Teaching and assessing undergraduate collaboration skills scaffolded through the biochemistry curriculum using collaboration rubrics and student learning contracts.

The Department of Chemistry and Biochemistry at St. Mary's College of Maryland has scaffolded collaboration skills throughout the Biochemistry curriculum and developed several assessment tools to evaluate these skills. Biochemistry I and II have used team contracts at the beginning of extensive team projects where students identify their strengths, review expectations, and plan for group communication. At the conclusion of each project, each student assesses their own contributions and team members for various parts of the project. A common collaboration rubric was also applied in Biochemistry I and II as well as in two other courses, General Chemistry II Lab and Physical Chemistry I Lab, for students to evaluate themself and team members using the following subcategories: quality of work, commitment, leadership, communication, and analysis. In Biochemistry I and II, we used this rubric for multiple assignments that are part of the projects in the lecture courses. In the General Chemistry II Lab, we provided elements of this rubric within an evaluation form that reflects these collaboration attributes after each lab experience, so students can assess and report privately on their experiences as part of their collaboration grade for the course. A similar collaboration rubric is completed by students for each team-based laboratory within Physical Chemistry I. We also demonstrate different ways that instructors can use the data from these assessment tools. In our department, we are using these tools to frame the importance of collaboration skills and collecting data to inform our teaching of these skills. Preliminary data suggest that our curriculum is successfully teaching students how to be good collaborators.

Chemistry and Dynamics of Supercritical Carbon Dioxide and Methane in the Slit Pores of Layered Silicates.

ConspectusIn the mid 2010s, high-pressure diffraction and spectroscopic tools opened a window into the molecular-scale behavior of fluids under the conditions of many CO2 sequestration and shale/tight gas reservoirs, conditions where CO2 and CH4 are present as variably wet supercritical fluids. Integrating high-pressure spectroscopy and diffraction with molecular modeling has revealed much about the ways that supercritical CO2 and CH4 behave in reservoir components, particularly in the slit-shaped micro- and mesopores of layered silicates (phyllosilicates) abundant in caprocks and shales. This Account summarizes how supercritical CO2 and CH4 behave in the slit pores of swelling phyllosilicates as functions of the H2O activity, framework structural features, and charge-balancing cation properties at 90 bar and 323 K, conditions similar to a reservoir at ∼1 km depth. Slit pores containing cations with large radii, low hydration energy, and large polarizability readily interact with CO2, allowing CO2 and H2O to adsorb and coexist in these interlayer pores over a wide range of fluid humidities. In contrast, cations with small radii, high hydration energy, and low polarizability weakly interact with CO2, leading to reduced CO2 uptake and a tendency to exclude CO2 from interlayers when H2O is abundant. The reorientation dynamics of confined CO2 depends on the interlayer pore height, which is strongly influenced by the cation properties, framework properties, and fluid humidity. The silicate structural framework also influences CO2 uptake and behavior; for example, smectites with increasing F-for-OH substitution in the framework take up greater quantities of CO2. Reactions that trap CO2 in carbonate phases have been observed in thin H2O films near smectite surfaces, including a dissolution-reprecipitation mechanism when the edge surface area is large and an ion exchange-precipitation mechanism when the interlayer cation can form a highly insoluble carbonate. In contrast, supercritical CH4 does not readily associate with cations, does not react with smectites, and is only incorporated into interlayer slit mesopores when (i) the pore has a z-dimension large enough to accommodate CH4, (ii) the smectite has low charge, and (iii) the H2O activity is low. The adsorption and displacement of CH4 by CO2 and vice versa have been studied on the molecular scale in one shale, but opportunities remain to examine behavioral details in this more complicated, slit-pore inclusive system.

Interlayer Cation Polarizability Affects Supercritical Carbon Dioxide Adsorption by Swelling Clays.

