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At least 379 records · Page 21

A Quantum Dot Nanobiosensor for Rapid Detection of Botulinum Neurotoxin Serotype E

Botulinum neurotoxins (BoNTs) are potent toxins produced by Clostridium bacteria that are responsible for the illness botulism and are listed as bioterrorism agents. BoNT serotype E (BoNT/E) is one of four BoNT serotypes that cause human botulism and is the second most frequent cause of foodborne botulism. Rapid detection and discrimination of BoNT serotypes implicated in human disease are critical for ensuring timely treatment of patients and identifying sources of toxins, but there have been few reported detection methods for BoNT/E and even fewer methods usable for BoNT serotyping. We report a nanobiosensor based on Forster resonance energy transfer (FRET) between semiconductor nanocrystals (quantum dots, QDs) and dark quencher-labeled peptide probes to detect biologically active BoNT/E in aqueous media. The peptide probes contain a specific cleavage site for active BoNT/E. QD photoluminescence, which changes intensity due to FRET when the peptide probe is cleaved, was used to indicate toxin presence and quantity. The detection of a BoNT/E light chain (LcE) and holotoxin was observed within 3 h. The limits of detection were 0.02 and 2 ng/mL for LcE and holotoxin, respectively. The nanobiosensor shows good specificity toward the target in tests with nontarget BoNT serotypes. The high sensitivity, simple operation, short detection time, and ability to be used in parallel with probes developed for other BoNT serotypes indicate that the nanobiosensor will be useful for rapid BoNT/E detection and serotype discrimination in food analysis.

botulinum neurotoxin E↗

RESURRECTION OF (3200) PHAETHON IN 2016

We present a study of the active asteroid (3200) Phaethon in the 2016 apparition using the Solar and Terrestrial Relations Observatory ( STEREO ) spacecraft and compare the results with data from the previous two perihelia in 2009 and 2012. Once again, Phaethon brightened by ∼2 mag soon after its perihelion passage, contradicting expectations from the phase function of a macroscopic monolithic body. Subsequently, a short antisolar tail of ∼0.°1 in length was formed within ∼1 day and quickly disappeared. No trail was seen. Our syndyne-synchrone analysis indicates that the tail was comprised of submicron to micron particles and can be approximated by a synchrone coinciding with the outburst. We estimate that the outburst has released a mass of ∼10{sup 4}–10{sup 5} kg, comparable to the two mass ejections in 2009 and 2012, and that the average mass-loss rate is ∼0.1–1 kg s{sup −1}. The forward-scattering effect hinted at low level activity of Phaethon prior to the outburst, which increased the effective cross section by merely ≲1 km{sup 2}. Without the forward-scattering enhancement, detecting such activity at side-scattering phase angles is very difficult. The forward-scattering effect also reinforces the idea that the ejected dust grains rather than gas emissions were responsible for the activity of Phaethon. Despite Phaethon's reactivation, it is highly unlikely that the Geminid meteoroid stream can be sustained by similar perihelion mass-loss events.

79 ASTRONOMY AND ASTROPHYSICS↗

Dual reporter transgene driven by 2.3Col1a1 promoter is active in differentiated osteoblasts

AIM: As quantitative and spatial analyses of promoter reporter constructs are not easily performed in intact bone, we designed a reporter gene specific to bone, which could be analyzed both visually and quantitatively by using chloramphenicol acetyltransferase (CAT) and a cyan version of green fluorescent protein (GFPcyan), driven by a 2.3-kb fragment of the rat collagen promoter (Col2.3). METHODS: The construct Col2.3CATiresGFPcyan was used for generating transgenic mice. Quantitative measurement of promoter activity was performed by CAT analysis of different tissues derived from transgenic animals; localization was performed by visualized GFP in frozen bone sections. To assess transgene expression during in vitro differentiation, marrow stromal cell and neonatal calvarial osteoblast cultures were analyzed for CAT and GFP activity. RESULTS: In mice, CAT activity was detected in the calvaria, long bone, teeth, and tendon, whereas histology showed that GFP expression was limited to osteoblasts and osteocytes. In cell culture, increased activity of CAT correlated with increased differentiation, and GFP activity was restricted to mineralized nodules. CONCLUSION: The concept of a dual reporter allows a simultaneous visual and quantitative analysis of transgene activity in bone.

