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At least 127 records · Page 7

A Rapid Method for Detecting Normal or Modified Plant and Algal Carbonic Anhydrase Activity Using Saccharomyces cerevisiae

In recent years, researchers have attempted to improve photosynthesis by introducing components from cyanobacterial and algal CO2-concentrating mechanisms (CCMs) into terrestrial C3 plants. For these attempts to succeed, we need to understand the CCM components in more detail, especially carbonic anhydrase (CA) and bicarbonate (HCO3−) transporters. Heterologous complementation systems capable of detecting carbonic anhydrase activity (i.e., catalysis of the pH-dependent interconversion between CO2 and HCO3−) or active HCO3− transport can be of great value in the process of introducing CCM components into terrestrial C3 plants. In this study, we generated a Saccharomyces cerevisiae CA knock-out (ΔNCE103 or ΔCA) that has a high-CO2-dependent phenotype (5% (v/v) CO2 in air). CAs produce HCO3− for anaplerotic pathways in S. cerevisiae; therefore, the unavailability of HCO3− for neutral lipid biosynthesis is a limitation for the growth of ΔCA in ambient levels of CO2 (0.04% (v/v) CO2 in air). ΔCA can be complemented for growth at ambient levels of CO2 by expressing a CA from human red blood cells. ΔCA was also successfully complemented for growth at ambient levels of CO2 through the expression of CAs from Chlamydomonas reinhardtii and Arabidopsis thaliana. The ΔCA strain is also useful for investigating the activity of modified CAs, allowing for quick screening of modified CAs before putting them into the plants. CA activity in the complemented ΔCA strains can be probed using the Wilbur–Anderson assay and by isotope exchange membrane-inlet mass spectrometry (MIMS). Other potential uses for this new ΔCA-based screening system are also discussed.

59 BASIC BIOLOGICAL SCIENCES↗

A separable Hartley-like transform in two or more dimensions

This letter introduces a discrete, separable, real-to-real transform, called the cas-cas transform. Theorems for the two-dimensional case are presented, and the cas-cas transform is compared to the Hartley transform as an alternative way to convolve two-dimensional arrays and compute two-dimensional power spectra.

Perkins, Michael G.↗

Radio observations of D I and fractionation

We report observations to detect the 327 MHz line of atomic deuterium in the primarily molecular clouds towards Cas A and Sgr A, making appropriate choices of spectral and spatial resolution. Our best results for Cas A, based on 1253 hr of observing with the 85 foot (26 m) Hat Creek telescope and 45 hr with the very large array (VLA), yield an upper limit for the fraction of deuterium in atomic form, D I, of 0.14. We present an approximate analytical analysis of deuterium fractionation in translucent clouds, which shows that gas-phase ion-molecule reactions, assisted by dust and HD line self-shielding, are efficient in converting deuterium to HD. We conclude that little atomic deuterium is present in the molecular clumps in the Cas A clouds and that much higher sensitivity observations would be required to detect 327 MHz line in molecular clouds. We have also attempted to detect the 72 GHz line of DCO(+) toward Cas A. The observed upper limit to the DCO(+)/HCO(+) ratio of approximately 0.03 is not much larger than our theoretical estimate, and the DCO(+) line should be detectable with existing instrumentation, unless the temperature of these clouds is much larger than usually assumed.

