Insulation for cryogenic tanks has reduced thickness and weight

Dual seal insulation, consisting of an inner layer of sealed-cell Mylar honeycomb core and an outer helium purge channel of fiber glass reinforced phenolic honeycomb core, is used as a thin, lightweight insulation for external surfaces of cryogenic-propellant tanks

Ultrahigh-throughput-directed enzyme evolution by absorbance-activated droplet sorting (AADS)

Ultrahigh-throughput screening, in which members of enzyme libraries compartmentalized in water-in-oil emulsion droplets are assayed, has emerged as a powerful format for directed evolution and functional metagenomics but is currently limited to fluorescence readouts. Here we describe a highly efficient microfluidic absorbance-activated droplet sorter (AADS) that extends the range of assays amenable to this approach. Using this module, microdroplets can be sorted based on absorbance readout at rates of up to 300 droplets per second (i.e., >1 million droplets per hour). To validate this device, we implemented a miniaturized coupled assay for NAD(+)-dependent amino acid dehydrogenases. The detection limit (10 μM in a coupled assay producing a formazan dye) enables accurate kinetic readouts sensitive enough to detect a minimum of 1,300 turnovers per enzyme molecule, expressed in a single cell, and released by lysis within a droplet. Sorting experiments showed that the AADS successfully enriched active variants up to 2,800-fold from an overwhelming majority of inactive ones at ∼100 Hz. To demonstrate the utility of this module for protein engineering, two rounds of directed evolution were performed to improve the activity of phenylalanine dehydrogenase toward its native substrate. Fourteen hits showed increased activity (improved >4.5-fold in lysate; kcat increased >2.7-fold), soluble protein expression levels (up 60%), and thermostability (Tm, 12 °C higher). The AADS module makes the most widely used optical detection format amenable to screens of unprecedented size, paving the way for the implementation of chromogenic assays in droplet microfluidics workflows.This research was funded by the Engineering and Physical Sciences Research Council (studentship to RH and an Impact Acceleration Account Partnership Development Award), the Biological and Biotechnological Research Council (BBSRC) and Johnson Matthey. SE and MF were supported by postdoctoral Marie-Curie fellowships

Ultrahigh-throughput-directed enzyme evolution by absorbance-activated droplet sorting (AADS)

This is the final version. Available from National Academy of Sciences via the DOI in this recordUltrahigh-throughput screening, in which members of enzyme libraries compartmentalized in water-in-oil emulsion droplets are assayed, has emerged as a powerful format for directed evolution and functional metagenomics but is currently limited to fluorescence readouts. Here we describe a highly efficient microfluidic absorbance-activated droplet sorter (AADS) that extends the range of assays amenable to this approach. Using this module, microdroplets can be sorted based on absorbance readout at rates of up to 300 droplets per second (i.e., >1 million droplets per hour). To validate this device, we implemented a miniaturized coupled assay for NAD+-dependent amino acid dehydrogenases. The detection limit (10 μM in a coupled assay producing a formazan dye) enables accurate kinetic readouts sensitive enough to detect a minimum of 1,300 turnovers per enzyme molecule, expressed in a single cell, and released by lysis within a droplet. Sorting experiments showed that the AADS successfully enriched active variants up to 2,800-fold from an overwhelming majority of inactive ones at ∼100 Hz. To demonstrate the utility of this module for protein engineering, two rounds of directed evolution were performed to improve the activity of phenylalanine dehydro-genase toward its native substrate. Fourteen hits showed increased activity (improved >4.5-fold in lysate; kcat increased >2.7-fold), soluble protein expression levels (up 60%), and thermostability (Tm, 12°C higher). The AADS module makes the most widely used optical detection format amenable to screens of unprecedented size, paving the way for the implementation of chromogenic assays in droplet microfluidics workflows.Biotechnology and Biological Sciences Research CouncilEuropean Research CouncilEngineering and Physical Sciences Research CouncilEuropean Commissio

A novel application of queueing theory on the Caulerpenyne secreted by invasive Caulerpa taxifolia (Vahl) C.Agardh (Ulvophyceae, Caulerpales): a preliminary study

Aquarium originated marine green alga Caulerpa taxifolia was introduced into the Mediterranean Sea accidentally in 1984. This invasion has been negatively affecting the sub-littoral ecosystem of the Mediterranean. One of the important reasons for its success in the Mediterranean is its secondary toxic metabolite called caulerpenyne (CPN). Furthermore, CPN has anti-proliferative and apoptotic activities, therefore, CPN can be considered as a potential native source in cancer therapy. For that reason, modeling this metabolite might be of importance. Increase and decrease of the CPN level before reaching the critical level can be expressed by means of a queueing system in which the number of ‘customers' increases and decreases. In fact, production of CPN shows fluctuations for many environmental reasons, which allow us to apply queueing theory. In the present study, the expected time to reach the maximum caulerpenyne level was analyzed and evaluated using queueing theory

