FmhA and FmhC of Staphylococcus aureus incorporate serine residues into peptidoglycan cross-bridges

Staphylococcal peptidoglycan is characterized by pentaglycine cross-bridges that are cross-linked between adjacent wall peptides by penicillin-binding proteins to confer robustness and flexibility. In Staphylococcus aureus, pentaglycine cross-bridges are synthesized by three proteins: FemX adds the first glycine, and the homodimers FemA and FemB sequentially add two Gly-Gly dipeptides. Occasionally, serine residues are also incorporated into the cross-bridges by enzymes that have heretofore not been identified. Here, we show that the FemA/FemB homologues FmhA and FmhC pair with FemA and FemB to incorporate Gly-Ser dipeptides into cross-bridges and to confer resistance to lysostaphin, a secreted bacteriocin that cleaves the pentaglycine cross-bridge. FmhA incorporates serine residues at positions 3 and 5 of the cross-bridge. In contrast, FmhC incorporates a single serine at position 5. Serine incorporation also lowers resistance toward oxacillin, an antibiotic that targets penicillin-binding proteins, in both methicillin-sensitive and methicillin-resistant strains of S. aureus. FmhC is encoded by a gene immediately adjacent to lytN, which specifies a hydrolase that cleaves the bond between the fifth glycine of cross-bridges and the alanine of the adjacent stem peptide. In this manner, LytN facilitates the separation of daughter cells. Cell wall damage induced upon lytN overexpression can be alleviated by overexpression of fmhC. Together, these observations suggest that FmhA and FmhC generate peptidoglycan cross-bridges with unique serine patterns that provide protection from endogenous murein hydrolases governing cell division and from bacteriocins produced by microbial competitors.

Beef veins record fluid overpressure during oil primary migration in source rocks

<p>Bedding-parallel, fibrous calcite veins (commonly referred to as “beefs”) are widely developed within Eocene, lacustrine, laminated organic-rich source rocks in the Dongying Depression, Bohai Bay Basin, East China. Based on the study of vein petrography and fluid inclusions features, we demonstrate the vein was the product of hydrocarbon generation and expulsion from organic-rich shales. Consequently, the primary inclusions in the fibrous calcites recorded the fluid conditions during maturation of these source rocks. In most cases, the calcite-hosted primary inclusion assemblages are composed of the two-phase (oil + gas) hydrocarbon inclusions, with or without coexisting aqueous inclusions. Less commonly, the assemblages are made up of inclusions with only liquid hydrocarbon (i.e., monophase, high-density petroleum inclusions). In addition, many bitumen-bearing oil inclusions could also be observed in the fibrous calcite veins. By modelling the isochores of two-phase oil inclusions and coexisting aqueous inclusions, in light of the burial history for the basin, we conclude the fluid overpressure up to approximately twice (2x) the hydrostatic value (i.e., ~0.5–0.6x lithostatic) are the most common during the hydrocarbon generation and primary migration. The highest degrees of overpressure are recorded by the rare monophase petroleum inclusions. The resulting isochores of these highest density inclusions project to pressures that overlap with the lithostatic gradient. Thus, the monophase inclusions indicate pressures approaching and in some cases exceeding lithostatic. Our results indicate that fluids present during hydrocarbon generation and expulsion in organic-rich shales were indeed overpressured, but that lithostatic pressures were not the norm and evidently not a prerequisite for vein dilation, which means the fluid pressures during dilation of horizontal veins are not necessarily equal to the overburden throughout the history of the opening. This suggests that at least some of the vein dilation is accommodated and offset by concomitant narrowing of the adjacent wall rock laminae, likely by scavenging (dissolution/reprecipitation) of CaCO<sub>3</sub> from the adjacent wall rock, owing to the positive pressure dependence of calcite solubility, and presence of organic acids as byproducts of hydrocarbon generation.</p>

Direct evidence for fluid overpressure during hydrocarbon generation and expulsion from organic-rich shales

Fluid overpressures are widely expected during hydrocarbon generation and expulsion from source rocks, yet direct evidence for this phenomenon is lacking in the case of organic-rich shales. Here we show that formation of bed-parallel fibrous calcite veins in mature laminated organic-rich shales in the Eocene Dongying depression, Bohai Bay Basin, east China, occurred in direct response to fluid overpressure due to hydrocarbon generation. The evidence for overpressure is recorded by coexisting primary aqueous and petroleum inclusions in the calcite fibers. Our results show that all analyzed fluid-inclusion assemblages record variable degrees of overpressure during vein dilation, ranging from only modestly in excess of hydrostatic, to approaching and perhaps exceeding lithostatic. Thus, our results indicate that fluid pressures during dilation of horizontal veins are not necessarily equal to the opposing force of overburden throughout the history of opening. This suggests that at least some of the vein dilation is accommodated by concomitant narrowing of the adjacent wall-rock laminae, likely by scavenging (dissolution and reprecipitation) of CaCO3 from the adjacent wall rock.

