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Journal of Fungi 8(3), 269 (2022). doi:10.3390/jof8030269 special issue: "Special Issue "Smut Fungi 2.0" / Special Issue Editors: Prof. Dr. Michael H. Perlin, Guest Editor; Prof. Dr. Barry J. Saville, Guest Editor; Prof. Dr. Jan Schirawski, Guest Editor", Published by MDPI, Basel

The plastic crisis requires drastic measures, especially for the plastics’ end-of-life. Mixed plastic fractions are currently difficult to recycle, but microbial metabolism might open new pathways. With new technologies for degradation of plastics to oligo- and monomers, these carbon sources can be used in biotechnology for the upcycling of plastic waste to valuable products, such as bioplastics and biosurfactants. We briefly summarize well-known monomer degradation pathways and computed their theoretical yields for industrially interesting products. With this information in hand, we calculated replacement scenarios of existing fossil-based synthesis routes for the same products. Thereby, we highlight fossil-based products for which plastic monomers might be attractive alternative carbon sources. Notably, not the highest yield of product on substrate of the biochemical route, but rather the (in-)efficiency of the petrochemical routes (i.e., carbon, energy use) determines the potential of biochemical plastic upcycling. Our results might serve as a guide for future metabolic engineering efforts towards a sustainable plastic economy., Metabolic Engineering, 71

Microbial activity is the driving force of the carbon cycle, including the digestion of biomass in the soil, oceans, and oil deposits. This natural diversity of microbial carbon sources poses challenges for humans. Contamination monitoring can be difficult in oil tanks and similar settings. To assess microbial activity in such industrial settings, off-gas analysis can be employed by considering growth and non-growth-associated metabolic activity. In this work, we describe the monitoring of CO

Liver international 42(3), 640-650 (2022). doi:10.1111/liv.15161, Published by Wiley-Blackwell, Oxford

Pseudomonas putida KT2440 has long been studied for its diverse and robust metabolisms, yet many genes and proteins imparting these growth capacities remain uncharacterized. Using pooled mutant fitness assays, we identified genes and proteins involved in the assimilation of 52 different nitrogen containing compounds. To assay amino acid biosynthesis, 19 amino acid drop- out conditions were also tested. From these 71 conditions, significant fitness phenotypes were elicited in 672 different genes including 100 transcriptional regulators and 112 transport-related proteins. We divide these conditions into 6 classes, and propose assimilatory pathways for the compounds based on this wealth of genetic data. To complement these data, we characterize the substrate range of three promiscuous aminotransferases relevant to metabolic engineering efforts in vitro. Furthermore, we examine the specificity of five transcriptional regulators, explaining some fitness data results and exploring their potential to be developed into useful synthetic biology tools. In addition, we use manifold learning to create an interactive visualization tool for interpreting our BarSeq data, which will improve the accessibility and utility of this work to other researchers.IMPORTANCEUnderstanding the genetic basis of P. putida’s diverse metabolism is imperative for us to reach its full potential as a host for metabolic engineering. Many target molecules of the bioeconomy and their precursors contain nitrogen. This study provides functional evidence linking hundreds of genes to their roles in the metabolism of nitrogenous compounds, and provides an interactive tool for visualizing these data. We further characterize several aminotransferases, lactamases, and regulators--which are of particular interest for metabolic engineering.

The sequence of the promoter used to drive expression of glcDEF operon in the engineered strain MFL185 in the original article was incorrect. As described here, we performed additional experiments that indicate expression of this operon was increased in MFL185, as intended. Ultimately, this error is immaterial with respect to the findings and conclusions reported in the original article.

