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At least 631 records · Page 35

Multiplexed Colorimetric Solid-Phase Extraction

Multiplexed colorimetric solid-phase extraction (MC-SPE) is an extension of colorimetric solid-phase extraction (C-SPE) an analytical platform that combines colorimetric reagents, solid phase extraction, and diffuse reflectance spectroscopy to quantify trace analytes in water. In CSPE, analytes are extracted and complexed on the surface of an extraction membrane impregnated with a colorimetric reagent. The analytes are then quantified directly on the membrane surface using a handheld diffuse reflectance spectrophotometer. Importantly, the use of solid-phase extraction membranes as the matrix for impregnation of the colorimetric reagents creates a concentration factor that enables the detection of low concentrations of analytes in small sample volumes. In extending C-SPE to a multiplexed format, a filter holder that incorporates discrete analysis channels and a jig that facilitates the concurrent operation of multiple sample syringes have been designed, enabling the simultaneous determination of multiple analytes. Separate, single analyte membranes, placed in a readout cartridge create unique, analyte-specific addresses at the exit of each channel. Following sample exposure, the diffuse reflectance spectrum of each address is collected serially and the Kubelka-Munk function is used to quantify each water quality parameter via calibration curves. In a demonstration, MC-SPE was used to measure the pH of a sample and quantitate Ag(I) and Ni(II).

Gazda, Daniel B.↗

Biogeochemical Effects of Rising Atmospheric CO2 on Terrestrial and Ocean Systems

Rising carbon dioxide (CO2) has decreased seawater pH at long-term observing stations around the world, including in the open ocean north of Oahu, Hawaii, near Alaska's Aleutian Islands, the Gulf of Maine shore, and on Gray's Reef in the southeastern United States. This ocean acidification process has already affected some marine species and altered fundamental ecosystem processes, and further effects are likely. While atmospheric CO rises at approximately the same rate all over the globe, its non-climate effects on land vary depending on climate and dominant species. In terrestrial ecosystems, rising atmospheric CO concentrations are expected to increase plant photosynthesis, growth, and water-use efficiency, though these effects are reduced when nutrients, drought or other factors limit plant growth. Rising CO would likely change carbon storage and influence terrestrial hydrology and biogeochemical cycling, but concomitant effects on vegetation composition and nutrient feedbacks are challenging to predict, making decadal forecasts uncertain. Consequences of rising atmospheric CO are expected to include difficult-to-predict changes in the ecosystem services that terrestrial and ocean systems provide to humans. For instance, ocean acidification resulting from rising CO has decreased the supply of larvae that sustains commercial shellfish production in the northwestern United States. In addition, CO fertilization (increases) plus warming (decreases) are changing terrestrial crop yields. Continued persistence of uptake of carbon by the land and ocean is uncertain. Climate and environmental change create complex feedbacks to the carbon cycle and it is not clear how feedbacks modulate future effects of rising CO on carbon sinks. These are several mechanisms that could reduce future sink capacity.

Biogeochemical↗

Follow-up Water Quality Analysis at the Little Patuxent River in Anne Arundel County, Maryland andImpact to the Health of the Chesapeake Bay

In accordance with the federal Clean Water Act, it is of utmost importance to identify impaired water bodies and subsequently implement Total Maximum Daily Loads (TMDL) as needed to achieve desirable water quality standards. One of these water bodies include the Little Patuxent River (LPR) in the Anne Arundel and Howard Counties in Maryland, which is the subject of this report. In the 1996, 1998, and then 2006 Maryland Integrated Report, the LPR was found to be impaired by nutrients, bacteria, suspended sediments, and cadmium (Cd). As a result, TMDLs were created to limit factors that were contributing to these problems (Maryland Department of the Environment 2011, December). But later in 2008 and 2009, water quality analyses (WQAs) found that the LPR’s condition had improved. From a Category 5 Cd and phosphorus classification in 1996 it was reduced to Category 2 in 2008 for Cd and in 2009 for phosphorus (Maryland Department of the Environment 2011, December). However, it has been more than a decade since the last WQA and the environmental conditions may have changed. This lack of testing could result in untracked, accumulated damages to the overall health of the LPR. We present testing results for the 13 site locations on the LPR to evaluate turbidity, pH, phosphates, nitrates, water temperature, dissolved oxygen (D.O.), coliform bacteria, and cadmium concentrations in sediments. We will also present next steps including remediation strategies.