Several strategies for mitigating the build-up of atmospheric carbon dioxide (CO2) bring wet supercritical CO2 (scCO2) in contact with phyllosilicates such as illites and smectites. While some work has examined the role of the charge-balancing cation and smectite framework features on CO2/smectite interactions, to our knowledge no one has examined how the polarizability of the charge-balancing cation influences these behaviors. In this paper, the scCO2 adsorption properties of Pb2+, Rb+, and NH4+ saturated smectite clays at variable relative humidity are studied by integrating in situ high-pressure X-ray diffraction (XRD), infrared spectroscopic titrations, and magic angle spinning nuclear magnetic resonance (MAS NMR) methods. The results are combined with previously published data for Na+, Cs+, and Ca2+ saturated versions of the same smectites to isolate the roles of the charge-balancing cations and perform two independent tests of the role of charge-balancing cation polarizability in determining the interlayer fluid properties and smectite expansion. Independent correlations developed for (i) San Bernardino hectorite (SHCa-1) and (ii) Wyoming montmorillonite (SWy-2) both show that cation polarizability is important in predicting the interlayer composition (mol% CO2 in the interlayer fluid and CO2/cation ratio in interlayer) and the expansion behavior for smectites in contact with wet and dry scCO2. In particular, this study shows that the charge-balancing cation polarizability is the most important cation-associated parameter in determining the expansion of the trioctahedral smectite, hectorite, when in contact with dry scCO2. While both independent tests show that cation polarizability is an important factor in smectite-scCO2 systems, the correlations for hectorite are different from those determined for montmorillonite. The root of these differences is likely associated with the roles of the smectite framework on adsorption, warranting follow-up studies with a larger number of unique smectite frameworks. Overall, the results show that the polarizability of the charge-balancing cation should be considered when preparing smectite clays (or industrial processes involving smectites) to capture CO2 and in predicting the behavior of caprocks over time.

A Mechanistic Exploration of Natural Organic Matter Aggregation and Surface Complexation in Smectite Mesopores.

Soil minerals and organic matter play critical roles in nutrient cycling and other life-essential biogeochemical processes, yet the structural and dynamical details of natural organic matter (NOM) film formation on smectites are not fully understood on the molecular scale. XRD of Suwannee River NOM-hectorite (a smectite clay) complexes shows that the humic and fulvic components of NOM bind predominantly at the external surfaces of packets of smectite platelets rather than in the interlayer slit pores, suggesting that the key behavior governing smectite-NOM interactions takes place in mesopores between smectite particles. New molecular dynamics modeling of a ∼110 ŠH2O-saturated smectite mesopore at near-neutral pH shows that model NOM molecules initially form small clusters of 2-3 NOM molecules near the center of the pore fluid. Formation of these clusters is driven by the hydrophobic mechanism, where aromatic/aliphatic regions associate with one another to minimize their interactions with H2O, and charge-balancing cations associated with the deprotonated carboxylate sites are located only at the outer surface of these clusters. Despite hydrophobicity driving the initial clustering, NOM clusters are formed more quickly when high-charge-density cations like Ca2+ are present vs low-charge-density cations like Cs+, as the former cations more effectively minimize the electrostatic repulsions between the negatively charged NOM molecules. Once the small hydrophobicity-driven NOM clusters form, the simulations show that Ca2+ promotes the aggregation of NOM clusters through tetradentate Ca2+ bridges involving carboxylate groups on two different NOM clusters. Importantly, our studies indicate that Ca2+ plays a crucial role in binding the NOM clusters to the smectite surface, which occurs through multiple quaternary complexes (Ob)-H2O-Ca2+-COO-NOM. In contrast, Cs+ never forms any coordination or acts like bridges between NOM molecules nor as ion bridges to the smectite surface. Additionally, we observe the formation of a metastable superaggregate involving all 16 NOM molecules several times in a Ca2+-bearing mesopore fluid. Superaggregates are never observed in the simulations involving Cs+. The modeling results are fully consistent with helium ion microscope images of NOM-hectorite complexes suggesting that NOM surface films develop when preformed NOM clusters interact with smectite surfaces. Overall, the binding of NOM clusters to the outer surfaces of smectite particles and the formation of large NOM aggregates at neutral pH occur through cation bridging, and cation bridging only occurs when high-charge-density cations like Ca2+ are present.

Understanding methane/carbon dioxide partitioning in clay nano- and meso-pores with constant reservoir composition molecular dynamics modeling.

