi-wire chambers filled with a mixture of methane gas and triethylamine vapor. Excellent pion/kaon separation is demonstrated.Comment: 75 pages, 57 figures, (updated July 26, 2005 to reflect reviewers comments), to be published in NIM

Alteration of ribosome function upon 5-fluorouracil treatment favors cancer cell drug-tolerance.

Mechanisms of drug-tolerance remain poorly understood and have been linked to genomic but also to non-genomic processes. 5-fluorouracil (5-FU), the most widely used chemotherapy in oncology is associated with resistance. While prescribed as an inhibitor of DNA replication, 5-FU alters all RNA pathways. Here, we show that 5-FU treatment leads to the production of fluorinated ribosomes exhibiting altered translational activities. 5-FU is incorporated into ribosomal RNAs of mature ribosomes in cancer cell lines, colorectal xenografts, and human tumors. Fluorinated ribosomes appear to be functional, yet, they display a selective translational activity towards mRNAs depending on the nature of their 5'-untranslated region. As a result, we find that sustained translation of IGF-1R mRNA, which encodes one of the most potent cell survival effectors, promotes the survival of 5-FU-treated colorectal cancer cells. Altogether, our results demonstrate that "man-made" fluorinated ribosomes favor the drug-tolerant cellular phenotype by promoting translation of survival genes