An Integrated Platform for in vivo Electrophysiology in Spatial Cognition Experiments.

Spatial cognition research requires behavioral paradigms that can distinguish between different navigational elements, such as allocentric (map-like) navigation and egocentric (e.g., body centered) navigation. To fill this need, we developed a flexible experimental platform that can be quickly modified without the need for significant changes to software and hardware. In this paper, we present this inexpensive and flexible behavioral platform paired with software which we are making freely available. Our behavioral platform serves as the foundation for a range of experiments, and though developed for assessing spatial cognition, it also has applications in the non-spatial domain of behavioral testing. There are two components of the software platform, 'Maze' and 'Stim Trigger'. Both programs can work in conjunction with electrophysiology acquisition systems, allowing for precise time stamping of neural events with behavior. The Maze program includes functionality for automatic reward delivery based on user defined zones. 'Stim Trigger' permits control of brain stimulation via any equipment that can be paired with an Arduino board. We seek to share our software and leverage the potential by expanding functionality in the future to meet the needs of a larger community of researchers.

Excess Heat Capacity in Mo/Au Transition Edge Sensor Bolometric Detectors.

Excess heat capacity in a bolometric detector has the consequence of increasing or leading to multiple device time constants. The Mo/Au bilayer transition edge sensor (TES) bolometric detectors initially fabricated for the high resolution mid-infrared spectrometer (HIRMES) exhibited two response thermalization scales, one of which is a few times longer than estimates based upon the properties of the bulk materials employed in the design. The relative contribution of this settling time to the overall time response of the detectors is roughly proportional to the pixel area, which ranges between ~0.3 and 2.6 mm2. Use of laser ablation to remove sections of the silicon membranes comprising the pixels results in a detector response with a smaller contribution from the secondary time constant. Additional information about the nature of this excess heat capacity is gleaned from glancing incidence x-ray diffraction, which reveals the presence of molybdenum silicides near the silicon surface which is a consequence of the bi-layer deposition. Quantitative analysis of the concentration of excess molybdenum, estimated with secondary ion mass spectroscopy, is commensurate to the additional heat capacity needed to explain the anomalous time response of the detectors.

Simple, compact, high-resolution monochromatic x-ray source for characterization of x-ray calorimeter arrays.

X-ray calorimeters routinely achieve very high spectral resolution, typically a few eV full width at half maximum (FWHM). Measurements of calorimeter line shapes are usually dominated by the natural linewidth of most laboratory calibration sources. This compounds the data acquisition time necessary to statistically sample the instrumental line broadening and can add systematic uncertainty if the intrinsic line shape of the source is not well known. To address these issues, we have built a simple, compact monochromatic x-ray source using channel cut crystals. A commercial x-ray tube illuminates a pair of channel cut crystals that are aligned in a dispersive configuration to select the Kα1 line of the x-ray tube anode material. The entire device, including the x-ray tube, can be easily hand-carried by one person and may be positioned manually or using a mechanical translation stage. The output monochromatic beam provides a collimated image of the anode spot with magnification of unity in the dispersion direction (typically 100 µm-200 µm for the x-ray tubes used here) and is unfocused in the cross-dispersion direction so that the source image in the detector plane appears as a line. We measured output count rates as high as 10 count/s/pixel for the Hitomi soft x-ray spectrometer, which had 819 µm square pixels. We implemented different monochromator designs for energies of 5.4 keV (one design) and 8.0 keV (two designs), which have effective theoretical FWHM energy resolution of 0.125 eV, 0.197 eV, and 0.086 eV, respectively; these are well-suited for optimal calibration measurements of state-of-the art x-ray calorimeters. We measured an upper limit for the energy resolution of our Cr Kα1 monochromator of 0.7 eV FWHM at 5.4 keV, consistent with the theoretical prediction of 0.125 eV.

Sub-Kelvin cooling for two kilopixel bolometer arrays in the PIPER receiver.

