A Potent Mimetic of the Siglec-8 Ligand 6’-Sulfo-Sialyl Lewisx

Siglecs are members of the immunoglobulin gene family containing sialic acid binding N-terminal domains. Among them, Siglec-8 is expressed on various cell types of the immune system such as eosinophils, mast cells and weakly on basophils. Cross-linking of Siglec-8 with monoclonal antibodies triggers apoptosis in eosinophils and inhibits degranulation of mast cells, making Siglec-8 a promising target for the treatment of eosinophil- and mast cell-associated diseases such as asthma. The tetrasaccharide 6’-sulfo-sialyl Lewis^x has been identified as a specific Siglec-8 ligand in glycan array screening. Here, we describe an extended study enlightening the pharmacophores of 6’-sulfo-sialyl Lewis^x and the successful development of a high-affinity mimetic. Retaining the neuraminic acid core, the introduction of a carbocyclic mimetic of the Gal moiety and a sulfonamide substituent in the 9-position gave a 20-fold improved binding affinity. Finally, the residence time, which usually is the Achilles tendon of carbohydrate/lectin interactions, could be improved.     

Supramolecular Assembly of Aminoethylene‐Lipopeptide PMO Conjugates into RNA Splice‐Switching Nanomicelles

Phosphorodiamidate morpholino oligomers (PMOs) are oligonucleotide analogs that can be used for therapeutic modulation of pre‐mRNA splicing. Similar to other classes of nucleic acid‐based therapeutics, PMOs require delivery systems for efficient transport to the intracellular target sites. Here, artificial peptides based on the oligo(ethylenamino) acid succinyl‐tetraethylenpentamine (Stp), hydrophobic modifications, and an azide group are presented, which are used for strain‐promoted azide‐alkyne cycloaddition conjugation with splice‐switching PMOs. By systematically varying the lead structure and formulation, it is determined that the type of contained fatty acid and supramolecular assembly have a critical impact on the delivery efficacy. A compound containing linolenic acid with three cis double bonds exhibits the highest splice‐switching activity and significantly increases functional protein expression in pLuc/705 reporter cells in vitro and after local administration in vivo. Structural and mechanistic studies reveal that the lipopeptide PMO conjugates form nanoparticles, which accelerate cellular uptake and that the content of unsaturated fatty acids enhances endosomal escape. In an in vitro Duchenne muscular dystrophy exon skipping model using H2K‐mdx52 dystrophic skeletal myotubes, the highly potent PMO conjugates mediate significant splice‐switching at very low nanomolar concentrations. The presented aminoethylene‐lipopeptides are thus a promising platform for the generation of PMO‐therapeutics with a favorable activity/toxicity profile.     

Automatic fog detection and estimation of visibility distance through use of an onboard camera

In this paper, we will present a technique for measuring visibility distances under foggy weather conditions using a camera mounted onboard a moving vehicle. Our research has focused in particular on the problem of detecting daytime fog and estimating visibility distances; thanks to these efforts, an original method has been developed, tested and patented. The approach consists of dynamically implementing Koschmieder's law. Our method enables computing the meteorological visibility distance, a measure defined by the International Commission on Illumination (CIE) as the distance beyond which a black object of an appropriate dimension is perceived with a contrast of less than 5%. Our proposed solution is an original one, featuring the advantage of utilizing a single camera and necessitating the presence of just the road and sky in the scene. As opposed to other methods that require the explicit extraction of the road, this method offers fewer constraints by virtue of being applicable with no more than the extraction of a homogeneous surface containing a portion of the road and sky within the image. This image preprocessing also serves to identify the level of compatibility of the processed image with the set of Koschmieder's model hypotheses. Nicolas Hautiére graduated from the École Nationale des Travaux Publics de l'État, France (2002). He received his M.S. and Ph.D. degree in computer vision, respectively, in 2002 and 2005 from Saint-Étienne University (France). From 2002, he is a researcher in the Laboratoire Central des Ponts et Chaussées (LCPC), Paris, France. His research interests include trafic engineering, computer vision, and pattern recognition. Jean-Philippe Tarel graduated from the École Nationale des Ponts et Chaussées, Paris, France (1991). He received his Ph.D. degree in Applied Mathematics from Paris IX-Dauphine University in 1996 and he was with the Institut National de Recherche en Informatique et Automatique (INRIA) from 1991 to 1996. From 1997 to 1998, he was a research associate at Brown University, USA. From 1999, he is a researcher in the Laboratoire Central des Ponts et Chaussées (LCPC), Paris, France, and from 2001 to 2003 in the INRIA. His research interests include computer vision, pattern recognition, and shape modeling. Jean Lavenant graduated from the École Nationale des Travaux Publics de l'État, Lyon, France (2001). He received the M.S. degree in Computer Vision from Jean Monnet university of Saint-Étienne in 2001. In 2001, he was a researcher in the Laboratoire Central des Ponts et Chaussées (LCPC). In 2002, he was a system engineer in Chicago (USA). He is currently an engineer for the french ministry of transports. Didier Aubert received the M.S. and Ph.D. degree, respectively, in 1985 and 1989 from the National Polytechnical Institut of Grenoble (INPG). From 1989--1990, he worked as a research scientist on the development of an automatic road following system for the NAVLAB at Carnegie Mellon University. From 1990–1994, he worked in the research department of a private company (ITMI). During this period he was the project leader of several projects dealing with computer vision. He is currently a researcher at INRETS since 1995 and works on Road traffic measurements, crowd monitoring, automated highway systems, and driving assistance systems for vehicles. He is an image processing expert for several companies, teaches at Universities (Paris VI, Paris XI, ENPC, ENST) and is at the editorial board of RTS (Research - Transport - Safety).     