The interactions among fluid species such as H2O, CO2, and CH4 confined in nano- and meso-pores in shales and other rocks is of central concern to understanding the chemical behavior and transport properties of these species in the earth's subsurface and is of special concern to geological C-sequestration and enhanced production of oil and natural gas. The behavior of CO2, and CH4 is less well understood than that of H2O. This paper presents the results of a computational modeling study of the partitioning of CO2 and CH4 between bulk fluid and nano- and meso-pores bounded by the common clay mineral montmorillonite. The calculations were done at 323 K and a total fluid pressure of 124 bars using a novel approach (constant reservoir composition molecular dynamics, CRC-MD) that uses bias forces to maintain a constant composition in the fluid external to the pore. This purely MD approach overcomes the difficulties in making stochastic particle insertion-deletion moves in dense fluids encountered in grand canonical Monte Carlo and related hybrid approaches. The results show that both the basal siloxane surfaces and protonated broken edge surfaces of montmorillonite both prefer CO2 relative to CH4 suggesting that methods of enhanced oil and gas production using CO2 will readily displace CH4 from such pores. This preference for CO2 is due to its preferred interaction with the surfaces and extends to approximately 20 Å from them.

Temperature dependent structure and dynamics in smectite interlayers: 23Na MAS NMR spectroscopy of Na-hectorite.

23Na MAS NMR spectroscopy of the smectite mineral hectorite acquired at temperatures from -120 °C to 40 °C in combination with the results from computational molecular dynamics (MD) simulations show the presence of complex dynamical processes in the interlayer galleries that depend significantly on their hydration state. The results indicate that site exchange occurs within individual interlayers that contain coexisting 1 and 2 water layer hydrates in different places. We suggest that the observed dynamical averaging may be due to motion of water volumes comparable to the dripplons recently proposed to occur in hydrated graphene interlayers (Yoshida et al. Nat. Commun., 2018, 9, 1496). Such motion would cause rippling of the T-O-T structure of the clay layers at frequencies greater than ∼25 kHz. For samples exposed to 0% relative humidity (R.H.), the 23Na spectra show the presence of two Na+ sites (probably 6 and 9 coordinated by basal oxygen atoms) that do not undergo dynamical averaging at any temperature from -120 °C to 40 °C. For samples exposed to R.H.s from 29% to 100% the spectra show the presence of three hydrated Na+ sites that undergo dynamical averaging beginning at -60 °C. These sites have different numbers of H2O molecules coordinating the Na+, and diffusion calculations indicate that they probably occur within the same individual interlayer. The average hydration state of Na+ increases with increasing R.H. and water content of the clay.

Tipping Point for Expansion of Layered Aluminosilicates in Weakly Polar Solvents: Supercritical CO2.

Layered aluminosilicates play a dominant role in the mechanical and gas storage properties of the subsurface, are used in diverse industrial applications, and serve as model materials for understanding solvent-ion-support systems. Although expansion in the presence of H2O is well-known to be systematically correlated with the hydration free energy of the interlayer cation, particularly in environments dominated by nonpolar solvents (i.e., CO2), uptake into the interlayer is not well-understood. Using novel high-pressure capabilities, we investigated the interaction of dry supercritical CO2 with Na-, NH4-, and Cs-saturated montmorillonite, comparing results with predictions from molecular dynamics simulations. Despite the known trend in H2O and that cation solvation energies in CO2 suggest a stronger interaction with Na, both the NH4- and Cs-clays readily absorbed CO2 and expanded, while the Na-clay did not. The apparent inertness of the Na-clay was not due to kinetics, as experiments seeking a stable expanded state showed that none exists. Molecular dynamics simulations revealed a large endothermicity to CO2 intercalation in the Na-clay but little or no energy barrier for the NH4- and Cs-clays. Indeed, the combination of experiment and theory clearly demonstrate that CO2 intercalation of Na-montmorillonite clays is prohibited in the absence of H2O. Consequently, we have shown for the first time that in the presence of a low dielectric constant, gas swelling depends more on the strength of the interaction between the interlayer cation and aluminosilicate sheets and less on that with solvent. The finding suggests a distinct regime in layered aluminosilicate swelling behavior triggered by low solvent polarizability, with important implications in geomechanics, storage, and retention of volatile gases, and across industrial uses in gelling, decoloring, heterogeneous catalysis, and semipermeable reactive barriers.