Aaban Syed↗

Modeling of Radiolytic Production of Oxidants Relevant to Corrosion of a Used Fuel Container in Deep Geologic Repository Environments - 17641

The Canadian used nuclear fuel disposal concept involves a deep geological repository using a multi-barrier system. A key barrier is the used fuel container (UFC). It consists of an inner vessel of carbon steel, which provides the structural strength to withstand repository loads, and an outer coating of copper acting as an external corrosion barrier. Relative to other design concepts, coated containers have thinner walls that provide less shielding for γ-radiation, raising the possibility of radiolysis-induced corrosion on the exterior of the container. Also, any trapped aerated water inside the container or in the bentonite clay may condense on the container surface, be exposed to γ-radiation and influence the container corrosion. Understanding the effect of radiation on container corrosion is one key factor to consider in the assessment of the integrity and longevity of the UFC. Humid air radiolysis produces HNO{sub 3} as the dominant oxidizing species. A conservative hemispherical water droplet model was developed to predict the accumulation of HNO{sub 3} and its impact on corrosion. Radiolysis of groundwater in the presence of Cl{sup -} mainly produces H{sub 2}O{sub 2} and HOCl. The concentration of these species is <10 mM and their sum remains almost constant. Radiation exposure tests showed radiation-induced corrosion on carbon steel and Cu. A systematic study of the Cu corrosion and the corrosion rate as a function of [H{sub 2}O{sub 2}], [HOCl], pH and γ-radiation is on-going. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Extraction Separation of Rare Earth Elements of the Medium Group with Isomolar Mixtures of Aliquat{sup ®}336–TBP and Cyanex{sup ®}572–TBP from Nitric Solutions

The results of extraction separation of the medium group of rare earth elements in the composition of the concentrate of JSC SMF with isomolar mixtures of Aliquat{sup ®}336–TBP and Cyanex{sup ®}572–TBP from nitric solutions are discussed. The influence of acidity of an aqueous solution on separation factors of representative pairs of REE and on the value of the capacity of mixtures of extractants is studied. The possibility of extraction separation on the lines of Sm–Gd and Eu–Gd (Aliquat{sup ®}336–TBP) and on the line of Eu–Sm (Cyanex{sup ®}572–TBP) is shown.

36 MATERIALS SCIENCE↗

Creation of a High-Yield AAV Vector Production Platform in Suspension Cells Using a Design-of-Experiment Approach

Recombinant adeno-associated virus (rAAV) vectors are a leading gene delivery platform, but vector manufacturing remains a challenge. New methods are needed to increase rAAV yields and reduce costs. Past efforts to improve rAAV production have focused on optimizing a single variable at a time, but this approach does not account for the interactions of multiple factors that contribute to vector generation. Here, we utilized a design-of-experiment (DOE) methodology to optimize rAAV production in a HEK293T suspension cell system. We simultaneously varied the transgene, packaging, and helper plasmid ratios, the total DNA concentration, and the cell density to systematically evaluate the impact of each variable across 52 conditions. The results revealed a unique set of parameters with a lower concentration of transgene plasmid, a higher concentration of packaging plasmid, and a higher cell density than previously described protocols. Using this DOE-optimized protocol, we achieved unpurified yields approaching 3 × 10 14 viral genomes (VGs)/L of cell culture. Additionally, we incorporated polyethylene glycol (PEG)-based virus precipitation, pH-mediated protein removal, and affinity chromatography to our downstream processing, enabling average purified yields of >1 × 10 14 VGs/L for rAAV-EGFPs across 13 serotypes and capsid variants.

