Mechanical Forces Guiding Staphylococcus aureus Cellular Invasion

Staphylococcus aureus can invade various types of mammalian cells, thereby enabling it to evade host immune defenses and antibiotics. The current model for cellular invasion involves the interaction between the bacterial cell surface located fibronectin (Fn)-binding proteins (FnBPA and FnBPB) and the α5β1 integrin in the host cell membrane. While it is believed that the extracellular matrix protein Fn serves as a bridging molecule between FnBPs and integrins, the fundamental forces involved are not known. Using single-cell and single-molecule experiments, we unravel the molecular forces guiding S. aureus cellular invasion, focusing on the prototypical three-component FnBPA-Fn-integrin interaction. We show that FnBPA mediates bacterial adhesion to soluble Fn via strong forces (∼1500 pN), consistent with a high-affinity tandem β-zipper, and that the FnBPA-Fn complex further binds to immobilized α5β1 integrins with a strength much higher than that of the classical Fn-integrin bond (∼100 pN). The high mechanical stability of the Fn bridge favors an invasion model in which Fn binding by FnBPA leads to the exposure of cryptic integrin-binding sites via allosteric activation, which in turn engage in a strong interaction with integrins. This activation mechanism emphasizes the importance of protein mechanobiology in regulating bacterial-host adhesion. We also find that Fn-dependent adhesion between S. aureus and endothelial cells strengthens with time, suggesting that internalization occurs within a few minutes. Collectively, our results provide a molecular foundation for the ability of FnBPA to trigger host cell invasion by S. aureus and offer promising prospects for the development of therapeutic approaches against intracellular pathogens

Search for Neutrinos in Super-Kamiokande Associated with the GW170817 Neutron-star Merger

We report the results of a neutrino search in Super-Kamiokande (SK) for coincident signals with the first detected gravitational wave (GW) produced by a binary neutron-star merger, GW170817, which was followed by a short gamma-ray burst, GRB170817A, and a kilonova/macronova. We searched for coincident neutrino events in the range from 3.5 MeV to ~100 PeV, in a time window ±500 s around the gravitational wave detection time, as well as during a 14-day period after the detection. No significant neutrino signal was observed for either time window. We calculated 90% confidence level upper limits on the neutrino fluence for GW170817. From the upward-going-muon events in the energy region above 1.6 GeV, the neutrino fluence limit is ${16.0}_{-0.6}^{+0.7}$ (${21.3}_{-0.8}^{+1.1}$) cm−2 for muon neutrinos (muon antineutrinos), with an error range of ±5° around the zenith angle of NGC4993, and the energy spectrum is under the assumption of an index of −2. The fluence limit for neutrino energies less than 100 MeV, for which the emission mechanism would be different than for higher-energy neutrinos, is also calculated. It is 6.6 × 107 cm−2 for anti-electron neutrinos under the assumption of a Fermi–Dirac spectrum with average energy of 20 MeV

A New Approach to Upper Eyelid Reconstruction

Background: Reconstruction of large defects of the upper eyelid is challenging because of its complex anatomy and specialized function. The aim of this work is to develop a single-stage reconstruction procedure based on a new approach. The technique consists of the advancement of an orbicularis oculi myocutaneous flap designed within the blepharoplasty skin excision pattern. Methods: After the tumor’s excision is completed with clear margins, the borders of the flap are incised down to the submuscular plane inside the classical pattern of upper eyelid blepharoplasty. Two myocutaneous triangles are excised on both sides of the flap to allow its advancement to cover the defect. When it is necessary to repair the posterior lamella, we harvest a mucochondral graft. Results: From 2012 to 2015, we performed upper eyelid reconstruction with this technique on six patients. The flap survived in all the patients, without total or partial necrosis. No patient required surgical revision. The results were aesthetically satisfying, and no tumor recurrence was noted. Conclusions: Our new approach to upper eyelid reconstruction maximizes the cosmetic outcome respecting the principles of radicality. This flap is better suited for lesions involving the median or paramedian eyelid border from the marginal zone up to the palpebral crease. The approach we propose is safe and versatile, and it ensures either a functional or a good aesthetic reconstruction. Level of Evidence IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266