The Libeň bridge in Prague is a cultural and technical heritage of concrete construction of the beginning from the twentieth century. The bridge was designed by architect Pavel Janák, the founder of cubism in architecture. According to experts, the Libeň bridge is the only example of cubist morphology application on the bridge structure. Evaluation of Libeň bridge structural condition is an up-to-date topic leading to decision whether this bridge should be repaired or replaced by a new bridge. This article deals with the static and dynamic load testing in comparison with following creation and validation of the FEA model for load carrying capacity assessment of the Libeň Bridge and possible way of reconstruction. Paper deals with crucial procedures for FEA model validation of this backfilled arched concrete structure with a focus on the static characteristic of the structure. Article deals with reconstruction of the vault part of the Libeň Bridge using R-UHPFRC in terms of static operation. It focuses on the two basic problems of the bridge, the solution to increase the bearing capacity of the vault arches and to increase the bearing capacity of the foundations under pillars. The concept of repair is based on extensive diagnostic surveys conducted in 2017. The concept of structural enhancement is based on adding new thin layer of R-UHPFRC (reinforced, ultra-high performance concrete) on the upper vault surface. Concrete bridge structures are exposed to extreme effects of aggressive environmental influences, especially chlorides, which leads to damaging of most exposed bridge components a long time before their service life. UHPFRC material is suitable for repairs of these structures and especially due to its impermeability and high resistance.

Non-alcoholic fatty liver disease (NAFLD) is a serious metabolic condition affecting millions of people worldwide. A “Western-style diet” has been shown to induce pediatric NAFLD with the potential disruption of skeletal muscle composition and metabolism. To determine the in vivo effect of a “Western-style diet” on pediatric skeletal muscle fiber type and fuel utilization, 28 juvenile Iberian pigs were fed either a control diet (CON) or a high-fructose, high-fat diet (HFF), with or without probiotic supplementation, for 10 weeks. The HFF diets increased the total triacylglycerol content of muscle tissue but decreased intramyocellular lipid (IMCL) content and the number of type I (slow oxidative) muscle fibers. HFF diets induced autophagy as assessed by LC3I and LC3II, and inflammation, as assessed by IL-1α. No differences in body composition were observed, and there was no change in insulin sensitivity, but HFF diets increased several plasma acylcarnitines and decreased expression of lipid oxidation regulators PGC1α and CPT1, suggesting disruption of skeletal muscle metabolism. Our results show that an HFF diet fed to juvenile Iberian pigs produces a less oxidative skeletal muscle phenotype, similar to a detraining effect, and reduces the capacity to use lipid as fuel, even in the absence of insulin resistance and obesity.

Frontiers in bioinformatics 1, 747428 (2021). doi:10.3389/fbinf.2021.747428, Published by Frontiers Media, Lausanne

Biotechnology has experienced innovations in analytics and data processing. As the volume of data and its complexity grows, new computational procedures for extracting information are developed. However, the rate of change outpaces the adaptation of biotechnology curricula, necessitating new teaching methodologies to equip biotechnologists with data analysis abilities. To simulate experimental data, we created a virtual organism simulator (silvio) by combining diverse cellular and sub-cellular microbial models. silvio was utilized to construct a computer-based instructional workflow with important steps during strain characterization and recombinant protein expression. The instructional workflow is provided as a Jupyter Notebook with comprehensive explanatory text of biotechnological facts and experiment simulations using silvio tools. The students conduct data analysis in Python or Excel. This instructional workflow was separately implemented in two distance courses for Master’s students in biology and biotechnology. The concept of using virtual organism simulations that generate coherent results across different experiments can be used to construct consistent and motivating case studies for biotechnological data literacy.

The elm zigzag sawfly, Aproceros leucopoda Takeuchi (Hymenoptera: Argidae), was reported for the first time in North America during the summer of 2020. Characteristic zigzag defoliation was reported in the province of Québec, Canada, on the community science website, iNaturalist. Field trips conducted to the site resulted in the collection of live specimens (a few larvae and a cocoon from which an adult emerged) and onsite observation of diagnostic defoliation and empty cocoons, confirming the presence of this exotic species in Canada. Subsequent inspection of elm trees by naturalists and scientists in the south of the province led to the conclusion that the species is more widely distributed than first expected and that the invasion is not localised to a small area. Preliminary genetic data pointed to a possible European origin of the Canadian population, but conclusive assignment to source will require examination of more specimens and the collection of reference sequences from different European and Asian populations. This is a good example of the importance of community science in the detection of new invasive species.

