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The wear behavior of SiC bonded diamond materials produced by liquid silicon infiltration in diamond preforms was investigated. The wear behavior in sand blasting tests (SiC abrasives, 5 bar pressure) was correlated with the microstructure. All SiC bonded diamond materials showed a wear, which was approximately 10 times less than the wear behavior of the reference SiC material.Systematic changed microstructures were created by increasing the infiltration temperature. As the infiltration temperature increases, a graphite layer is formed at the diamond-SiC interface. At the highest infiltration temperature (1670 °C), the layer thickness reaches approx. 580 nm. The results show that wear resistance is not negatively affected by the graphite layer. On the contrary, for materials with a graphite layer thickness of up to 70 nm, the wear resistance increases by up to 30 %. The wear increases again only at the highest infiltration temperature. However, this is probably caused more by the internal damage to the diamonds and not by the graphite layer at the interface.

Materials with high thermal conductivity are required in a wide range of thermal management applications. Silicon carbide (SiC) bonded diamond materials, which can be produced without pressure, are a possible candidate for such applications. They exhibit thermal conductivities >650 W/(m·K) depending on the diamond content, diamond grain size and residual silicon content. It is difficult to experimentally determine the influence of these factors on the thermal conductivity separately, as changing one of these parameters affects the other parameters. Therefore, a method for generating digital microstructures of SiC- bonded diamond materials was developed and the thermal conductivity was calculated as a function of the grain size These data were compared with real structures.

Analysis of the microstructure of electro-porcelain is important to better understand its influence on mechanical, electrical and aging behavior. The microstructures of two electro-porcelain materials with low and high quartz contents were analyzed with respect to the distribution of quartz. Using an adequate evaluation of EDS (energy dispersive spectroscopy) mapping data, a reproducible analysis of the size distribution and quantity of quartz was achieved. The method allows the analysis of large areas (12.5 mm2). Therefore, the probability of occurrence for a few large quartz grains could be determined. Independent of the overall amount of quartz in the materials, a wide distribution of the grain size was observed. The size of the large detected quartz grains in both materials was very similar. Around the large quartz particles, microcrack systems with lengths of several 100 µm were observed. They are linearly correlated with the equivalent circle diameter of the quartz grains. The evaluation of the cracks allowed us to determine the critical size below which no cracks around the quartz particles are formed. This size is approximately 10 µm.

ABSTRACT Invasion of host cells is an important feature of Staphylococcus aureus. The main internalization pathway involves binding of the bacteria to host cells, e.g., endothelial cells, via a fibronectin (Fn) bridge between S. aureus Fn binding proteins and α5β1-integrin, followed by phagocytosis. The secreted extracellular adherence protein (Eap) has been shown to promote this cellular uptake pathway of not only S. aureus, but also of bacteria otherwise poorly taken up by host cells, such as Staphylococcus carnosus. The exact mechanisms are still unknown. Previously, we demonstrated that Eap induces platelet activation by stimulation of the protein disulfide isomerase (PDI), a catalyst of thiol-disulfide exchange reactions. Here, we show that Eap promotes PDI activity on the surface of endothelial cells, and that this contributes critically to Eap-driven staphylococcal invasion. PDI-stimulated β1-integrin activation followed by increased Fn binding to host cells likely accounts for the Eap-enhanced uptake of S. aureus into non-professional phagocytes. Additionally, Eap supports the binding of S. carnosus to Fn-α5β1 integrin, thereby allowing its uptake into endothelial cells. To our knowledge, this is the first demonstration that PDI is crucial for the uptake of bacteria into host cells. We describe a hitherto unknown function of Eap—the promotion of an enzymatic activity with subsequent enhancement of bacterial uptake—and thus broaden mechanistic insights into its importance as a driver of bacterial pathogenicity. IMPORTANCE Staphylococcus aureus can invade and persist in non-professional phagocytes, thereby escaping host defense mechanisms and antibiotic treatment. The intracellular lifestyle of S. aureus contributes to the development of infection, e.g., in infective endocarditis or chronic osteomyelitis. The extracellular adherence protein secreted by S. aureus promotes its own internalization as well as that of bacteria that are otherwise poorly taken up by host cells, such as Staphylococcus carnosus. In our study, we demonstrate that staphylococcal uptake by endothelial cells requires catalytic disulfide exchange activity by the cell-surface protein disulfide isomerase, and that this critical enzymatic function is enhanced by Eap. The therapeutic application of PDI inhibitors has previously been investigated in the context of thrombosis and hypercoagulability. Our results add another intriguing possibility: therapeutically targeting PDI, i.e., as a candidate approach to modulate the initiation and/or course of S. aureus infectious diseases.