The Primordial Inflation Polarization Explorer (PIPER) is a balloon-borne telescope mission to search for inflationary gravitational waves from the early universe. PIPER employs two 32 × 40 arrays of superconducting transition-edge sensors, which operate at 100 mK. An open bucket Dewar of liquid helium maintains the receiver and telescope optics at 1.7 K. We describe the thermal design of the receiver and sub-Kelvin cooling with a continuous adiabatic demagnetization refrigerator (CADR). The CADR operates between 70 and 130 mK and provides ≈10 µW cooling power at 100 mK, nearly five times the loading of the two detector assemblies. We describe electronics and software to robustly control the CADR, overall CADR performance in flightlike integrated receiver testing, and practical considerations for implementation in the balloon float environment.

Analysis and calibration techniques for superconducting resonators.

A method is proposed and experimentally explored for in-situ calibration of complex transmission data for superconducting microwave resonators. This cryogenic calibration method accounts for the instrumental transmission response between the vector network analyzer reference plane and the device calibration plane. Once calibrated, the observed resonator response is analyzed in detail by two approaches. The first, a phenomenological model based on physically realizable rational functions, enables the extraction of multiple resonance frequencies and widths for coupled resonators without explicit specification of the circuit network. In the second, an ABCD-matrix representation for the distributed transmission line circuit is used to model the observed response from the characteristic impedance and propagation constant. When used in conjunction with electromagnetic simulations, the kinetic inductance fraction can be determined with this method with an accuracy of 2%. Datasets for superconducting microstrip and coplanar-waveguide resonator devices were investigated and a recovery within 1% of the observed complex transmission amplitude was achieved with both analysis approaches. The experimental configuration used in microwave characterization of the devices and self-consistent constraints for the electromagnetic constitutive relations for parameter extraction are also presented.

Micro-Spec: an ultracompact, high-sensitivity spectrometer for far-infrared and submillimeter astronomy.

High-performance, integrated spectrometers operating in the far-infrared and submillimeter ranges promise to be powerful tools for the exploration of the epochs of reionization and initial galaxy formation. These devices, using high-efficiency superconducting transmission lines, can achieve the performance of a meter-scale grating spectrometer in an instrument implemented on a 4 inch silicon wafer. Such a device, when combined with a cryogenic telescope in space, provides an enabling capability for studies of the early universe. Here, the optical design process for Micro-Spec (µ-Spec) is presented, with particular attention given to its two-dimensional diffractive region, where the light of different wavelengths is focused on the different detectors. The method is based on the stigmatization and minimization of the light path function in this bounded region, which results in an optimized geometrical configuration. A point design with an efficiency of ~90% has been developed for initial demonstration and can serve as the basis for future instruments. Design variations on this implementation are also discussed, which can lead to lower efficiencies due to diffractive losses in the multimode region.

Properties of a variable-delay polarization modulator.

We investigate the polarization modulation properties of a variable-delay polarization modulator (VPM). The VPM modulates polarization via a variable separation between a polarizing grid and a parallel mirror. We find that in the limit where the wavelength is much larger than the diameter of the metal wires that comprise the grid, the phase delay derived from the geometric separation between the mirror and the grid is sufficient to characterize the device. However, outside of this range, additional parameters describing the polarizing grid geometry must be included to fully characterize the modulator response. In this paper, we report test results of a VPM at wavelengths of 350 µm and 3 mm. Electromagnetic simulations of wire grid polarizers were performed and are summarized using a simple circuit model that incorporates the loss and polarization properties of the device.

Modeling the intensity and polarization response of planar bolometric detectors.

Far-infrared bolometric detectors are used extensively in ground-based and space-borne astronomy, and thus it is important to understand their optical behavior precisely. We have studied the intensity and polarization response of free-space bolometers and shown that when the size of the absorber is reduced below a wavelength, the response changes from being that of a classical optical detector to that of a few-mode antenna. We have calculated the modal content of the reception patterns and found that for any volumetric detector having a side length of less than a wavelength, three magnetic and three electric dipoles characterize the behavior. The size of the absorber merely determines the relative strengths of the contributions. The same formalism can be applied to thin-film absorbers, where the induced current is forced to flow in a plane. In this case, one magnetic and two electric dipoles characterize the behavior. The ability to model easily the intensity, polarization, and straylight characteristics of electrically small detectors will be of great value when designing high-performance polarimetric imaging arrays.