Heiles, Carl↗

A study of coronal X-ray emission from short-period Algol binaries

A study of X-ray emission from five short-period Algol-type binaries based on observations with Advanced Satellite for Cosmology and Astrophysics (ASCA) and ROSAT is presented. We have observed RZ Cas with both satellites, and beta Per, U Cep, delta Lib, and TW Dra with ROSAT. Significant intensity variations are seen in the X-ray emission from RZ Cas, U Cep, TW Dra, and delta Lib. These variations seem unrelated to the eclipsing behavior of these systems and are probably due to either rotational modulation of compact active regions on the surfaces of the chromospherically active secondary components or to flaring activity in the systems. The spectra of all but one of the systems require the presence of at least two discrete plasma components with different temperatures (0.6 - 0.7 keV, and approximately 2 keV) and the abundances of the medium-Z elements 20% - 50% of the solar photospheric values. The high resolving power and signal-to-noise ratio of the ASCA spectra allow us to individually constrain the coronal abundances of O, Ne, Mg, Si, S, and Fe in RZ Cas. We demonstrate that, if we use the elemental abundances and temperatures obtained from the analysis of their ASCA spectra as (fixed) inputs, to fit the ROSAT PSPC spectra well requires the presence of a third component (kT approximately 0.2 - 0.3 keV) in RZ Cas and beta Per. A continuous emission measure model of the power-law type (EM(T) variesas (T/T(sub max)(sup alpha)) generally gives a poor fit to the ASCA and ROSAT data on most sources. Circumstellar or circumbinary absorbing matter seems to be present in some of these systems, as indicated by the variable total column density needed to fit their X-ray spectra.

Singh, K. P.↗

The NAS Computational Aerosciences Archive

In order to further the state-of-the-art in computational aerosciences (CAS) technology, researchers must be able to gather and understand existing work in the field. One aspect of this information gathering is studying published work available in scientific journals and conference proceedings. However, current scientific publications are very limited in the type and amount of information that they can disseminate. Information is typically restricted to text, a few images, and a bibliography list. Additional information that might be useful to the researcher, such as additional visual results, referenced papers, and datasets, are not available. New forms of electronic publication, such as the World Wide Web (WWW), limit publication size only by available disk space and data transmission bandwidth, both of which are improving rapidly. The Numerical Aerodynamic Simulation (NAS) Systems Division at NASA Ames Research Center is in the process of creating an archive of CAS information on the WWW. This archive will be based on the large amount of information produced by researchers associated with the NAS facility. The archive will contain technical summaries and reports of research performed on NAS supercomputers, visual results (images, animations, visualization system scripts), datasets, and any other supporting meta-information. This information will be available via the WWW through the NAS homepage, located at http://www.nas.nasa.gov/, fully indexed for searching. The main components of the archive are technical summaries and reports, visual results, and datasets. Technical summaries are gathered every year by researchers who have been allotted resources on NAS supercomputers. These summaries, together with supporting visual results and references, are browsable by interested researchers. Referenced papers made available by researchers can be accessed through hypertext links. Technical reports are in-depth accounts of tools and applications research projects performed by NAS staff members and collaborators. Visual results, which may be available in the form of images, animations, and/or visualization scripts, are generated by researchers with respect to a certain research project, depicting dataset features that were determined important by the investigating researcher. For example, script files for visualization systems (e.g. FAST, PLOT3D, AVS) are provided to create visualizations on the user's local workstation to elucidate the key points of the numerical study. Users can then interact with the data starting where the investigator left off. Datasets are intended to give researchers an opportunity to understand previous work, 'mine' solutions for new information (for example, have you ever read a paper thinking "I wonder what the helicity density looks like?"), compare new techniques with older results, collaborate with remote colleagues, and perform validation. Supporting meta-information associated with the research projects is also important to provide additional context for research projects. This may include information such as the software used in the simulation (e.g. grid generators, flow solvers, visualization). In addition to serving the CAS research community, the information archive will also be helpful to students, visualization system developers and researchers, and management. Students (of any age) can use the data to study fluid dynamics, compare results from different flow solvers, learn about meshing techniques, etc., leading to better informed individuals. For these users it is particularly important that visualization be integrated into dataset archives. Visualization researchers can use dataset archives to test algorithms and techniques, leading to better visualization systems, Management can use the data to figure what is really going on behind the viewgraphs. All users will benefit from fast, easy, and convenient access to CFD datasets. The CAS information archive hopes to serve as a useful resource to those interested in computational sciences. At present, only information that may be distributed internationally is made available via the archive. Studies are underway to determine security requirements and solutions to make additional information available. By providing access to the archive via the WWW, the process of information gathering can be more productive and fruitful due to ease of access and ability to manage many different types of information. As the archive grows, additional resources from outside NAS will be added, providing a dynamic source of research results.