Non-NASA Center↗

Imatinib mesylate elicits extracellular signal-related kinase (ERK) activation and enhances the survival of γ-irradiated epithelial cells

Highlights: • Imatinib mesylate induced ERK activation within 30 min. • Imatinib mesylate reduced radiation-induced apoptosis and enhanced cell survival. • Imatinib mesylate protects epithelial-derived cells regardless of malignancy status. Imatinib mesylate, commercially known as Gleevec/Glivec, is the first targeted anticancer drug that inhibits activity of the tyrosine kinases, c-ABL, c-KIT, and PDGFR. A number of studies have shown that treatment with imatinib mesylate elicits extracellular signal-related kinase (ERK) activation, which, in turn, has been shown to confer radioresistance. Here, we investigated whether treatment with imatinib mesylate protects skin-derived epithelial cells, including normal keratinocytes, immortalized HaCaT and A431 cancer cell lines, from the effects of γ-radiation. ERK activation was detected 30 min after imatinib mesylate treatment in all three cell lines. In cells exposed to γ-irradiation in the presence of imatinib mesylate, this activation of ERK was associated with a reduction in radiation-induced apoptosis and enhanced cell survival. Similar effects of imatinib mesylate treatment were observed following γ-irradiation of a three-dimensional human skin culture system that reproduces a fully differentiated epithelium. Collectively, our findings provide the evidence of a protective effect of imatinib mesylate against the effects of γ-irradiation on epithelial-derived cells, regardless of their malignancy status.

60 APPLIED LIFE SCIENCES↗

Maturation of Rhodobacter capsulatus Multicopper Oxidase CutO Depends on the CopA Copper Efflux Pathway and Requires the cutF Product

Copper (Cu) is an essential cofactor required for redox enzymes in all domains of life. Because of its toxicity, tightly controlled mechanisms ensure Cu delivery for cuproenzyme biogenesis and simultaneously protect cells against toxic Cu. Many Gram-negative bacteria contain extracytoplasmic multicopper oxidases (MCOs), which are involved in periplasmic Cu detoxification. MCOs are unique cuproenzymes because their catalytic center contains multiple Cu atoms, which are required for the oxidation of Cu 1+ to the less toxic Cu 2+ . Hence, Cu is both substrate and essential cofactor of MCOs. Here, we investigated the maturation of Rhodobacter capsulatus MCO CutO and its role in periplasmic Cu detoxification. A survey of CutO activity of R. capsulatus mutants with known defects in Cu homeostasis and in the maturation of the cuproprotein cbb 3 -type cytochrome oxidase ( cbb 3 -Cox) was performed. This revealed that CutO activity is largely independent of the Cu-delivery pathway for cbb 3 -Cox biogenesis, except for the cupric reductase CcoG, which is required for full CutO activity. The most pronounced decrease of CutO activity was observed with strains lacking the cytoplasmic Cu chaperone CopZ, or the Cu-exporting ATPase CopA, indicating that CutO maturation is linked to the CopZ-CopA mediated Cu-detoxification pathway. Our data demonstrate that CutO is important for cellular Cu resistance under both aerobic and anaerobic growth conditions. CutO is encoded in the cutFOG operon, but only CutF, and not CutG, is essential for CutO activity. No CutO activity is detectable when cutF or its putative Cu-binding motif are mutated, suggesting that the cutF product serves as a Cu-binding component required for active CutO production. Bioinformatic analyses of CutF-like proteins support their widespread roles as putative Cu-binding proteins for several Cu-relay pathways. Our overall findings show that the cytoplasmic CopZ-CopA dependent Cu detoxification pathway contributes to providing Cu to CutO maturation, a process that strictly relies on cutF .