Molecular dynamics modeling of carbon dioxide, water and natural organic matter in Na-hectorite.

Molecular dynamics (MD) modeling of systems containing a Na-exchanged smectite clay (hectorite) and model natural organic matter (NOM) molecules along with pure H2O, pure CO2, or a mixture of H2O and CO2 provides significant new insight into the molecular scale interactions among silicate surfaces, dissolved cations and organic molecules, H2O and CO2 relevant to geological C-sequestration strategies. The simulations for systems containing H2O show the following results; (1) Na(+) does not bridge between NOM molecules and the clay surface at protonation states comparable to near neutral pH conditions. (2) In systems without CO2 the NOM molecules retain charge balancing cations and drift away from the silicate surface. (3) In systems containing both H2O and CO2, the NOM molecules adopt equilibrium positions at the H2O-CO2 interface with the more hydrophilic structural elements facing the H2O and the more hydrophobic ones facing the CO2. In systems with only CO2, NOM and Na(+) ions are pinned to the clay surface with the hydrophilic structural elements of the NOM pointed toward the clay surface. Dynamically, in systems with only CO2, Na(+) diffusion is nearly eliminated, and in systems with a thin water film on the clay surface diffusion perpendicular the surface is greatly reduced relative to the system with bulk water. Energetically, the results for the systems with only H2O show that hydration of the net charge neutral Na-NOM molecule outweighs the sum of its Coulombic and dispersive interactions with the net charge-neutral Na-clay particle and the interactions of the water molecules with the hydrophobic structural elements of the NOM. The aggregation of NOM molecules in solution appears to be driven not by Na(+) bridging between the molecules but by hydrophobic interactions between them. In contrast, for the systems with only CO2 the interaction between the Na-NOM molecules and the CO2 is outweighed by the interaction of NOM with the clay particle. With both H2O and CO2 present, the energetic interactions leading to the hydration of the Na-clay surface and the hydrophilic structural elements of the Na-NOM molecule and the hydrophobic interactions between the CO2 and the hydrophobic aromatic and aliphatic structural elements of the NOM can both be satisfied, leading to the Na-NOM molecules migrating away from the surface and residing at the H2O-CO2 interface. The MD results suggest some alternative explanations for the previously observed (23)Na NMR behavior of Na-hectorite at elevated temperatures and CO2 pressures.

Cation exchange at the mineral-water interface: H3O+/K+ competition at the surface of nano-muscovite.

This article describes a (39)K nuclear magnetic resonance (NMR) spectroscopic study of K+ displacement at the muscovite/water interface as a function of aqueous phase pH. (39)K NMR spectra and T 2 relaxation data for nanocrystalline muscovite wet with a solid/solution weight ratio of 1 at pH 1, 3, and 5.5 show substantial liquid-like K+ only at pH 1. At pH 3 and 5.5, all K+ appears to be associated with muscovite as inner- or outer-sphere complexes, indicating that H(3)O+ does not displace basal surface K+ beyond the (39)K detection limit under these conditions. In our pH 1 mixture, only approximately 1/3 of the initial basal surface K+ population is located more than 3-4 A from the surface. (29)Si and (27)Al MAS NMR spectra and SEM images show no evidence of dissolution during the (39)K experiments, consistent with the liquid-like (39)K fraction originating from displaced basal surface K+. Assuming no muscovite dissolution or interlayer exchange, the K+/H(3)O+ ratio relevant to the solution/surface exchange equilibrium is controlled by the total amount of K+ on the surface and H(3)O+ in solution (K+(surf)/H(3)O+(aq)). These parameters, in turn, depend on the basal surface area, solution pH, and the solid/solution ratio. The results here are consistent with significant displacement of surface K+ only under conditions where the initial K+(surf)/H(3)O+(aq). ratio is less than approximately 1. Computational molecular models of the muscovite/water interface should account for both K+ and H(3)O+ in the near-surface region.

High-field (75)As NMR study of arsenic oxysalts.