Miceli, Kristina D.↗

Air Quality Study Using Satellites - Current Capability and Future Plans

Satellite instruments have had great success in monitoring the stratospheric ozone and in understanding the processes that control its daily to decadal scale variations. This field is now reaching its zenith with a number of satellite instruments from the US, Europe and Canada capping several decades of active research in this field. The primary public policy imperative of this research was to make reliable prediction of increases in biologically active surface UV radiation due to human activity. By contrast retrieval from satellite data of atmospheric constituents and photo-chemically active radiation that affect air quality is a new and growing field that is presenting us with unique challenges in measurement and data interpretation. A key distinction compared to stratospheric sensors is the greatly enhanced role of clouds, aerosols, and surfaces (CAS) in determining the quality and quantity of useful data that is available for air quality research. In our presentation we will use data from several sensors that are currently flying on the A-train satellite constellation, including OMI, MODIS, CLOUDSAT, and CALIPSO, to highlight that CAS can have both positive and negative effects on the information content of satellite measurements. This is in sharp contrast to other fields of remote sensing where CAS are usually considered an interference except in those cases when they are the primary subject of study. Our analysis has revealed that in the reflected wavelengths one often sees much further down into the atmosphere, through most cirrus, than one does in the emitted wavelengths. The lower level clouds provide a nice background against which one can track long-range transport of trace gases and aerosols. In addition, differences in trace gas columns estimated over cloudy and adjacent clear pixels can be used to measure boundary layer trace gases. However, in order to take full advantage of these features it will be necessary to greatly advance our understanding of how CAS affect the radiation at wavelengths that are used to derive the atmospheric constituents that affect air quality as well as the radiation that controls the photolysis of chemically active trace gases. We will discuss how we are using these new insights to design future satellite missions to study air quality.

Bhartia, Pawan K.↗

Generic, Extensible, Configurable Push-Pull Framework for Large-Scale Science Missions

The push-pull framework was developed in hopes that an infrastructure would be created that could literally connect to any given remote site, and (given a set of restrictions) download files from that remote site based on those restrictions. The Cataloging and Archiving Service (CAS) has recently been re-architected and re-factored in its canonical services, including file management, workflow management, and resource management. Additionally, a generic CAS Crawling Framework was built based on motivation from Apache s open-source search engine project called Nutch. Nutch is an Apache effort to provide search engine services (akin to Google), including crawling, parsing, content analysis, and indexing. It has produced several stable software releases, and is currently used in production services at companies such as Yahoo, and at NASA's Planetary Data System. The CAS Crawling Framework supports many of the Nutch Crawler's generic services, including metadata extraction, crawling, and ingestion. However, one service that was not ported over from Nutch is a generic protocol layer service that allows the Nutch crawler to obtain content using protocol plug-ins that download content using implementations of remote protocols, such as HTTP, FTP, WinNT file system, HTTPS, etc. Such a generic protocol layer would greatly aid in the CAS Crawling Framework, as the layer would allow the framework to generically obtain content (i.e., data products) from remote sites using protocols such as FTP and others. Augmented with this capability, the Orbiting Carbon Observatory (OCO) and NPP (NPOESS Preparatory Project) Sounder PEATE (Product Evaluation and Analysis Tools Elements) would be provided with an infrastructure to support generic FTP-based pull access to remote data products, obviating the need for any specialized software outside of the context of their existing process control systems. This extensible configurable framework was created in Java, and allows the use of different underlying communication middleware (at present, both XMLRPC, and RMI). In addition, the framework is entirely suitable in a multi-mission environment and is supporting both NPP Sounder PEATE and the OCO Mission. Both systems involve tasks such as high-throughput job processing, terabyte-scale data management, and science computing facilities. NPP Sounder PEATE is already using the push-pull framework to accept hundreds of gigabytes of IASI (infrared atmospheric sounding interferometer) data, and is in preparation to accept CRIMS (Cross-track Infrared Microwave Sounding Suite) data. OCO will leverage the framework to download MODIS, CloudSat, and other ancillary data products for use in the high-performance Level 2 Science Algorithm. The National Cancer Institute is also evaluating the framework for use in sharing and disseminating cancer research data through its Early Detection Research Network (EDRN).

