Correction to "Rational Chemical Design of Molecular Glue Degraders".

[This corrects the article DOI: 10.1021/acscentsci.2c01317.].

Interlaboratory comparison using inactivated SARS-CoV-2 variants as a feasible tool for quality control in COVID-19 wastewater monitoring.

Wastewater-based SARS-CoV-2 epidemiology (WBE) has proven as an excellent tool to monitor pandemic dynamics supporting individual testing strategies. WBE can also be used as an early warning system for monitoring the emergence of novel pathogens or viral variants. However, for a timely transmission of results, sophisticated sample logistics and analytics performed in decentralized laboratories close to the sampling sites are required. Since multiple decentralized laboratories commonly use custom in-house workflows for sample purification and PCR-analysis, comparative quality control of the analytical procedures is essential to report reliable and comparable results. In this study, we performed an interlaboratory comparison at laboratories specialized for PCR and high-throughput-sequencing (HTS)-based WBE analysis. Frozen reserve samples from low COVID-19 incidence periods were spiked with different inactivated authentic SARS-CoV-2 variants in graduated concentrations and ratios. Samples were sent to the participating laboratories for analysis using laboratory specific methods and the reported viral genome copy numbers and the detection of viral variants were compared with the expected values. All PCR-laboratories reported SARS-CoV-2 genome copy equivalents (GCE) for all spiked samples with a mean intra- and inter-laboratory variability of 19 % and 104 %, respectively, largely reproducing the spike-in scheme. PCR-based genotyping was, in dependence of the underlying PCR-assay performance, able to predict the relative amount of variant specific substitutions even in samples with low spike-in amount. The identification of variants by HTS, however, required >100 copies/ml wastewater and had limited predictive value when analyzing at a genome coverage below 60 %. This interlaboratory test demonstrates that despite highly heterogeneous isolation and analysis procedures, overall SARS-CoV-2 GCE and mutations were determined accurately. Hence, decentralized SARS-CoV-2 wastewater monitoring is feasible to generate comparable analysis results. However, since not all assays detected the correct variant, prior evaluation of PCR and sequencing workflows as well as sustained quality control such as interlaboratory comparisons are mandatory for correct variant detection.

Performance and cost-benefit analysis of anaerobic moving bed biofilm reactor for pretreatment of textile wastewater.

Performance of an anaerobic moving bed biofilm reactor (AnMBBR) was evaluated for pretreatment of real textile desizing wastewater at organic loading rate (OLR) of 1±0.05 to 6.3±0.37 kgCOD/m3/d. After OLR optimization, the performance of AnMBBR was evaluated for biodegradation of reactive dyes. AnMBBR was operated under a mesophilic temperature range of 30 to 36 °C, while the oxidation-reduction potential (ORP) and pH were in the range of 504 to 594 (-mV) and 6.98 to 7.28, respectively. By increasing the OLR from 1±0.05 to 6.3±0.37 kgCOD/m3/d, COD and BOD5 removal was decreased from 84 to 39% and 89 to 49%, respectively. While the production of biogas was increased from 0.12 to 0.83 L/L·d up to an optimum OLR of 4.9±0.43 kgCOD/m3/d. With increase in the dye concentration in the feed, COD, BOD5, color removal and biogas production reduced from 56, 63, 70% and 0.65 L/L·d to 34, 43, 41% and 0.08 L/L·d, respectively. Based on the data obtained, a cost-benefit analysis of AnMBBR was also investigated for the pretreatment of real textile desizing wastewater. Cost estimation of anaerobic pretreatment of textile desizing wastewater indicated a net profit of 21.09 million PKR/yr (114,000 €/yr) and a potential payback period of 2.54 years.

Rational Chemical Design of Molecular Glue Degraders.

