Introductory lectures to loop quantum gravity

We give a standard introduction to loop quantum gravity, from the ADM variables to spin network states. We include a discussion on quantum geometry on a fixed graph and its relation to a discrete approximation of general relativity.Comment: Based on lectures given at the 3eme Ecole de Physique Theorique de Jijel, Algeria, 26 Sep -- 3 Oct, 2009. 52 pages, many figures. v2 minor corrections. To be published in the proceeding

Asymptotics of lowest unitary SL(2,C) invariants on graphs

We describe a technique to study the asymptotics of SL(2,C) invariant tensors associated to graphs, with unitary irreps and lowest SU(2) spins, and apply it to the Lorentzian EPRL-KKL (Engle, Pereira, Rovelli, Livine; Kaminski, Kieselowski, Lewandowski) model of quantum gravity. We reproduce the known asymptotics of the 4-simplex graph with a different perspective on the geometric variables and introduce an algorithm valid for any graph. On general grounds, we find that critical configurations are not just Regge geometries, but a larger set corresponding to conformal twisted geometries. These can be either Euclidean or Lorentzian, and include curved and flat 4d polytopes as subsets. For modular graphs, we show that multiple pairs of critical points exist, and there exist critical configurations of mixed signature, Euclidean and Lorentzian in different subgraphs, with no 4d embedding possible.Comment: 40 Pages + 5 Appendices. 11 Figures. v2: Refined presentation of the general algorithm, additional minor amendments. v3: paragraph added in section 5 about curved embedding

Isolation and characterization of a putative collagen receptor from Staphylococcus aureus strain Cowan 1.

In a previous study we demonstrated that cells of Staphylococcus aureus strain Cowan bind 125I-collagen in a receptor-ligand type of interaction (Speziale, P., Raucci, G., Visai, L., Switalski, L.M., Timpl, R., and Hook, M. (1986) J. Bacteriol. 167, 77-81). In the present communication we report on the isolation and preliminary characterization of a putative collagen receptor from a lysate of S. aureus strain Cowan. Antibodies raised against a collagen receptor positive strain inhibit the binding of 125I-collagen to bacterial cells, whereas antibodies raised against a collagen receptor negative strain were without effect. Solubilized cell surface components did not exhibit any measurable affinity for collagen-Sepharose. However, the inhibitory effect of the antibodies against bacterial cells was neutralized by the lysate from a receptor-positive but not receptor-negative strain. A collagen receptor assay was designed based on this observation and used to develop a receptor purification protocol involving anion exchange chromatography, ammonium sulfate precipitation, and gel chromatography. Using this procedure a protein with an apparent Mr of 135,000 was purified. This protein which was present on a collagen receptor-positive strain but not on a receptor-negative strain could completely neutralize the inhibitory activity of the antibodies raised against S. aureus strain Cowan. Furthermore, antibodies raised against the 135-kDa protein inhibited the binding of collagen to bacteria, and this protein is tentatively identified as a collagen receptor

Mechanostability of the Fibrinogen Bridge between Staphylococcal Surface Protein ClfA and Endothelial Cell Integrin αVβ3

Binding of the Staphylococcus aureus surface protein clumping factor A (ClfA) to endothelial cell integrin αVβ3 plays a crucial role during sepsis, by causing endothelial cell apoptosis and loss of barrier integrity. ClfA uses the blood plasma protein fibrinogen (Fg) to bind to αVβ3 but how this is achieved at the molecular level is not known. Here we investigate the mechanical strength of the three-component ClfA-Fg-αVβ3 interaction on living bacteria, by means of single-molecule experiments. We find that the ClfA-Fg-αVβ3 ternary complex is extremely stable, being able to sustain forces (∼800 pN) that are much stronger than those of classical bonds between integrins and the Arg-Gly-Asp (RGD) tripeptide sequence (∼100 pN). Adhesion forces between single bacteria and αVβ3 are strongly inhibited by an anti-αVβ3 antibody, the RGD peptide, and the cyclic RGD peptide cilengitide, showing that formation of the complex involves RGD-dependent binding sites and can be efficiently inhibited by αVβ3 blockers. Collectively, our experiments favor a binding mechanism involving the extraordinary elasticity of Fg. In the absence of mechanical stress, RGD572-574 sequences in the Aα chains mediate weak binding to αVβ3, whereas under high mechanical stress exposure of cryptic Aα chain RGD95-97 sequences leads to extremely strong binding to the integrin. Our results identify an unexpected and previously undescribed force-dependent binding mechanism between ClfA and αVβ3 on endothelial cells, which could represent a potential target to fight staphylococcal bloodstream infections

FbsA-driven fibrinogen polymerization: a bacterial Deceiving Strategy.