59 BASIC BIOLOGICAL SCIENCES↗

UHF and S-band radar observations of electrically active storm cells

The UHF-band and S-band research radars of NASA Goddard's atmospheric sciences research facility at Wallop's Island, Virginia are used to study the correlation between the location of radar-detected electrical activity in a storm cell and its precipitation reflectivity structure. The technique used to make the measurements is discussed and some typical results are presented. The existence of a bimodal distribution of lightning activity as a function of altitude is reported and supported by the data. The vertical structure of the electrical activity may be evidence of K and/or J processes. The technique used is very valuable, and provides high-resolution radar data of both reflectivities and flash density.

Gerlach, J. C.↗

ShadeLab/PAPER_Howe_2023_switchgrass_MetaT

The raw data (metagenomes and metatranscriptomes) for this study are available in the Joint Genomes Institute Genome Portal (https://genome.jgi.doe.gov/portal/ Project ID 503249) with projects designated by year and product type. The MAG genomes analyzed in this paper are available on NCBI, as bioproject PRJNA800073. Plants and microorganisms form beneficial associations. Understanding plant-microbe interactions will inform microbiome management to enhance crop productivity and resilience to stress. Here, we apply a genome-centric approach to identify ecologically important leaf microbiome members on field-grown switchgrass and miscanthus and to quantify their activities for switchgrass over two growing seasons. We integrate metagenome and metatranscriptome sequencing from 192 leaf samples collected over representative time points in crop phenology. We curated 40 medium- and high-quality metagenome-assembled-genomes (MAGs) and focused analysis on seasonal transcript recruitment to them. Classes represented by these focal MAGs (Actinomycetia, Alpha- and Gamma- Proteobacteria, and Bacteroidota) were active and had increases in transcripts for short-chain dehydrogenase, molybdopterin oxidoreductase, and polyketide cyclase in the late season. The majority of MAGs had activated stress-associated pathways, including trehalose metabolism, indole acetic acid degradation, betaine biosynthesis, and reactive oxygen species degradation, suggesting direct engagement with the host environment. We also detected seasonally activated biosynthetic pathways for terpenes (carotenoids and isoprenoids) and for various non-ribosomal peptide pathways that were poorly annotated. Overall, this study overcame laboratory and bioinformatic challenges associated with field-based leaf metatranscriptome analysis to inform both general and likely specialized activities of these phyllosphere populations. These activities collectively support that leaf-associated bacterial populations are seasonally dynamic, responsive to host cues, and interactively engage in feedback with the plant. This analysis represented quality filtering of metagenomes and metatranscriptomes (data-preparation folder), metagenome assemblies (metagenome-assembly folder) and metagenome-assembled genome binning, curation, refinement, annotation (mag-evaluation folder). Abundances of sequencing libraries were calculated based on reads mapped (mapping folder). Additionallly, analysis of our annotated results are also included (analysis folder).

Howe, Adina↗

Simultaneous S- and X-band uplink-downlink performance at DSS 13

The Deep Space Station 13 26-meter antenna with the second generation S/X feedcone was tested to determine the dual S- and X-band (2.1 to 2.3 GHz and 7.1 to 8.5 GHz) transmit and receive performance. Measurements were conducted using the 20 kW transmitters at S- and X-band while simultaneously receiving S- and X-band. This system proved to be very quiet compared with the other DSN antennas. Under normal tracking configurations, no noise burst or intermodulation product (IMP) activity was detectable to the -175 dBm level. To prove the instrumentation's ability to detect such phenomena, an IMP generator was introduced onto the system with positive, verifiable results. The IMP occurred at the -162 dBm level, accompanied by moderate noise burst activity, and was readily repeatable. The measurement also showed the possible need for additional fourth channel filtering in the system to reduce the effect of the transmitter power on the low noise amplifiers.