Targeted protein degradation with molecular glue degraders has arisen as a powerful therapeutic modality for eliminating classically undruggable disease-causing proteins through proteasome-mediated degradation. However, we currently lack rational chemical design principles for converting protein-targeting ligands into molecular glue degraders. To overcome this challenge, we sought to identify a transposable chemical handle that would convert protein-targeting ligands into molecular degraders of their corresponding targets. Using the CDK4/6 inhibitor ribociclib as a prototype, we identified a covalent handle that, when appended to the exit vector of ribociclib, induced the proteasome-mediated degradation of CDK4 in cancer cells. Further modification of our initial covalent scaffold led to an improved CDK4 degrader with the development of a but-2-ene-1,4-dione ("fumarate") handle that showed improved interactions with RNF126. Subsequent chemoproteomic profiling revealed interactions of the CDK4 degrader and the optimized fumarate handle with RNF126 as well as additional RING-family E3 ligases. We then transplanted this covalent handle onto a diverse set of protein-targeting ligands to induce the degradation of BRD4, BCR-ABL and c-ABL, PDE5, AR and AR-V7, BTK, LRRK2, HDAC1/3, and SMARCA2/4. Our study undercovers a design strategy for converting protein-targeting ligands into covalent molecular glue degraders.

Early Detection of SARS-CoV-2 Omicron BA.4 and BA.5 in German Wastewater.

Wastewater-based SARS-CoV-2 epidemiology (WBE) has been established as an important tool to support individual testing strategies. The Omicron sub-variants BA.4/BA.5 have spread globally, displacing the preceding variants. Due to the severe transmissibility and immune escape potential of BA.4/BA.5, early monitoring was required to assess and implement countermeasures in time. In this study, we monitored the prevalence of SARS-CoV-2 BA.4/BA.5 at six municipal wastewater treatment plants (WWTPs) in the Federal State of North Rhine-Westphalia (NRW, Germany) in May and June 2022. Initially, L452R-specific primers/probes originally designed for SARS-CoV-2 Delta detection were validated using inactivated authentic viruses and evaluated for their suitability for detecting BA.4/BA.5. Subsequently, the assay was used for RT-qPCR analysis of RNA purified from wastewater obtained twice a week at six WWTPs. The occurrence of L452R carrying RNA was detected in early May 2022, and the presence of BA.4/BA.5 was confirmed by variant-specific single nucleotide polymorphism PCR (SNP-PCR) targeting E484A/F486V and NGS sequencing. Finally, the mutant fractions were quantitatively monitored by digital PCR, confirming BA.4/BA.5 as the majority variant by 5 June 2022. In conclusion, the successive workflow using RT-qPCR, variant-specific SNP-PCR, and RT-dPCR demonstrates the strength of WBE as a versatile tool to rapidly monitor variants spreading independently of individual test capacities.

Wastewater surveillance allows early detection of SARS-CoV-2 omicron in North Rhine-Westphalia, Germany.

Wastewater-based epidemiology (WBE) has demonstrated its importance to support SARS-CoV-2 epidemiology complementing individual testing strategies. Due to their immune-evasive potential and the resulting significance for public health, close monitoring of SARS-CoV-2 variants of concern (VoC) is required to evaluate the regulation of early local countermeasures. In this study, we demonstrate a rapid workflow for wastewater-based early detection and monitoring of the newly emerging SARS-CoV-2 VoCs Omicron in the end of 2021 at the municipal wastewater treatment plant (WWTP) Emschermuendung (KLEM) in the Federal State of North-Rhine-Westphalia (NRW, Germany). Initially, available primers detecting Omicron-related mutations were rapidly validated in a central laboratory. Subsequently, RT-qPCR analysis of purified SARS-CoV-2 RNA was performed in a decentral PCR laboratory in close proximity to KLEM. This decentralized approach enabled the early detection of K417N present in Omicron in samples collected on 8th December 2021 and the detection of further mutations (N501Y, Δ69/70) in subsequent biweekly sampling campaigns. The presence of Omicron in wastewater was confirmed by next generation sequencing (NGS) in a central laboratory with samples obtained on 14th December 2021. Moreover, the relative increase of the mutant fraction of Omicron was quantitatively monitored over time by dPCR in a central PCR laboratory starting on 12th December 2021 confirming Omicron as the dominant variant by the end of 2021. In conclusions, WBE plays a crucial role in surveillance of SARS-CoV-2 variants and is suitable as an early warning system to identify variant emergence. In particular, the successive workflow using RT-qPCR, RT-dPCR and NGS demonstrates the strength of WBE as a versatile tool to monitor variant spreading.