The frequency of vascular calcification in patients with osteoarthritis (OA) and calcium pyrophosphate crystal deposition disease (CPPD) has not yet been studied, and the role of calcium crystals (basic and pyrophosphates) in the development of calcification is also unknown.Objective. Determine the presence and degree of calcification of the coronary vessels in patients with calcium pyrophosphate crystal deposition disease and osteoarthritis of the knee joints with no clinical signs of cardiovascular diseases.Materials and methods. One-stage, single-center study, performed by the “case – control” method. The main group – 20 patients with CPPD, the comparison group – 20 patients with OA of the knee joints. Inclusion criteria: age from 18 to 65 years; absence of clinical signs of cardiovascular disease at the time of examination and indications of a history of cardiovascular accidents. Exclusion criteria: unsigned informed consent; pregnancy; breastfeeding; other rheumatic disease; cancer; high and very high cardiovascular risk on the SCORE scale. The survey included an assessment of anthropometric data, blood pressure (BP), lipid profile, serum levels of glucose, creatinine, uric acid, C-reactive protein, vitamin D, osteoprotegerin, parathyroid hormone, and the levels of magnesium, phosphorus, and total calcium were studied. All patients underwent multispiral computed tomography with determination of calcium count and the number of affected arteries. To calculate the coronary score, the A.S. Agatston et al.Results and discussion. Most of the parameters in the compared groups did not differ. When assessing the calcification of the coronary arteries according to the A.S. Agatston et al. 9 (45%) patients with CPPD and 8 (40%) patients with OA had a coronary calcium score >1. Quantitative indicators of calcium score can correspond to coronary artery stenosis ≥20% in 8 (40%) patients with CPPD and in 5 (25%) patients with OA according to J.A. Rumberger et al. The serum level of osteoprotegerin was significantly higher in patients with a calcium score ≥27 according to J.A. Rumberger et al. (p=0.04). Calcification was detected in 9 (56%) of 16 patients with serum vitamin D levels 30 ng/ml.Conclusions. In patients with an initially low cardiovascular risk, the probability of a combination of chondrocalcinosis and cardiovascular calcification is 45%, in OA it is 40%. The risk factors for coronary calcification in patients with CPPD and OA should be studied further.

The physical motivations and performance of the TAIGA (Tunka Advanced Instrument for cosmic ray physics and Gamma Astronomy) project are presented. The TAIGA observatory addresses ground-based gamma-ray astronomy at energies from a few TeV to several PeV, as well as cosmic ray physics from 100 TeV to several EeV and astroparticle physics. The pilot TAIGA complex locates in the Tunka valley, ~50 km West from the southern tip of the lake Baikal. It includes integrating air Cherenkov TAIGA-HiSCORE array with 120 wide-angle optical stations distributed over on area 1 square kilometer about and three the 4-m class Imaging Atmospheric Cherenkov Telescopes of the TAIGA-IACT array. The latter array has a shape of triangle with side lengths of about 300m, 400m and 500m. The expected integral sensitivity of the 1 km2 TAIGA detector will be about 2,5 × 10-13 TeV cm-2 sec-1 for detection of E ≥ 100 TeV gamma-rays in 300 hours of source observations. The combination of the wide angle Cherenkov array and IACTs could offer a cost effective-way to build a really large (up to 10 km2) array for very high energy gamma-ray astronomy. The reconstruction of a given EAS energy, incoming direction and the core position, based on the TAIGA-HiSCORE data, allows one to increase the distance between the relatively expensive IACTs up to 600-800 m. These, together with the surface and underground electron/Muon detectors will be used for selection of gamma-ray induced EAS. Present status of the project, together with the current array description and the first experimental results and plans for the future will be reported.