59 BASIC BIOLOGICAL SCIENCES↗

Characterization of soil and postlaunch pad debris from Cape Canaveral launch complex and analysis of soil interaction with aqueous HCl

Soil samples were fractionated and analyzed in order to assess the physical and chemical interactions of entrained soil with solid-rocket exhaust clouds. The sandy soil consisted primarily of quartz (silica) particles, 30 to 500 microns in diameter, and also contained seashell fragments. Differential and cumulative soil-mass size distributions are presented along with mineralogy, elemental compositions, and solution pH histories. About 90 percent of the soil mass consisted of particles 165 microns in diameter. Characteristic reaction times in aqueous HC1 slurries varied from a few minutes to several days, and capacities for reaction under acidic conditions varied from 10 to 40 g HCl/kg soil, depending on particle size. Airborne lifetimes of particles 165 microns are conservatively 30 min, and this major grouping is predicted to represent a small short-term chemical sink for up to 5% of the total HC1. The smaller and more minor fractions, below a 165 micron diameter, may act as giant cloud condensation nuclei over much longer airborne lifetimes. Finally, the demonstrated time dependency of neutralization is a complicating factor; it can influence the ability to deduce in-cloud HCl scavenging with reaction and can affect the accuracy of measured chemical compositions of near-field wet deposition.

Pellett, G. L.↗

A portable fiber optic sensor for the luminescent sensing of cobalt ions using carbon dots

Cobalt is critical to energy-relevant technologies, and demand for cobalt will increase significantly with growing global adoption of renewables. However, supply chain stability is threatened by economic and geopolitical factors, incentivizing domestic cobalt production from alternative resources such as coal, coal utilization byproducts (e.g., ash, acid mine drainage) and electronic waste. Rapid, inexpensive, and portable characterization techniques are needed to reduce production costs associated with cobalt prospecting and process monitoring. Here, in this research, we develop a compact, portable fiber optic luminescent probe for cobalt using phosphorus and nitrogen co-doped carbon dots as the sensing material. The carbon dot emission overlaps well with the cobalt absorption band at ~510 nm, leading to a selective decrease in emission as a function of cobalt concentration. The system responds nearly instantly to the presence of cobalt, with detection limits of 0.7 and 3.5 ppm in water and pH 1.68 buffer, respectively, providing comparable performance to a commercial spectrometer at a significantly lower cost. Moreover, the sensor is selective for cobalt in the presence of 13 of the most common metal ions encountered in coal utilization byproducts and is responsive to cobalt when spiked into an acid mine drainage leachate sample, highlighting the sensor's potential for real-world deployment in challenging matrices. In addition, integrating the carbon dots with a filter paper substrate produced ‘test strip’ sensors that exhibited a selective and sensitive visual response to cobalt using a handheld UV lamp. Taken together, the sensing system represents a significant step forward in the development of low-cost practical sensors for high-value metals in complex streams.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Comparison of the Microbial Diversity and Abundance Between the Freshwater Land-Locked Lakes of Schirmacher Oasis and the Perennially Ice-Covered Lake Untersee in East Antarctica

Extreme conditions such as low temperature, dryness, and constant UV-radiation in terrestrial Antarctica are limiting factors of the survival of microbial populations. The objective of this study was to investigate the microbial diversity and enumeration between the open water lakes of Schirmacher Oasis and the permanently ice-covered Lake Untersee. The lakes in Schirmacher Oasis possessed abundant and diverse group of microorganisms compared to the Lake Untersee. Furthermore, the microbial diversity between two lakes in Schirmacher Oasis (Lake L27C and L47) was compared by culture-based molecular approach. It was determined that L27Chad a richer microbial diversity representing 5 different phyla and 7 different genera. In contrast L47 consisted of 4 different phyla and 6 different genera. The difference in microbial community could be due to the wide range of pH between L27C (pH 9.1) and L47 (pH 5.7). Most of the microbes isolated from these lakes consisted of adaptive biological pigmentation. Characterization of the microbial community found in the freshwater lakes of East Antarctica is important because it gives a further glimpse into the adaptation and survival strategies found in extreme conditions.

