Nonribosomal Peptides Produced by Minimal and Engineered Synthetases with Terminal Reductase Domains

Nonribosomal peptide synthetases (NRPSs) use terminal reductase domains for 2-electron reduction of the enzyme-bound thioester releasing the generated peptides as C-terminal aldehydes. Herein, we reveal the biosynthesis of a pyrazine that originates from an aldehyde-generating minimal NRPS termed ATRed in entomopathogenic Xenorhabdus indica. Reductase domains were also investigated in terms of NRPS engineering and, although no general applicable approach was deduced, we show that they can indeed be used for the production of similar natural and unnatural pyrazinones.     

Assessment of extreme and metocean conditions in the Maldives for OTEC applications

The Maldives is a group of tropical atolls, considered globally to be one of the most desirable holiday destinations. There is an urgent requirement to decrease their dependency on fossil fuels that are currently the main source of energy, and a number of renewable energy alternatives are being evaluated. Among these, due to the favorable oceanographic and bathymetric conditions, ocean thermal energy conversion (OTEC) systems represent a viable opportunity for clean and reliable power. However, the stresses the OTEC platform will need to endure during adverse environmental conditions are not well defined. The magnitude of these stresses will then have a direct influence on the design of the OTEC device. In order to overcome this uncertainty, this paper uses hindcast data sets from global weather and ocean models to assess the metocean conditions of the Maldives, with particular reference to extreme conditions. After selecting a suitable location for the deployment of the devices, return values calculated using the peaks‐over‐threshold (POT) methodology are estimated for wind, waves, and currents. The 100‐year return value for the significant wave height is found to be 4.5 m, with a joint occurrence of energy periods between 7.5 and 8.5 seconds, whereas the 100‐year return wind has a velocity of 17.8 m/s and the 100‐year return current of 1.9 m/s. The directionality of these extreme events is also considered, showing the southern and western sub‐quadrants as the prevailing sources, which provides essential information for positioning of the platform. Additional evaluations of tropical revolving storms (TRS) and variations in temperature and salinity patterns are also provided over a 1500‐m water column; temperature varies by approximately 24°C, and salinity by around 2 ppt, showing the suitability of OTEC platforms in the Maldives. This work is therefore of interest to offshore renewable energy stakeholders interested in developing a project in the Maldives or those conducting an analogous analysis in other locations.     

Detection and mitigation of monitor identification attacks in collaborative intrusion detection systems

Collaborative defensive approaches such as collaborative intrusion detection system (CIDS) have emerged as a response to the continuous increase in the sophistication of cyberattacks. Such systems utilize a plethora of heterogeneous monitors to create a holistic picture of the monitored network. A number of research institutes deploy CIDSs that publish their alert data publicly over the Internet. This is important for researchers and security administrators, as such systems provide a source of real‐world alert data for experimentation. However, a class of identification attacks exists, namely probe‐response attacks (PRAs), which can significantly reduce the benefits of a CIDS. In particular, such attacks allow an adversary to detect the network location of the monitors of a CIDS. This article discusses the state of the art, with an emphasis on our previous and ongoing work, with regard to the detection and the mitigation of PRAs. We compare the most promising defensive mechanisms with respect to their effectiveness and the possible negative effects they might introduce to the CIDS. Finally, we provide a thorough discussion of research gaps and possible future directions for the field.     

Non-proportional loading in sequentially linear analysis for 3D stress states

Sequentially linear analysis (SLA), a non-incremental-iterative approach towards finite element simulation of quasi-brittle materials, is based on sequentially identifying a critical integration point in the model, to reduce its strength and stiffness, and the associated critical load multiplier (λ_crit), to scale the linear analysis results. In this article, two novel methods are presented to enable SLA simulations for non-proportional loading situations in a three-dimensional fixed smeared crack framework. In the first approach, the cubic function in the load multiplier is analytically solved for real roots using trigonometric solutions or the Cardano method. In the second approach, the load multiplier is expressed as a function of the inclination of a potential damage plane and is deduced using a constrained optimization approach. The first method is preferred over the second for the validation studies due to computational efficiency and accuracy reasons. A three-point bending beam test, with and without prestress, and an RC slab tested in shear, with and without axial loads, are used as benchmarks. The proposed solution method shows good agreement with the experiments in terms of force-displacement curves and damage evolution.     