Cydistinae are a rare monogeneric beetle lineage from Asia with a convoluted history of classification, historically placed in various groups within the series Elateriformia. However, their position has never been rigorously tested. To resolve this long-standing puzzle, we are the first to present sequences of two nuclear and two mitochondrial markers for four species of Cydistinae to determine their phylogenetic position. We included these sequences in two rounds of analyses: one including a broad Elateriformia dataset to test placement at the superfamily/family level, and a second, including a richer, targeted sampling of presumed close relatives. Our results strongly support Cydistinae as sister to Phengodidae in a clade with Rhagophthalmidae. Based on our molecular phylogenetic results and examination of morphological characters, we hereby transfer the formerly unplaced Cydistinae into Phengodidae and provide diagnoses for the newly circumscribed Phengodidae, Cydistinae and Cydistus. Since both Phengodidae and Rhagophthalmidae have bioluminescent larvae and strongly neotenic females, similar features can be hypothesized for Cydistinae. Additionally, Cydistus minor is transferred to the new genus Microcydistus.

Poly-γ-glutamic acid (γ-PGA), which is produced by several Bacillus species, is a chiral biopolymer composed of D- and L-glutamate monomers and has various industrial applications. However, synthesized γ-PGA exhibits great structural diversity, and the structure must be controlled to broaden its industrial use. The biochemical pathways for γ-PGA production suggest that the polymer properties molecular weight (MW) and stereochemical composition are influenced by (1) the affinity of γ-PGA synthetase for the two alternative glutamate enantiomers and (2) glutamate racemase activity; hence, the availability of the monomers. In this study, we report tailor-made γ-PGA synthesis with B. subtilis by combining PGA synthetase and glutamate racemase genes from several Bacillus strains. The production of structurally diverse γ-PGA was thereby achieved. Depending on the PGA synthetase and glutamate racemase origins, the synthesized γ-PGA contained 3–60% D-glutamate. The exchange of PGA synthetase changed the MW from 40 to 8500 kDa. The results demonstrate the production of low-, medium-, and high-MW γ-PGA with the same microbial chassis.

Pollution from ethylene glycol, and plastics containing this monomer, represent a significant environmental problem. The investigation of its microbial metabolism therefore provides insights into the environmental fate of this pollutant and also enables its utilization as a carbon source for microbial biotechnology. Here, we reveal the genomic and metabolic basis of ethylene glycol metabolism in Pseudomonas putida KT2440. Although this strain cannot grow on ethylene glycol as sole carbon source, it can be used to generate growth-enhancing reducing equivalents upon co-feeding with acetate. Mutants that utilize ethylene glycol as sole carbon source were isolated through adaptive laboratory evolution. Genomic analysis of these mutants revealed a central role of the transcriptional regulator GclR, which represses the glyoxylate carboligase pathway as part of a larger metabolic context of purine and allantoin metabolism. Secondary mutations in a transcriptional regulator encoded by PP_2046 and a porin encoded by PP_2662 further improved growth on ethylene glycol in evolved strains, likely by balancing fluxes through the initial oxidations of ethylene glycol to glyoxylate. With this knowledge, we reverse engineered an ethylene glycol utilizing strain and thus revealed the metabolic and regulatory basis that are essential for efficient ethylene glycol metabolism in P. putida KT2440.

The insect order Hymenoptera originated during the Permian nearly 300 Mya. Ancestrally herbivorous hymenopteran lineages today make up the paraphyletic suborder ‘Symphyta’, which encompasses c. 8200 species with very diverse host-plant associations. We use phylogeny-based statistical analyses to explore the drivers of diversity dynamics within the ‘Symphyta’, with a particular focus on the hypothesis that diversification of herbivorous insects has been driven by the explosive radiation of angiosperms during and after the Cretaceous. Our ancestral-state estimates reveal that the first symphytans fed on gymnosperms, and that shifts onto angiosperms and pteridophytes – and back – have occurred at different time intervals in different groups. Trait-dependent analyses indicate that average net diversification rates do not differ between symphytan lineages feeding on angiosperms, gymnosperms or pteridophytes, but trait-independent models show that the highest diversification rates are found in a few angiosperm-feeding lineages that may have been favoured by the radiations of their host taxa during the Cenozoic. Intriguingly, lineages-through-time plots show signs of an early Cretaceous mass extinction, with a recovery starting first in angiosperm-associated clades. Hence, the oft-invoked assumption of herbivore diversification driven by the rise of flowering plants may overlook a Cretaceous global turnover in insect herbivore communities during the rapid displacement of gymnosperm- and pteridophyte-dominated floras by angiosperms.