Foster, Brian M.↗

Assessing Helicopter Pilots’ Detect and Avoid and Collision Avoidance Performance With ACAS Xr

The latest variant of the Federal Aviation Administration’s Airborne Collision Avoidance System (ACAS X) is being designed for both crewed and uncrewed rotorcraft. Referred to as ACAS Xr, the system joins a suite of other ACAS X variants poised to replace the Traffic Alert and Collision Avoidance System (TCAS II). ACAS Xr is tuned to support current-day helicopter platforms as well as electric Vertical Takeoff and Landing (eVTOL) vehicles that are still under development. Given this flexibility, ACAS Xr may be used by helicopter crews currently in operation or by remotely-operated eVTOL aircraft in the emerging Advanced Air Mobility (AAM) market. To cover the range of potential uses, two distinct configurations are being proposed for ACAS Xr: Collision Avoidance System (CAS) and Detect and Avoid (DAA). Under the CAS configuration, ACAS Xr provides minimal caution-level alerting but issues directive warning-level alerting and guidance. The DAA configuration, by contrast, provides caution-level alerting and guidance, in addition to the warning-level alerting and guidance. The current study was performed as part of the National Aeronautics and Space Administration’s AAM project. Six helicopter pilots were recruited to fly a variety of scripted traffic scenarios in a full-motion, crewed eVTOL simulator. Participants flew 60 encounters over two days, reacting to pre-recorded intruder aircraft that were scripted to fly into the participant’s aircraft from different approach angles, relative altitudes, and during different phases of flight. The pilots flew half of the encounters with the CAS configuration and half with the DAA configuration. Within each block of 30 encounters, pilots experienced 10 conflicts while in cruise, 10 in hover, and 10 while on approach to a heliport. Results showed that pilot response times were consistently under 5 seconds for RAs and under 10 seconds for DAA alerts, when present, during all three phases of flight. Unsurprisingly, the DAA configuration was associated with lower rates of en-route and high-severity losses of DAA well clear compared to the CAS configuration in all phases of flight except for the terminal area. Rates of losses of DAA well clear were found to be substantially higher in the Hover scenario, compared to Cruise. Pilots failed to fully comply with RAs at a rate of 0.10-0.18 in all conditions except for the DAA configuration in the Hover scenario, which was associated with a higher non-compliance rate of 0.4 due to Descend RAs issued at low altitudes. The implications of these results with regards to the ongoing development of ACAS Xr is discussed.

air taxis↗

Assessing Helicopter Pilots’ Detect and Avoid and Collision Avoidance Performance with ACAS Xr

The latest variant of the Federal Aviation Administration’s Airborne Collision Avoidance System (ACAS X) is being designed for both crewed and uncrewed rotorcraft. Referred to as ACAS Xr, the system joins a suite of other ACAS X variants poised to replace the Traffic Alert and Collision Avoidance System (TCAS II). ACAS Xr is tuned to support current-day helicopter platforms as well as electric Vertical Takeoff and Landing (eVTOL) vehicles that are still under development. Given this flexibility, ACAS Xr may be used by helicopter crews currently in operation or by remotely-operated eVTOL aircraft in the emerging Advanced Air Mobility (AAM) market. To cover the range of potential uses, two distinct configurations are being proposed for ACAS Xr: Collision Avoidance System (CAS) and Detect and Avoid (DAA). Under the CAS configuration, ACAS Xr provides minimal caution-level alerting but issues directive warning-level alerting and guidance. The DAA configuration, by contrast, provides caution-level alerting and guidance, in addition to the warning-level alerting and guidance. The current study was performed as part of the National Aeronautics and Space Administration’s AAM project. Six helicopter pilots were recruited to fly a variety of scripted traffic scenarios in a full-motion, crewed eVTOL simulator. Participants flew 60 encounters over two days, reacting to pre-recorded intruder aircraft that were scripted to fly into the participant’s aircraft from different approach angles, relative altitudes, and during different phases of flight. The pilots flew half of the encounters with the CAS configuration and half with the DAA configuration. Within each block of 30 encounters, pilots experienced 10 conflicts while in cruise, 10 in hover, and 10 while on approach to a heliport. Results showed that pilot response times were consistently under 5 seconds for RAs and under 10 seconds for DAA alerts, when present, during all three phases of flight. Unsurprisingly, the DAA configuration was associated with lower rates of en-route and high-severity losses of DAA well clear compared to the CAS configuration in all phases of flight except for the terminal area. Rates of losses of DAA well clear were found to be substantially higher in the Hover scenario, compared to Cruise. Pilots failed to fully comply with RAs at a rate of 0.10-0.18 in all conditions except for the DAA configuration in the Hover scenario, which was associated with a higher non-compliance rate of 0.4 due to Descend RAs issued at low altitudes. The implications of these results with regards to the ongoing development of ACAS Xr is discussed.