Freiley, A. J.↗

Assessing the Expansion of Ground-Motion Sensing Capability in Smart Cities via Internet Fiber-Optic Infrastructure

Monitoring ground motion in smart cities can improve the public safety by providing critical insights on natural and anthropogenic hazards, for example, earthquakes, landslides, explosions, infrastructure failures, and so forth. Although seismic activity is typically measured using dedicated point sensors (e.g., geophones and accelerometers), techniques such as distributed acoustic sensing have demonstrated the utility of using fiber-optic cable to detect seismic activity over comparable distances. In this article, we present the results of a study that quantifies the expansion in an area monitored for low-amplitude ground-motion events by augmenting existing point sensors with the internet fiber-optic cable infrastructure. Here we begin by describing our methodology, which utilizes geospatial data on point sensors and internet optical fiber deployed in metropolitan statistical areas (MSAs) in the United States. We extend these data to identify the area that can be monitored by (1) considering the observed seismic noise data in target locations, (2) applying the model from Wilson et al. (2021) to understand the potential coverage area gains using optical fiber sensing, and (3) optimizing the selection of fiber segments to maximize coverage and minimize deployment costs. We implement our methodology in ArcGIS to assess the additional area that can be monitored for low-amplitude ground-motion events (i.e., magnitude >0.5) by utilizing internet fiber-optic cables in the 100 most populous MSAs in the United States. We find that the addition of internet fiber-based sensors in MSAs would increase the area monitored on average by over an order of magnitude from 1% to 12%, if the subset of fiber cable segments that maximize coverage and minimize deployment costs is chosen even if only 20% of all fibers are used.

58 GEOSCIENCES↗

Long-distance communication in Arabidopsis involving a self-activating G protein

In plant cells, heterotrimeric G protein signaling mediates development, biotic/abiotic stress responsiveness, hormone signaling, and extracellular sugar sensing. The amount of sugar in plant cells fluctuates from nanomolar to high millimolar concentrations over time depending on changes in the light environment. Arabidopsis thaliana Regulator of G Signaling protein 1 (AtRGS1) modulates G protein activation and detects the concentration and the exposure time of sugars. This is called dose–duration reciprocity in sugar sensing and occurs through AtRGS1 internalization which is directly proportional to G protein activation. One source of sugars is from CO 2 fixation by photosynthesis. Through a simple set of experiments, we show that sugars made in cotyledons that are undergoing photomorphogenesis activate G signaling in cells distal to the nascent photosynthesis center. This occurs with sufficient speed to enable distal cells to monitor changes in photosynthetic activity in the leaves.

photosynthesis↗

The 3-(bromoacetamido)-propylamine hydrochloride: A novel sulfhydryl reagent and its future potential in the configurational study of S1-myosin

Configurational study of S1-Myosin is an important step towards understanding force generation in muscle contraction. Previously reported NMR studies were corroborated. A new compound was synthesized, 3-(Bromoacetamido)-propylamine hydrochloride. Its potential as a sulfhydryl reagent provides an indirect but elegant approach towards future structural elucidation of S1-Myosin. The preliminary investigation has shown that this compound, BAAP, reacted with S1 in the absence of MgADP. The modified enzyme had a 2-fold increase in CaATPase activity and no detectable K-EDTA ATPase activity. Reaction of BAAP with S1 in the presence of MgADP resulted in a modified enzyme which retained a Ca-ATPase activity that was about 60 percent of the unmodified S1 and had essentially zero K-EDTA ATPase activity. Sulfhydryl titration indicated that about 1.5 and 3.5 SH groups per S1 molecule were blocked by BAAP in the absence and presence of MgADP, respectively. When coupled to a carboxyl group of EDTA, the resulting reagent could become a useful SH reagent in which chelated paramagnetic or luminescent lanthanide ions can be exploited to probe S1 conformation.