Steerable Microinvasive Probes for Localized Drug Delivery to Deep Tissue

Enhanced understanding of neuropathologies has created a need for more advanced tools. Current neural implants result in extensive glial scarring and are not able to highly localize drug delivery due to their size. Smaller implants reduce surgical trauma and improve spatial resolution, but such a reduction requires improvements in device design to enable accurate and chronic implantation in subcortical structures. Flexible needle steering techniques offer improved control over implant placement, but often require complex closed‐loop control for accurate implantation. This study reports the development of steerable microinvasive neural implants (S‐MINIs) constructed from borosilicate capillaries (OD = 60 µm, ID = 20 µm) that do not require closed‐loop guidance or guide tubes. S‐MINIs reduce glial scarring 3.5‐fold compared to prior implants. Bevel steered needles are utilized for open‐loop targeting of deep‐brain structures. This study demonstrates a sinusoidal relationship between implant bevel angle and the trajectory radius of curvature both in vitro and ex vivo. This relationship allows for bevel‐tipped capillaries to be steered to a target with an average error of 0.23 mm ± 0.19 without closed‐loop control. Polished microcapillaries present a new microinvasive tool for chronic, predictable targeting of pathophysiological structures without the need for closed‐loop feedback and complex imaging.     

High intensity pulsed electric fields applied to egg white: effect on Salmonella Enteritidis inactivation and protein denaturation

High-intensity electric fields have been successfully applied to the destruction of Salmonella Enteritidis in diaultrafiltered egg white. The effects of electric field strength (from 20 to 35 kV x cm(-1)), pulse frequency (from 100 to 900 Hz), pulse number (from 2 to 8), temperature (from 4 to 30 degrees C), pH (from 7 to 9), and inoculum size (from 10(3) to 10(7) CFU x ml(-1)) were tested through a multifactorial experimental design. Experimental results indicate that, for Salmonella inactivation, the electric field intensity is the dominant factor with a strongly positive effect, strengthened by its positive interaction with pulse number. Pulse number, temperature, and pH have also significant positive effects but to a lesser extent. In the most efficient conditions, the pulsed electric field (PEF) treatment is capable of 3.5 log10 reduction in viable salmonellae. Simultaneously, the measure of surface hydrophobicity does not indicate any increase after PEF treatment. These results suggest that no protein denaturation occurs, unlike what is observed after comparable heat treatment in terms of Salmonella inactivation (55 degrees C for 15 min).     

G?rungserscheinungen

Ohne Zusammenfassung     

Identification of Entry Inhibitors against Delta and Omicron Variants of SARS-CoV-2

Entry inhibitors against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are urgently needed to control the outbreak of coronavirus disease 2019 (COVID-19). This study developed a robust and straightforward assay that detected the molecular interaction between the receptor-binding domain (RBD) of viral spike protein and the angiotensin-converting enzyme 2 (ACE2) receptor in just 10 min. A drug library of 1068 approved compounds was used to screen for SARS-CoV2 entry inhibition, and 9 active drugs were identified as specific pseudovirus entry inhibitors. A plaque reduction neutralization test using authentic SARS-CoV-2 virus in Vero E6 cells confirmed that 2 of these drugs (Etravirine and Dolutegravir) significantly inhibited the infection of SARS-CoV-2. With molecular docking, we showed that both Etravirine and Dolutegravir are preferentially bound to primary ACE2-interacting residues on the RBD domain, implying that these two drug blocks may prohibit the viral attachment of SARS-CoV-2. We compared the neutralizing activities of these entry inhibitors against different pseudoviruses carrying spike proteins from alpha, beta, gamma, and delta variants. Both Etravirine and Dolutegravir showed similar neutralizing activities against different variants, with EC50 values between 4.5 to 5.8 nM for Etravirine and 10.2 to 22.9 nM for Dolutegravir. These data implied that Etravirine and Dolutegravir may serve as general spike inhibitors against dominant viral variants of SARS-CoV-2.