Huang, Jonathan↗

Magnetic Ordering in Gold Nanoclusters

Here, several research groups have observed magnetism in monolayer-protected gold-cluster samples, but the results were often contradictory and thus a clear understanding of this phenomenon is still missing. We used Au 25 (SCH 2 CH 2 Ph) 18 0 , which is a paramagnetic cluster that can be prepared with atomic precision and whose structure is known precisely. Previous magnetometry studies only detected paramagnetism. We used samples representing a range of crystallographic orders and studied their magnetic behaviors by electron paramagnetic resonance (EPR). As a film, Au 25 (SCH 2 CH 2 Ph) 18 0 displays paramagnetic behavior but, at low temperature, ferromagnetic interactions are detectable. One or few single crystals undergo physical reorientation with the applied field and display ferromagnetism, as detected through hysteresis experiments. A large collection of microcrystals is magnetic even at room temperature and shows distinct paramagnetic, superparamagnetic, and ferromagnetic behaviors. Simulation of the EPR spectra shows that both spin-orbit coupling and crystal distortion are important to determine the observed magnetic behaviors. DFT calculations carried out on single cluster and periodic models predict values of spin6orbit coupling and crystal6splitting effects in agreement with the EPR derived quantities. Magnetism in gold nanoclusters is thus demonstrated to be the outcome of a very delicate balance of factors. To obtain reproducible results, the samples must be (i) controlled for composition and thus be monodispersed with atomic precision, (ii) of known charge state, and (iii) well defined also in terms of crystallinity and experimental conditions. This study highlights the efficacy of EPR spectroscopy to provide a molecular understanding of these phenomena

36 MATERIALS SCIENCE↗

Morphological and crystallographic controls in the replacement of calcite and aragonite by cerussite and otavite

Mineral replacement reactions are essential solid-fluid interactions in natural and industrial processes including metasomatism, diagenesis, and metamorphism, and sequestration of metal toxins from polluted water. In this paper, we explore the morphological evolution of aragonite and calcite (polymorphs of CaCO 3 ) during their replacement by cerussite (PbCO 3 , an isomorph of aragonite) and otavite (CdCO 3 , an isomorph of calcite) in acidic aqueous solutions (with initial pH 3 to 4). Observations using scanning electron and synchrotron-based transmission X-ray microscopies reveal the formation of pseudomorphic shells (~5 to 10 µm thick) of cerussite and otavite when Pb 2+ is reacted with either calcite or aragonite and when Cd 2+ is reacted with aragonite. The formation of pore space at the substrate-precipitate interface and within the precipitate is found to be key in promoting chemical exchange between the dissolving phase and the solution. In contrast, minimal reactivity is observed when Cd 2+ is reacted with calcite. These results demonstrate that the lack of substrate-precipitate epitaxy (either because of inconsistent symmetries or due to large lattice constant mismatch) is a critical factor that enables replacement of primary CaCO 3 minerals by Pb- or Cd-bearing secondary minerals while the presence of epitaxial relationships inhibit those reactions. When calcite and aragonite coexist, the replacement of aragonite by cerussite and otavite occurs preferentially over calcite. We postulate that the relative reactivity of calcite and aragonite to Pb 2+ and Cd 2+ can be determined by interplay between morphological and crystallographic properties of CaCO 3 materials as well as solution chemistry. Our results demonstrate that coupled dissolution-precipitation processes of carbonate minerals play an important role in controlling mobility and bioavailability of lead and cadmium in environments where acidic metal-contaminated water interacts with carbonate-rich rocks and soils.