Sharma, Prasanta↗

Market Assessment of Forward-Looking Turbulence Sensing Systems

In recognition of the importance of turbulence mitigation as a tool to improve aviation safety, NASA's Aviation Safety Program developed a Turbulence Detection and Mitigation Sub-element. The objective of this effort is to develop highly reliable turbulence detection technologies for commercial transport aircraft to sense dangerous turbulence with sufficient time warning so that defensive measures can be implemented and prevent passenger and crew injuries. Current research involves three forward sensing products to improve the cockpit awareness of possible turbulence hazards. X-band radar enhancements will improve the capabilities of current weather radar to detect turbulence associated with convective activity. LIDAR (Light Detection and Ranging) is a laser-based technology that is capable of detecting turbulence in clear air. Finally, a possible Radar-LIDAR hybrid sensor is envisioned to detect the full range of convective and clear air turbulence. To support decisions relating to the development of these three forward-looking turbulence sensor technologies, the objective of this study was defined as examination of cost and implementation metrics. Tasks performed included the identification of cost factors and certification issues, the development and application of an implementation model, and the development of cost budget/targets for installing the turbulence sensor and associated software devices into the commercial transport fleet.

Kauffmann, Paul↗

Do Molecular Fingerprints Identify Diverse Active Drugs in Large-Scale Virtual Screening? (No)

Computational approaches for small-molecule drug discovery now regularly scale to the consideration of libraries containing billions of candidate small molecules. One promising approach to increased the speed of evaluating billion-molecule libraries is to develop succinct representations of each molecule that enable the rapid identification of molecules with similar properties. Molecular fingerprints are thought to provide a mechanism for producing such representations. Here, we explore the utility of commonly used fingerprints in the context of predicting similar molecular activity. We show that fingerprint similarity provides little discriminative power between active and inactive molecules for a target protein based on a known active—while they may sometimes provide some enrichment for active molecules in a drug screen, a screened data set will still be dominated by inactive molecules. We also demonstrate that high-similarity actives appear to share a scaffold with the query active, meaning that they could more easily be identified by structural enumeration. Furthermore, even when limited to only active molecules, fingerprint similarity values do not correlate with compound potency. In sum, these results highlight the need for a new wave of molecular representations that will improve the capacity to detect biologically active molecules based on their similarity to other such molecules.

59 BASIC BIOLOGICAL SCIENCES↗

Sennoside A protects mitochondrial structure and function to improve high-fat diet-induced hepatic steatosis by targeting VDAC1

Highlights: • SA inhibited VDAC1 expression in livers of HFD-induced hepatic steatosis mice. • SA suppressed the ATP surplus by inhibiting mitochondrial complex I in livers of HFD-induced hepatic steatosis mice. • SA protected mitochondrial function and structure in livers of HFD-induced hepatic steatosis mice. • SA improved HFD-induced hepatic steatosis and hepatomegaly. Mitochondrial dysfunction is mainly associated with high-fat-diet (HFD)-induced hepatic steatosis. Sennoside A (SA), a commonly used clinical stimulant laxative, is reported to improve energy metabolism and insulin resistance. However, the effect and mechanism of SA on HFD-induced hepatic steatosis remain largely unknown. The aim of this study was to determine the effect and mechanism of SA on HFD-induced hepatic steatosis in mice. We examined the liver and body weight of mice to evaluate the physical changes in the liver. Hematoxylin and eosin (H&E) and oil red O staining were used to detect the lipid accumulation. The mitochondrial structure and function were tested by transmission electron microscopy and the Seahorse XF24 Analyzer. Furthermore, mitochondrial complexes I, II, and IV and voltage-dependent anion channel 1 (VDAC1) protein activity were detected to understand the mechanism of the protective effect on mitochondria. As a result, damage to the structure and function in the hepatic mitochondria of HFD-induced hepatic steatosis was observed in mice. The structural damage was in the form of loss of cristae, mitochondrial swelling, vacuolization and even rupturing of the outer mitochondrial membrane (OMM). Functional alterations were found by activation of complex I and deficiency in complexes II and IV. The VDAC1 activity and the total ATP in the liver tissue was increased under hepatic steatosis conditions. The above effects were reversed by SA. These data suggest that inhibition of VDAC1 may be an underlying mechanism of SA for protecting mitochondria in HFD-induced hepatic steatosis in mice. Thus, VDAC1 may be a promising target for treating fatty liver disease.