We show that FbsA, a cell wall protein of the bacterium Streptococcus agalactiae, promotes large-scale aggregation of human plasma fibrinogen, leading to the formation of a semiflexible polymerlike network. This extensive aggregation process takes place not only in solution, but also on FbsA-functionalized colloidal particles, and leads to the formation of a thick layer on the bacterial cell wall itself, which becomes an efficient mask against phagocytosis

Conformational changes in the fibronectin binding MSCRAMMs are induced by ligand binding

Bacterial adherence to host tissue involves specific microbial surface adhesins of which a subfamily termed microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) specifically recognize extracellular matrix components. We now report on the biophysical characterization of recombinant fibronectin binding MSCRAMMs originating from several different species of Gram-positive bacteria. The far-UV CD spectra (190-250 nm) of recombinant forms of the ligand binding domain of the MSCRAMMs, in a phosphate-buffered saline solution at neutral pH, were characteristic of a protein containing little or no regular secondary structure. The intrinsic viscosity of this domain was found to be the same in the presence or absence of 6 M guanidine hydrochloride, indicating that the native and denatured conformations are indistinguishable. On addition of fibronectin NH2 terminus as ligand to the recombinant adhesin there is a large change in the resulting far-UV CD difference spectra. At a 4.9 M excess of the NH2 terminus the difference spectra shifted to what was predominately a beta-sheet conformation, as judged by comparison with model far-UV CD spectra. The fibronectin NH2-terminal domain undergoes a minute but reproducible blue-shift of its intrinsic tryptophan fluorescence on addition of rFNBD-A, which contains no tryptophan residues. Since this result indicates that there is no large change in the environment of the tryptophan residues of the NH2 terminus on binding, the large shift in secondary structure observed by CD analysis is attributed to induction of a predominately beta-sheet secondary structure in the adhesin on binding to fibronectin NH2 terminus

Fibronectin-binding protein B variation in Staphylococcus aureus

BACKGROUND: Fibronectin binding proteins A and B (FnBPA and FnBPB) mediate adhesion of S. aureus to fibrinogen, elastin and fibronectin. We previously identified seven different isotypes of FnBPA based on divergence in the fibrinogen- and elastin-binding A domains. The variation created differences in antigenicity while ligand binding functions were retained. Here, FnBPB variation was examined in both human and bovine isolates and compared to that of FnBPA. RESULTS: Seven different fnbB allelic variants were identified. Some strains that cluster by phylogenetic analysis contain different fnbB variants, whereas more divergent strains contain the same fnbB variant. The phylogeny of fnbB alleles does not match the phylogeny of fnbA alleles. Some FnBPA and FnBPB isotypes that are specified by human S. aureus strains are also found in bovine strains. The seven fnbB allelic variants encode seven distinct isotypes of the FnBPB A domain that are 61 to 85% identical in amino acid sequence. Variant amino acid residues were mapped on a three-dimensional model of the FnBPB A domain and were predicted to be surface-exposed. They are responsible for the antigenic diversity detected with polyclonal antibody and a monoclonal antibody raised against isotype I. Ligand binding by recombinant FnBPB N23 isotypes was compared by ELISA-based solid phase assays and surface plasmon resonance. Each bound to immobilized fibrinogen, elastin and fibronectin dose-dependently and saturably with similar affinities. Binding to fibronectin was surprising because the A domains do not contain any known motifs that mediate binding to fibronectin. This raises the possibility that the A domain of FnBPB contains a novel fibronectin binding motif that binds fibronectin by a novel mechanism. CONCLUSIONS: Seven different isoforms of FnBPB A domain retain ligand-binding functions but are antigenically distinct. The variation in FnBPA and FnBPB occurs in human and bovine S. aureus strains and may act as an immune evasion mechanism. All seven isotypes of FnBPB are capable of binding fibronectin though none contain any known fibronectin-binding motifs. These results have implications for the development of vaccines or immunotherapeutics that target FnBPB

A Comparison of 2 Mitral Annuloplasty Rings for Severe Ischemic Mitral Regurgitation: Clinical and Echocardiographic Outcomes.