air taxis↗

Novel adaptive immune systems in pristine Antarctic soils

Antarctic environments are dominated by microorganisms, which are vulnerable to viral infection. Although several studies have investigated the phylogenetic repertoire of bacteria and viruses in these poly-extreme environments with freezing temperatures, high ultra violet irradiation levels, low moisture availability and hyper-oligotrophy, the evolutionary mechanisms governing microbial immunity remain poorly understood. Using genome-resolved metagenomics, we test the hypothesis that Antarctic poly-extreme high-latitude microbiomes harbour diverse adaptive immune systems. Our analysis reveals the prevalence of prophages in bacterial genomes (Bacteroidota and Verrucomicrobiota), suggesting the significance of lysogenic infection strategies in Antarctic soils. Furthermore, we demonstrate the presence of diverse CRISPR-Cas arrays, including Class 1 arrays (Types I-B, I-C, and I-E), alongside systems exhibiting novel gene architecture among their effector cas genes. Notably, a Class 2 system featuring type V variants lacks CRISPR arrays, encodes Cas1 and Cas2 adaptation module genes. Phylogenetic analysis of Cas12 effector proteins hints at divergent evolutionary histories compared to classified type V effectors and indicates that TnpB is likely the ancestor of Cas12 nucleases. Our findings suggest substantial novelty in Antarctic cas sequences, likely driven by strong selective pressures. These results underscore the role of viral infection as a key evolutionary driver shaping polar microbiomes.

59 BASIC BIOLOGICAL SCIENCES↗

Chapter 7 - CRISPR-Based Tools for Microbial Cell Factories

The development of microbial chassis for the production of a variety of biochemicals and biofuels is a growing area of research. How to efficiently manipulate genetic information to achieve optimal production of these compounds is a key area of focus in the field. In recent years, clustered regularly interspaced palindromic repeats (CRISPR) and its associated proteins (Cas) have become a popular strategy for gene editing and regulation in many organisms due to its versatility and efficacy. Here, we describe methods developed utilizing CRISPR-Cas systems for engineering microbial cell factories (e.g., bacteria and yeast) at the single gene to genome scale for mutagenesis and transcriptional regulation of target genes. Finally, we provide a perspective on the challenges and opportunities for the applications of advanced CRISPR-Cas-based tools for engineering microbial cell factories.