ABSTRACT Belonging to a group of membrane proteins, bacterial lipoproteins (LPPs) are defined by a unique lipid structure at their N-terminus providing the anchor in the bacterial cell membrane. In Gram-positive bacteria, LPPs play a key role in host immune activation triggered through a Toll-like receptor 2 (TLR2)-mediated action resulting in macrophage stimulation and subsequent tissue damage demonstrated in in vivo experimental models. Yet the physiologic links between LPP activation, cytokine release, and any underlying switches in cellular metabolism remain unclear. In this study, we demonstrate that Staphylococcus aureus Lpl1 not only triggers cytokine production but also confers a shift toward fermentative metabolism in bone marrow-derived macrophages (BMDMs). Lpl1 consists of di- and tri-acylated LPP variants; hence, the synthetic P2C and P3C, mimicking di-and tri-acylated LPPs, were employed to reveal their effect on BMDMs. Compared to P3C, P2C was found to shift the metabolism of BMDMs and the human mature monocytic MonoMac 6 (MM6) cells more profoundly toward the fermentative pathway, as indicated by lactate accumulation, glucose consumption, pH reduction, and oxygen consumption. In vivo, P2C caused more severe joint inflammation, bone erosion, and lactate and malate accumulation than P3C. These observed P2C effects were completely abrogated in monocyte/macrophage-depleted mice. Taken together, these findings now solidly confirm the hypothesized link between LPP exposure, a macrophage metabolic shift toward fermentation, and ensuing bone destruction. IMPORTANCE Osteomyelitis caused by S. aureus is a severe infection of the bone, typically associated with severe bone function impairment, therapeutic failure, high morbidity, invalidity, and occasionally even death. The hallmark of staphylococcal osteomyelitis is the destruction of the cortical bone structures, yet the mechanisms contributing to this pathology are hitherto poorly understood. One bacterial membrane constituent found in all bacteria is bacterial lipoproteins (LPPs). Previously, we have shown that injection of purified S. aureus LPPs into wild-type mouse knee joints caused a TLR2-dependent chronic destructive arthritis but failed to elicit such effect in monocyte/macrophage-depleted mice. This observation stirred our interest in investigating the interaction of LPPs and macrophages and analyzing the underlying physiological mechanisms. This ascertainment of LPP-induced changes in the physiology of macrophages provides an important clue in the understanding of the mechanisms of bone disintegration, opening novel avenues to manage the course of S. aureus disease.

The technology of multi-wire sawing is well established in the production of silicon wafers but can also be applied in the production of ceramic substrates. In this study, the influence of the Al2O3-grain size of the alumina ceramic on the efficiency of the multi-wire slurry process was investigated. The grain size of HIPed alumina ceramics was changed by heat treatment processes at 1350 °C and 1400 °C. A B4C slurry was used for the investigation of the cutting of high purity alumina ceramic. With increasing grain size of the ceramic, the efficiency of the sawing process increases. The analysis of the as-cut surface morphology of the substrates shows a change in material removal from trans- to intergranular micro-fracture with increasing grain size. Furthermore, grain coarsening leads to substrates with increased roughness values and reduced biaxial strength.

High quality α/β-Sialon materials were reproducible prepared using cost effective silicon nitride powders and an aqueous processing route. The influence of the powder quality (SN-E10, Silzot HQ, SicoNide P95H) on sintering and phase formation was investigated. With all 3 powders dense Sialon materials with nominal composition RxSi12-(m + n)Alm + nOnN16-n and m = 0.5 and n = 1 and 4 wt% excess rare earth additives could be prepared. The material based on Silzot HQ exhibits the highest α-Sialon content and subsequently the highest hardness due to the lower oxygen content of the starting powder.The microstructural analysis using XRD, FESEM including EDX- and EBSD-mapping reveal, that the α-Sialon grains are formed on existing α-Si3N4 grains of the starting powder. Anisotropic grain growth of the α-Sialon takes place therefore the differences in the microstructure depend strongly on the oxygen content but not from the α/β-content of the starting powders.

Traditionally, Staphylococcus aureus

The potential combinations of favorable properties give metal–ceramic laminates (MCLs) a high degree of application flexibility. However, the different thermal expansion coefficients (CTEs) and shrinkage rates of the metals and ceramics during the co-sintering process often lead to large internal stresses that cause undesired deformation or even production failures. In practice, the identification of manufacturable MCLs relies on the “trial and error” principle, which usually requires a long development period. Therefore, there is a great demand for analytic and numerical methods that allow the prediction of the deformation and manufacturability of MCLs during the co-sintering process. The main objective of this study is to investigate the curvature and stress distribution in the MCLs (steel 17-4PH/ ceramic 3Y-TZP) based on the analytic solution and finite element (FE) simulation. To achieve this, the Young’s moduli (E) and shear moduli (G) at high temperatures and the CTEs of both materials were measured. In addition, the curvatures and stress distributions of the two-layer and three-layer laminates were obtained based on the analytic method and FE simulation, which were in very good agreement. Furthermore, the influence of the CTE, Young’s modulus, height ratio, and interface on the curvature were studied. The results showed that the CTE and height ratio have a higher influence on the curvature in comparison to the Young’s modulus. The interface prevents the curvature significantly by assuming it to be a cohesive surface in the FE simulation. This provides hints to avoid delamination during the manufacturing process.

The density, microstructure, and ionic conductivity of solid electrolyte Li1.3Al0.3Ti1.7(PO4)3 (LATP) ceramics prepared by cold sintering using liquid and solid sintering additives are studied. The effects of both liquid (water and water solutions of acetic acid and lithium hydroxide) and solid (lithium acetate) additives on densification are investigated. The properties of cold-sintered LATP are compared to those of conventionally sintered LATP. The materials cold-sintered at temperatures 140–280 °C and pressures 510–600 MPa show relative density in the range of 90–98% of LATP’s theoretical value, comparable or higher than the density of conventionally sintered ceramics. With the relative density of 94%, a total ionic conductivity of 1.26 × 10−5 S/cm (room temperature) is achieved by cold sintering at the temperature of 200 °C and uniaxial pressure of 510 MPa using water as additive. The lower ionic conductivities of the cold-sintered ceramics compared to those prepared by conventional sintering are attributed to the formation of amorphous secondary phases in the intergranular regions depending on the type of additives used and on the processing conditions selected.