Early Detection of SARS-CoV-2 Omicron BA.4/5 in German wastewater

Wastewater-based SARS-CoV-2 epidemiology (WBE) has been established as an important tool to support individual testing strategies. Omicron sub-variants BA.4/5 have spread globally displacing the predeceasing variants. Due to the severe transmissibility and immune escape potential of BA.4/5, early monitoring was required to asses and implement countermeasures in time. In this study, we monitored the prevalence of SARS-CoV-2 BA.4/5 at six municipal wastewater treatment plants (WWTPs) in the Federal State of North-Rhine-Westphalia (NRW, Germany) in May and June 2022. Initially, L452R-specific primers/probes originally designed for SARS-CoV-2 Delta detection were validated using inactivated authentic viruses and evaluated for their suitability to detect BA.4/5. Subsequently, the assay was used for RT-qPCR analysis of RNA purified from wastewater obtained twice a week at six WWTPs. The occurrence of L452R carrying RNA was detected in early May 2022 and the presence of BA.4/5 was confirmed by variant-specific single nucleotide polymorphism PCR (SNP-PCR) targeting E484A/F486V. Finally, the mutant fractions were quantitatively monitored by digital PCR confirming BA.4/5 as the majority variant by 5th June 2022. In conclusions, the successive workflow using RT-qPCR, variant-specific SNP-PCR, and RT-dPCR demonstrates the strength of WBE as a versatile tool to rapidly monitor variant spreading independent of individual test capacities.

Targeted Printing of Cells: Evaluation of ADA-PEG Bioinks for Drop on Demand Approaches.

A novel approach, in the context of bioprinting, is the targeted printing of a defined number of cells at desired positions in predefined locations, which thereby opens up new perspectives for life science engineering. One major challenge in this application is to realize the targeted printing of cells onto a gel substrate with high cell survival rates in advanced bioinks. For this purpose, different alginate-dialdehyde-polyethylene glycol (ADA-PEG) inks with different PEG modifications and chain lengths (1-8 kDa) were characterized to evaluate their application as bioinks for drop on demand (DoD) printing. The biochemical properties of the inks, printing process, NIH/3T3 fibroblast cell distribution within a droplet and shear forces during printing were analyzed. Finally, different hydrogels were evaluated as a printing substrate. By analysing different PEG chain lengths with covalently crosslinked and non-crosslinked ADA-PEG inks, it was shown that the influence of Schiff's bases on the viscosity of the corresponding materials is very low. Furthermore, it was shown that longer polymer chains resulted in less stable hydrogels, leading to fast degradation rates. Several bioinks highly exhibit biocompatibility, while the calculated nozzle shear stress increased from approx. 1.3 and 2.3 kPa. Moreover, we determined the number of cells for printed droplets depending on the initial cell concentration, which is crucially needed for targeted cell printing approaches.

Functional Mesostructured Electrospun Polymer Nonwovens with Supramolecular Nanofibers.

