Structural characterization of EnpA D,L-Endopeptidase from Enterococcus faecalis prophage provides insights into substrate specificity of M23 peptidases

The best-characterized members of the M23 family are glycyl-glycine hydrolases, such as lysostaphin (Lss) from Staphylococcus simulans or LytM from Staphylococcus aureus. Recently, enzymes with broad specificities were reported, such as EnpACD from Enterococcus faecalis, that cleaves D,L peptide bond between the stem peptide and a cross-bridge. Previously, the activity of EnpACD was demonstrated only on isolated peptidoglycan fragments. Herein we report conditions in which EnpACD lyses bacterial cells live with very high efficiency demonstrating great bacteriolytic potential, though limited to a low ionic strength environment. We have solved the structure of the EnpACD H109A inactive variant and analyzed it in the context of related peptidoglycan hydrolases structures to reveal the bases for the specificity determination. All M23 structures share a very conserved β-sheet core which constitutes the rigid bottom of the substrate-binding groove and active site, while variable loops create the walls of the deep and narrow binding cleft. A detailed analysis of the binding groove architecture, specificity of M23 enzymes and D,L peptidases demonstrates that the substrate groove, which is particularly deep and narrow, is accessible preferably for peptides composed of amino acids with short side chains or subsequent L and D-isomers. As a result, the bottom of the groove is involved in interactions with the main chain of the substrate while the side chains are protruding in one plane towards the groove opening. We concluded that the selectivity of the substrates is based on their conformations allowed only for polyglycine chains and alternating chirality of the amino acids

Transvenous Lead Extraction in Patients with Cardiac Implantable Device: The Impact of Systemic and Local Infection on Clinical Outcomes. An ESC‐EHRA ELECTRa (European Lead Extraction Controlled) Registry Substudy

Background: Infections of cardiac implantable devices (CIEDI) have poor outcomes despite improvement in lead extraction (TLE) procedures. Methods: To explore the influence of CIEDI on the outcomes of TLE and the differences between patients with systemic (Sy) vs. local (Lo) CIEDI, we performed a sub‐analysis of the EORP ELECTRa (European Lead Extraction ConTRolled) Registry. Results: Among 3555 patients enrolled by 73 centers in 19 Countries, the indication for TLE was CIEDI in 1850: 1170 with Lo‐CIEDI and 680 with Sy‐CIEDI. Patients with CIEDI had a worse in‐hospital prognosis in terms of major complications (3.57% vs. 1.71%; p = 0.0007) and mortality (2.27% vs. 0.49%; p < 0.0001). Sy‐CIEDI was an independent predictor of in‐hospital death (H.R. 2.14; 95%CI 1.06–4.33. p = 0.0345). Patients with Sy‐CIEDI more frequently had an initial CIED implant and a higher prevalence of comorbidities, while subjects with Lo‐CIEDI had a higher prevalence of previous CIED procedures. Time from signs of CIEDI and TLE was longer for Lo‐CIEDI despite a shorter pre‐TLE antibiotic treatment. Conclusions: Patients with CIEDI have a worse in‐hospital prognosis after TLE, especially for patients with Sy‐CIEDI. These results raise the suspicion that in a relevant group of patients CIEDI can be systemic from the beginning without progression from Lo‐CIEDI. Future research is needed to characterize this subgroup of patients

Two-site recognition of Staphylococcus aureus peptidoglycan by lysostaphin SH3b

Lysostaphin is a bacteriolytic enzyme targeting peptidoglycan, the essential component of the bacterial cell envelope. It displays a very potent and specific activity toward staphylococci, including methicillin-resistant Staphylococcus aureus. Lysostaphin causes rapid cell lysis and disrupts biofilms, and is therefore a therapeutic agent of choice to eradicate staphylococcal infections. The C-terminal SH3b domain of lysostaphin recognizes peptidoglycans containing a pentaglycine crossbridge and has been proposed to drive the preferential digestion of staphylococcal cell walls. Here we elucidate the molecular mechanism underpinning recognition of staphylococcal peptidoglycan by the lysostaphin SH3b domain. We show that the pentaglycine crossbridge and the peptide stem are recognized by two independent binding sites located on opposite sides of the SH3b domain, thereby inducing a clustering of SH3b domains. We propose that this unusual binding mechanism allows synergistic and structurally dynamic recognition of S. aureus peptidoglycan and underpins the potent bacteriolytic activity of this enzyme