The Hertz/VPM polarimeter: design and first light observations.

We present first results of Hertz/VPM, the first submillimeter polarimeter employing the dual Variable-delay Polarization Modulator (dual-VPM). This device differs from previously used polarization modulators in that it operates in translation, rather than mechanical rotation. We discuss the basic theory behind this device and its potential advantages over the commonly used half-wave plate. The dual-VPM was tested both at the Submillimeter Telescope Observatory and in the laboratory. In each case we present a detailed description of the setup. We discovered that properties of the VPM wire grids (diameter and spacing) caused behavior that differs from theoretical predictions for ideal wire grid performance. By modifying the polarimeter settings to compensate for this behavior, we found that the dual-VPM system is robust, operating with high efficiency and low instrumental polarization. This device is well suited for air- and space-borne applications.

Interferometric polarization control.

We develop the Jones and Mueller matrices for structures that allow control of the path length difference between two linear orthogonal polarizations and consider the effect of placing multiple devices in series. Specifically, we find that full polarization modulation (measurement of Stokes Q, U, and V) can be achieved by placing two such modulators in series if the relative angles of the beam-splitting grids with respect to the analyzer orientation are appropriately chosen. Such a device has several potential advantages over a spinning wave plate modulator for measuring astronomical polarization in the far infrared through millimeter: (i) The use of small, linear motions eliminates the need for cryogenic rotational bearings; (ii) the phase flexibility allows measurement of circular as well as linear polarization; and (iii) this architecture allows for both multiwavelength and broadband modulation. We also present initial laboratory results.

Tracing the first stars with fluctuations of the cosmic infrared background.

The deepest space- and ground-based observations find metal-enriched galaxies at cosmic times when the Universe was less than 1 Gyr old. These stellar populations had to be preceded by the metal-free first stars, known as 'population III'. Recent cosmic microwave background polarization measurements indicate that stars started forming early--when the Universe was < or =200 Myr old. It is now thought that population III stars were significantly more massive than the present metal-rich stellar populations. Although such sources will not be individually detectable by existing or planned telescopes, they would have produced significant cosmic infrared background radiation in the near-infrared, whose fluctuations reflect the conditions in the primordial density field. Here we report a measurement of diffuse flux fluctuations after removing foreground stars and galaxies. The anisotropies exceed the instrument noise and the more local foregrounds; they can be attributed to emission from population III stars, at an era dominated by these objects.

Detection of simple mutations and polymorphisms in large genomic regions.

We have developed a novel technology that makes it possible to detect simple nucleotide polymorphisms directly within a sample of total genomic DNA. It allows, in a single Southern blot experiment, the determination of sequence identity of genomic regions with a combined length of hundreds of kilobases. This technology does not require PCR amplification of the target DNA regions, but exploits preparative size-fractionation of restriction-digested genomic DNA and a newly discovered property of the mismatch-specific endonuclease CEL I to cleave heteroduplex DNA with a very high specificity and sensitivity. We have used this technique to detect various simple mutations directly in the genomic DNA of isogenic pairs of recombinant Pseudomonas aeruginosa, Escherichia coli and Salmonella isolates. Also, by using a cosmid DNA library and genomic fractions as hybridization probes, we have compared total genomic DNA of two clinical P.aeruginosa clones isolated from the same patient, but exhibiting divergent phenotypes. The mutation scan correctly detected a GA insertion in the quorum-sensing regulator gene rhlR and, in addition, identified a novel intragenomic polymorphism in rrn operons, indicating very high stability of the bacterial genomes under natural non-mutator conditions.

