TCR Convergence in Individuals Treated With Immune Checkpoint Inhibition for Cancer.

Tumor antigen-driven selection may expand T cells having T cell receptors (TCRs) of shared antigen specificity but different amino acid or nucleotide sequence in a process known as TCR convergence. Substitution sequencing errors introduced by TCRβ (TCRB) repertoire sequencing may create artifacts resembling TCR convergence. Given the anticipated differences in substitution error rates across different next-generation sequencing platforms, the choice of platform could be consequential. To test this, we performed TCRB sequencing on the same peripheral blood mononuclear cells (PBMC) from individuals with cancer receiving anti-CTLA-4 or anti-PD-1 using an Illumina-based approach (Sequenta) and an Ion Torrent-based approach (Oncomine TCRB-LR). While both approaches found similar TCR diversity, clonality, and clonal overlap, we found that Illumina-based sequencing resulted in higher TCR convergence than with the Ion Torrent approach. To build upon this initial observation we conducted a systematic comparison of Illumina-based TCRB sequencing assays, including those employing molecular barcodes, with the Oncomine assay, revealing differences in the frequency of convergent events, purportedly artifactual rearrangements, and sensitivity of detection. Finally, we applied the Ion Torrent-based approach to evaluate clonality and convergence in a cohort of individuals receiving anti-CTLA-4 blockade for cancer. We found that clonality and convergence independently predicted response and could be combined to improve the accuracy of a logistic regression classifier. These results demonstrate the importance of the sequencing platform in assessing TCRB convergence

Actuated Hybrid Mirror Telescope

The figure depicts the planned Actuated Hybrid Mirror Telescope (AHMT), which is intended to demonstrate a new approach to the design and construction of wide-aperture spaceborne telescopes for astronomy and Earth science. This technology is also appropriate for Earth-based telescopes. The new approach can be broadly summarized as using advanced lightweight mirrors that can be manufactured rapidly at relatively low cost. More specifically, it is planned to use precise replicated metallic nanolaminate mirrors to obtain the required high-quality optical finishes. Lightweight, dimensionally stable silicon carbide (SiC) structures will support the nanolaminate mirrors in the required surface figures. To enable diffraction- limited telescope performance, errors in surface figures will be corrected by use of mirror-shape-control actuators that will be energized, as needed, by a wave-front-sensing and control system. The concepts of nanolaminate materials and mirrors made from nanolaminate materials were discussed in several previous NASA Tech Briefs articles. Nanolaminates constitute a relatively new class of materials that can approach theoretical limits of stiffness and strength. Nanolaminate mirrors are synthesized by magnetron sputter deposition of metallic alloys and/or compounds on optically precise master surfaces to obtain optical-quality reflector surfaces backed by thin shell structures. As an integral part of the deposition process, a layer of gold that will constitute the reflective surface layer is deposited first, eliminating the need for a subsequent and separate reflective-coating process. The crystallographic textures of the nanolaminate will be controlled to optimize the performance of the mirror. The entire deposition process for making a nanolaminate mirror takes less than 100 hours, regardless of the mirror diameter. Each nanolaminate mirror will be bonded to its lightweight SiC supporting structure. The lightweight nanolaminate mirrors and SiC supporting structures will be fabricated from reusable master molds. The mirror-shape-control actuators will be low-power, high-capacitance lead magnesium niobate electrostrictive actuators that will be embedded in the SiC structures. The mode of operation of these actuators will be such that once power was applied, they will change in length and once power was removed, they will maintain dimensional stability to nanometer precision. This mode of operation will enable the use of low-power, minimally complex electronic control circuitry. The wave-front-sensing and control system will be designed and built according to a two-stage architecture. The first stage will be implemented by a Shack- Hartmann (SH) sensor subsystem, which will provide a large capture range. The second, higher-performance stage will be implemented by an image-based wave-front-sensing subsystem that will include a phase-retrieval camera (PRC), and will utilize phase retrieval and other techniques to measure wavefront error directly. Phase retrieval is a process in which multiple images of an unresolved object are iterated to estimate the phase of the optical system that acquired the images. The combination of SH and phase-retrieval sensors will afford the virtues of both a dynamic range of 105 and an accuracy of <10 nm

Standard-Level Herbivory in an Old-Growth Conifer Forest Canopy

Herbivory is an important ecological process in forest canopies but is difficult to measure, especially for whole stands. We used the Wind River Canopy Crane in Washington State to access 101 randomly-located sample points throughout the forest canopy. This provided a relatively quick and convenient way to estimate herbivory for a whole stand. The overall level of herbivory was estimated at 1.6% of leaf area. The distribution was strongly skewed to the lower canopy where broad-leafed species experienced higher levels of herbivory. Herbivory averaged 0.3% in conifers and 13.5% in broad-leafed species. Fully half of the sample points had no detectable herbivory. Herbivory in this old-growth conifer forest is among the lowest levels published for forests around the globe and may reflect the general levels of herbivory in temperate coniferous forests during nonoutbreak conditions. Our whole-stand estimate is the first attempt at measuring herbivory for an entire forest stand in the Pacific Northwest