BIOMASS FUELS↗

Convergent Aeronautics Solutions Project

NASA is committed to transforming our aviation system to best meet demands and opportunities of the future. With a vision of safe, efficient, flexible, and environmentally sustainable air transportation, the NASA Aeronautics Research Mission Directorate is conducting research and development to address future needs of the aviation community, the Nation, and the world. While our NASA Aeronautics vision and strategy reaches into the next 25 years and beyond, we recognize that our vision and strategy must be responsive to new discoveries and emerging markets. For this reason, we are empowering our research community to redefine the future of aviation by dreaming up convergent/transformative ideas and studying if those ideas are possible. By modeling the new NASA Aeronautics' Convergent Aeronautics Solutions (CAS) Project after the venture capital community, we created opportunities for teams of intrapreneurs to mature their ideas into concepts through rapid feasibility studies. The CAS Project expects teams to consider the complexities and potential benefits of multi-disciplinary solutions and to leverage technology advances from outside the field of aeronautics. We also expect teams to explore their concepts in a rapid, iterative manner that allows them to learn and adjust their research approach. Within a year or two, teams are responsible for reporting on the feasibility of their concept. The findings inform NASA Aeronautics strategic planning and further investment. This presentation will give an overview of CAS.

Transformative↗

Co(II) Substitution Enhances the Esterase Activity of a de Novo Designed Zn(II) Carbonic Anhydrase

Abstract Carbonic Anhydrases (CAs) have been a target forde novoprotein designers due to the simplicity of the active site and rapid rate of the reaction. The first reported mimic contained a Zn(II) bound to three histidine imidazole nitrogens and an exogenous water molecule, hence closely mimicking the native enzymes’ first coordination sphere. Co(II) has served as an alternative metal to interrogate CAs due to its d 7 electronic configuration for more detailed solution characterization. We present here the Co(II) substituted [Co(II)(H 2 O/OH − )] N (TRIL2WL23H) 3 n+ that behaves similarly to native Co(II) substituted human‐CAs. Like the Zn(II) analogue, the cobalt‐derivative at slightly basic pH is incapable of hydrolyzing p‐nitrophenylacetate (pNPA); however, as the pH is increased a significant activity develops, which at pH values above 10 eventually yields a catalytic efficiency that exceeds that of the [Zn(II)(OH − )] N (TRIL2WL23H) 3 + peptide complex. X‐ray absorption analysis is consistent with an octahedral species at pH 7.5 that converts to a 5‐coordinate species by pH 11. UV‐vis spectroscopy can monitor this transition, giving a pK a for the conversion of 10.3. We assign this conversion to the formation of a 5‐coordinate Co(II)(N imid ) 3 (OH)(H 2 O) species. The pH dependent kinetic analysis indicates the maximal rate (k cat ), and thus the catalytic efficiency (k cat /K m ), follow the same pH profile as the spectroscopic conversion to the pentacoordinate species. This correlation suggests that the chemically irreversible ester hydrolysis corresponds to the rate determining process.

Chemistry↗

Spectroscopy-based isotopic (δ 13 C) analysis for high spatial resolution of carbon exchange in the rhizosphere

The rhizosphere is a highly dynamic zone bridging plant roots with needed nutrient resources in soil. While the rhizosphere may be small, it has a disproportionally large impact on plant success and biomass production. A suite of rhizosphere-hosted microbial and geochemical interactions facilitate nutrient acquisition by plant roots, and, in turn, the roots stimulate these processes by supplying organic carbon into the rhizosphere. The small physical dimensions of the rhizosphere, however, can constrain efforts to elucidate key carbon exchange processes and their spatial extent and localization. We present a method for spatially resolved δ 13 C analysis of rhizosphere samples by coupling laser ablation (LA) sampling with isotopic analysis using capillary absorption spectroscopy (CAS) which differs from conventional mass spectrometer (MS) approaches. The CAS system has high sensitivity (requires fewer nanomoles of CO 2 per analysis) than comparable MS systems, which enables reduced sample size requirements to thereby improve spatial resolution (from 25 μm to as low as a projected 5 μm spatial resolution). We demonstrate the utility of CAS using rhizosphere samples from switchgrass plants exposed to 13 CO 2 . As a result, this technique will provide a capability for tracking the extent and spatial distribution of root exudate into the rhizosphere at highly detailed spatial scales.