Controversies regarding the choice of annuloplasty rings for treatment of ischemic mitral regurgitation still exist. Aim of the study is to compare early performance of 2 different rings in terms of rest and exercise echocardiographic parameters (transmitral gradient, systolic pulmonary artery pressure, and mitral valve area), clinical outcomes, and recurrence of mitral regurgitation. From January 2008 till December 2013, prospectively collected data of patients who underwent coronary artery bypass grafting and undersizing mitral valve annuloplasty for severe chronic ischemic mitral regurgitation at our Institution were reviewed. A total of 93 patients were identified; among them 44 had semirigid Memo 3D ring implanted (group A) whereas 49 had a rigid profile 3D ring (group B). At 6 months, recurrent ischemic mitral regurgitation, equal or more than moderate, was observed in 4 and 6 patients in the group A and B, respectively (P = 0.74). Group A showed certain improved valve geometric parameters such as posterior leaflet angle, tenting area, and coaptation depth. Transmitral gradient was significantly higher at rest in the group B (P < 0.0001). During exercise, significant increase of transmitral gradient and systolic pulmonary artery pressure was observed in group B (P < 0.0001). Mitral valve area was not statistically significantly smaller at rest in between groups (P = 0.09); however, it significantly decreased with exercise in group B (P = 0.01). At midterm follow-up, patients in group B were more symptomatic. In patients with chronic ischemic mitral regurgitation, use of semirigid Memo 3D ring when compared to the rigid Profile 3D may be associated with early improved mitral valve geometrical conformation and hemodynamic profile, particularly during exercise. No difference was observed between both groups in recurrent mitral regurgitation.Peer reviewe

Sequence diversity in the A domain of Staphylococcus aureus fibronectin-binding protein A

<p>Abstract</p> <p>Background</p> <p>Fibronectin-binding protein A (FnBPA) mediates adhesion of <it>Staphylococcus aureus </it>to fibronectin, fibrinogen and elastin. We previously reported that <it>S. aureus </it>strain P1 encodes an FnBPA protein where the fibrinogen/elastin-binding domain (A domain) is substantially divergent in amino acid sequence from the archetypal FnBPA of <it>S. aureus </it>NCTC8325, and that these variations created differences in antigenicity. In this study strains from multilocus sequence types (MLST) that spanned the genetic diversity of <it>S.aureus </it>were examined to determine the extent of FnBPA A domain variation within the <it>S. aureus </it>population and its effect on ligand binding and immuno-crossreactivity.</p> <p>Results</p> <p>Seven different isotype forms (I – VII) of the FnBPA A domain were identified which were between 66 to 76% identical in amino acid sequence in any pair-wise alignment. The <it>fnbA </it>allelic variants in strains of different multilocus sequence type were identified by DNA hybridization using probes specific for sequences encoding the highly divergent N3 sub-domain of different isotypes. Several isotypes were not restricted to specific clones or clonal complexes but were more widely distributed. It is highly likely that certain <it>fnbA </it>genes have been transferred horizontally. Residues lining the putative ligand-binding trench were conserved, which is consistent with the ability of each A domain isotype to bind immobilized fibrinogen and elastin by the dock-latch-lock mechanism. Variant amino acid residues were mapped on a three-dimensional model of the FnBPA A domain and were predicted to be surface-exposed. Polyclonal antibodies raised against the recombinant isotype I A domain bound that protein with a 4 – 7 fold higher apparent affinity compared to the A domains of isotypes II – VII, while some monoclonal antibodies generated against the isotype I A domain showed reduced or no binding to the other isotypes.</p> <p>Conclusion</p> <p>The FnBPA A domain occurs in at least 7 different isotypes which differ antigenically and exhibit limited immuno-crossreactivity, yet retain their ligand-binding functions. Antigenic variation of the FnBPA A domain may aid <it>S. aureus </it>to evade the host's immune responses. These findings have implications for the development of vaccines or immunotherapeutics that target FnBPA.</p