60 APPLIED LIFE SCIENCES↗

Hyper Spectral Anomaly Detection

Anomaly detection is a common machine learning (ML) task with growing importance in the fields of imaging, quality assurance, and multiple security related disciplines. Anomaly detection is more difficult than traditional machine learning methods due to the inherent unlabeled nature of the datasets. Existing anomaly detection architectures commonly face challenges with explainability, retaining information related to the relational structure of the data, and false positive rates. Hyperspectral Imaging Anomaly Detection (HSI) is a statistical model that employs vertex and edge weighted graphs to preserve the data’s relationships on different topographical scales. The model is able to generalize from anomaly detection in 2D images to novel datasets related to cyber-security. Furthermore, the use of multi-spectral and other filtering methods results in fewer false positives and increases the explainability of model predictions. When applying HSI to cyber-security datasets, we are able to successfully detect malicious activity with a relatively high degree of accuracy.

97 - MATHEMATICS AND COMPUTING↗

Impacts of active satellite sensors' low-level cloud detection limitations on cloud radiative forcing in the Arctic

Previous studies revealed that satellites sensors with the best detection capability identify 25 %–40 % and 0 %–25 % fewer clouds below 0.5 and between 0.5–1.0 km, respectively, over the Arctic. Quantifying the impacts of cloud detection limitations on the radiation flux are critical especially over the Arctic Ocean considering the dramatic changes in Arctic sea ice. In this study, the proxies of the space-based radar, CloudSat, and lidar, CALIPSO (Cloud-Aerosol Lidar and Infrared Pathfinder Satellite Observations), cloud masks are derived based on simulated radar reflectivity with QuickBeam and cloud optical thickness using retrieved cloud properties from surface-based radar and lidar during the Surface Heat Budget of the Arctic Ocean (SHEBA) experiment. Limitations in low-level cloud detection by the space-based active sensors, and the impact of these limitations on the radiation fluxes at the surface and the top of the atmosphere (TOA), are estimated with radiative transfer model Streamer. The results show that the combined CloudSat and CALIPSO product generally detects all clouds above 1 km, while detecting 25 % (9 %) fewer in absolute values below 600 m (600 m to 1 km) than surface observations. These detection limitations lead to uncertainties in the monthly mean cloud radiative forcing (CRF), with maximum absolute monthly mean values of 2.5 and 3.4 Wm –2 at the surface and TOA, respectively. Cloud information from only CALIPSO or CloudSat lead to larger cloud detection differences compared to the surface observations and larger CRF uncertainties with absolute monthly means larger than 10.0 Wm –2 at the surface and TOA. The uncertainties for individual cases are larger – up to 30 Wm –2 . These uncertainties need to be considered when radiation flux products from CloudSat and CALIPSO are used in climate and weather studies.

54 ENVIRONMENTAL SCIENCES↗

Magnetic field observations of evolved stars

A search has been made for Zeeman broadening in high resolution spectra of eight G and K type, evolved stars which have active chromospheres and coronae. No Zeeman broadening was found. It is likely that the fraction of the stellar surface that contains magnetic fields is similar in active giants and dwarfs, based on their comparable chromospheric line strengths and plage surface brightnesses (deduced from Mg II/C IV ratios) and on their comparable requirements for coronal confinement. Thus, since Zeeman broadening is detectable on active dwarfs, the nondetections of Zeeman broadening on these evolved stars are probably due to systematically lower field strengths. That the lower field strengths do not result in lower chromospheric surface brightnesses on giants is consistent with the theoretial 'slow mode' MHD wave generation rates of Ulmschneider and Stein (1982). The low field strengths can be explained by requiring that the fields be in equipartition with either the thermal or turbulent energy densities, both of which are reduced in late-type evolved stars compared with the sun.

Marcy, G. W.↗