Arsenic is an important environmental hazard, but there have been few NMR investigations of its molecular scale structure and dynamics, due principally to the large quadrupole moment of (75)As and consequent large quadrupole couplings. We examine here the potential of existing, single-field solid-state NMR technology to investigate solids containing arsenate and arsenite oxyanions. The results show that current techniques have significant potential for arsenates that do not contain both protonated H(x)AsO4-(3-x) groups and structural water molecules, but that the quadrupole couplings for the arsenites examined here are large enough that interpretation of the spectra is difficult, even at 21.1T. Compounds that contain both structural H(2)O molecules and protonated arsenate groups do not yield resolvable signal, likely a result of T(2) effects related to a combination of strong quadrupolar interactions and proton exchange. Spin-echo experiments at 11.7 and 14.1T were effective for Li(3)AsO(4) and CsH(2)AsO(4), as were whole-pattern spikelet experiments for arsenate oxide (As(2)O(5)) at 17.6 and 21.1T. The central transition resonance of Ca(3)(AsO(4))(2).8H(2)O is approximately 6 MHz broad and required a non-conventional, histogram-style spikelet method at high field to improve acquisition efficiency. This approach reduces the acquisition time due to the sensitivity enhancement of the spikelet sequence and a reduction in the number of frequency increments required to map the resonance. Despite the large quadrupole couplings, we have identified a correlation between the (75)As isotropic chemical shift and the electronegativity of the next-nearest neighbor cation in arsenate compounds.

NMR study of strontium binding by a micaceous mineral.

The nature of strontium binding by soil minerals directly affects the transport and sequestration/remediation of radioactive strontium species released from leaking high-level nuclear waste storage tanks. However, the molecular-level structure of strontium binding sites has seldom been explored in phyllosilicate minerals by direct spectroscopic means and is not well-understood. In this work, we use solid-state NMR to analyze strontium directly and indirectly in a fully strontium-exchanged synthetic mica of nominal composition Na(4)Mg(6)Al(4)Si(4)O(20)F(4). Thermogravimetric analysis, X-ray diffraction analysis, and NMR evidence supports that heat treatment at 500 degrees C for 4 h fully dehydrates the mica, creating a hydrogen-free interlayer. Analysis of the strontium NMR spectrum of the heat-treated mica shows a single strontium environment with a quadrupolar coupling constant of 9.02 MHz and a quadrupolar asymmetry parameter of 1.0. These quadrupolar parameters are consistent with a highly distorted and asymmetric coordination environment that would be produced by strontium cations without water in the coordination sphere bound deep within the ditrigonal holes. Evidence for at least one additional strontium environment, where proton-strontium couplings may occur, was found via a (1)H-(87)Sr transfer of populations by double resonance NMR experiment. We conclude that the strontium cations in the proton-free interlayer are observable by (87)Sr NMR and bound through electrostatic interactions as nine coordinate inner-sphere complexes sitting in the ditrigonal holes. Partially hydrated strontium cations invisible to direct (87)Sr NMR are also present and located on the external mica surfaces, which are known to hydrate upon exposure to atmospheric moisture. These results demonstrate that modern pulsed NMR techniques and high fields can be used effectively to provide structural details of strontium binding by phyllosilicate minerals.

High-field QCPMG NMR of strontium nuclei in natural minerals.

The only stable NMR-active isotope of strontium, (87)Sr, is a spin-9/2 quadrupolar nucleus that has a low gyromagnetic ratio, a low natural abundance, and a large nuclear electric quadrupole moment. In this work, we utilize the quadrupolar Carr-Purcell-Meiboom-Gill (QCPMG) pulse sequence and a 21.14 T NMR spectrometer at the Pacific Northwest National Laboratory to characterize the strontium sites in the natural minerals strontianite (SrCO(3)) and celestine (SrSO(4)). QCPMG at 21.14 T was found to provide sensitivity enhancements of roughly two orders of magnitude over Hahn-echo experiments at an 11.74 T magnetic field. We extracted the quadrupolar parameters for the strontium nuclei through iterative simulations of the experimental spectra with the SIMPSON program by Bak, Rasmussen, and Nielsen. The data show that the quadrupolar parameters of (87)Sr appear to be highly sensitive to the symmetry of the strontium coordination environment and can thus provide information about the strontium binding environment in complex systems.