Emerging antibiotic resistance is a major global health threat. The analysis of nucleic acid sequences linked to susceptibility phenotypes facilitates the study of genetic antibiotic resistance determinants to inform molecular diagnostics and drug development. We collected genetic data (11,087 newly-sequenced whole genomes) and culture-based resistance profiles (10,991 out of the 11,087 isolates comprehensively tested against 22 antibiotics in total) of clinical isolates including 18 main species spanning a time period of 30 years. Species and drug specific resistance patterns were observed including increased resistance rates for Acinetobacter baumannii to carbapenems and for Escherichia coli to fluoroquinolones. Species-level pan-genomes were constructed to reflect the genetic repertoire of the respective species, including conserved essential genes and known resistance factors. Integrating phenotypes and genotypes through species-level pan-genomes allowed to infer gene–drug resistance associations using statistical testing. The isolate collection and the analysis results have been integrated into GEAR-base, a resource available for academic research use free of charge at https://gear-base.com. Keywords: Antibiotic resistance, Whole-genome sequencing, Bacteria, Pan-genome

The Staphylococcus aureus-related complex is formed by the Staphylococcus aureus, Staphylococcus schweitzeri, Staphylococcus argenteus, Staphylococcus roterodami and Staphylococcus singaporensis. Within this complex, S. schweitzeri is the only species mainly found in African wildlife, but it is rarely detected as a colonizer in humans or as a contaminant of fomites. The few detections in humans are most likely spillover events after contact with wildlife. However, since S. schweitzeri can be misidentified as S. aureus using culture-based routine techniques, it is likely that S. schweitzeri is under-reported in humans. The low number of isolates in humans, though, is consistent with the fact that the pathogen has typical animal adaptation characteristics (e.g., growth kinetics, lack of immune evasion cluster and antimicrobial resistance); however, evidence from selected in vitro assays (e.g., host cell invasion, cell activation, cytotoxicity) indicate that S. schweitzeri might be as virulent as S. aureus. In this case, contact with animals colonized with S. schweitzeri could constitute a risk for zoonotic infections. With respect to antimicrobial resistance, all described isolates were found to be susceptible to all antibiotics tested, and so far no data on the development of spontaneous resistance or the acquisition of resistance genes such the mecA/mecC cassette are available. In summary, general knowledge about this pathogen, specifically on the potential threat it may incur to human and animal health, is still very poor. In this review article, we compile the present state of scientific research, and identify the knowledge gaps that need to be filled in order to reliably assess S. schweitzeri as an organism with global One Health implications.

Several methods to isolate monocytes from whole blood have been previously published, with different advantages and disadvantages. For the purpose of cytokine release assessment upon external stimulation, the use of monocyte preparations consisting of non-activated cells is prerequisite. Affinity-isolated monocyte preparations from peripheral blood mononuclear cells (PBMCs), obtained via positive or negative selection using magnetic beads, released pro-inflammatory cytokines such as TNF-α and IL-6 even without adding external stimuli, hindering any assessment of an effect of bacterial lipoproteins on cell stimulation. Hence, the cell preparation protocol was modified by adding a quiescence step on repellent surface culture plates, dampening any monocyte pre-activation. This protocol now provides a robust method to prepare silent yet fully activatable, pure monocyte populations for further use in stimulus-elicited activation experiments.

When one thinks of the Gram+ cell wall, the peptidoglycan (PG) scaffold in particular comes to mind. However, the cell wall also consists of many other components, for example those that are covalently linked to the PG: the wall teichoic acid and the cell wall proteins tethered by the sortase. In addition, there are completely different molecules that are anchored in the cytoplasmic membrane and span the cell wall. These are lipoteichoic acids and bacterial lipoproteins (Lpp). The latter are in the focus of this review. Lpp are present in almost all bacteria. They fulfill a wealth of different tasks. They represent the window to the outside world by recognizing nutrients and incorporating them into the bacterial cell via special transport systems. Furthermore, they perform very diverse and special tasks such as acting as chaperonin, as cyclomodulin, contributing to invasion of host cells or uptake of plasmids via conjugation. All these functions are taken over by the protein part. Nevertheless, the lipid part of the Lpp plays an as important role as the protein part. It is the released lipoproteins and derived lipopeptides that massively modulate our immune system and ultimately play an important role in immune tolerance or non-tolerance. All these varied activities of the Lpp are considered in this review article.

Diamond bonded materials can be produced without pressure by infiltration with liquid Si. They are very wear-resistant and show high chemical resistance in water, acids and alkaline solutions. However, there are no reliable data on the oxidation resistance of these materials. The oxidation resistance has been investigated by thermal analysis and by long-term ageing at 900–1000 ​°C up to 100 ​h. It is shown that not only the surface diamonds are oxidized, but the oxidation of the diamond reaches deep into the volume of the materials. The analysis of the mass changes and the corrosion depth shows that the oxidation rate is probably determined by the diffusion of the reaction gases. The application limits regarding the temperature of the material under oxidizing conditions are therefore in a similar range as those of diamond materials produced under high pressure.

Given that binding and internalization of bacteria to host cells promotes infections and invasion, we aimed at characterizing how various S. aureus isolates adhere to and are internalized by different white blood cells. In particular, the role of genetic determinants on the association kinetics should be unveiled. A flow cytometric (FACS) whole blood assay with fluorescently labelled isolates was applied to 56 clinical S. aureus isolates. This phenotypic data was then linked to previously obtained genotyping data (334 genes) with the help of a redescription mining algorithm. Professional phagocytes showed a time-dependent increase of bacterial adhesion and internalization. Isolates showing higher affinity to granulocytes were associated with lower binding to monocytes. In contrast binding activity between S. aureus and lymphocytes could be subdivided into two phases. Preliminary binding (phase 1) was highest directly after co-incubation and was followed by S. aureus detachment or by sustained binding of a small lymphocyte subset (phase 2). Strain-dependent low granulocyte binding was observed for clonal complex 5 (CC5) isolates (MRSA), as compared to CC30 and CC45 (MSSA). S. aureus isolates associated with low granulocyte phagocytosis were characterized by the presence (cap8, can) and the absence (sasG, lukD, isdA, splA, setC) of specific hybridization signals.

