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ZusammenfassungDieses Review fokussiert die Anatomie der spinalen Übergänge und bringt sie in Zusammenhang mit ihrer manualtherapeutischen Bedeutung. Eingehende anatomische Kenntnisse dieser stärker beweglichen Wirbelsäulenbereiche sind grundlegend für eine korrekte Diagnostik und Therapie. Gerade durch biomechanische Veränderungen der Wirbelsäulenabschnitte zeigen sich an diesen Übergängen oft pathologische Veränderungen, aber auch Möglichkeiten, angegliederte oder weiter entfernte Bereiche und/oder Organe manualtherapeutisch zu behandeln. Die Zusammenhänge sollen hier zusammengefasst dargestellt werden.

The potential of polyketide synthase (PKS) domains for chemoenzymatic synthesis can often not be tapped due to their low stability and activity in vitro. In this proof-of-principle study, the immobilisation of the heterocycle-forming PKS domain AmbDH3 as a cross-linked enzyme aggregate (CLEA) is described. The AmbDH3-CLEA showed good activity recovery, stability and recyclability. Repetitive reactions on the semi-preparative scale were performed with high conversion and isolated yield. Similar to that observed for the free enzyme, the aggregate retained substrate tolerance and the ability for kinetic resolution. This first example of a successful enzymatic PKS domain immobilisation demonstrates that cross-linking can in principle be applied to this type of enzyme to increase its applicability for chemoenzymatic synthesis.

Biosynthetic pathways of natural products contain many enzymes that contribute to the rapid assembly of molecular complexity. Enzymes that form complex structural elements with multiple stereocenters, like chiral saturated oxygen heterocycles (CSOH), are of particular interest for a synthetic application, as their use promises to significantly simplify access to these elements. Here, the biocatalytic characterization of AmbDH3, an enzyme that catalyzes intramolecular oxa-Michael addition (IMOMA) is reported. This reaction essentially gives access to various types of CSOH with adjacent stereocenters, but it is not yet part of the repertoire of preparative biocatalysis. An in-depth study on the synthetic utility of AmbDH3 was performed, which made extensive use of complex synthetic precursor surrogates. The enzyme exhibited stability and broad substrate tolerance in in vitro experiments, which was in agreement with the results of molecular modeling. Its selectivity profile enabled kinetic resolution of chiral tetrahydropyrans (THPs) under control of up to four stereocenters. A systematic optimization of the reaction conditions enabled gram-scale conversions yielding preparative amounts of chiral THP. The synthetic utility of AmbDH3 was finally demonstrated by its successful application in the key step of a chemoenzymatic total synthesis to the THP-containing phenylheptanoid (−)-centrolobine. These results highlight the synthetic potential of AmbDH3 and related IMOMA cyclases as a biocatalytic alternative that further develops the available chemical-synthetic IMOMA methodology. FP7 People: Marie-Curie Actions

Ultrashort laser pulses in the deep ultraviolet (DUV) based on nonlinear-optical conversion of NIR solid state sources promise advantages for the machining of wide-bandgap materials such as compound semiconductors like indium / aluminum / gallium nitride (InAlGaN, AlN, GaN), silicon carbide (SiC), zinc sulfide (ZnS), or boron nitride (BN). We present systems providing few-picosecond pulses at 257.5 nm with high average power exceeding 20 W of DUV obtained by frequency quadrupling of our proven TruMicro Series of industrial ultrafast amplifiers. High beam quality and decent lifetime of the DUV optics are demonstrated.

Oxygen-containing heterocycles are a common structural motif of polyketide natural products and contribute significantly to their biological activity. However, there are only limited reports of the responsible cyclases. Here, we report structural and mechanistic investigations on AmbDH3, a polyketide synthase domain with dual activity as dehydratase (DH) and pyran-forming cyclase in ambruticin biosynthesis. AmbDH3 is similar to monofunctional DH domains, using H51 and D215 for dehydration. V173 was confirmed as a diagnostic residue for cyclisation activity by a mutational study and enzymatic in vitro experiments. Similar motifs were observed in the seemingly monofunctional AmbDH2, which also shows an unexpected cyclase activity. Our results pave the way for mining of hidden cyclases in biosynthetic pathways. They also open interesting prospects for the generation of novel biocatalysts for chemoenzymatic synthesis and pyran-polyketides by combinatorial biosynthesis.