Schodkowa funkcja bazowa w asymptotycznej homogenizacji systemu eliptycznego

The problem of asymptotic homogenization for one-dimensional stationary elliptic system was studied. The value of effective conductivity coefficient of homogeneous elliptic macroscopic system was determined for the basic cell approximated by a "stepped" function. The conditions to obtain given conductivity coefficient, in the considered homogenization problem, were shown. Results of numerical calculations were provided to illustrate discussed issues.W pracy rozważa się problem asymptotycznej homogenizacji dla jednowymiarowego stacjonarnego systemu eliptycznego. Dokonano aproksymacji "schodkowej" funkcji bazowej i wyznaczono przybliżoną wartość efektywnego współczynnika przewodności eliptycznego jednorodnego układu makroskopowego. Przedstawiono również jak w omawianym procesie homogenizacji uzyskać z góry zadany efektywny współczynnik przewodności. Rozważania zostały zilustrowane wynikami obliczeń numerycznych

Fractional-order models of the supercapacitors in the form of RC ladder networks

In the paper, mathematical models of the supercapacitors are investigated. The models are based on electrical circuits in the form of RC ladder networks. The elementary cell of the network may consist of resistances and capacitances that are connected in series or parallel. The dynamic behavior of the circuit is described using fractional-order differential equations and its properties are analyzed. The identification procedure with quadratic performance index is performed in time domain to identify the parameters of the supercapacitor models. The results of numerical simulations are compared with the results measured experimentally in the physical system. In addition, an example from the automotive industry is used for an experimental evaluation of the theoretical analysis and to present a perspective on the applicability of the approach for other industrial projects

Fractional-order models of the supercapacitors in the form of RC ladder networks

In the paper, mathematical models of the supercapacitors are investigated. The models are based on electrical circuits in the form of RC ladder networks. The elementary cell of the network may consist of resistances and capacitances that are connected in series or parallel. The dynamic behavior of the circuit is described using fractional-order differential equations and its properties are analyzed. The identification procedure with quadratic performance index is performed in time domain to identify the parameters of the supercapacitor models. The results of numerical simulations are compared with the results measured experimentally in the physical system. In addition, an example from the automotive industry is used for an experimental evaluation of the theoretical analysis and to present a perspective on the applicability of the approach for other industrial projects

Membrane processes database: MemData

Zazwyczaj dobór membranowych technik rozdziału dokonywany jest przez inżynierów na podstawie ich doświadczenia i analizy literatury. Jest to spowodowane znikomą wiedzą na temat permcatywności składników, w szczególności w mieszaninach wieloskładnikowych. Aby usprawnić wyszukiwanie stworzono bazę danych procesów membranowych MemData, która zawiera uporządkowane szeroko dostępne dane. W pracy przedstawiono strukturę MemData i przykłady jej zastosowania.The membrane separation technique is selected by engineers mainly on basis of their experience and literature survey. It is caused by small availability of information on the prediction of component permeabilities in multicom-ponent mixtures. In order to facilitate the search, a membrane database, MemData, was created by collecting and adequate structuring the available data. In this paper the structure of MemData along with illustrative examples of its using are presented

Evaluation of lactuca germplasm for resistance to the northern root-knot nematode

Meloidogyne hapla Chitwood, the northern root-knot nematode, is a major pest on lettuce (Lactuca spp.) in the northeastern United States. In order to identify resistant germplasm, 494 lettuce accessions (443 L. sativa, 36 L. serriola, 7 L. virosa. and 8 L. saligna) were evaluated in the greenhouse in two trials. Six-week-old Lactuca spp. seedlings were inoculated with 5000 M. hapla eggs per plant and evaluated after 12 weeks. Galling severity and fresh root weight were measured. Of the 443 L. sativa accessions, only two, PI 508476 and PI 419140, repeatedly exhibited a root galling index of ≤2, indicating up to 20% galling, in two screening trials. Four wild Lactuca accessions, L. saligna PI 281876, L. saligna PI 491000, L. saligna 261653, and L. virosa PI 273579, were also considered highly resistant, suggesting that these accessions may be useful in hybrid breeding studies. © Taylor & Francis Group, LLC