58 GEOSCIENCES↗

Supercharged cellulases show superior thermal stability and enhanced activity towards pretreated biomass and cellulose

Non-productive binding of cellulolytic enzymes to various plant cell wall components, such as lignin and cellulose, necessitates high enzyme loadings to achieve efficient conversion of pretreated lignocellulosic biomass to fermentable sugars. Protein supercharging was previously employed as one of the strategies to reduce non-productive binding to biomass. However, various questions remain unanswered regarding the hydrolysis kinetics of supercharged enzymes towards pretreated biomass substrates and the role played by enzyme interactions with individual cell wall polymers such as cellulose and xylan. In this study, CBM2a (from Thermobifida fusca ) fused with endocellulase Cel5A (from T. fusca ) was used as the model wild-type enzyme and CBM2a was supercharged using Rosetta, to obtain eight variants with net charges spanning -14 to +6. These enzymes were recombinantly expressed in E. coli , purified from cell lysates, and their hydrolytic activities were tested against pretreated biomass substrates (AFEX and EA treated corn stover). Although the wild-type enzyme showed greater activity compared to both negatively and positively supercharged enzymes towards pretreated biomass, thermal denaturation assays identified two negatively supercharged constructs that perform better than the wild-type enzyme (~3 to 4-fold difference in activity) upon thermal deactivation at higher temperatures. To better understand the causal factor of reduced supercharged enzyme activity towards AFEX corn stover, we performed hydrolysis assays on cellulose-I/xylan/pNPC, lignin inhibition assays, and thermal stability assays. Altogether, these assays showed that the negatively supercharged mutants were highly impacted by reduced activity towards xylan whereas the positively supercharged mutants showed dramatically reduced activity towards cellulose and xylan. It was identified that a combination of impaired cellulose binding and lower thermal stability was the cause of reduced hydrolytic activity of positively supercharged enzyme sub-group. Overall, this study demonstrated a systematic approach to investigate the behavior of supercharged enzymes and identified supercharged enzyme constructs that show superior activity at elevated temperatures. Future work will address the impact of parameters such as pH, salt concentration, and assay temperature on the hydrolytic activity and thermal stability of supercharged enzymes.

09 BIOMASS FUELS↗

Extraction of lithium from battery recycling wastewater using synergistic D2EHPA and TBP

The recycling of spent lithium-ion batteries (LIBs) poses significant challenges, including the generation of large volumes of chemically complex wastewater. The composition of this wastewater is influenced by both the intrinsic chemistry of the batteries and the specific recycling processes employed. Notably, this wastewater contains economically valuable components, such as lithium, which can be recovered. Here, in this study, a solvent extraction (SX) process was investigated as a method to recover lithium from battery recycling wastewater, especially from anode-washing stage. Initially, various commercial extractants were evaluated, including di(2-ethylhexyl)phosphoric acid (D2EHPA), mono-2-ethylhexyl (2-ethylhexyl)phosphonate (PC88A), bis(2,4,4-trimethylpentyl)phosphinic acid (Cyanex 272), 2-hydroxy-5-nonylacetophenone oxime (LIX 84-I), tri-butyl phosphate (TBP), and their combinations. Among these, D2EHPA + TBP demonstrated a synergism to advance and maximize the lithium extraction. Subsequently, the effects of key parameters, including D2EHPA concentration, TBP concentration, contact time, initial pH, and aqueous-to-organic (A/O) phase ratio, were systematically investigated and optimized. A two-stage SX approach was employed to enhance lithium recovery. Under the optimized conditions of 30.0 vol% D2EHPA, 10.0 vol% TBP, 10 min of contact time, and a 1:1 A/O phase ratio, a lithium extraction efficiency of more than 88% in a two-stage solvent extraction was achieved. Lithium was subsequently stripped from the loaded organic solution using sulfuric acid (H 2 SO 4 ). Using 2.0 M H 2 SO 4 , lithium stripping was achieved after two counter-current stripping stages at an organic-to-aqueous (O/A) phase ratio of 6:1. This stripping process enriched the lithium concentration by a factor of four compared to the original lithium concentration in the anode-washing wastewater. The recyclability of the synergistic D2EHPA + TBP system was also evaluated over four extraction-stripping cycles. The results demonstrated that the system maintained high extraction and stripping efficiencies.