Currently, 31P NMR is the only analytical method that quantitatively determines the average chain length of long inorganic polyphosphate (>80 P-subunits). In this study, an enzyme assay is presente...

Besides enzymatic conversions, many eukaryotic metabolic pathways also involve transport proteins that shuttle molecules between subcellular compartments, or into the extracellular space. Fungal itaconate production involves two such transport steps, involving an itaconate transport protein (Itp), and a mitochondrial tricarboxylate transporter (Mtt). The filamentous ascomycete Aspergillus terreus and the unicellular basidiomycete Ustilago maydis both produce itaconate, but do so via very different molecular pathways, and under very different cultivation conditions. In contrast, the transport proteins of these two strains are assumed to have a similar function. This study aims to investigate the roles of both the extracellular and mitochondrial transporters from these two organisms by expressing them in the corresponding U. maydis knockouts and monitoring the extracellular product concentrations. Both transporters from A. terreus complemented their corresponding U. maydis knockouts in mediating itaconate production. Surprisingly, complementation with At_MfsA from A. terreus led to a partial switch from itaconate to (S)-2-hydroxyparaconate secretion. Apparently, the export protein from A. terreus has a higher affinity for (S)-2-hydroxyparaconate than for itaconate, even though this species is classically regarded as an itaconate producer. Complementation with At_MttA increased itaconate production by 2.3-fold compared to complementation with Um_Mtt1, indicating that the mitochondrial carrier from A. terreus supports a higher metabolic flux of itaconic acid precursors than its U. maydis counterpart. The biochemical implications of these differences are discussed in the context of the biotechnological application in U. maydis and A. terreus for the production of itaconate and (S)-2-hydroxyparaconate.

Eosinophilic esophagitis (EoE) is detected frequently in dysphagia and noncardiac chest pain. Management of patients with EoE may be complicated because EoE is associated frequently with esophageal motility disorders. We present the rare case of esophageal achalasia (EA) associated with eosinophilic infiltration and a literature review. A patient with dysphagia and eosinophilic infiltration referred to our clinic underwent standardized diagnostic work-up including symptom questionnaire, esophagogastroduodenoscopy (EGD) with esophageal biopsies, barium swallow, high-resolution esophageal manometry, and combined intraluminal 24-hour pH-impedance testing (pH/MII). The patient had an Eckardt score of 8. EGD and mucosal biopsies showed typical EoE with 15 eosinophil leucocytes per high-power field. Barium swallow revealed typical sign of achalasia. HREM indicated EA type 2 according to the Chicago classification. PH/MII was normal. Oral and systemic corticoid therapy were without effect. After successful treatment by pneumatic dilation of the cardia, symptoms relieved and eosinophilic infiltration returned to normal. The results suggest that the patient had primary EA associated with eosinophilic infiltration and that the combined occurrence of these rare diseases is not just a coincidence. Die eosinophile Ösophagitis (EoE) ist häufig Ursache von Schluckstörungen und nichtkardialen Brustschmerzen (NCCP). Das Management von Patienten mit EoE kann kompliziert werden, da die EoE häufig mit Ösophagusmotilitätsstörungen assoziiert sind. Wir stellen den seltenen Fall einer Achalasie (EA) assoziiert mit einer eosinophilen Infiltration und eine Literaturübersicht vor. Ein Patient mit Dysphagie und eosinophiler Infiltration wurde in unserer Klinik vorgestellt und durch einen standardisierten Symptomenfragebogen, Ösophagogastroduodenoskopie (EGD) mit Biopsien, Röntgen-Breischluck, hochauflösende Ösophagusmanometrie (HREM) und kombinierte 24-Std.-pH-Metrie-Impedanzmessung untersucht (pH/MII). Der Patient hatte einen Eckardt-Score von 8. EGD und die Mukosabiopsien zeigten die typischen Befunde einer EoE mit 15 eosinophilen Leukozyten pro HPF. Der Röntgen-Breischluck erwies die typischen Befunde einer EA. Die HREM konnte eine EA Typ 2 nach der Chicago-Klassifikation nachweisen. Die PH/MII war unauffällig. Die lokale und systemische Kortisontherapie waren ohne Effekt. Nach der pneumatischen Kardiadilatation waren die Beschwerden rückläufig und die eosinophilen Infiltrate normalisiert. Die Ergebnisse lassen vermuten, dass bei dem Patienten primär eine EA mit assoziierter eosinophiler Infiltration vorlag und dass das Zusammentreffen dieser beiden seltenen Erkrankungen nicht zufällig ist.

