Rapid, efficient, and economical synthesis of PET tracers in a droplet microreactor: application to O-(2-[18F]fluoroethyl)-L-tyrosine ([18F]FET).

BackgroundConventional scale production of small batches of PET tracers (e.g. for preclinical imaging) is an inefficient use of resources. Using O-(2-[18F]fluoroethyl)-L-tyrosine ([18F]FET), we demonstrate that simple microvolume radiosynthesis techniques can improve the efficiency of production by consuming tiny amounts of precursor, and maintaining high molar activity of the tracers even with low starting activity.ProceduresThe synthesis was carried out in microvolume droplets manipulated on a disposable patterned silicon "chip" affixed to a heater. A droplet of [18F]fluoride containing TBAHCO3 was first deposited onto a chip and dried at 100 °C. Subsequently, a droplet containing 60 nmol of precursor was added to the chip and the fluorination reaction was performed at 90 °C for 5 min. Removal of protecting groups was accomplished with a droplet of HCl heated at 90 °C for 3 min. Finally, the crude product was collected in a methanol-water mixture, purified via analytical-scale radio-HPLC and formulated in saline. As a demonstration, using [18F]FET produced on the chip, we prepared aliquots with different molar activities to explore the impact on preclinical PET imaging of tumor-bearing mice.ResultsThe microdroplet synthesis exhibited an overall decay-corrected radiochemical yield of 55 ± 7% (n = 4) after purification and formulation. When automated, the synthesis could be completed in 35 min. Starting with < 370 MBq of activity, ~ 150 MBq of [18F]FET could be produced, sufficient for multiple in vivo experiments, with high molar activities (48-119 GBq/μmol). The demonstration imaging study revealed the uptake of [18F]FET in subcutaneous tumors, but no significant differences in tumor uptake as a result of molar activity differences (ranging 0.37-48 GBq/μmol) were observed.ConclusionsA microdroplet synthesis of [18F]FET was developed demonstrating low reagent consumption, high yield, and high molar activity. The approach can be expanded to tracers other than [18F]FET, and adapted to produce higher quantities of the tracer sufficient for clinical PET imaging

Response Rates of Online Surveys in Published Research: A Meta-analysis

The response rates of online surveys were often examined in the literature by comparing to other modes of surveys. Questions regarding what constitutes a respectable response rate for online surveys in research remained unanswered. To fill in the knowledge gap, we conducted a comprehensive search, screened 8672 studies, and examined 1071 online survey response rates reported in education-related research. Our analyses showed the number of online surveys in published research grew steadily across the years. The average online survey response rate is 44.1%. Our results indicate that sending an online survey to more participants did not generate a higher response rate. Instead, sending surveys to a clearly defined and refined population positively impacts the online survey response rate. In addition, pre-contacting potential participants, using other types of surveys in conjunction with online surveys, and using phone calls to remind participants about the online survey could also yield a higher response rate. The use of incentives did not show a significant impact on the response rate of online surveys. Other factors that impacted the rates included the funding status of a project, and the age and occupation of the participants. Concrete suggestions for reviewing and improving the online survey response rates are provided

Non-Abelian Medium Effects in Quark-Gluon Plasma

Based on the kinetic theory, the non-Abelian medium property of hot Quark-Gluon Plasma is investigated. The nonlinearity of the plasma comes from two aspects: The nonlinear wave-wave interaction and self-interaction of color field. The non-Abelian color permittivity is obtained by expanding the kinetic equations to third order. As an application, the nonlinear Landau damping rate and the nonlinear eigenfrequency shift are calculated in the longwave length limit.Comment: 12 pages(Revtex), no figure

Who\u27s accessing emergency food services?

Introduction: Last year, Chittenden Emergency Food Shelf provided 1,260,517 pounds of food to over 11,000 people each month via groceries, hot meals and home delivery, supplying an average of almost 40% of food for families. CEFS seeks to improve their services and offerings by better understanding the demographics, food preference, and needs of the clients they serve. Our goal was to collect demographic and utilization data to identify areas where CEFS could enhance services and improve client access to healthful food.https://scholarworks.uvm.edu/comphp_gallery/1208/thumbnail.jp

Freshwater Composition and Connectivity of the Connecticut River Plume During Ambient Flood Tides

