Langerin-Heparin Interaction: Two Binding Sites for Small and Large Ligands as revealed by a combination of NMR Spectroscopy and Cross-Linking Mapping Experiments

Langerin is a C-type lectin present on Langerhans cells that mediates capture of pathogens in a carbohydrate-dependent manner, leading to subsequent internalization and elimination in the cellular organelles called Birbeck granules. This mechanism mediated by langerin was shown to constitute a natural barrier for HIV-1 particle transmission. Besides interacting specifically with high mannose and fucosylated neutral carbohydrate structures, langerin has the ability to bind sulfated carbohydrate ligands as 6-sulfated galactosides in the Ca2+ dependent binding site. Very recently langerin was demonstrated to interact with sulfated glycosaminoglycans (GAGs), in a Ca2+ independent way, resulting in the proposal of a new binding site for GAGs. Based on those results, we have conducted a structural study of the interactions of small heparin (HEP) like oligosaccharides with langerin in solution. Heparin-bead cross-linking experiments, an approach specifically designed to identify HEP/HS binding sites in proteins were first carried out and experimentally validated the previously proposed model for the interaction of Lg ECD with 6 kDa HEP. High-resolution NMR studies of a set of 8 synthetic HEP-like trisaccharides harboring different sulfation patterns demonstrated that all of them bound to langerin in a Ca2+ dependent way. The binding epitopes were determined by STD NMR and the bound conformations by transferred NOESY experiments. These experimental data were combined with docking and molecular dynamics and resulted in the proposal of a binding mode characterized by the coordination of calcium by the two equatorial hydroxyl groups OH3 and OH4 at the non-reducing end. The binding also includes the carboxylate group at the adjacent iduronate residue. Such epitope is shared by all the 8 ligands, explaining the absence of any impact on binding from their differences in substitution pattern. Finally, in contrast to the small trisaccharides, we demonstrated that a longer HEP-like hexasaccharide, bearing an additional O-sulfate group at the non-reducing end, which precludes binding to the Ca2+ site, interacts with langerin in the previously identified Ca2+ independent binding site

Statistical analysis of post mortem DNA damage-derived miscoding lesions in Neandertal mitochondrial DNA

Background. We have analysed the distribution of post mortem DNA damage derived miscoding lesions from the datasets of seven published Neandertal specimens that have extensive cloned sequence coverage over the mitochondrial DNA (mtDNA) hypervariable region 1 (HVS1). The analysis was restricted to C → T and G → A miscoding lesions (the predominant manifestation of post mortem damage) that are seen at a frequency of more than one clone among sequences from a single PCR, but do not represent the true endogenous sequence. Findings. The data indicates an extreme bias towards C → T over G → A miscoding lesions (observed ratio of 67:2 compared to an expected ratio of 7:2), implying that the mtDNA Light strand molecule suffers proportionally more damage-derived miscoding lesions than the Heavy strand. Conclusion. The clustering of Cs in the Light strand as opposed to the singleton pattern of Cs in the Heavy strand could explain the observed bias, a phenomenon that could be further tested with non-PCR based approaches. The characterization of the HVS1 hotspots will be of use to future Neandertal mtDNA studies, with specific regards to assessing the authenticity of new positions previously unknown to be polymorphic

Efficiency of Industrially Relevant Atropisomeric Diphosphines in Copper-Catalyzed 1,4-Asymmetric Conjugate Addition of Dialkylzincs to Cyclic or Acyclic Enones or Dienones

International audienceIndustrially relevant atropisomeric diphosphines such as 2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP), 6,6'-bis(diphenylphosphino)-2,2',3,3'-tetrahydro-5,5'-bi-1,4-benzodioxin (SYNPHOS), and 5,5'-bis(diphenylphosphino)-2,2,2',2'-tetrafluoro-4,4'-bi-1,3-benzodioxole (DIFLUORPHOS) have demonstrated their efficiency in the copper-catalyzed asym. conjugate addn. of various dialkylzincs to α-aryl enones, α-aryl dienones, and cyclic dienones. Excellent 1,4- or 1,6-regioselectivities and enantioselectivities (up to 97% ee) were attained, even with challenging sterically hindered Michael acceptors

CD148 is a membrane protein tyrosine phosphatase present in all hematopoietic lineages and is involved in signal transduction on lymphocytes

Producción CientíficaEvidence is presented showing that a protein tyrosine phosphatase different from CD45 is present on the membrane of human hematopoietic cells. The molecule recognized by the monoclonal antibody 143-41, which has been classified as CD148 in the VI International Workshop on Leukocyte Differentiation Antigens, was immunopurified and sequenced. The sequence obtained from N-terminus as well as from two different CNBr-digested peptides showed a close identity with a previously described tyrosine phosphatase named HPTP-eta/DEP-1. CD148 is present on all hematopoietic lineages, being expressed with higher intensity on granulocytes than on monocytes and lymphocytes. Interestingly, whereas it is clearly present on peripheral blood lymphocytes, it is poorly expressed on different lymphoid cell lines of T and B origin. When this protein tyrosine phosphatase was cocrosslinked with CD3, an inhibition of the normally observed calcium mobilization was observed. This inhibition correlates with a decrease in phospholipase C-gamma (PLC-gamma) phosphorylation and is similar to the one observed with CD45. In addition, it is shown that the crosslinking of the CD148 alone is also able to induce an increase in [Ca2+]i. This increase is abolished in the presence of genistein and by cocrosslinking with CD45. These data, together with the induction of tyrosine phosphorylation on several substrates, including PLC-gamma, after CD148 crosslinking, suggest the involvement of a tyrosine kinase-based signaling pathway in this process. In conclusion, the data presented show that CD148 corresponds to a previously described protein tyrosine phosphatase HPTP-eta/DEP-1 and that this molecule is involved in signal transduction in lymphocytes