Restoration of Apollo Data by the NSSDC and the PDS Lunar Data Node

The Lunar Data Node (LDN), under the auspices of the Geosciences Node of the Planetary Data System (PDS), is restoring Apollo data archived at the National Space Science Data Center. The Apollo data were arch ived on older media (7 -track tapes. microfilm, microfiche) and in ob solete digital formats, which limits use of the data. The LDN is maki ng these data accessible by restoring them to standard formats and archiving them through PDS. The restoration involves reading the older m edia, collecting supporting data (metadata), deciphering and understa nding the data, and organizing into a data set. The data undergo a pe er review before archive at PDS. We will give an update on last year' s work. We have scanned notebooks from Otto Berg, P.1. for the Lunar Ejecta and Meteorites Experiment. These notebooks contain information on the data and calibration coefficients which we hope to be able to use to restore the raw data into a usable archive. We have scanned Ap ollo 14 and 15 Dust Detector data from microfilm and are in the proce ss of archiving thc scans with PDS. We are also restoring raw dust de tector data from magnetic tape supplied by Yosio Nakamura (UT Austin) . Seiichi Nagihara (Texas Tech Univ.) and others in cooperation with NSSDC are recovering ARCSAV tapes (tapes containing raw data streams from all the ALSEP instruments). We will be preparing these data for archive with PDS. We are also in the process of recovering and archivi ng data not previously archived, from the Apollo 16 Gamma Ray Spectro meter and the Apollo 17 Infrared Spectrometer

Nanolaminate Mirrors With "Piston" Figure-Control Actuators

Efforts are under way to develop a special class of thin-shell curved mirrors for high-resolution imaging in visible and infrared light in a variety of terrestrial or extraterrestrial applications. These mirrors can have diameters of the order of a meter and include metallic film reflectors on nanolaminate substrates supported by multiple distributed piezoceramic gpiston h-type actuators for micron-level figure control. Whereas conventional glass mirrors of equivalent size and precision have areal mass densities between 50 and 150 kg/sq m, the nanolaminate mirrors, including not only the reflector/ shell portions but also the actuators and the backing structures needed to react the actuation forces, would have areal mass densities that may approach .5 kg/m2. Moreover, whereas fabrication of a conventional glass mirror of equivalent precision takes several years, the reflector/shell portion of a nanolaminate mirror can be fabricated in less than a week, and its actuation system can be fabricated in 1 to 2 months. The engineering of these mirrors involves a fusion of the technological heritage of multisegmented adaptive optics and deformable mirrors with more recent advances in metallic nanolaminates and in mathematical modeling of the deflections of thin, curved shells in response to displacements by multiple, distributed actuators. Because a nanolaminate shell is of the order of 10 times as strong as an otherwise identical shell made of a single, high-strength, non-nanolaminate metal suitable for mirror use, a nanolaminate mirror can be made very thin (typically between 100 and 150 m from the back of the nanolaminate substrate to the front reflecting surface). The thinness and strength of the nanolaminate are what make it possible to use distributed gpiston h-type actuators for surface figure control with minimal local concentrated distortion (called print-through in the art) at the actuation points

Cell Wall N-Linked Mannoprotein Biosynthesis Requires Goa1p, a Putative Regulator of Mitochondrial Complex I in Candida Aabicans

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. The Goa1p of Candida albicans regulates mitochondrial Complex I (CI) activities in its role as a putative CI accessory protein. Transcriptional profiling of goa1Δ revealed a down regulation of genes encoding β-oligomannosyl transferases. Herein, we present data on cell wall phenotypes of goa1Δ (strain GOA31). We used transmission electron microscopy (TEM), GPC/MALLS, and NMR to compare GOA31 to a gene-reconstituted strain (GOA32) and parental cells. We note by TEM a reduction in outer wall fibrils, increased inner wall transparency, and the loss of a defined wall layer close to the plasma membrane. GPC-MALLS revealed a reduction in high and intermediate Mw mannan by 85% in GOA31. A reduction of β-mannosyl but not α-mannosyl linkages was noted in GOA31 cells. β-(1,6)-linked glucan side chains were branched about twice as often but were shorter in length for GOA31. We conclude that mitochondrial CI energy production is highly integrated with cell wall formation. Our data also suggest that not all cell wall biosynthetic processes are dependent upon Goa1p even though it provides high levels of ATP to cells. The availability of both broadly conserved and fungal-specific mutants lacking CI subunit proteins should be useful in assessing functions of fungal-specific functions subunit proteins