The Connecticut River plume interacts with the strong tidal currents of the ambient receiving waters in eastern Long Island Sound. The plume formed during ambient flood tides is studied as an example of tidal river plumes entering into energetic ambient tidal environments in estuaries or continental shelves. Conservative passive freshwater tracers within a high-resolution nested hydrodynamic model are applied to determine how source waters from different parts of the tidal cycle contribute to plume composition and interact with bounding plume fronts. The connection to source waters can be cut off only under low-discharge conditions, when tides reverse surface flow through the mouth after max ambient flood. Upstream plume extent is limited because ambient tidal currents arrest the opposing plume propagation, as the tidal internal Froude number exceeds one. The downstream extent of the tidal plume always is within 20 km from the mouth, which is less than twice the ambient tidal excursion. Freshwaters in the river during the preceding ambient ebb are the oldest found in the new flood plume. Connectivity with source waters and plume fronts exhibits a strong upstream-to- downstream asymmetry. The arrested upstream front has high connectivity, as all freshwaters exiting the mouth immediately interact with this boundary. The downstream plume front has the lowest overall connectivity, as interaction is limited to the oldest waters since younger interior waters do not overtake this front. The offshore front and inshore boundary exhibit a downstream progression from younger to older waters and decreasing overall connectivity with source waters. Plume-averaged freshwater tracer concentrations and variances both exhibit an initial growth period followed by a longer decay period for the remainder of the tidal period. The plume-averaged tracer variance is increased by mouth inputs, decreased by entrainment, and destroyed by internal mixing. Peak entrainment velocities for younger waters are higher than values for older waters, indicating stronger entrainment closer to the mouth. Entrainment and mixing time scales (1–4 h at max ambient flood) are both shorter than half a tidal period, indicating entrainment and mixing are vigorous enough to rapidly diminish tracer variance within the plume

A Database of Teleseismic Shear-Wave Splitting Measurements for the Ordos Block and Adjacent Areas

The Ordos block of the north China craton and its surrounding regions are affected by the India-Eurasia collision to the southwest and the subduction of the Pacific beneath the Eurasian plates to the east. To provide the foundation for delineating lithospheric deformation and asthenospheric flow beneath this tectonically diverse region, we have created a database of individual shear-wave splitting (SWS) parameters by applying a uniform set of data processing procedures. After automatic data processing and manual checking, a total of 16,228 pairs of well-defined PKS, SKKS, and SKS splitting parameters (fast orientations and splitting times) are obtained from 1023 broadband seismic stations that recorded data between 2007 and 2019. Along the western and southern margins of the Ordos block, the observed seismic anisotropy is attributable to mantle flow deflected by the relatively thick lithospheric root of the Ordos block. A clear back-Azimuthal dependence of the fast orientations is observed at some of the stations located in the Alxa block, Ordos block, and Sichuan basin, indicating possible existence of complex anisotropic structures. The new SWS database can be employed by researchers in various fields to study lithospheric deformation and asthenospheric flow beneath the Ordos block and surrounding regions

Recognition of nonproline N-terminal residues by the Pro/N-degron pathway

Eukaryotic N-degron pathways are proteolytic systems whose unifying feature is their ability to recognize proteins containing N-terminal (Nt) degradation signals called N-degrons, and to target these proteins for degradation by the 26S proteasome or autophagy. GID4, a subunit of the GID ubiquitin ligase, is the main recognition component of the proline (Pro)/N-degron pathway. GID4 targets proteins through their Nt-Pro residue or a Pro at position 2, in the presence of specific downstream sequence motifs. Here we show that human GID4 can also recognize hydrophobic Nt-residues other than Pro. One example is the sequence Nt-IGLW, bearing Nt-Ile. Nt-IGLW binds to wild-type human GID4 with a K_d of 16 μM, whereas the otherwise identical Nt-Pro–bearing sequence PGLW binds to GID4 more tightly, with a K_d of 1.9 μM. Despite this difference in affinities of GID4 for Nt-IGLW vs. Nt-PGLW, we found that the GID4-mediated Pro/N-degron pathway of the yeast Saccharomyces cerevisiae can target an Nt-IGLW–bearing protein for rapid degradation. We solved crystal structures of human GID4 bound to a peptide bearing Nt-Ile or Nt-Val. We also altered specific residues of human GID4 and measured the affinities of resulting mutant GID4s for Nt-IGLW and Nt-PGLW, thereby determining relative contributions of specific GID4 residues to the GID4-mediated recognition of Nt-Pro vs. Nt-residues other than Pro. These and related results advance the understanding of targeting by the Pro/N-degron pathway and greatly expand the substrate recognition range of the GID ubiquitin ligase in both human and yeast cells