3-Hydroxy decanoic acid (3-HDA), derivable from glucose or xylose waste-streams, was successfully upgraded electrochemically into a drop-in oxygenate with promising fuel characteristics. Properties such as a cetane number of 63 together with a suitable flash point and cold filter plugging point qualify the mixture for high blending ratios with fossil diesel or Fischer–Tropsch diesel. The individual C9 oxygenates are otherwise only accessible by hydroformylation. Under optimised conditions, anodic decarboxylation of 3-HDA allowed near full conversion and over 95% total yield after 30 minutes. Further, direct electrochemical 3-HDA conversion of a fermentation broth was demonstrated. This allows for a substantial reduction of unit operations and energy requirements, highlighting the potential of integrating electrochemistry and biotechnology into future bio-refineries.

The question of what determines whether cells are big or small has been the focus of many studies because it is thought that such determinants underpin the coupling of cell growth with cell division. In contrast, what determines the overall pattern of how cell size is distributed within a population of wild type or mutant cells has received little attention. Knowing how cell size varies around a characteristic pattern could shed light on the processes that generate such a pattern and provide a criterion to identify its genetic basis. Here, we show that cell size values of wild typeSaccharomyces cerevisiaecells fit a gamma distribution, in haploid and diploid cells, and under different growth conditions. To identify genes that influence this pattern, we analyzed the cell size distributions of all single-gene deletion strains inSaccharomyces cerevisiae.We found that yeast strains which deviate the most from the gamma distribution are enriched for those lacking gene products functioning in gene expression, especially those in transcription or transcription-linked processes. We also show that cell size is increased in mutants carrying altered activity substitutions in Rpo21p/Rpb1, the largest subunit of RNA polymerase II (Pol II). Lastly, the size distribution of cells carrying extreme altered activity Pol II substitutions deviated from the expected gamma distribution. Our results are consistent with the idea that genetic defects in widely acting transcription factors or Pol II itself compromise both cell size homeostasis and how the size of individual cells is distributed in a population.

An efficient route to obtain aromatic products based on biomass feedstock is a challenge for future biorefineries. Isobutyraldehyde is a promising feedstock available via biotechnological routes based on both carbohydrates and the direct bioconversion of CO2. Herein, we report an efficient process for the aromatisation of isobutyraldehyde over zeolite catalysts in a continuous fixed bed reactor to provide value-added aromatic compounds with a yield of 93%. Benzene, toluene and xylenes are the major compounds formed with 79% yield and a productivity of 65 mmol gcat−1 h−1. Zeolite Y, Beta and Mordenite and ZSM-5 of different modules were studied. Comprehensive catalyst characterisation using XRD, NH3-TPD, N2-physisorption, and ICP-OES enabled establishing structure–performance relationships with a major role of zeolite structure, density of strong acid sites and mesoporosity, respectively. HZSM-5 with Si/Al ratios of 15 to 49 proved effective at the aromatisation possessing a comparable density of strong acid sites of 0.37–0.52 mmol g−1. Superior stability was observed for HZSM-5 zeolites with higher mesopore volumes. The reaction is suggested to mainly proceed via catalytic cracking to C1–C4 alkanes/alkenes, which undergo further oligomerisation, cyclisation, and aromatisation to form the observed alkyl-aromatics. Emphasizing the potential of the concept, long term continuous operation was carried out by alternating 10 hours time on steam operation and catalyst regeneration via in situ calcination.