Measuring Low-Order Aberrations in a Segmented Telescope

The in-focus PSF optimizer (IPO) is an algorithm for use in monitoring and controlling the alignment of the segments of a segmented-mirror astronomical telescope. IPO is so named because it computes wave-front aberrations of the telescope from digitized pointspread functions (PSFs) measured in infocus images. Inasmuch as distant astronomical objects that behave optically as point sources can typically be seen in almost any astronomical image, the main benefit afforded by IPO may be to enable maintenance of mirror-segment alignments without detracting from valuable scientific-observation time. IPO evolved from prescription-retrieval type algorithms. Prescription retrieval uses in-focus and out-of-focus PSFs to infer the state of an imaging optical system. The state, in this context, refers to the positions, orientations, and low-order figure errors of the optical elements in the system. Both prescription- retrieval and IPO use an iterative, nonlinear, least-squares optimizer to compute the optimal state parameters such that a digital computer-generated model image matches the digitized image acquired from the real system. The difference between IPO and prescription- retrieval algorithms is that IPO is specifically designed to utilize infocus images only. Although the restriction to in-focus images limits IPO to calculating only the lowest-order wave front aberrations, it also causes the resulting computation to take much less time because fewer degrees of freedom are included in the optimization process. In the prescription retrieval software developed at JPL, the model images are generated using the ray-trace/physical optics program, MACOS. IPO, on the other hand, uses a linear sensitivity matrix to compute the exit-pupil wave front from the system parameters; the wave front is then converted into a complex pupil field, which is then propagated to the image plane via a fast Fourier transform. This approach is computationally faster and requires less computer memory than is needed for prescription retrieval

Developmental Cryogenic Active Telescope Testbed, a Wavefront Sensing and Control Testbed for the Next Generation Space Telescope

As part of the technology validation strategy of the next generation space telescope (NGST), a system testbed is being developed at GSFC, in partnership with JPL and Marshall Space Flight Center (MSFC), which will include all of the component functions envisioned in an NGST active optical system. The system will include an actively controlled, segmented primary mirror, actively controlled secondary, deformable, and fast steering mirrors, wavefront sensing optics, wavefront control algorithms, a telescope simulator module, and an interferometric wavefront sensor for use in comparing final obtained wavefronts from different tests. The developmental. cryogenic active telescope testbed (DCATT) will be implemented in three phases. Phase 1 will focus on operating the testbed at ambient temperature. During Phase 2, a cryocapable segmented telescope will be developed and cooled to cryogenic temperature to investigate the impact on the ability to correct the wavefront and stabilize the image. In Phase 3, it is planned to incorporate industry developed flight-like components, such as figure controlled mirror segments, cryogenic, low hold power actuators, or different wavefront sensing and control hardware or software. A very important element of the program is the development and subsequent validation of the integrated multidisciplinary models. The Phase 1 testbed objectives, plans, configuration, and design will be discussed

New Insights into the Structure of (1→3,1→6)-β-D-Glucan Side Chains in the Candida glabrata Cell Wall

β-glucan is a (1→3)-β-linked glucose polymer with (1→6)-β-linked side chains and a major component of fungal cell walls. β-glucans provide structural integrity to the fungal cell wall. The nature of the (1–6)-β-linked side chain structure of fungal (1→3,1→6)-β-D-glucans has been very difficult to elucidate. Herein, we report the first detailed structural characterization of the (1→6)-β-linked side chains of Candida glabrata using high-field NMR. The (1→6)-β-linked side chains have an average length of 4 to 5 repeat units spaced every 21 repeat units along the (1→3)-linked polymer backbone. Computer modeling suggests that the side chains have a bent curve structure that allows for a flexible interconnection with parallel (1→3)-β-D-glucan polymers, and/or as a point of attachment for proteins. Based on these observations we propose new approaches to how (1→6)-β-linked side chains interconnect with neighboring glucan polymers in a manner that maximizes fungal cell wall strength, while also allowing for flexibility, or plasticity

Mannan Molecular Substructures Control Nanoscale Glucan Exposure in Candida

Cell wall mannans of Candida albicans mask β-(1,3)-glucan from recognition by Dectin-1, contributing to innate immune evasion. Glucan exposures are predominantly single receptor-ligand interaction sites of nanoscale dimensions. Candida species vary in basal glucan exposure and molecular complexity of mannans. We used super-resolution fluorescence imaging and a series of protein mannosylation mutants in C. albicans and C. glabrata to investigate the role of specific N-mannan features in regulating the nanoscale geometry of glucan exposure. Decreasing acid labile mannan abundance and α-(1,6)-mannan backbone length correlated most strongly with increased density and nanoscopic size of glucan exposures in C. albicans and C. glabrata, respectively. Additionally, a C. albicans clinical isolate with high glucan exposure produced similarly perturbed N-mannan structures and elevated glucan exposure geometry. Thus, acid labile mannan structure influences the nanoscale features of glucan exposure, impacting the nature of the pathogenic surface that triggers immunoreceptor engagement, aggregation, and signaling. Graus et al. find that N-mannan structural features regulated by Candida mannosyltransfersases control glucan exposure. Loss of mannan increased the frequency and size of glucan exposures and changed multivalent receptor engagement. Changes to mannan structure in a bloodstream isolate are associated with elevated glucan exposure at the nanoscale