Synthesis and evaluation of 2-amino-4-[(18)F]fluoro-2-methylbutanoic acid (FAMB): relationship of amino acid transport to tumor imaging properties of branched fluorinated amino acids.

Radiolabeled amino acids represent a promising class of tumor imaging agents, and the determination of the optimal characteristics of these tracers remains an area of active investigation. A new (18)F-labeled branched amino acid, 2-amino-4-[(18)F]fluoro-2-methylbutanoic acid (FAMB), has been prepared in 36% decay-corrected yield using no-carrier-added [(18)F]fluoride. In vitro uptake assays with rat 9L gliosarcoma cells suggest that [(18)F]FAMB was transported primarily via the L type amino acid transport system. In vivo studies with [(18)F]FAMB demonstrated tumor to normal brain ratios of 14:1 in rats with intracranial 9L gliosarcoma tumors at 60 minutes after injection. Comparison of [(18)F]FAMB with structurally related (18)F-labeled branched amino acids demonstrated that A type transport in vitro was positively correlated with the tumor to brain ratios observed in vivo.

Efficacy of adenoviral p53 delivery with SCH58500 in the intracranial 9l and RG2 models.

Malignant gliomas remain incurable entities that provide fertile ground for experimental therapy. The observation that impaired p53 expression is present in a proportion of these tumors suggests that reconstitution of this ability may impart some degree of tumor control. In this investigation, the 9L and RG2 intracranial rodent tumor models are utilized to assess SCH58500, an adenoviral p53 delivery system. The RG2 tumors demonstrate a greater propensity for transfection with this vector in vitro than the 9L tumors. In vivo, little tumor transfection beyond the immediate area of the needle tract used for direct SCH58500 injection was observed in either tumor type. Intracarotid injection resulted in no tumor transfection. Even at high concentrations of SCH58500 or control virus, injections resulted in no apparent toxicity in terms of weight gain, eating habits or activity in normal animals. The intratumoral administration of SCH58500 enhances the survival of animals with established 9L tumors. Both SCH58500 and its control viral construct not containing the p53 gene enhance survival in animals with RG2 tumors. None of the injected viral constructs caused an alteration in the markers used to detect the character of the white cell infiltrate in either of these tumors. Utilization of SCH58500 provides measurable efficacy in these preclinical brain tumor models without significant toxicity.

Radiolabeled amino acids for tumor imaging with PET: radiosynthesis and biological evaluation of 2-amino-3-[18F]fluoro-2-methylpropanoic acid and 3-[18F]fluoro-2-methyl-2-(methylamino)propanoic acid.

Novel radiopharmaceuticals, including amino acids, that target neoplasms through their altered metabolic states have shown promising results in preclinical and clinical studies. Two fluorinated analogues of alpha-aminoisobutyric acid, 2-amino-3-fluoro-2-methylpropanoic acid (FAMP) and 3-fluoro-2-methyl-2-(methylamino)propanoic acid (N-MeFAMP), have been radiolabeled with fluorine-18, characterized in amino acid uptake assays, and evaluated in vivo in normal rats and a rodent tumor model. The key steps in the syntheses of both radiotracers involved the preparation of cyclic sulfamidate precursors. Radiosyntheses of both [18F]FAMP and [18F]N-MeFAMP via no-carrier-added nucleophilic substitution provided high yields (>78% decay-corrected) in high radiochemical purity (>99%). Amino acid transport assays using 9L gliosarcoma cells demonstrated that both compounds are substrates for the A type amino acid transport system, with [18F]N-MeFAMP showing higher specificity than [18F]FAMP for A type transport. Tissue distribution studies in normal Fischer rats and Fischer rats implanted intracranially with 9L gliosarcoma tumor cells were also performed. At 60 min postinjection, the tumor vs normal brain ratio of radioactivity was 36:1 in animals receiving [18F]FAMP and 104:1 in animals receiving [18F]N-MeFAMP. On the basis of these studies, both [18F]FAMP and [18F]N-MeFAMP are promising imaging agents for the detection of intracranial neoplasms via positron emission tomography.