The TAIGA astroparticle observatory is progressing with the deployment of new detector stations. The Tunka-Grande — scintillation counter array of the observatory expands with the new TAIGA-Muon stations. Several simulation studies were conducted for optimisation of the new station positioning and performance. Extensive air showers induced by gamma quanta or a proton in the range from 100 TeV to 1 PeV at a zenith angle of 0° were used for these studies. Based on the developed simulation, the capabilities of identification of high energy extensive air showers were studied. The soil thickness, the detector and station positions, the lowest measurable energy range of the cosmic rays, and different methods of air shower identification were investigated.

High-throughput screening of cell-secreted proteins is essential for various biotechnological applications. In this article, we show a microfluidic approach to perform the analysis of cell-secreted proteins in nanoliter droplet arrays by two complementary methods, fluorescence microscopy and mass spectrometry. We analyzed the secretion of the enzyme phytase, a phosphatase used as an animal feed additive, from a low number of yeast cells. Yeast cells were encapsulated in nanoliter volumes by droplet microfluidics and deposited on spatially defined spots on the surface of a glass slide mounted on the motorized stage of an inverted fluorescence microscope. During the following incubation for several hours to produce phytase, the droplets can be monitored by optical microscopy. After addition of a fluorogenic substrate at a defined time, the relative concentration of phytase was determined in every droplet. Moreover, we demonstrate the use of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) to monitor the multistep conversion of the native substrate phytic acid by phytase secreted in 7 nL droplets containing 50-100 cells. Our method can be adapted to various other protocols. As the droplets are easily accessible, compounds such as assay reagents or matrix molecules can be added to all or to selected droplets only, or part of the droplet volume could be removed. Hence, this platform is a versatile tool for questions related to cell secretome analysis.

Volume 86, no. 21, 2020, e01665-20, . Readers should note that when the authors created the deletion mutant of PP\_2675, they deleted the entire region between the gene’s start and stop codons. However, this also deleted the DNA that codes for the first 14

Mental health treatment in schools has the potential to improve youth treatment access. However, school-specific barriers can make implementing evidence-based interventions difficult. Task-shifting (i.e., training lay staff to implement interventions) and computer-assisted interventions may mitigate these barriers. This paper reports on a qualitative examination of facilitators and barriers of a school-based implementation of a computer-assisted intervention for anxious youth (Camp Cope-A-Lot; CCAL). Participants (N = 45) included school staff in first through fourth grades. Providers attended a training in CCAL and received weekly, hour-long group consultation calls for three months. In the second year, the sustainability of CCAL use was assessed. Qualitative interviews were conducted after the first year (initial implementation) and second year (sustainability). Interviews were analyzed using the Consolidated Framework for Implementation Research domains to classify themes. Although participants reported that CCAL included useful skills, they expressed concerns about recommended session length (45 minutes) and frequency (weekly). Time burden of consultation calls was also a barrier. School staff facilitated implementation by enabling flexible scheduling for youth to be able to participate in the CCAL program. However, the sustainability of the program was limited due to competing school/time demands. Results suggest that even with computer assisted programs, there is a need to tailor interventions and implementation efforts to account for the time restrictions experienced by school-based service providers. Optimal fit between the intervention and specific school is important to maintain the potential benefits of computer-assisted treatments delivered by lay service providers in schools.