New sodium-based battery concepts require solid electrolytes as ion conducting separators. Besides NaSICON and β-Al2O3 in the Na2O-R2O3-SiO2 system (R = rare earth), a rarely noticed glass-ceramic solid electrolyte with the composition Na5RSi4O12 (N5-type) exists. The present study addresses the investigation of the ionic conductivity of Na5RSi4O12 solid electrolytes sintered from pre-crystallized glass-ceramic powders. The sintering behavior (optical dilatometry), the microstructure (SEM/EDX), and phase composition (XRD), as well as electrochemical properties (impedance spectroscopy), were investigated. To evaluate the effect of the ionic radii, Y, Sm and Gd rare elements were chosen. All compositions were successfully synthesized to fully densified compacts having the corresponding conducting N5-type phase as the main component. The densification behavior was in agreement with the melting point, which decreased with increasing ionic radii and specific cell volume. Alternatively, the ionic conductivities of N5-phases decreased from Y to Gd and Sm containing samples. The highest ionic conductivity of 1.82 × 10−3 S cm−1 at 20 °C was obtained for Na5YSi4O12 composition. The impact of grain boundaries and bulk conductivity on measured values is discussed. A powder-based synthesis method of this glass-ceramic solid electrolyte using different rare earth elements opens possibilities for optimizing ionic conductivity and scalable technological processing by tape casting.

Abstract Clonal clusters and gene repertoires of Staphylococcus aureus are essential to understand disease and are well characterized in industrialized countries but poorly analysed in developing regions. The objective of this study was to compare the molecular-epidemiologic profiles of S. aureus isolates from Sub-Saharan Africa and Germany. S. aureus isolates from 600 staphylococcal carriers and 600 patients with community-associated staphylococcal disease were characterized by DNA hybridization, clonal complex (CC) attribution, and principal component (PCA)-based gene repertoire analysis. 73% of all CCs identified representing 77% of the isolates contained in these CCs were predominant in either African or German region. Significant differences between African versus German isolates were found for alleles encoding the accessory gene regulator type, enterotoxins, the Panton-Valentine leukocidin, immune evasion gene cluster, and adhesins. PCA in conjunction with silhouette analysis distinguished nine separable PCA clusters, with five clusters primarily comprising of African and two clusters of German isolates. Significant differences between S. aureus lineages in Africa and Germany may be a clue to explain the apparent difference in disease between tropical/(so-called) developing and temperate/industrialized regions. In low-resource countries further clinical-epidemiologic research is warranted not only for neglected tropical diseases but also for major bacterial infections.

Wear-resistant, super hard ceramic composites based on cubic boron nitride (cBN) are of great interest to industry. However, cBN is metastable under sintering conditions at normal pressure and converts into the soft hexagonal BN (hBN). Therefore, efforts are being made to avoid this process. Besides short sintering times, the use of coated cBN-particles is a way to minimize this process. Therefore, the thermal stability of TiN coated cBN powders in high purity argon and nitrogen atmospheres up to temperatures of 1600 °C was investigated by thermogravimetry, X-ray phase analysis, scanning electron microscopy and Raman spectroscopy. The TiN coating was prepared by the atomic layer deposition (ALD)-method. The investigations showed that the TiN layer reacts in Ar at T ≥ 1200 °C with the cBN and forms a porous TiB2 layer. No reaction takes place in nitrogen up to temperatures of 1600 °C. Nevertheless, the 20 and 50 nm thin coatings also undergo a recrystallization process during heat treatment up to temperatures of 1600 °C.

Dense, high-entropy carbide cobalt-bonded hardmetals with two different compositions, namely (Hf-Ta-Ti-Nb-V)C-19.2 vol% Co and (Ta-Ti-Nb-V-W)C-19.2 vol% Co, were successfully manufactured by gas pressure sintering (SinterHIP) at 1400 °C and 100 bar Ar pressure. The microstructure of these hardmetals consists of a rigid skeletal carbide phase embedded in a tough Co binder phase. EDS mappings showed that the high-entropy carbide phase did not decompose and that a typical hardmetal microstructure was realized. Only in the case of the (Hf-Ta-Ti-Nb-V)C-Co hardmetal was some undissolved TaC and HfO2, as well as some clustered vanadium titanium carbide phase, found, resulting in a split-up of the HEC phase into two very similar HEC phases. This resulted in a reduced hardness to fracture toughness ratio for this composition. Measurements of magnetic saturation polarization showed values between 57.5% and 70% of theoretical magnetic saturation polarization, indicating marginal dissolution of the carbide-forming metal elements in the binder phase. The hardness value HV10 for (Hf-Ta-Ti-Nb-V)C-19.2 vol% Co was 1203 HV10 and 1432 HV10 for (Ta-Ti-Nb-V-W)C-19.2 vol% Co.

Silicon nitride–zirconia–graphene composites with high graphene content (5 wt.% and 30 wt.%) were sintered by gas pressure sintering (GPS). The effect of the multilayer graphene (MLG) content on microstructure and fracture mechanism is investigated by multi-scale and in-situ microscopy. Multi-scale microscopy confirms that the phases disperse evenly in the microstructure without obvious agglomeration. The MLG flakes well dispersed between ceramic matrix grains slow down the phase transformation from α to β-Si3N4, subsequent needle-like growth of β-Si3N4 rods and the densification due to the reduction in sintering additives particularly in the case with 30 wt.% MLG. The size distribution of Si3N4 phase shifts towards a larger size range with the increase in graphene content from 5 to 30 wt.%, while a higher graphene content (30 wt.%) hinders the growth of the ZrO2 phase. The composite with 30 wt.% MLG has a porosity of 47%, the one with 5 wt.% exhibits a porosity of approximately 30%. Both Si3N4/MLG composites show potential resistance to contact or indentation damage. Crack initiation and propagation, densification of the porous microstructure, and shift of ceramic phases are observed using in-situ transmission electron microscopy. The crack propagates through the ceramic/MLG interface and through both the ceramic and the non-ceramic components in the composite with low graphene content. However, the crack prefers to bypass ceramic phases in the composite with 30 wt.% MLG.