Functional, hierarchically mesostructured nonwovens are of fundamental importance because complex fiber morphologies increase the active surface area and functionality allowing for the effective immobilization of metal nanoparticles. Such complex functional fiber morphologies clearly widen the property profile and enable the preparation of more efficient and selective filter media. Here, the realization of hierarchically mesostructured nonwovens with barbed wire-like morphology is demonstrated by combining electrospun polystyrene fibers, decorated with patchy worm-like micelles, with solution-processed supramolecular short fibers composed of 1,3,5-benzenetricarboxamides with peripheral N,N-diisopropylaminoethyl substituents. The worm-like micelles with a patchy microphase-separated corona are prepared by crystallization-driven self-assembly of a polyethylene based triblock terpolymer and deposited on top of the polystyrene fibers by coaxial electrospinning. The micelles are designed in a way that their patches promote the directed self-assembly of the 1,3,5-benzenetricarboxamide and the fixation of the supramolecular nanofibers on the supporting polystyrene fibers. Functionality of the mesostructured nonwoven is provided by the peripheral N,N-diisopropylaminoethyl substituents of the 1,3,5-benzenetricarboxamide and proven by the effective immobilization of individual palladium nanoparticles on the supramolecular nanofibers. The preparation of hierarchically mesostructured nonwovens and their shown functionality demonstrate that such systems are attractive candidates to be used for example in filtration, selective separation and heterogenous catalysis.

Deubiquitinase-targeting chimeras for targeted protein stabilization.

Many diseases are driven by proteins that are aberrantly ubiquitinated and degraded. These diseases would be therapeutically benefited by targeted protein stabilization (TPS). Here we present deubiquitinase-targeting chimeras (DUBTACs), heterobifunctional small molecules consisting of a deubiquitinase recruiter linked to a protein-targeting ligand, to stabilize the levels of specific proteins degraded in a ubiquitin-dependent manner. Using chemoproteomic approaches, we discovered the covalent ligand EN523 that targets a non-catalytic allosteric cysteine C23 in the K48-ubiquitin-specific deubiquitinase OTUB1. We showed that a DUBTAC consisting of our EN523 OTUB1 recruiter linked to lumacaftor, a drug used to treat cystic fibrosis that binds ΔF508-cystic fibrosis transmembrane conductance regulator (CFTR), robustly stabilized ΔF508-CFTR protein levels, leading to improved chloride channel conductance in human cystic fibrosis bronchial epithelial cells. We also demonstrated stabilization of the tumor suppressor kinase WEE1 in hepatoma cells. Our study showcases covalent chemoproteomic approaches to develop new induced proximity-based therapeutic modalities and introduces the DUBTAC platform for TPS.

Structure-Property Relationships of Polyamide 12 Grades Exposed to Rapid Crack Extension.

Thermoplastic materials have established a reputation for long-term reliability in low-pressure gas and water distribution pipe systems. However, occasional Slow Crack Growth (SCG) and Rapid Crack Propagation (RCP) failures still occur. SCG may initiate only a small leak, but it has the potential to trigger RCP, which is much rarer but more catastrophic and destructive. RCP can create a long, straight or meandering axial crack path at speeds of up to hundreds of meters per second. It is driven by internal (residual) and external (pressure) loads and resisted by molecular and morphological characteristics of the polymer. The safe installation and operation of a pipe throughout its service lifetime therefore requires knowledge of its resistance to RCP, particularly when using new materials. In this context, the RCP resistance of five different polyamide (PA) 12 grades was investigated using the ISO 13477 Small-Scale Steady State (S4) test. Since these grades differed not only in molecular weight but also in their use of additives (impact modifiers and pigments), structure-property relationships could be deduced from S4 test results. A new method is proposed for correlating these results more efficiently to evaluate each grade using the crack arrest lengths from individual S4 test specimens.

[Excited Delirium Syndrome (ExDS): emergence and emergency management]. / Erregungssturm ­ Entstehung und Notfallbehandlung.

The Excited Delirium Syndrome (ExDS) is a state of maximum psychophysiological excitation far beyond mental agitation. Patients themselves are at an acute risk and put others at a high risk. We present data from eleven patients and a short literature review. Results: 9 of 11 patients were acutely intoxicated (alcohol, magic mushrooms, THC, ecstacy, or "flakka"). Eight had a history of substance abuse and 4 of other mental illness. None of the patients responded to attempts at verbal de-escalation. Seven had significant injuries at the time of admission to the hospital. The aggressive "psychiatric" presentation of ExDS can lead to vital risks being overlooked (hyperexcitation, acidosis, rhabdomyolysis, hypoxemia, and cardiovascular decompensation), which may even be increased by severe methods of restraint (hogtie; positional asphyxia). Early recognition of ExDS and its risks, comprehensive management and the prevention of inappropriate treatment will help to reduce fatal outcomes.