The competitive advantage of today’s evermore complex production systems is characterized by effective and efficient maintenance. In order to ensure the availability of spare parts and maintenance personnel while operating at reasonable costs, the coordination and planning of spare parts supply chains based on machine condition monitoring information provided by intelligent maintenance systems (IMS) has become more important. Therefore, this paper provides decision support for the coordination of an IMS-enabled spare parts supply chain. It explains designing a tactical collaborative planning concept for planning the maintenance capacities of the involved actors based on IMS-supported forecast information.

Machine condition monitoring is a maintenance strategy, which enables real-time diagnostics and prognostics of machine conditions. One major problem of machine fault diagnostics is that data presenting faulty behavior is significantly underrepresented resulting in poor classifier accuracy for the faulty classes. Since misclassification of faulty behavior results in unplanned machine breakdowns and thus economic loss, improved fault diagnostics classifiers handling data imbalance are of importance. This paper addresses the problem of binary and multi-class imbalanced data sets. Based on the structural KDD process for data mining, required steps for imbalanced data sets are defined. The KDD process is evaluated through a real-world industrial case including data sampling and the development of a support vector machine classifier.

The Internet of Things concept is increasingly being used in projects from different areas. Often, projects encompass applications from different domains that need to share information for a common purpose. These types of systems can become complex because they involve different domains and it can be difficult to share information without misinterpreting certain information. Ontologies can be a way to describe information to be shared between different domains. The present work aims to propose a tool that facilitates the creation of Internet of Things systems using ontologies and through this model generate code automatically to pre-automate the integration of devices. A tool has been developed that facilitates the extraction of information from the ontology and it has been validated in the case study.

Summary The invasion of polarized epithelial cells by Salmonella enterica requires the cooperative activity of the Salmonella pathogenicity island (SPI) 1-encoded type III secretion system (T3SS) and the SPI4-encoded giant non-fimbrial adhesin SiiE. SiiE is a highly repetitive protein composed of 53 bacterial Ig (BIg) domains and mediates binding to the apical side of polarized epithelial cells. We analysed the binding properties of SiiE and observed lectin-like activity. SiiE-dependent cell invasion can be ablated by chemical or enzymatic deglycosylation. Lectin blockade experiments revealed that SiiE binding is specific for glycostructures with terminal N-acetyl-glucosamine (GlcNAc) and/or α 2,3-linked sialic acid. In line with these data, we found that SiiE-expressing Salmonella bind to the GlcNAc polymer chitin. Various recombinant SiiE fragments were analysed for host cell binding. We observed that C-terminal portions of SiiE bind to the apical side of polarized cells and the intensity of binding increases with the number of BIg domains present in the recombinant proteins. Based on these results, we propose that SiiE mediates multiple interactions per molecule with glycoproteins and/or glycosylated phospholipids present in the apical membrane of polarized epithelial cells. Thisintimate binding enables the subsequent function of the SPI1-T3SS, resulting in host cell invasion.

Optical tweezers-assisted dynamic force spectroscopy is employed to investigate specific receptor-ligand interactions on the level of single binding events. In particular, we analyze binding of the phosphorylation-specific monoclonal antibody (mAb) HPT-101 to synthetic tau-peptides with two potential phosphorylation sites (Thr231 and Ser235), being the most probable markers for Alzheimer's disease. Whereas the typical interpretation of enzyme-linked immunosorbent assay (ELISA) suggests that this monoclonal antibody binds exclusively to the double-phosphorylated tau-peptide, we show here by DFS that the specificity of only mAb HPT-101 is apparent. In fact, binding occurs also to each sort of monophosphorylated peptide. Therefore, we characterize the unbinding process by analyzing the measured rupture force distributions, from which the lifetime of the bond without force τ0, its characteristic length xts, and the free energy of activation ΔG are extracted for the three mAb/peptide combinations. This information is used to build a simple theoretical model to predict features of the unbinding process for the double-phosphorylated peptide purely based on data on the monophosphorylated ones. Finally, we introduce a method to combine binding and unbinding measurements to estimate the relative affinity of the bonds. The values obtained for this quantity are in accordance with ELISA, showing how DFS can offer important insights about the dynamic binding process that are not accessible with this common and widespread assay.