Synthesis of syn- and anti-1-amino-3-[18F]fluoromethyl-cyclobutane-1-carboxylic acid (FMACBC), potential PET ligands for tumor detection.

syn- and anti-1-amino-3-[18F]fluoromethyl-cyclobutane-1-carboxylic acid (FMACBC, 16 and 17), analogues of anti-1-amino-3-[18F]fluorocyclobutyl-1-carboxylic acid (FACBC), were prepared to evaluate the contributions of C-3 substitution and configuration on the uptake of these radiolabeled amino acids in a rodent model of brain tumors. Radiofluorinated targets [18F]16 and [18F]17 were prepared by no-carrier-added radiofluorination from their corresponding methanesulfonyl esters 12 and 13, respectively, with decay-corrected radiochemical yields of 30% for [18F]16 and 20% for [18F]17. In amino acid transport assays performed in vitro using 9L gliosarcoma cells, both [18F]16 and [18F]17 were substrates for L type amino acid transport, while [18F]17 but not [18F]16 was a substrate for A type transport. Biodistribution studies in normal Fischer rats with [18F]16 and [18F]17 showed high uptake of radioactivity (>2.0% dose/g) in the pancreas while other tissues studied, including liver, heart, lung, kidney, blood, muscle, and testis, showed relatively low uptake of radioactivity (<1.0% dose/g). In rats implanted intracranially with 9L gliosarcoma cells, the retention of radioactivity in tumor tissue was high at 5, 60, and 120 min after intravenous injection of [18F]16 and [18F]17 while the uptake of radioactivity in brain tissue contralateral to the tumor remained low (<0.3% dose/g). Ratios of tumor uptake to normal brain uptake for [18F]16 were 7.5:1, 7:1, and 5:1 at 5, 60, and 120 min, respectively, while for [18F]17 the ratios were 7.5:1, 9:1, and 9:1 at the same time points. This work demonstrates that like anti-[18F]FACBC, [18F]16 and [18F]17 are excellent candidates for imaging brain tumors.

Differential expression between pilocytic and anaplastic astrocytomas: identification of apolipoprotein D as a marker for low-grade, non-infiltrating primary CNS neoplasms.

Fibrillary astrocytoma, the most common primary central nervous system neoplasm, is infiltrating, rapidly proliferating, and almost invariably fatal. This contrasts with the biologically distinct pilocytic astrocytoma, which is circumscribed, often cystic, slowly proliferating, and associated with a favorable long-term outcome. Diagnostic markers for distinguishing pilocytic astrocytomas from infiltrating anaplastic astrocytomas are currently not available. To identify genes that might either serve as markers or explain these distinct biologic behaviors, cDNA microarray analysis was used to compare the expression of 7,073 genes (nearly one quarter of the human genome) between these 2 types of astrocytoma. Messenger RNAs pooled from 3 pilocytic astrocytomas and from 4 infiltrating anaplastic astrocytomas were compared. Apolipoprotein D (apoD), which expressed 8.5-fold higher in pilocytic astrocytomas, showed the greatest level of differential expression and emerged as a potential marker for pilocytic tumors. By immunohistochemistry, 10 of 13 pilocytic astrocytomas stained positively for apoD, while none of 21 infiltrating astrocytomas showed similar staining. ApoD immunostaining was also seen in 9 of 14 of gangliogliomas, 4 of 5 subependymal giant cell astrocytomas (SEGAs), and a single pleomorphic xanthoastrocytomas (PXAs). By in situ hybridization, pilocytic astrocytomas, in contrast with infiltrating astrocytomas, showed widespread increased apoD expression. SAGE analysis using the NCBI database showed a higher level of expression of apoD RNA in pilocytic astrocytoma than in any of the other 94 neoplastic and non-neoplastic tissues in the database. ApoD is associated with decreased proliferation in some cell lines, and is the protein found in highest concentration in cyst fluid from benign cystic disease of the breast. ApoD might play a role in either decreased proliferation or cyst formation in pilocytic astrocytomas, gangliogliomas, SEGAs, and PXAs.