Chemokine gene expression in rat pancreatic acinar cells is an early event associated with acute pancreatitis

BACKGROUND & AIMS: The molecular mechanisms underlying pancreatitis are largely unknown. The goal of this study was to identify an early genetic event that correlated with pancreatitis. METHODS: Differential display of messenger RNAs (mRNAs) was conducted on normal pancreas vs. those of animals with secretagogue-induced pancreatitis. Northern blots from normal animals and animals with experimental acute pancreatitis were probed with cloned complementary DNAs for chemokines. Pancreatitis was induced with cerulein and by retrograde injection of bile salts. Immunocytochemistry was used to identify the source of chemokine expression. Pyrrolidine dithiocarbamate was tested for effects on chemokine expression and pancreatitis. RESULTS: A differentially amplified band was consistently observed early after cerulein hyperstimulation. This band was identified as a portion of the mob-1 gene, an alpha-chemokine. Northern analysis indicated that mRNAs for mob-1 and another chemokine, mcp-1, were induced after cerulein hyperstimulation in vivo. mob-1 mRNA was also induced by retrograde injection of bile salts and by cerulein in acinar cells in vitro. mob-1 protein was localized to exocrine cells in pancreata of diseased animals. Pyrrolidine dithiocarbamate inhibited both chemokine gene expression and early inflammatory characteristics of pancreatitis. CONCLUSIONS: Chemokines are induced in acinar cells by treatments that induce pancreatitis and may play an important role in the early stages of the disease.     

Driving Mechanisms of Delamination Related Reliability Problems in Exposed Pad Packages

Exposed pad packages were introduced in the late 1980s and early 1990s because of their excellent thermal and electrical performance. Despite these advantages, the exposed pad packages experience a lot of thermo-hygro-mechanical related reliability problems during qualification and testing. Examples are die lift, which occurs predominantly after moisture sensitivity level conditions, and die-attach to leadframe delamination leading to downbond stitch breaks during temperature cycling. In this chapter, nonlinear finite element (FE) models using fracture mechanics based J-integral calculations are used to assess the reliability problems of the exposed pad package family. Using the parametric FE models any geometrical and material effects can be explored to their impact on the occurrence diepad delamination, and dielift. For instance the impact of diepad size is found to be of much less importance as the impact of die thickness is. Using the fracture mechanics approach, the starting location for the delamination from thermo-hygro-mechanical point of view is deducted. The results indicate that when diepad delamination is present, cracks are likely to grow beneath the die and dielift will occur. The interaction between dielift and other failure modes, such as lifted ball bonds, are not found to be very significant. The FE models are combined with simulation-based optimization methods to deduct design guidelines for optimal reliability of the exposed pad family.     

Subcutaneous soft tissue tumours at the site of implanted microchips in mice

An experiment using 4279 CBA/J mice of two generations was carried out to investigate the influence of parental preconceptual exposure to X-ray radiation or to chemical carcinogens. Microchips were implanted subcutaneously in the dorsolateral back for unique identification of each animal. The animals were kept for lifespan under standard laboratory conditions. In 36 mice a circumscribed neoplasm occurred in the area of the implanted microchip. Females were significantly more frequently affected than male mice. An influence of age or different treatment on the s.c. tumour incidence in two mice generations could not be observed. Macroscopically, firm, pale white nodules up to 25 mm in diameter with the microchip in its center were found. Microscopically, soft tissue tumours such as fibrosarcoma and malignant fibrous histiocytoma were detected.