59 BASIC BIOLOGICAL SCIENCES↗

Hexagonal Lattices of HIV Capsid Proteins Explored by Simulations Based on a Thermodynamically Consistent Model

HIV capsid proteins (CAs) may self-assemble into a variety of shapes under in vivo and in vitro conditions. Here, we employed simulations based on a residue-level coarse-grained (CG) model with full conformational flexibility to investigate hexagonal lattices, which are the underlying structural pattern for CA aggregations. Facilitated by enhanced sampling simulations to rigorously calculate CA dimerization and polymerization affinities, we calibrated our model to reproduce the experimentally measured affinities. Here using the calibrated model, we performed unbiased simulations on several large systems consisting of 1512 CA subunits, allowing reversible binding and unbinding of the CAs in a thermodynamically consistent manner. In one simulation, a preassembled hexagonal CA sheet developed spontaneous curvatures reminiscent of those observed in experiments, and the edges of the sheet exhibited local curvatures larger than those of the interior. In other simulations starting with randomly distributed CAs at different concentrations, existing CA assemblies grew by binding free capsomeres to the edges and by merging with other assemblies. At high CA concentrations, rapid establishment of predominant aggregates was followed by much slower adjustments toward more regular hexagonal lattices, with increasing numbers of intact CA hexamers and pentamers being formed. Our approach of adapting a general CG model to specific systems by using experimental binding data represents a practical and effective strategy for simulating and elucidating intricate protein aggregations.

59 BASIC BIOLOGICAL SCIENCES↗

3D Printed Parahydrophobic Surfaces as Multireaction Platforms

We report Parahydrophobic surfaces (PHSs) composed of arrays of cubic μ-pillars with a double scale of roughness and variable wettability were systematically obtained in one step and a widely accessible stereolithographic Formlabs 3D printer. The wettability control was achieved by combining the geometrical parameters (H = height and P = pitch) and the surface modification with fluoroalkyl silane compounds. Homogeneous distribution of F and Si atoms onto the pillars was observed by XPS and SEM-EDAX. A nano-roughness on the heads of the pillars was achieved without any post-treatment. The smallest P values lead to surfaces with static contact angles (CAs) >150° regardless of the H utilized. Interestingly, the relationship 0.6 ≤ H/P ≤ 2.6 obtained here was in good agreement with the H/P values reported for nano- and submicron pillars. Furthermore, experimental CAs, advancing and receding CAs, were consistent with the theoretical prediction from the Cassie–Baxter model. Structures covered with perfluorodecyltriethoxysilane with high H and short P lead to PHSs. Conversely, structures covered with perfluorodecyltrimethoxysilane exhibited a superhydrophobic behavior. Finally, several aqueous reactions, such as precipitation, coordination complex, and nanoparticle synthesis, were carried out by placing the reactive agents as microdroplets on the parahydrophobic pillars, demonstrating the potential application as chemical multi-reaction array platforms for a large variety of relevant fields in microdroplet manipulation, microfluidics systems, and health monitoring, among others.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Clades of huge phages from across Earth’s ecosystems

Bacteriophages typically have small genomes and depend on their bacterial hosts for replication. Here we sequenced DNA from diverse ecosystems and found hundreds of phage genomes with lengths of more than 200 kilobases (kb), including a genome of 735 kb, which is—to our knowledge—the largest phage genome to be described to date. Thirty-five genomes were manually curated to completion (circular and no gaps). Expanded genetic repertoires include diverse and previously undescribed CRISPR–Cas systems, transfer RNAs (tRNAs), tRNA synthetases, tRNA-modification enzymes, translation-initiation and elongation factors, and ribosomal proteins. The CRISPR–Cas systems of phages have the capacity to silence host transcription factors and translational genes, potentially as part of a larger interaction network that intercepts translation to redirect biosynthesis to phage-encoded functions. In addition, some phages may repurpose bacterial CRISPR–Cas systems to eliminate competing phages. We phylogenetically define the major clades of huge phages from human and other animal microbiomes, as well as from oceans, lakes, sediments, soils and the built environment. We conclude that the large gene inventories of huge phages reflect a conserved biological strategy, and that the phages are distributed across a broad bacterial host range and across Earth’s ecosystems.

59 BASIC BIOLOGICAL SCIENCES↗