D2EHPA↗

Cycling phosphorus on the Archean Earth: Part II. Phosphorus limitation on primary production in Archean ecosystems

Several lines of evidence point to low rates of net primary production (NPP) in Archean oceans. However, whether Archean NPP was limited by electron donors or nutrients, particularly phosphorus (P), and how these factors might have changed over a billion years of recorded Archean history, remains contentious. One major challenge is to understand quantitatively the biogeochemical cycling of P on the early Earth. In Part I of this series (Hao et al., 2020), we estimated the weathering flux of P to the oceans as a function of temporally increasing continental emergence and elevation through Archean time. In Part II, we conduct thermodynamic and kinetic simulations to understand key processes of P cycling within the Archean ocean, including seafloor weathering, recycling of organic P, the solubility and precipitation of secondary phosphate minerals, and the burial diagenesis of P precipitates. Our calculations suggest low solubilities of apatite minerals in Archean seawater, primarily due to nearly neutral pH and high levels of Ca. This low solubility, in turn, implies a negligible contribution of apatite dissolution to P bioavailability in Archean seawater. We also simulate the solubility limits of common secondary P-bearing minerals, showing that vivianite would have been the least soluble P mineral in ferruginous Archean seawater (0.1–0.3 µM), even at moderate supersaturation states (Ω = 100 or 1000). If vivianite precipitation was kinetically favorable by microbial activities and mineral adsorption, the sinking flux of P as vivianite in Archean seawater could have reached the modern sinking flux, implying that vivianite precipitation was a potentially major sink for P in Archean oceans. During burial diagenesis, however, vivianite in porewater would have become less stable than Ca-phosphates of lower solubility. At elevated temperatures (>100 °C) associated with burial diagenesis and low-grade metamorphism, vivianite is predicted to react irreversibly with calcite to form apatite. Optimistic assumptions about the recycling efficiency of P on the Archean Earth lead us to estimate that by the end of the eon the total flux of P (continental weathering + recycling) could have supported NPP at levels up to 7% of the modern. The total flux of P would have been much lower on the early and middle Archean Earth, whereas fluxes of electron donors could have been higher, suggesting very low productivity and P-limitation of marine ecosystems during much of the eon. Comparing our estimates of NPP as limited by P supply with the estimate by Ward et al. (2019), in which NPP was limited by electron donors and metabolic efficiency, there could have been a transition between P-limited productivity in the early to middle Archean to electron donor-limitation closer to the eon’s end (assuming no oxygenic photosynthesis). Once oxygenic photosynthesis reached ecological significance, probably near the end of the Archean, our estimated flux of P would allow rapid oxidation of atmosphere.

Vivianite↗

Single-phase and two-phase anaerobic digestion of fruit and vegetable waste: Comparison of start-up, reactor stability and process performance

Highlights: • Single-phase and two-phase systems were compared for fruit and vegetable waste digestion. • Single-phase digestion produced a methane yield of 0.45 m{sup 3} CH{sub 4}/kg VS and 83% VS removal. • Substrate solubilization was high in acidification conditions at 7.0 kg VS/m{sup 3} d and pH 5.5–6.2. • Energy yield was lower by 33% for two-phase system compared to the single-phase system. • Simple and straight-forward operation favored single phase process over two-phase process. - Abstract: Single-phase and two-phase digestion of fruit and vegetable waste were studied to compare reactor start-up, reactor stability and performance (methane yield, volatile solids reduction and energy yield). The single-phase reactor (SPR) was a conventional reactor operated at a low loading rate (maximum of 3.5 kg VS/m{sup 3} d), while the two-phase system consisted of an acidification reactor (TPAR) and a methanogenic reactor (TPMR). The TPAR was inoculated with methanogenic sludge similar to the SPR, but was operated with step-wise increase in the loading rate and with total recirculation of reactor solids to convert it into acidification sludge. Before each feeding, part of the sludge from TPAR was centrifuged, the centrifuge liquid (solubilized products) was fed to the TPMR and centrifuged solids were recycled back to the reactor. Single-phase digestion produced a methane yield of 0.45 m{sup 3} CH{sub 4}/kg VS fed and VS removal of 83%. The TPAR shifted to acidification mode at an OLR of 10.0 kg VS/m{sup 3} d and then achieved stable performance at 7.0 kg VS/m{sup 3} d and pH 5.5–6.2, with very high substrate solubilization rate and a methane yield of 0.30 m{sup 3} CH{sub 4}/kg COD fed. The two-phase process was capable of high VS reduction, but material and energy balance showed that the single-phase process was superior in terms of volumetric methane production and energy yield by 33%. The lower energy yield of the two-phase system was due to the loss of energy during hydrolysis in the TPAR and the deficit in methane production in the TPMR attributed to COD loss due to biomass synthesis and adsorption of hard COD onto the flocs. These results including the complicated operational procedure of the two-phase process and the economic factors suggested that the single-phase process could be the preferred system for FVW.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