During metal cutting, high temperatures of several hundred-degree Celsius occur locally at the cutting edge, which greatly impacts tool wear and life. Not only the cutting parameters, but also the tool material’s properties influence the arising cutting temperature which in turn alters the mechanical properties of the tool. In this study, the hardness and thermal conductivity of cemented tungsten carbides were investigated in the range between room temperature and 1000 °C. The occurring temperatures close to the cutting edge were measured with two color pyrometry. The interactions between cemented carbide tool properties and cutting process parameters, including cutting edge rounding, are discussed. The results show that cemented carbides with higher thermal conductivities lead to lower temperatures during cutting. As a result, the effective hardness at the cutting edge can be strongly influenced by the thermal conductivity. The differences in hardness measured at room temperature can be equalized or evened out depending on the combination of hardness and thermal conductivity. This in turn has a direct influence on tool wear. Wear is also influenced by the softening of the workpiece, so that higher cutting temperatures can lead to less wear despite the same effective hardness.

Multi-wire sawing has emerged as the leading technology in wafer production for a variety of semiconductor materials. This study investigates the process stability and efficiency of conventional semiconductor multi-wire slurry saws in routinely machining translucent, high-density alumina ceramics. The brittle and fine-grained translucent alumina ceramics with extreme hardness and wear resistance represents a major challenge for the process. The alumina ceramic substrates are used for sensor applications, energy storage technology and applications in power electronics. An ideal adaptation of the sawing process parameters to the workpiece properties guarantees the efficiency of the slurry sawing process and the quality of the ceramic wafers. An indicator of the efficiency and cutting ability of the sawing process is the size of the bow of the wire web. The first time was shown that the wire bow can be used for the characterization of the sawing processes for hard and brittle technical ceramics. It was found that a longer workpiece length, a higher number of wafers and stronger abrasive wear lead to an increased size of the bow. The rocking frequency has no measurable influence on the size of the bow. Knowledge of these relations is an extremely valuable tool in the sawing process development for hard and brittle technical ceramics.

The extracellular adherence protein (Eap) of Staphylococcus aureus is a secreted protein known to exert a number of adhesive and immunomodulatory properties. Here we describe the intrinsic DNA binding activity of this multifunctional secretory factor. By using atomic force microscopy, we provide evidence that Eap can bind and aggregate DNA. While the origin of the DNA substrate (e.g., eukaryotic, bacterial, phage, and artificial DNA) seems to not be of major importance, the DNA structure (e.g., linear or circular) plays a critical role with respect to the ability of Eap to bind and condense DNA. Further functional assays corroborated the nature of Eap as a DNA binding protein, since Eap suppressed the formation of “neutrophil extracellular traps” (NETs), composed of DNA-histone scaffolds, which are thought to function as a neutrophil-mediated extracellular trapping mechanism. The DNA binding and aggregation activity of Eap may thereby protect S. aureus against a specific anti-microbial defense reaction from the host.

Background: The emergence of community-acquired Staphylococcus aureus infections is increasingly recognized as life threating problem worldwide. In Manhiça district, southern Mozambique, S. aureus is the leading cause of community-acquired bacteremia in neonates.Methods: Eighty-four S. aureus isolates from children less than 5 years admitted to Manhiça District Hospital from 2001 to 2009 were randomly selected and genetically characterized by DNA microarray and spa typing. Antimicrobial susceptibility was determined by VITEK 2.Results: Thirty-eight different spa types and 14 clonal complexes (CC) were identified. Spa-type t084 (n = 10; 12%) was the most predominant while CC8 (n = 18; 21%) and CC15 (n = 14; 16%) were the most frequent CCs. Mortality tended to be higher among children infected with CC45 (33.3%, 1/3) and CC8 (27.8%, 5/18). The majority of isolates possessed the accessory gene regulator I (45%) and belonged to either capsule type 8 (52%) or 5 (47%). Panton valentine leukocidin (PVL) encoding genes were detected in 30%. Antibiotic resistance was high for penicillin (89%), tetracycline (59%) and Trimethoprim Sulfamethoxazole (36%) while MRSA was uncommon (8%).Conclusions: Although MRSA were uncommon, we found high genetic diversity of methicillin susceptible S. aureus causing bacteremia in Mozambican children, associated with high resistance to the most available antibiotics in this community. Some CCs are likely to be more lethal indicating the need for prompt recognition and appropriate treatment.

Many bacteria regulate the expression of virulence factors via carbon catabolite responsive elements. In Gram-positive bacteria, the predominant mediator of carbon catabolite repression is the catabolite control protein A (CcpA). Hyperglycemia is a widespread disorder that predisposes individuals to an array of symptoms and an increased risk of infections. In hyperglycemic individuals, the bacterium Staphylococcus aureus causes serious, life-threatening infections. The importance of CcpA in regulating carbon catabolite repression in S. aureus suggests it may be important for infections in hyperglycemic individuals. To test this suggestion, hyperglycemic non-obese diabetic (NOD; blood glucose level ≥20 mM) mice were challenged with the mouse pathogenic S. aureus strain Newman and the isogenic ccpA deletion mutant (MST14), and the effects on infectivity were determined. Diabetic NOD mice challenged with the ccpA deletion mutant enhanced the symptoms of infection in an acute murine pneumonia model relative to the parental strain. Interestingly, when diabetic NOD mice were used in footpad or catheter infection models, infectivity of the ccpA mutant decreased relative to the parental strain. These differences greatly diminished when normoglycemic NOD mice (blood glucose level ≤ 10 mM) were used. These data suggest that CcpA is important for infectivity of S. aureus in hyperglycemic individuals.