Summary The giant non-fimbrial adhesin SiiE is essential to establish intimate contact between Salmonella enterica and the apical surface of polarized epithe- lial cells. SiiE is secreted by a type I secretion system (T1SS) encoded by Salmonella Pathogenic- ity Island 4 (SPI4). We identified SiiA and SiiB as two regulatory proteins encoded by SPI4. Mutant strains in siiA or siiB still secrete SiiE, but are highly reduced in adhesion to, and invasion of polarized cells. SiiA and SiiB are inner membrane proteins with one and three transmembrane (TM) helices respectively. TM2 and TM3 of SiiB are similar to members of the ExbB/TolQ family, while the TM of SiiA is similar to MotB and a conserved aspartate residue in this TM is essential for SPI4- encoded T1SS function. Co-immunoprecipitation, bacterial two-hybrid and FRET demonstrate homo- and heterotypic protein interactions for SiiA and SiiB. SiiB, but not SiiA also interacts with the SPI4- T1SS ATPase SiiF. The integrity of the Walker A box in SiiF was required for SiiB-SiiF interaction and SiiF dimer formation. Based on these data, we describe SiiA and SiiB as new, exclusively virulence-associated members of the Mot/Exb/Tol family of membrane proteins. Both proteins are involved in a novel mechanism of controlling SPI4- T1SS-dependent adhesion, most likely by forma- tion of a proton-conducting channel.

Summary SiiE from Salmonella enterica is a giant 5,559-residue-long nonfimbrial adhesin that is secreted by a type 1 secretion system (T1SS) and initiates bacterial adhesion to polarized host cells. Structural insight has been gained into the 53 bacterial Ig-like (BIg) domains of SiiE, which account for 94% of the entire SiiE sequence. The crystal structure of a fragment comprising BIg domains 50 to 52 of SiiE reveals the BIg domain architecture and highlights two types of SiiE-specific Ca 2+ -binding sites. Sequence homology considerations suggest that full-length SiiE interacts with more than 100 Ca 2+ ions. Molecular dynamics simulations and single-molecule imaging indicate that Ca 2+ binding confers SiiE with a rigid 200 nm rod-like habitus that is required to reach out beyond the Salmonella lipopolysaccharide layer and to promote adhesion to host cells. The crystal structure suggests plausible routes for the establishment of the initial contact between Salmonella and host cells.

In patients with chronic cough, nearly 40% of the population does not experience definitive improvement of their cough despite correctly applying the anatomic diagnosis. In many of these patients with refractory cough, laryngeal symptoms are frequent. The region of the larynx/pharynx is configured as a bridge between the esophagus and the upper and lower respiratory tract. The association of reflux in patients with chronic cough and symptoms such as globus pharyngis, itchiness or the need to clear one's throat have recently been given attention due to the possibility of joint therapeutic intervention of the gastroesophageal reflux and larynx, both with new medications as well as with laryngeal rehabilitation therapies, with observed benefits in the disappearance of chronic cough in cases that had been previously labeled as refractory.