Vaginal Submucosal Dendritic Cells, but Not Langerhans Cells, Induce Protective Th1 Responses to Herpes Simplex Virus-2

Herpes simplex virus (HSV) type 2 infection occurs primarily at the genital mucosal surfaces and is a leading cause of ulcerative lesions. Despite the availability of animal models for HSV-2 infection, little is known regarding the mechanism of immune induction within the vaginal mucosa. Here, we examined the cell types responsible for the initiation of protective Th1 immunity to HSV-2. Intravaginal inoculation of HSV-2 led to a rapid recruitment of submucosal dendritic cells (DCs) to the infected epithelium. Subsequently, CD11c+ DCs harboring viral peptides in the context of MHC class II molecules emerged in the draining lymph nodes and were found to be responsible for the stimulation of IFNγ secretion from HSV-specific CD4+ T cells. Other antigen-presenting cells including B cells and macrophages did not present viral peptides to T cells in the draining lymph nodes. Next, we assessed the relative contribution to immune generation by the Langerhans cells in the vaginal epithelium, the submucosal CD11b+ DCs, and the CD8α+ lymph node DCs. Analysis of these DC populations from the draining lymph nodes revealed that only the CD11b+ submucosal DCs, but not Langerhans cell–derived or CD8α+ DCs, presented viral antigens to CD4+ T cells and induced IFNγ secretion. These results demonstrate a previously unanticipated role for submucosal DCs in the generation of protective Th1 immune responses to HSV-2 in the vaginal mucosa, and suggest their importance in immunity to other sexually transmitted diseases

Recognition of nonproline N-terminal residues by the Pro/N-degron pathway

Eukaryotic N-degron pathways are proteolytic systems whose unifying feature is their ability to recognize proteins containing N-terminal (Nt) degradation signals called N-degrons, and to target these proteins for degradation by the 26S proteasome or autophagy. GID4, a subunit of the GID ubiquitin ligase, is the main recognition component of the proline (Pro)/N-degron pathway. GID4 targets proteins through their Nt-Pro residue or a Pro at position 2, in the presence of specific downstream sequence motifs. Here we show that human GID4 can also recognize hydrophobic Nt-residues other than Pro. One example is the sequence Nt-IGLW, bearing Nt-Ile. Nt-IGLW binds to wild-type human GID4 with a K_d of 16 μM, whereas the otherwise identical Nt-Pro–bearing sequence PGLW binds to GID4 more tightly, with a K_d of 1.9 μM. Despite this difference in affinities of GID4 for Nt-IGLW vs. Nt-PGLW, we found that the GID4-mediated Pro/N-degron pathway of the yeast Saccharomyces cerevisiae can target an Nt-IGLW–bearing protein for rapid degradation. We solved crystal structures of human GID4 bound to a peptide bearing Nt-Ile or Nt-Val. We also altered specific residues of human GID4 and measured the affinities of resulting mutant GID4s for Nt-IGLW and Nt-PGLW, thereby determining relative contributions of specific GID4 residues to the GID4-mediated recognition of Nt-Pro vs. Nt-residues other than Pro. These and related results advance the understanding of targeting by the Pro/N-degron pathway and greatly expand the substrate recognition range of the GID ubiquitin ligase in both human and yeast cells

Planned Missing Data Designs & Small Sample Size: How Small is Too Small?

Utilizing planned missing data (PMD) designs (ex. 3-form surveys) enables researchers to ask participants fewer questions during the data collection process. An important question, however, is just how few participants are needed to effectively employ planned missing data designs in research studies. This paper explores this question by using simulated three-form planned missing data to assess analytic model convergence, parameter estimate bias, standard error bias, mean squared error (MSE), and relative efficiency (RE).Three models were examined: a one-time point, cross-sectional model with 3 constructs; a two-time point model with 3 constructs at each time point; and a three-time point, mediation model with 3 constructs over three time points. Both full-information maximum likelihood (FIML) and multiple imputation (MI) were used to handle the missing data. Models were found to meet convergence rate and acceptable bias criteria with FIML at smaller sample sizes than with MI