Exact Solution of Return Hysteresis Loops in One Dimensional Random Field Ising Model at Zero Temperature

Minor hysteresis loops within the main loop are obtained analytically and exactly in the one-dimensional ferromagnetic random field Ising-model at zero temperature. Numerical simulations of the model show excellent agreement with the analytical results

Critical Behaviour of Non-Equilibrium Phase Transitions to Magnetically Ordered States

We describe non-equilibrium phase transitions in arrays of dynamical systems with cubic nonlinearity driven by multiplicative Gaussian white noise. Depending on the sign of the spatial coupling we observe transitions to ferromagnetic or antiferromagnetic ordered states. We discuss the phase diagram, the order of the transitions, and the critical behaviour. For global coupling we show analytically that the critical exponent of the magnetization exhibits a transition from the value 1/2 to a non-universal behaviour depending on the ratio of noise strength to the magnitude of the spatial coupling.Comment: 4 pages, 5 figure

MaRCoS, an open-source electronic control system for low-field MRI

Every magnetic resonance imaging (MRI) device requires an electronic control system that handles pulse sequences and signal detection and processing. Here we provide details on the architecture and performance of MaRCoS, a MAgnetic Resonance COntrol System developed by an open international community of low-field MRI researchers. MaRCoS is inexpensive and can handle cycle-accurate sequences without hard length limitations, rapid bursts of events, and arbitrary waveforms. It can also be easily adapted to meet further specifications required by the various academic and private institutions participating in its development. We describe the MaRCoS hardware, firmware and software that enable all of the above, including a Python-based graphical user interface for pulse sequence implementation, data processing and image reconstruction.Comment: 10 pages, 4 figure

Real-Time Position Reconstruction with Hippocampal Place Cells

Brain–computer interfaces (BCI) are using the electroencephalogram, the electrocorticogram and trains of action potentials as inputs to analyze brain activity for communication purposes and/or the control of external devices. Thus far it is not known whether a BCI system can be developed that utilizes the states of brain structures that are situated well below the cortical surface, such as the hippocampus. In order to address this question we used the activity of hippocampal place cells (PCs) to predict the position of an rodent in real-time. First, spike activity was recorded from the hippocampus during foraging and analyzed off-line to optimize the spike sorting and position reconstruction algorithm of rats. Then the spike activity was recorded and analyzed in real-time. The rat was running in a box of 80 cm × 80 cm and its locomotor movement was captured with a video tracking system. Data were acquired to calculate the rat's trajectories and to identify place fields. Then a Bayesian classifier was trained to predict the position of the rat given its neural activity. This information was used in subsequent trials to predict the rat's position in real-time. The real-time experiments were successfully performed and yielded an error between 12.2 and 17.4% using 5–6 neurons. It must be noted here that the encoding step was done with data recorded before the real-time experiment and comparable accuracies between off-line (mean error of 15.9% for three rats) and real-time experiments (mean error of 14.7%) were achieved. The experiment shows proof of principle that position reconstruction can be done in real-time, that PCs were stable and spike sorting was robust enough to generalize from the training run to the real-time reconstruction phase of the experiment. Real-time reconstruction may be used for a variety of purposes, including creating behavioral–neuronal feedback loops or for implementing neuroprosthetic control

Validity of quasi-degenerate neutrino mass models and their predictions on baryogenesis

Quasi-degenerate neutrino mass models (QDN) which can explain the current data on neutrino masses and mixings,are studied. In the first part, we study the effect of CP-phases on QDN mass matrix obeying $\mu-\tau$ symmetry in normal hierarchical (QD-NH) and inverted hierarchical (QD-IH) patterns.The numerical predictions are consistent with observed data on solar mixing angle, absolute neutrino mass parameter consistent with neutrinoless double beta decay mass parameter and sum of three absolute neutrino masses from cosmological bound.The neutrino mass matrix is parameterized using only two unknown parameters. The second part deals with the estimation of observed baryon asymmetry of the universe. The prediction is nearly consistent with observation with flavoured thermal leptogenesis scenario. QD-NH model appears to be more favourable than those of QD-IH models.The present analysis shows that the three absolute neutrino masses may exhibit quasi-degenerate pattern in nature. They are far from discrimination at the moment.Comment: 17 pages, no figure, poster presentation in the 25th International Conference on neutrino physics and Astrophysics, Kyoto, Japan, July,201