The novel IMPDH inhibitor VX-497 prolongs skin graft survival and improves graft versus host disease in mice.

VX-497 is the first inosine-5'-monophosphate dehydrogenase (IMPDH) inhibitor generated in a structure-based drug design program specifically addressing the tolerability problems of currently available immunosuppressive drugs. The pharmacological activity of the compound has been examined in murine skin transplantation and graft versus host disease (GVHD) models. In the skin transplant study, trunk skin grafts from Balb/c mice were grafted onto C57Bl/6 mice. Mice were administered vehicle or VX-497 twice daily until day 10. Mean survival of skin grafts on vehicle-treated animals was 9.9 +/- 0.9 days. Graft survival was prolonged significantly in animals treated with VX-497 to 13.2 +/- 1.2 (p < 0.001, Kaplan Meier Log-Rank test) days in the 50 mg/kg group and 13.9 +/- 1.0 (p < 0.001) days in the 85 mg/kg group. In the GVHD study, 150 x 10(6) nonadherent splenocytes from B6 mice were injected intravenously into the F1 hybrid strain B6DBA/2. Groups of animals (n = 6) were administered vehicle or 50 or 100 mg/kg VX-497 b.i.d for 8 days. Animals were sacrificed and spleen weights and interferon-gamma (IFN-gamma) serum levels were determined by enzyme-linked immunosorbent assay. In addition, spontaneous spleen cell proliferation was measured using a 3H-thymidine uptake assay. Isografted F1 animals served as controls. GVHD developed in the vehicle-treated allografted F1 mice and treatment with VX-497 improved all manifestations of the disease significantly. The 2.9-fold increase in spleen weight in allografted animals was reduced to a 1.6-fold increase in the VX-497-treated mice. Serum IFN-gamma levels were increased 54-fold in the vehicle group while there was a 7.4-fold increase in VX-497-treated animals. Spontaneous spleen cell proliferation was increased 9.9-fold in the absence of VX-497 and there was a 3.5-fold increase in its presence. Thus, VX-497 has been shown to be effective in both a skin transplantation and a GVHD model in the mouse. The demonstrated pharmacological activity of VX-497 in these murine transplantation models warrants further evaluation of the drug in transplantation indications.

A tripeptide sequence within the nascent DaaP protein is required for mRNA processing of a fimbrial operon in Escherichia coli.

The biogenesis of F1845 fimbriae, a member of the Dr family of Escherichia coli adhesins, is regulated by endonucleolytic cleavage of the daaABCDPE primary transcript and differential stability of the resulting cleavage products. Processing of daa mRNA is dependent upon translation of a small open reading frame, designated daaP, which flanks the daa processing site. Here, we demonstrate that daa mRNA processing is directly coupled to daaP translation. Cleavage of the daaA-E mRNA was shown to require the tripeptide Gly-Pro-Pro (GPP), encoded by daaP codons 49-51 downstream of the processing site. Processing also required active translation through RNA located upstream of the processing site; however, processing did not depend on the amino acid sequence encoded by the region of daaP upstream of the processing site. Finally, determination of the processing site was shown to involve its location relative to the codons encoding the GPP tripeptide. These data show that translation of daaP is required in cis to promote RNA processing. These data suggest a model involving interaction of the nascent GPP tripeptide portion of the DaaP polypeptide with the ribosome, triggering cleavage of the associated mRNA at a fixed distance upstream. A model of active involvement of the ribosome in this process is proposed.

Benchmarking graduate health administration programs: stakeholders' perceptions.