Hierarchical Superstructures by Combining Crystallization-Driven and Molecular Self-Assembly.

Combining the unique corona structure of worm-like patchy micelles immobilized on a polymer fiber with the molecular self-assembly of 1,3,5-benzenetricarboxamides (BTAs) leads to hierarchical superstructures with a fir-tree-like morphology. For this purpose, worm-like patchy micelles bearing pendant, functional tertiary amino groups in one of the corona patches were prepared by crystallization-driven self-assembly and immobilized on a supporting polystyrene fiber by coaxial electrospinning. The obtained patchy fibers were then immersed in an aqueous solution of a tertiary amino-functionalized BTA to induce patch-mediated molecular self-assembly to well-defined fir-tree-like superstructures upon solvent evaporation. Interestingly, defined superstructures are obtained only if the pendant functional groups in the surface patches match with the peripheral substituents of the BTA, which is attributed to a local increase in BTA concentration at the polymer fibers' surface.

Visualization of the epimysium and fascia thoracolumbalis at the lumbar spine using MRI.

BACKGROUND: The fascia thoracolumbalis (FTL) is an important component for stabilization and motion control of the lumbar spine. It coordinates the traction forces of the autochthonous muscles of the back (AM) and connects them to the muscles of the abdominal wall, shoulder, and buttocks. OBJECTIVES: The aim of our study was to describe the assessment of the normal FTL and epimysium of the AM in MRI and to identify patterns associated with pathological changes in the lumbar spine. MATERIAL AND METHODS: A total of 33 patients were retrospectively evaluated: 15 patients had no pathology at the lumbar spine; six patients had previous hemilaminectomy, three had spondylodesis, two had ventrolisthesis, and seven had scoliosis. The thickness of the FTL and EM was measured, and the adhesion of both structures was assessed. RESULTS: The fascial thickness at the levels of the lumbar vertebral bodies LVB 3 was 1.8, of LVB 4 it was 2.0, of LVB 5 it was 2.1, and at the sacral vertebra SVB 1 it was 1.8 mm. Fascial adhesions together with thickening of the EM occurred at the level of LVB 4 in 36% of the cases independently of the underlying disorder. Only thickening of the EM was seen in 48% of cases at the level of SVB 1. By contrast, adhesion of the FTL without epimysial changes occurred in 36% of cases at the level of LVB 3. CONCLUSION: Thickening and adhesions at the EM and FTL occurred both postoperatively and in the case of scoliosis. Furthermore, lipomatous and muscular herniation could be detected in the FTL postoperatively. Epimysial and fascial alterations may be imaging manifestations of chronic myofascial back pain and should be included in radiological assessments.

Thickness sensor for drying paints using THz spectroscopy.

Optical terahertz technology has, despite its exciting properties, such as transparency of visibly opaque materials, 30 years after its technological breakthrough, not found a widespread application with societal relevance. Main causes are its maturity and costs. If, however, the uniqueness of both THz radiation and time-domain spectroscopy is used to close a technological gap in the right market sector, we here show that successful applications are in reach. We have chosen the automotive industry, where the optimization of coatings applied in the paint shop is of longstanding concern for this most expensive unit of the car production line. Here we report on the development of a THz-based sensor system. We study the light-matter interaction of drying polymer coatings and use advanced novel signal processing algorithms to determine the state of matter of drying paints. This very first sensor system for the inspection of wet coatings that accurately predicts the eventual dry thickness without requiring paint-type calibration. Industrial robustness is guaranteed by a bespoke optical beam design. Our sensor demonstrator outperforms state-of-the-art thickness sensors applied on dry coatings by a factor three and is expected to become a game changer for the paint shop, where today's scrap rates worldwide are as high as one car body per minute.