NASA SMART Probe: Breast Cancer Application

There is evidence in breast cancer and other malignancies that the physiologic environment within a tumor correlates with clinical outcome. We are developing a unique percutaneous Smart Probe to be used at the time of needle biopsy of the breast. The Smart Probe will simultaneously measure multiple physiologic parameters within a breast tumor. Direct and indirect measurements of tissue oxygen levels, blood flow, pH, and tissue fluid pressure will be analyzed in real-time. These parameters will be interpreted individually and collectively by innovative neural network techniques using advanced intelligent software. The goals are 1) develop a pecutaneous Smart Probe with multiple sensor modalities and applying advanced Information Technologies to provide real time diagnostic information of the tissue at tip of the probe, 2) test the percutaneous Smart Probe in women with benign and malignant breast masses who will be undergoing surgical biopsy, 3) correlate probe sensor data with benign and malignant status of breast masses, 4) determine whether the probe can detect physiologic differences within a breast tumor, and its margins, and in adjacent normal breast tissue, 5) correlate probe sensor data with known prognostic factors for breast caner, including tumor size, tumor grade, axillary lymph node metastases, estrogen receptor and progesterone receptor status.

Mah, Robert W.↗

Development of an optical Zn 2+ probe based on a single fluorescent protein

Various fluorescent probes have been developed to reveal the biological functions of intracellular labile Zn 2+ . Here we present Green Zinc Probe (GZnP), a novel genetically encoded Zn 2+ sensor design based on a single fluorescent protein (single-FP). The GZnP sensor is generated by attaching two zinc fingers (ZF) of the transcription factor Zap1 (ZF1 and ZF2) to the two ends of a circularly permuted green fluorescent protein (cpGFP). Formation of ZF folds induces interaction between the two ZFs, which induces a change in the cpGFP conformation, leading to an increase in fluorescence. A small sensor library is created to include mutations in the ZFs, cpGFP and linkers between ZF and cpGFP to improve signal stability, sensor brightness and dynamic range based on rational protein engineering and computational design by Rosetta. Using a cell-based library screen, we identify sensor GZnP1 which demonstrates a stable maximum signal, decent brightness (QY = 0.42 at apo state), as well as specific and sensitive response to Zn 2+ in HeLa cells (F max /F min = 2.6, K d = 58 pM, pH 7.4). The subcellular localizing sensors mito-GZnP1 (in mitochondria matrix) and Lck-GZnP1 (on plasma membrane) display sensitivity to Zn 2+ (F max /F min = 2.2). In conclusion, this sensor design provides freedom to be used in combination with other optical indicators and optogenetic tools for simultaneous imaging and advancing our understanding of cellular Zn 2+ function.

59 BASIC BIOLOGICAL SCIENCES↗

Galactose inhibits auxin-induced growth of Avena coleoptiles by two mechanisms

Galactose inhibits auxin-induced growth of Avena coleoptiles by at least two mechanisms. First, it inhibits auxin-induced H(+)-excretion needed for the initiation of rapid elongation. Galactose cannot be doing so by directly interfering with the ATPase since fusicoccin-induced H(+)-excretion is not affected. Secondly, galactose inhibits long-term auxin-induced growth, even in an acidic (pH 4.5) solution. This may be due to an inhibition of cell wall synthesis. However, galactose does not reduce the capacity of walls to be loosened by H+, given exogenously or excreted in response to fusicoccin.

NASA Discipline Plant Biology↗