The purpose of this study was to identify and compare among key stakeholders the factors in graduate health care administration education that are perceived to be important for ranking, or benchmarking, based on the opinions of those stakeholders, i.e., program directors, faculty, graduate students, and accrediting agency commissioners. We used an original survey to obtain stakeholders' perceptions and opinions about important process and outcome measures. We sent it to all ACEHSA-accredited graduate health care administration programs in the United States, Canada, and Puerto Rico; to full-time faculty members in each program; to three current graduate students in each program, and to all ACEHSA commissioners. We performed frequency of responses, Analysis of Variances (ANOVA) tests, and Dunnett T3 tests. A response rate of 32 percent (n = 156) was achieved for all stakeholders. A total of 67 percent of all respondents reported that benchmarking graduate health care administration programs was important. The study results revealed a significant difference between populations on the importance of evaluating certain process and outcome measures related to curriculum, research, student characteristics, and resources. However, most of the stakeholders reported that curriculum, faculty, and graduate student performance were the key quality indicators of a program. The results of this study provide preliminary information for health care administration programs to begin to develop an educational benchmarking effort.

IS1414, an Escherichia coli insertion sequence with a heat-stable enterotoxin gene embedded in a transposase-like gene.

Enteroaggregative Escherichia coli (EAEC) heat-stable enterotoxin 1 (EAST1) was originally discovered in EAEC but has also been associated with enterotoxigenic E. coli (ETEC). Multiple genomic restriction fragments from each of three ETEC strains of human origin showed homology with an EAST1 gene probe. A single hybridizing fragment was detected on the plasmid of ETEC strain 27D that also encodes heat-stable enterotoxin Ib and colonization factor antigen I. We isolated and characterized this fragment, showing that it (i) carries an allele of astA nearly identical to that originally reported from EAEC 17-2 and (ii) expressed enterotoxic activity. Sequence analysis of the toxin coding region revealed that astA is completely embedded within a 1,209-bp open reading frame (ORF1), whose coding sequence is on the same strand but in the -1 reading frame in reference to the toxin gene. In vitro expression of the predicted M(r)- approximately 46,000 protein product of ORF1 was demonstrated. ORF1 is highly similar to transposase genes of IS285 from Yersinia pestis, IS1356 from Burkholderia cepacia, and ISRm3 from Rhizobium meliloti. It is bounded by 30-bp imperfect inverted repeat sequences and flanked by 8-bp direct repeats. Based on these structural features, pathognomonic of a regular insertion sequence, this element was designated IS1414. Preliminary experiments to show IS1414 translocation were unsuccessful. Overlapping genes of the type suggested by the IS1414 core region have heretofore not been described in bacteria. It seems to offer a most efficient mechanism for intragenomic and horizontal dissemination of EAST1.

Structural and functional lesions in brush border of human polarized intestinal Caco-2/TC7 cells infected by members of the Afa/Dr diffusely adhering family of Escherichia coli.

Diffusely adhering Escherichia coli (DAEC) strains expressing F1845 fimbrial adhesin or Dr hemagglutinin belonging to the Afa/Dr family of adhesins infect cultured polarized human intestinal cells through recognition of the brush border-associated decay-accelerating factor (DAF; CD55) as a receptor. The wild-type Afa/Dr DAEC strain C1845 has been shown to induce brush border lesions by an adhesin-dependent mechanism triggering apical F-actin rearrangements. In the present study, we undertook to further characterize cell injuries following the interaction of wild-type Afa/Dr DAEC strains C1845 and IH11128 expressing fimbrial F1845 adhesin and Dr hemagglutinin, respectively, with polarized, fully differentiated Caco-2/TC7 cells. In both cases, bacterium-cell interaction was followed by rearrangement of the major brush border-associated cytoskeletal proteins F-actin, villin, and fimbrin, proteins which play a pivotal role in brush border assembly. In contrast, distribution of G-actin, actin-depolymerizing factor, and tubulin was not modified. Using draE mutants, we found that a mutant in which cysteine replaces aspartic acid at position 54 conserved binding capacity but failed to induce F-actin disassembly. Accompanying the cytoskeleton injuries, we found that the distribution of brush border-associated functional proteins sucrase-isomaltase (SI), dipeptidylpeptidase IV (DPPIV), glucose transporter SGLT1, and fructose transporter GLUT5 was dramatically altered. In parallel, SI and DPPIV enzyme activity decreased.