In-situ hot forging directed energy deposition-arc of CuAl8 alloy

Funding Information: Authors acknowledge the Portuguese Fundação para a Ciência e a Tecnologia ( FCT - MCTES ) for its financial support via the project UID/EMS/00667/2019 (UNIDEMI). VD acknowledges Portuguese Fundação para a Ciência e a Tecnologia ( FCT - MCTES ) for funding the PhD grant SFRH/BD/139454/2018 . TAR acknowledges Portuguese Fundação para a Ciência e a Tecnologia ( FCT - MCTES ) for funding the PhD grant SFRH/BD/144202/2019 . Funding of CENIMAT/i3N by national funds through the Portuguese Fundação para a Ciência e a Tecnologia, I.P., within the scope of Multiannual Financing of R&D Units , reference UIDB/50025/2020–2023 is also acknowledge. This activity has received funding from the European Institute of Innovation and Technology (EIT) Raw Materials through the project Smart WAAM: Microstructural Engineering and Integrated Non-Destructive Testing. This body of the European Union receives support from the European Union's Horizon 2020 research and innovation programme. Parts of this research were carried out at PETRA III at DESY, a member of the Helmholtz Association. The research leading to this result has been supported by the project CALIPSOplus under the Grant Agreement 730872 from the EU Framework Programme for Research and Innovation HORIZON 2020 . This project has received funding from the EU-H2020 research and innovation programme under grant agreement No 654360 having benefitted from the access provided by PETRA III at DESY in Hamburg, Germany within the framework of the NFFA-Europe Transnational Access Activity. The authors acknowledge support by OCAS NV and GUARENTEED via Joachim Antonissen. Funding Information: Authors acknowledge the Portuguese Fundação para a Ciência e a Tecnologia (FCT - MCTES) for its financial support via the project UID/EMS/00667/2019 (UNIDEMI). VD acknowledges Portuguese Fundação para a Ciência e a Tecnologia (FCT - MCTES) for funding the PhD grant SFRH/BD/139454/2018. TAR acknowledges Portuguese Fundação para a Ciência e a Tecnologia (FCT - MCTES) for funding the PhD grant SFRH/BD/144202/2019. Funding of CENIMAT/i3N by national funds through the Portuguese Fundação para a Ciência e a Tecnologia, I.P. within the scope of Multiannual Financing of R&D Units, reference UIDB/50025/2020–2023 is also acknowledge. This activity has received funding from the European Institute of Innovation and Technology (EIT) Raw Materials through the project Smart WAAM: Microstructural Engineering and Integrated Non-Destructive Testing. This body of the European Union receives support from the European Union's Horizon 2020 research and innovation programme. Parts of this research were carried out at PETRA III at DESY, a member of the Helmholtz Association. The research leading to this result has been supported by the project CALIPSOplus under the Grant Agreement 730872 from the EU Framework Programme for Research and Innovation HORIZON 2020. This project has received funding from the EU-H2020 research and innovation programme under grant agreement No 654360 having benefitted from the access provided by PETRA III at DESY in Hamburg, Germany within the framework of the NFFA-Europe Transnational Access Activity. The authors acknowledge support by OCAS NV and GUARENTEED via Joachim Antonissen. Remark: The supplementary material is temporarily available in the Drive folder here: https://drive.google.com/drive/folders/1SFFlhJlmL5p3IkQis8cB6UVWva3wozGi?usp=sharing. Publisher Copyright: © 2022 Elsevier B.V.CuAl8 alloy finds applications in industrial components, where a good anti-corrosion and anti-wearing properties are required. The alloy has a medium strength and a good toughness with an elongation to fracture at room temperature of about 40%. Additionally, it has a good electrical conductivity, though lower than that of pure Al or pure Cu. Despite these characteristics, additive manufacturing of the CuAl8 alloy was not yet reported. In this work, the direct energy deposition-arc (DED-arc) with and without in-situ hot forging was used to determine the microstructure evolution and mechanical properties. No internal defects were seen on the parts produced. Hot forging combined with DED-arc was seen to reduce and homogenize the grain size, improve mechanical strength and isotropy of mechanical properties. Moreover, the use of this novel DED-arc variant was seen to reduce the magnitude of residual stresses throughout the fabricated part. We highlight that this alloy can be processed by DED-arc, and the hot forging operation concomitant with the material deposition has beneficial effects on the microstructure refinement and homogenization.publishersversionpublishe

Arnol'd Tongues and Quantum Accelerator Modes

The stable periodic orbits of an area-preserving map on the 2-torus, which is formally a variant of the Standard Map, have been shown to explain the quantum accelerator modes that were discovered in experiments with laser-cooled atoms. We show that their parametric dependence exhibits Arnol'd-like tongues and perform a perturbative analysis of such structures. We thus explain the arithmetical organisation of the accelerator modes and discuss experimental implications thereof.Comment: 20 pages, 6 encapsulated postscript figure

Deterministic ratchets: route to diffusive transport

The rectification efficiency of an underdamped ratchet operated in the adiabatic regime increases according to a scaling current-amplitude curve as the damping constant approaches a critical threshold; below threshold the rectified signal becomes extremely irregular and eventually its time average drops to zero. Periodic (locked) and diffusive (fully chaotic) trajectories coexist on fine tuning the amplitude of the input signal. The transition from regular to chaotic transport in noiseless ratchets is studied numerically.Comment: 9 pages, 5 figures, to be published in Phys. Rev.