Detection of SARS-CoV-2 in raw and treated wastewater in Germany - Suitability for COVID-19 surveillance and potential transmission risks.

Wastewater-based monitoring of the spread of the new SARS-CoV-2 virus, also referred to as wastewater-based epidemiology (WBE), has been suggested as a tool to support epidemiology. An extensive sampling campaign, including nine municipal wastewater treatment plants, has been conducted in different cities of the Federal State of North Rhine-Westphalia (Germany) on the same day in April 2020, close to the first peak of the corona crisis. Samples were processed and analysed for a set of SARS-CoV-2-specific genes, as well as pan-genotypic gene sequences also covering other coronavirus types, using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Additionally, a comprehensive set of chemical reference parameters and bioindicators was analysed to characterize the wastewater quality and composition. Results of the RT-qPCR based gene analysis indicate the presence of SARS-CoV-2 genetic traces in different raw wastewaters. Furthermore, selected samples have been sequenced using Sanger technology to confirm the specificity of the RT-qPCR and the origin of the coronavirus. A comparison of the particle-bound and the dissolved portion of SARS-CoV-2 virus genes shows that quantifications must not neglect the solid-phase reservoir. The infectivity of the raw wastewater has also been assessed by viral outgrowth assay with a potential SARS-CoV-2 host cell line in vitro, which were not infected when exposed to the samples. This first evidence suggests that wastewater might be no major route for transmission to humans. Our findings draw attention to the need for further methodological and molecular assay validation for enveloped viruses in wastewater.

Photoregulated Gradient Structure and Programmable Mechanical Performances of Tough Hydrogels with a Hydrogen-Bond Network.

Gradient materials exist widely in natural living organisms, affording fascinating biological and mechanical properties. However, the synthetic gradient hydrogels are usually mechanically weak or only have relatively simple gradient structures. Here, we report on tough nanocomposite hydrogels with designable gradient network structure and mechanical properties by a facile post-photoregulation strategy. Poly(1-vinylimidazole-co-methacrylic acid) hydrogels containing gold nanorods (AuNRs) are in a glassy state and show typical yielding and forced elastic deformation at room temperature. The gel slightly contracts its volume when the temperature is above the glass-transition temperature that results in a collapse of the chain segments and formation of denser intra- and interchain hydrogen bonds. Consequently, the mechanical properties of the gels are enhanced, when the temperature returns to room temperature. The mechanical performances of hydrogels can also be locally tuned by near-infrared light irradiation due to the photothermal effect of AuNRs. Hydrogels with arbitrary two-dimensional gradients can be facilely developed by site-specific photoirradiation. The treated and untreated regions with different stiffness and yielding stress possess construct behaviors in stretching or twisting deformations. A locally reinforced hydrogel with the kirigami structure becomes notch-insensitive and exhibits improved strength and stretchability because the treated regions ahead the cuts have better resistance to crack advancement. These tough hydrogels with programmable gradient structure and mechanics should find applications as structural elements, biological devices, etc.

[COVID-19 Concerns and Worries in Patients with Mental Illness]. / Psychisch krank in Krisenzeiten: Subjektive Belastungen durch COVID-19.

OBJECTIVE:  This study examines the impact of the COVID-19 pandemic and the lock-down on patients with mental illness. METHODS:  Patients in inpatient or outpatient psychiatric treatment received a questionnaire, examining psychological distress and psychiatric care during the COVID-19 pandemic. RESULTS:  More than half of the patients indicated that the state of emergency had a negative impact on their mental illness. Severely ill patients were more affected. CONCLUSION:  Patients with mental illness are a particularly vulnerable group in the current crisis. Psychiatric and psychotherapeutic care needs to be adapted accordingly; the specific burden and distress needs to be examined actively in patients from all diagnostic groups.