Unspecific adhesion of bacteria is usually the first step in the formation of biofilms on abiotic surfaces, yet it is unclear up to now which forces are governing this process. Alongside long-ranged van der Waals and electrostatic forces, short-ranged hydrophobic interaction plays an important role. To characterize the forces involved during approach and retraction of an individual bacterium to and from a surface, single cell force spectroscopy is applied: A single cell of the apathogenic species Staphylococcus carnosus isolate TM300 is used as bacterial probe. With the exact same bacterium, hydrophobic and hydrophilic surfaces can be probed and compared. We find that as far as 50 nm from the surface, attractive forces can already be recorded, an indication of the involvement of long-ranged forces. Yet, comparing the surfaces of different surface energy, our results corroborate the model that large, bacterial cell wall proteins are responsible for adhesion, and that their interplay with the short-ranged hydrophobic interaction of the involved surfaces is mainly responsible for adhesion. The ostensibly long range of the attraction is a result of the large size of the cell wall proteins, searching for contact via hydrophobic interaction. The model also explains the strong (weak) adhesion of S. carnosus to hydrophobic (hydrophilic) surfaces.

BACKGROUND:Multiresistant organisms pose a threat for patients and care recipients. Control interventions need to be tailored to region, the type of institution considered, and risk factors. The German state of Saarland is ideally suited to study colonisation epidemiology throughout its various health and care institutions. After conclusion of a large admission prevalence study in acute care hospitals, we now performed a methicillin-resistant Staphylococcus aureus (MRSA) point prevalence study in Saarland long term care facilities (LTCF), allowing for a direct comparison with respect of MRSA prevalence and associated risk factors between these two institutional types located within a confined region. METHODOLOGY AND PRINCIPAL FINDINGS:Of all LTCF of the region, 65/136 participated in the study performed between 09/2013 and 07/2014. Overall, complete microbiological specimen and questionnaires of 2,858 of 4,275 (66.8%) LTCF residents were obtained. 136/2,858 (4.8%) screened residents revealed MRSA carrier status. Multivariate risk factor analysis yielded ulcer/deep soft tissue infection, urinary tract catheter, and MRSA history with multiple MRSA decolonisation cycles to be independently associated with MRSA carrier status. CONCLUSION:As already known from previous studies, colonisation with MRSA is common in LTCF residents even in an area with relatively low MRSA prevalence. This found prevalence can now be related to the acute care admission prevalence (2.2%) as well as to the admission prevalence in acute care geriatric departments (7.6%). The common clonal attribution (spa type) of MRSA isolates prevalent in the LTCF population as well as in the acute care admission population points towards a close relationship between both types of institutions. However, the ostensible absence of risk factors such as "previous hospitalisation" in conjunction with newly identified factors such as "multiple decolonisation cycles" refers to MRSA colonisation risks independent of contact with acute care facilities. Overall, this large LTCF point prevalence study allows data-based, region-tailored decisions on MRSA screening policies and provides a basis for additional preventative measures.

The emergence of bacteria that are resistant to many currently used drugs emphasizes the need to discover and develop new antibiotics that are effective against such multi-resistant strains. Kendomycin is a novel polyketide that has a unique quinone methide ansa structure and various biological properties. This compound exhibits strong antibacterial activity against Gram-negative and Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). Despite the promise of kendomycinin in several therapeutic areas, its mode of action has yet to be identified.In this study, we used a multidisciplinary approach to gain insight into the antibacterial mechanism of this compound.The antibacterial activity of kendomycin appears to be bacteriostatic rather than bactericidal. Kendomycin inhibited the growth of the MRSA strain COL at a low concentration (MIC of 5 μg/mL). Proteomic analysis and gene transcription profiling of kendomycin-treated cells indicated that this compound affected the regulation of numerous proteins and genes involved in central metabolic pathways, such as the tricarboxylic acid (TCA) cycle (SdhA) and gluconeogenesis (PckA and GapB), cell wall biosynthesis and cell division (FtsA, FtsZ, and MurAA), capsule production (Cap5A and Cap5C), bacterial programmed cell death (LrgA and CidA), the cellular stress response (ClpB, ClpC, ClpP, GroEL, DnaK, and GrpE), and oxidative stress (AhpC and KatA). Electron microscopy revealed that kendomycin strongly affected septum formation during cell division. Most kendomycin-treated cells displayed incomplete septa with abnormal morphology.Kendomycin might directly or indirectly affect the cell division machinery, protein stability, and programmed cell death in S. aureus. Additional studies are still needed to obtain deeper insight into the mode of action of kendomycin.

Bulk B-C-N materials were synthesized under static high thermobaric conditions (20 GPa and 2,000 °C) in a multianvil apparatus from a polymer derived t-BC1.97N ceramic. The bulk samples were characterised using X-ray synchrotron radiation and analytical transmission electron microscopy in combination with electron energy loss spectroscopy. Polycrystalline B-C-N materials with a cubic type structure were formed under the applied reaction conditions, but the formation of a ternary cubic diamond-like c-BC2N compound, could not be unambiguously confirmed.