By employing single particle tracking on the basis of fast video microscopy, we directly measure the equilibrium short and long time diffusion coefficients of blank and DNA-grafted (molecular weight of 250, 1000 and 4000 base pairs (bp)) tracer colloids in dilute λ-DNA solutions. We compare these results to recently reported microfluidic mobilities under non-equilibrium steady state flow and to the predictions of quantitative theoretical models. A pronounced discrepancy between the measured equilibrium and non-equilibrium mobilities is observed for colloids grafted with 4000 bp DNA molecules, revealing the inapplicability of the Stokes–Einstein relation under these conditions. We demonstrate that this deviation may be interpreted as a strong micro-shear-thinning effect that is caused by the formation of entanglements between the grafted DNA brush and λ-DNA coils from the suspension.

Background This study aimed to assess the performance of two prognostic models—the European Society of Cardiology (ESC) model and the simplified Pulmonary Embolism Severity Index (sPESI)—in predicting short-term mortality in patients with pulmonary embolism (PE). Methods We compared the test characteristics of the ESC model and the sPESI for predicting 30-day outcomes in a cohort of 526 patients with objectively confirmed PE. The primary end point of the study was all-cause mortality. The secondary end point included all-cause mortality, nonfatal symptomatic recurrent VTE, or nonfatal major bleeding. Results Overall, 40 of 526 patients died (7.6%; 95% CI, 5.3%–9.9%) during the first month of follow-up. The sPESI classified fewer patients as low risk (31% [165 of 526], 95% CI, 27%–35%) compared with the ESC model (39% [207 of 526], 95% CI, 35% to 44%; P P Conclusions Both the sPESI and the ESC model successfully predict 30-day mortality after acute symptomatic PE, but exclusion of an adverse early outcome does not appear to require routine imaging procedures or laboratory biomarker testing.

On the basis of a brief review of four common image recognition algorithms for microspheres made of polystyrene or melamine resin, we present a new microsphere localization method for low-contrast silica beads under white light illumination. We compare both the polystyrene and silica procedures with respect to accuracy and precision by means of an optical tweezers setup providing CMOS video microscopy capability. By that we demonstrate that our new silica algorithm achieves a relative position uncertainty of less than ±1 nm for micron-sized microspheres, significantly exceeding the precision of the other silica approaches studied. Second, we present an advancement of our single microsphere tracking method to scenarios where two polystyrene, melamine resin or silica microspheres are in close-to-contact proximity. While the majority of the analysis algorithms studied generate artefacts due to interference effects under these conditions, we show that our new approach yields accurate and precise results.

Summary Salmonella enterica deploys the giant non-fimbrial adhesin SiiE to adhere to the apical side of polar- ized epithelial cells. The establishment of close contact is a prerequisite for subsequent invasion mediated by translocation of effector proteins of the Salmonella Pathogenicity Island 1 (SPI1)- encoded type III secretion system (T3SS). Although SiiE is secreted into the culture medium, the adhesin is retained on the bacterial envelope in the phase of highest bacterial invasiveness. To dissect the structural requirements for secretion, retention and adhesive properties, comprehensive deletional and functional analyses of various domains of SiiE were performed. We observed that b-sheet and coiled-coil domains in the N-terminal moiety of SiiE are required for the control of SiiE retention on the surface and co-ordinated release. These results indicate a novel molecular mecha- nism for the control of surface display of a T1SS- secreted adhesin that acts cooperatively with the SPI1-T3SS.

Optical tweezers accomplished with fast position detection enable one to carry out Brownian motion analysis of single DNA-grafted (grafting density: ∼1000 molecules per particle, molecular weight: 4000 bp) colloids in media of varying NaCl concentration. By that the effective hydrodynamic radius of the colloid under study is determined and found to be strongly dependent on the conformation of the grafted DNA chains. Our results compare well both with recent measurements of the pair interaction potential between DNA-grafted colloids (Kegler et al . Phys Rev Lett 2008; 100:118302) and with microfluidic studies (Gutsche et al . Microfluid Nanofluid 2006; 2:381-386). The observed scaling of the brush height with the ion concentration is in full accord with the theoretical predictions by Pincus, Zhulina, Birshtein and Borisov.