The enzymatic degradation of polyethylene terephthalate (PET) results in a hydrolysate consisting almost exclusively of its two monomers, ethylene glycol and terephthalate. To biologically valorize the PET hydrolysate, microbial upcycling into high-value products is proposed. Fatty acid derivatives hydroxyalkanoyloxy alkanoates (HAAs) represent such valuable target molecules. HAAs exhibit surface-active properties and can be exploited in the catalytical conversion to drop-in biofuels as well as in the polymerization to bio-based poly(amide urethane). This chapter presents the genetic engineering methods of pseudomonads for the metabolization of PET monomers and the biosynthesis of HAAs with detailed protocols concerning product purification.

Metabolic engineering 66, 167-178 (2021). doi:10.1016/j.ymben.2021.03.011, Published by Academic Press, Orlando, Fla.

9 p.-2 fig., This article introduces the EU Horizon 2020 research project MIX-UP, "Mixed plastics biodegradation and upcycling using microbial communities". The project focuses on changing the traditional linear value chain of plastics to a sustainable, biodegradable based one. Plastic mixtures contain five of the top six fossil-based recalcitrant plastics [polyethylene (PE), polyurethane (PUR), polypropylene (PP), polyethylene terephthalate (PET), polystyrene (PS)], along with upcoming bioplastics polyhydroxyalkanoate (PHA) and polylactate (PLA) will be used as feedstock for microbial transformations. Consecutive controlled enzymatic and microbial degradation of mechanically pre-treated plastics wastes combined with subsequent microbial conversion to polymers and value-added chemicals by mixed cultures. Known plastic-degrading enzymes will be optimised by integrated protein engineering to achieve high specific binding capacities, stability, and catalytic efficacy towards a broad spectrum of plastic polymers under high salt and temperature conditions. Another focus lies in the search and isolation of novel enzymes active on recalcitrant polymers. MIX-UP will formulate enzyme cocktails tailored to specific waste streams and strives to enhance enzyme production significantly. In vivo and in vitro application of these cocktails enable stable, self-sustaining microbiomes to convert the released plastic monomers selectively into value-added products, key building blocks, and biomass. Any remaining material recalcitrant to the enzymatic activities will be recirculated into the process by physicochemical treatment. The Chinese–European MIX-UP consortium is multidisciplinary and industry-participating to address the market need for novel sustainable routes to valorise plastic waste streams. The project's new workflow realises a circular (bio)plastic economy and adds value to present poorly recycled plastic wastes where mechanical and chemical plastic recycling show limits., This project has received funding from the European Union's Horizon 2020 research and innovation programme under Grant Agreement No. 870294. In addition, this project is funded by the National Natural Science Foundation of China (Grant Numbers 31961133017, 31961133018, 31961133019).

mSystems 6(2), (2021). doi:10.1128/mSystems.00625-20, Published by American Society for Microbiology, Washington, DC

Frontiers in genetics 11, 637738 (2021). doi:10.3389/fgene.2020.637738, Published by Frontiers Media, Lausanne

In times of ever-increasing demand for chemicals and the subsequent increase in CO2 in the atmosphere, we have to intensify our efforts to establish a circular (bio) economy [...]

Dascillidae are a species-poor beetle group with a scarce fossil record. Here, we describe Baltodascillus serraticornis gen. et sp. nov. based on a well-preserved specimen from Eocene Baltic amber. It differs from all known Dascillidae by its reduced mandibles. After studying the specimen using light microscopy and X-ray microtomography, we tentatively place this genus in the poorly defined subfamily Karumiinae based on the large eyes, serrate antennae, and lack of prosternal process. This is the first representative of the Dascillidae formally described from Baltic amber and the first described fossil member of the subfamily Karumiinae. We briefly discuss the problematic higher classification of Dascillidae, along with the morphology and biogeography of the group.

Biotechnology & bioengineering (2021). doi:10.1002/bit.27822, Published by Wiley, New York, NY [u.a.]