Cubic boron nitride (c-BN) composites produced at high pressures and temperatures are widely used as cutting tool materials. The advent of new, effective pressure-assisted densification methods, such as spark plasma sintering (SPS), has stimulated attempts to produce these composites at low pressures. Under low-pressure conditions, however, transformation of c-BN to the soft hexagonal BN (h-BN) phase can occur, with a strong deterioration in hardness and wear. In the present work, the influence of secondary phases (B2O3, Si3N4, and oxide glasses) on the transformation of c-BN was studied in the temperature range between 1100 °C and 1575 °C. The different heat treated c-BN particles and c-BN composites were analyzed by SEM, X-ray diffraction, and Raman spectroscopy. The transformation mechanism was found to be kinetically controlled solution–diffusion–precipitation. Given a sufficiently low liquid phase viscosity, the transformation could be observed at temperatures as low as 1200 °C for the c-BN–glass composites. In contrast, no transformation was found at temperatures up to 1575 °C when no liquid oxide phase is present in the composite. The results were compared with previous studies concerning the c-BN stability and the c-BN phase diagram.

Staphylococcus aureus is a major human pathogen that causes a range of infections from acute invasive to chronic and difficult-to-treat. Infection strategies associated with persisting S. aureus infections are bacterial host cell invasion and the bacterial ability to dynamically change phenotypes from the aggressive wild-type to small colony variants (SCVs), which are adapted for intracellular long-term persistence. The underlying mechanisms of the bacterial switching and adaptation mechanisms appear to be very dynamic, but are largely unknown. Here, we analyzed the role and the crosstalk of the global S. aureus regulators agr, sarA and SigB by generating single, double and triple mutants, and testing them with proteome analysis and in different in vitro and in vivo infection models. We were able to demonstrate that SigB is the crucial factor for adaptation in chronic infections. During acute infection, the bacteria require the simultaneous action of the agr and sarA loci to defend against invading immune cells by causing inflammation and cytotoxicity and to escape from phagosomes in their host cells that enable them to settle an infection at high bacterial density. To persist intracellularly the bacteria subsequently need to silence agr and sarA. Indeed agr and sarA deletion mutants expressed a much lower number of virulence factors and could persist at high numbers intracellularly. SigB plays a crucial function to promote bacterial intracellular persistence. In fact, ΔsigB-mutants did not generate SCVs and were completely cleared by the host cells within a few days. In this study we identified SigB as an essential factor that enables the bacteria to switch from the highly aggressive phenotype that settles an acute infection to a silent SCV-phenotype that allows for long-term intracellular persistence. Consequently, the SigB-operon represents a possible target to develop preventive and therapeutic strategies against chronic and therapy-refractory infections.

We applied whole-genome sequencing to reconstruct the spatial and temporal dynamics underpinning the expansion of Clostridium difficile ribotype 027 in Germany. Based on re-sequencing of genomes from 57 clinical C. difficile isolates, which had been collected from hospitalized patients at 36 locations throughout Germany between 1990 and 2012, we demonstrate that C. difficile genomes have accumulated sequence variation sufficiently fast to document the pathogen's spread at a regional scale. We detected both previously described lineages of fluoroquinolone-resistant C. difficile ribotype 027, FQR1 and FQR2. Using Bayesian phylogeographic analyses, we show that fluoroquinolone-resistant C. difficile 027 was imported into Germany at least four times, that it had been widely disseminated across multiple federal states even before the first outbreak was noted in 2007, and that it has continued to spread since.

UnlabelledChronic obstructive pulmonary disease (COPD) is a progressive, inflammatory lung disease that affects a large number of patients and has significant impact. One hallmark of the disease is the presence of bacteria in the lower airways.ObjectiveThe aim of this study was to analyze the detailed structure of microbial communities found in the lungs of healthy individuals and patients with COPD. Nine COPD patients as compared and 9 healthy individuals underwent flexible bronchoscopy and BAL was performed. Bacterial nucleic acids were subjected to terminal restriction fragment (TRF) length polymorphism and clone library analysis. Overall, we identified 326 T-RFLP band, 159 in patients and 167 in healthy controls. The results of the TRF analysis correlated partly with the data obtained from clone sequencing. Although the results of the sequencing showed high diversity, the genera Prevotella, Sphingomonas, Pseudomonas, Acinetobacter, Fusobacterium, Megasphaera, Veillonella, Staphylococcus, and Streptococcus constituted the major part of the core microbiome found in both groups. A TRF band possibly representing Pseudomonas sp. monoinfection was associated with a reduction of the microbial diversity. Non-cultural methods reveal the complexity of the pulmonary microbiome in healthy individuals and in patients with COPD. Alterations of the microbiome in pulmonary diseases are correlated with disease.

BACKGROUND: The screening of hospital admission patients for methicillin resistant Staphylococcus aureus (MRSA) is of undisputed value in controlling and reducing the overall MRSA burden; yet, a concerted parallel universal screening intervention throughout all hospitals of an entire German Federal State has not yet been performed. METHODOLOGY/PRINCIPAL FINDINGS: During a four-week period, all 24 acute care hospitals of the State of Saarland participated in admission prevalence screening. Overall, 436/20,027 screened patients revealed MRSA carrier status (prevalence, 2.2/100 patients) with geriatrics and intensive care departments associated with highest prevalence (7.6/100 and 6.3/100, respectively). Risk factor analysis among 17,975 admission patients yielded MRSA history (OR, 4.3; CI₉₅ 2.7-6.8), a skin condition (OR, 3.2; CI₉₅ 2.1-5.0), and/or an indwelling catheter (OR, 2.2; CI₉₅ 1.4-3.5) among the leading risks. Hierarchical risk factor ascertainment of the six risk factors associated with highest odd's ratios would require 31% of patients to be laboratory screened to allow for detection of 67% of all MRSA positive admission patients in the State. CONCLUSIONS/SIGNIFICANCE: State-wide admission prevalence screening in conjunction with risk factor ascertainment yields important information on the distribution of the MRSA burden for hospitals, and allows for data-based decisions on local or institutional MRSA screening policies considering risk factor prevalence and expected MRSA identification rates.