Homocysteine-Enhanced Proteolytic and Fibrinolytic Processes in Thin Intraluminal Thrombus and Adjacent Wall of Abdominal Aortic Aneurysm: Study In Vitro

Homocysteine (Hcy) may affect the pathogenesis of abdominal aortic aneurysms (AAAs) through enhancement of proteolysis and an impaired coagulation/fibrinolysis system. Intensified haemostatic capacity may promote local proteolytic degradation of the aortic wall. This study aimed to examine the effects of Hcy on haemostatic and proteolytic processes in samples of thick and thin fragments of the ILT and underlying walls.Subjects and Methods. Thirty-six patients who underwent AAA surgery were enrolled. Aneurysm tissue sections were incubated with DL-Hcy (100 and 500μmol/L) in a series of experiments and analyzed for concentration/activity of proteolytic and haemostatic markers by enzyme-linked immunosorbent assay.Results. Incubation of wall underlying thin ILT segments (B) with DL-Hcy resulted in an increase of active MMP-2 levels compared to control tissue (9.54 ± 5.88 versus 7.44 ± 4.48, p=0.011). DL-Hcy also induced t-PA and plasminogen concentration increases in thin thrombus sections (B1) compared to control tissue (respectively: 1.39 ± 1.65 versus 0.84 ± 0.74, p=0.024; 11.64 ± 5.05 versus 10.34 ± 5.52, p=0.018). In contrast, wall adjacent to thick thrombus segments (A) showed decreases in MMP-2 and TF activities compared to control (respectively, 5.89 ± 3.39 versus 7.26 ± 5.49, p=0.046; 67.13 ± 72.59 versus 114.46 ± 106.29, p=0.007). In thick ILT sections (A1), DL-Hcy decreased MMP-2 activity and t-PA and plasminogen concentrations compared to control tissue (respectively, 2.53 ± 2.02 versus 3.28 ± 2.65, p=0.006; 0.67 ± 0.57 versus 0.96 ± 0.91, p=0.021; 9.25 ± 4.59 versus 12.63 ± 9.56, p=0.017). In addition, analysis revealed positive correlations at all sites between activities/concentrations of MMP-2, TF, and PAI-1 measured in control tissues and after incubation with DL-Hcy.Conclusions. These data indicate the potential for excess Hcy to enhance damage of arterial wall in thinner AAA segments as a result of the increased activity of MMP-2 and fibrinolytic factors.

Reconstruction of the tower in defensive walls in Świebodzice

Fortification of Świebodzice, as presented in the eighteenthcentury drawings of Friedrich Bernhard Wernher, comprised one ring of walls, three gates in the form of towers with a passage on the ground floor, a rectangular gatehouse matching the height of the adjacent wall, and at least 8 rectangular towers. The entire fortification was surrounded by a moat. These architectural forms are in accordance with the oldest view over the city presented in the seventeenth-century oil painting [1]. The walls were crowned with rectangular battlements. Elements known from iconography were confirmed during research and inventory works initiated in 2007. The project, which was an aim of revitalizing the part of defensive walls along Piłsudskiego street in Świebodzice, was not only to make this relic accessible to the public (there is an observation deck in the defensive tower), but also to repair deformations formed as a result of the first restoration and because of the didactic exposition of wooden elements in the wall crowning (these elements were popular in medieval fortifications).

Ultrastructure of internal jugular vein defective valves

Objectives To study the ultrastructure of intraluminal defects found in the internal jugular vein by using a scanning electron microscopy. Methods Using a scanning electron microscopy, intraluminal septa and/or defective valves blocking the flow in the distal internal jugular vein of seven patients were studied together with the adjacent wall and compared with control specimen. Results The internal jugular veins’ wall showed a significant derangement of the endothelial layer as compared to controls. Surprisingly, no endothelial cells were found in the defective cusps, and the surface of the structure is covered by a fibro-reticular lamina. Conclusions Although the lack of endothelial cells in the internal jugular vein intraluminal obstacles is a further abnormality found in course of chronic cerebrospinal venous insufficiency, our investigation cannot clarify whether this finding is primary or caused by progressive loss of endothelium in relation to altered haemodynamic forces and/or to a past post-thrombotic/inflammatory remodelling.

Kinematics of a Free Particle Moving Between Two Parallel Walls

The understanding of some physical phenomena involved in the transport of free particles such as fibers during injection processes is an important issue. To answer some of the questions arising in such problems, we study here numerically the quasi-steady kinematics of a free cylindrical solid particle moving in a Newtonian fluid confined between two parallel plane walls taking the hydrodynamic interactions into account. This is achieved by the use of the resistance matrix technique relating the kinematics of the particle to the forces and the torques exerted on the particle and to the dissipation induced by the motion of this particle. Our approach is confirmed by asymptotical developments and by a comparison with other authors in some cases. The solutions of three practical problems are given. In the first one, the sedimentation of the particle is studied. It is found that the maximum settling velocity of the free particle is obtained at a position off the symmetry plane. The cylinder is observed to rotate counter intuitively against the direction of rolling along the adjacent wall. Moreover the angular velocity has an influence on the settling velocity when the concentration is very high. The second problem concerns the transport of a neutrally buoyant cylindrical particle in a Poiseuille flow. This study reveals that there are relative translational and angular velocities between the free particle and the undisturbed fluid particle contrary to the commonly admitted hypothesis used in several models and numerical codes. Finally the third problem is a combination of the two previous situations: the transport of a non-neutrally buoyant particle in a Poiseuille flow. Depending on the ratio of the buoyancy forces to the viscous ones, different solutions are possible and exposed. Other problems can also be solved with this approach which is less time-consuming than complex methods such as DNS.