The main advantage of nebulized drugs is that they are deposited directly into the respiratory tract and thus higher drug concentrations can be achieved in the bronchial tree and pulmonary bed with fewer adverse effects than when the systemic route is used. The effectiveness of nebulization depends on many factors, including the characteristics of the drug to be nebulized (particle size, form, density, and surface tension), the anatomy of the airways, the patient's inhalation technique and the nebulization system employed, among others. The factors determining the particle size produced by a nebulizer include both the characteristics of the solution and the flow velocity of the nebulization system. The higher the flow velocity, the smaller the aerosol particles. The parameter most widely used to measure aerosolized particle size is the mass median aerodynamic diameter. Particles of between 1 and 5 μm are those with the greatest probability of reaching the site of infection in the bronchial tree and of achieving the desired therapeutic effect. Nebulization systems or apparatus are composed of a nebulization chamber where the liquid to be nebulized is introduced and aerosol and an energy source required to make the nebulizer work are generated. There are three types of nebulization systems: ultrasonic, jet and mesh nebulizers. Of the three, the most effective are mesh nebulizers. These nebulizers are also the most compact, quietest and most rapid, which translates into better patient compliance.

Resumen Introduccion y objetivos Evaluar la incidencia de hipertension pulmonar tromboembolica cronica (HPTEC) sintomatica y asintomatica en una cohorte de pacientes con tromboembolia de pulmon (TEP), y las variables predictoras de su aparicion. Metodos Estudio prospectivo de cohorte de 110 pacientes consecutivos diagnosticados de TEP en un hospital universitario terciario, y seguidos durante 24 meses. Todos los pacientes fueron sometidos a una ecocardiografia transtoracica (ETT) al final del seguimiento. En los pacientes sintomaticos con una presion sistolica pulmonar estimada (PAP) ≥40 mmHg se realizo un cateterismo cardiaco derecho para confirmar la HPTEC. En los pacientes asintomaticos se repitio la ETT a los 6 meses de la primera y, si la PAP se mantenia por encima de 40 mmHg, se indico un cateterismo cardiaco. Resultados Se confirmo HPTEC en 10 pacientes de la serie (9,1%; intervalo de confianza [IC] 95%, 3,7 a 14,5%). Todos los pacientes con HPTEC presentaron sintomas sugestivos de la enfermedad. La edad (riesgo relativo [RR] ajustado 1,2 por cada ano; IC 95%, 1,0 a 1,3; p=0,03) y el antecedente de TEP (RR 5,7; IC 95%, 1,5 a 22,0; p=0,01) se asociaron de forma estadisticamente significativa al diagnostico de HPTEC. Conclusiones La incidencia de HPTEC es mas frecuente de lo descrito previamente en la literatura. La asociacion entre las recurrencias tromboembolicas y la HPTEC sugiere la necesidad de optimizar la duracion y la intensidad del tratamiento anticoagulante en los pacientes con TEP. No se detectaron episodios de HPTEC en pacientes asintomaticos.

3 MPI fur Entwicklungsbiologie, Tubingen, Germany. Summary Pathogenicity Islands play a major role in the pathogenesis of infections by Salmonella enterica. The molecular function of Salmonella Pathogenicity Island 4 (SPI4) is largely unknown, but recent work indicated a role of SPI4 for Salmonella pathogenesis in certain animal models. We analysed the virulence functions of SPI4 and observed that SPI4 is contrib- uting to intestinal inflammation in a mouse model. On a cellular level, SPI4 mediates adhesion to epithelial cells. We demonstrate the function of SPI4-encoded proteins as a type I secretion system (T1SS) and identify SiiE as the substrate protein of the T1SS. SiiE is secreted into the culture medium but med- iates contact-dependent adhesion to epithelial cell surfaces. SiiE is a very large non-fimbrial adhesin of 600 kDa and consists of 53 repeats of Ig domains. Our study describes the first T1SS-secreted protein that functions as a non-fimbrial adhesin in binding to eukaryotic cells. The SPI4-encoded T1SS and SiiE might functionally resemble the type I fimbrial adhesins.