Results on a systematic study on the effects of ion implantation on the near-surface mechanical and structural properties of boron suboxide (B6O) prepared by uniaxial hot pressing are reviewed. 150 keV fluorine ions at fluences of up to 5.0×1016 ions/cm2 were implanted into the ultrahard ceramic material at room temperature and characterized using Raman spectroscopy, atomic force microscopy, and scanning electron microscopy with energy-dispersive X-ray spectroscopy. Evidence of ion-beam-assisted nucleation of novel clustered BxOyFz particles by ion implantation is revealed. In addition, obtained results also reveal that fluorine implantation into the B6O specimen leads to an overall degradation of near-surface mechanical properties with increasing fluorine fluence. Implications of these observations in the creation of amorphous near-surface layers by high-dose ion implantation are discussed in this paper.

As genotyping of S. aureus is important for epidemiologic research and for hygiene management, methods are required for standardized fast and easily applicable evaluation of closely related epidemic strains with high prevalence in hospitals. In this single centre matched control study we compared a new commercially available DNA microarray (IdentiBAC) with standard spa-typing for S. aureus genotyping. Included in the study was a subgroup of 46 MRSA and matched 46 MSSA nasal isolates of the Saarland University Medical Center collected during a state-wide admission prevalence screening. Microarray (MA) and also spa-typing could easily differentiate the genetically diverse MSSA group. However, due to the predominance of CC5/t003 in the MRSA group a sufficient subtyping required analysis of more complex genetic profiles as was shown here by the MA comprising a total number of 334 different hybridization probes. The genetic repertoire of the MRSA group was characterized by more virulence genes as compared to the MSSA group. The standard evaluation of MA results by the original software into CCs, agr-, SCCmec- and capsule-types was substituted in the present study by implementation of multivariate subtyping of closely related CC5 isolates using three different bioinformatic methods (splits graph, cluster dendrogram, and principal component analysis). Each method used was applicable for standardized and highly discriminative subtyping with high concordance. We propose that the identified S. aureus subtypes with characteristic virulence gene profiles are presumably associated also with virulence and pathogenicity in vivo; however, this remains to be analyzed in future studies. MA was superior to spa-typing for epidemiologic and presumably also provide functional respectively virulence associated characterization of S. aureus isolates. This is of specific importance for the hospital setting. In future, MA could become a new standard test for S. aureus typing in combination with multivariate bioinformatic analysis.

The Vickers microhardness measurements of boron suboxide (B6O) ceramics prepared by uniaxial hot-pressing was investigated at indentation test loads in the range from 0.10 to 2.0 kgf. Results from the investigation indicate that the measured microhardness exhibits an indentation load dependence. Based on the results, we present a comprehensive model intercomparison study of indentation size effects (ISEs) in the microhardness measurements of hot-pressed B6O discussed using existing models, that is, the classical Meyer's law, Li and Bradt's proportional specimen resistance model (PSR), the modified proportional specimen resistance model (MPSR), and Carpinteri's multifractal scaling law (MFSL). The best correlation between literature-cited load-independent Vickers microhardness values, the measured values, and applied models was achieved in the case of the MPSR and the MFSL models.

BACKGROUND: Pandemic community-acquired methicillin-resistant Staphylococcus aureus isolates (CA-MRSA) predominantly encode the Panton-Valentine leukocidin (PVL), which can be associated with severe infections. Reports from non-indigenous Sub-Saharan African populations revealed a high prevalence of PVL-positive isolates. The objective of our study was to investigate the S. aureus carriage among a remote indigenous African population and to determine the molecular characteristics of the isolates, particularly those that were PVL-positive. METHODOLOGY/PRINCIPAL FINDINGS: Nasal S. aureus carriage and risk factors of colonization were systematically assessed in remote Gabonese Babongo Pygmies. Susceptibility to antibiotics, possession of toxin-encoding genes (i.e., PVL, enterotoxins, and exfoliative toxins), S. aureus protein A (spa) types and multi-locus sequence types (MLST) were determined for each isolate. The carriage rate was 33%. No MRSA was detected, 61.8% of the isolates were susceptible to penicillin. Genes encoding PVL (55.9%), enterotoxin B (20.6%), exfoliative toxin D (11.7%) and the epidermal cell differentiation inhibitor B (11.7%) were highly prevalent. Thirteen spa types were detected and were associated with 10 STs predominated by ST15, ST30, ST72, ST80, and ST88. CONCLUSIONS: The high prevalence of PVL-positive isolates among Babongo Pygmies demands our attention as PVL can be associated with necrotinzing infection and may increase the risk of severe infections in remote Pygmy populations. Many S. aureus isolates from Babongo Pygmies and pandemic CA-MRSA-clones have a common genetic background. Surveillance is needed to control the development of resistance to antibiotic drugs and to assess the impact of the high prevalence of PVL in indigenous populations.

Lightweight linear cellular composite materials on basis of austenite stainless TRIP- (TRansformation Induced Plasticity-) steel as matrix with reinforcements of MgO partially stabilized zirconia (Mg-PSZ) are described. Two-dimensional cellular materials for structural applications are conventionally produced by sheet expansion or corrugation processes. The presented composites are fabricated by a modified ceramic extrusion powder technology. Characterization of the microstructure in as-received and deformed conditions was carried out by optical and scanning electron microscopy. Magnetic balance measurements and electron backscatter diffraction (EBSD) were used to identify the deformation-induced martensite evolution in the cell wall material. The honeycomb composite samples exhibit an increased strain hardening up to a certain engineering compressive strain and an extraordinary high specific energy absorption per unit mass and unit volume, respectively. Based on improved property-to-weight ratio such linear cellular structures will be of interest as crash absorbers or stiffened core materials for aerospace, railway, or automotive applications.