Potent spinal parenchymal AAV9-mediated gene delivery by subpial injection in adult rats and pigs.

Effective in vivo use of adeno-associated virus (AAV)-based vectors to achieve gene-specific silencing or upregulation in the central nervous system has been limited by the inability to provide more than limited deep parenchymal expression in adult animals using delivery routes with the most clinical relevance (intravenous or intrathecal). Here, we demonstrate that the spinal pia membrane represents the primary barrier limiting effective AAV9 penetration into the spinal parenchyma after intrathecal AAV9 delivery. We develop a novel subpial AAV9 delivery technique and AAV9-dextran formulation. We use these in adult rats and pigs to show (i) potent spinal parenchymal transgene expression in white and gray matter including neurons, glial and endothelial cells after single bolus subpial AAV9 delivery; (ii) delivery to almost all apparent descending motor axons throughout the length of the spinal cord after cervical or thoracic subpial AAV9 injection; (iii) potent retrograde transgene expression in brain motor centers (motor cortex and brain stem); and (iv) the relative safety of this approach by defining normal neurological function for up to 6 months after AAV9 delivery. Thus, subpial delivery of AAV9 enables gene-based therapies with a wide range of potential experimental and clinical utilizations in adult animals and human patients

The Ionized Gas and Nuclear Environment in NGC 3783 V. Variability and Modeling of the Intrinsic Ultraviolet Absorption

We present results on the location, physical conditions, and geometry of the outflow in the Seyfert 1 galaxy NGC 3783 from a study of the variable intrinsic UV absorption. Based on 18 observations with HST/STIS and 6 observations with FUSE, we find: 1) The absorption from the lowest-ionization species in each of the three strong kinematic components varied inversely with the continuum flux, indicating the ionization structure responded to changes in the photoionizing flux over the weekly timescales sampled by our observations. 2) A multi- component model with an unocculted NLR and separate BLR and continuum line-of-sight covering factors predicts saturation in several lines, consistent with the lack of observed variability. 3) Column densities for the individual metastable levels are measured from the resolved C III *1175 absorption complex observed in one component. Based on our computed metastable level populations, the electron density of this absorber is ~3x10^4 cm^-3. Photoionization modeling results place it at ~25 pc from the central source. 4) Using time-dependent calculations, we are able to reproduce the detailed variability observed in this absorber, and derive upper limits on the distances for the other components of 25-50 pc. 5) The ionization parameters derived for the higher ionization UV absorbers are consistent with the modeling results for the lowest-ionization X-ray component, but with smaller total column density. They have similar pressures as the three X-ray ionization components. These results are consistent with an inhomogeneous wind model for the outflow in NGC 3783. 6) Based on the predicted emission-line luminosities, global covering factor constraints, and distances derived for the UV absorbers, they may be identified with emission- line gas observed in the inner NLR of AGNs. (abridged)Comment: 30 pages, 18 figures (7 color), emulateapj, accepted for publication in The Astrophysical Journa

The Ionized Gas and Nuclear Environment in NGC 3783. I. Time-Averaged 900 ks Chandra Grating Spectroscopy

We present results from a 900 ks exposure of NGC 3783 with the High-Energy Transmission Grating Spectrometer on board the Chandra X-ray Observatory. The resulting X-ray spectrum has the best combination of signal-to-noise and resolution ever obtained for an AGN. This spectrum reveals absorption lines from H-like and He-like ions of N, O, Ne, Mg, Al, Si, and S. There are also possible absorption lines from H-like and He-like Ar and Ca. We also identify inner-shell absorption from lower-ionization ions such as Si_VII-Si_XII and S_XII-S_XIV. The iron absorption spectrum is very rich; L-shell lines of Fe_XVII-Fe_XXIV are detected, strong complex of M-shell lines, and probable resonance lines from Fe_XXV. The absorption lines are blueshifted relative to the systemic velocity by a mean velocity of -590+-150 km/s. We resolve many of the absorption lines, and their mean FWHM is 820+-280 km/s. We do not find correlations between the velocity shifts or the FWHMs with the ionization potentials of the ions. Most absorption lines show asymmetry, having more extended blue wings than red wings. In O_VII we have resolved this asymmetry to be from an additional absorption system at ~ -1300 km/s. The two X-ray absorption systems are consistent in velocity shift and FWHM with the ones identified in the UV lines of C IV, N V, and H I. Equivalent width measurements for all lines are given and column densities are calculated for several ions. We resolve the narrow Fe_K\alpha line at 6398.2+-3.3 eV to have a FWHM of 1720+-360 km/s, which suggests that this narrow line may be emitted from the outer part of the broad line region or the inner part of the torus. We also detect a `Compton shoulder' redward of the narrow Fe_K\alpha line which indicates that it arises in cold, Compton-thick gas.Comment: 19 pages, 12 figures (2 in color), emulateapj5, accepted for publication in The Astrophysical Journal Supplement

myTAI: evolutionary transcriptomics with R.

MOTIVATION: Next Generation Sequencing (NGS) technologies generate a large amount of high quality transcriptome datasets enabling the investigation of molecular processes on a genomic and metagenomic scale. These transcriptomics studies aim to quantify and compare the molecular phenotypes of the biological processes at hand. Despite the vast increase of available transcriptome datasets, little is known about the evolutionary conservation of those characterized transcriptomes. RESULTS: The myTAI package implements exploratory analysis functions to infer transcriptome conservation patterns in any transcriptome dataset. Comprehensive documentation of myTAI functions and tutorial vignettes provide step-by-step instructions on how to use the package in an exploratory and computationally reproducible manner. AVAILABILITY AND IMPLEMENTATION: The open source myTAI package is available at https://github.com/HajkD/myTAI and https://cran.r-project.org/web/packages/myTAI/index.html. CONTACT: [email protected]. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online

AIAA Design, Build, Fly Team - MULLET Competition Aircraft 2021-2022

MULLET, the Medical Unmanned Low-Level Electric Transport, is Embry-Riddle Aeronautical University Daytona Beach’s aircraft for the 2021–2022 AIAA Design, Build, Fly competition. This UAV was designed to perform four missions, including a ground mission and three flight missions. Mission 1 is a deployment flight that demonstrates the aircraft’s flight capability; Mission 2 is a staging flight for the transportation of vaccine syringes; Mission 3 is a delivery flight for the transportation and deployment of vaccine vial packages; and the Ground Mission is a demonstration of the ability to rapidly prepare the aircraft for flight. The aircraft was designed, manufactured, and flown by a team of 40 undergraduate aerospace engineering students. The design process comprised three phases: conceptual, preliminary, and detail design. Initially, the conceptual design focused on analyzing the requirements with a scoring analysis to select the optimal payload that maximized the mission scores. After the aircraft and subsystem configurations were selected, the weight, wing, tail, and propulsion system were sized during the preliminary design. A detail design then focused on the aircraft’s structural characteristics and systems integration. The manufacturing process followed with the goal of fabricating the aircraft to the designed specifications and weight. A detailed schedule was developed and was continuously refined to manufacture each aircraft iteration in a timely manner, enabling rapid prototyping throughout the design, build, and fly process. Finally, a testing plan was established to evaluate a series of test objectives essential to the aircraft’s mission performance

The Ionized Gas and Nuclear Environment in NGC 3783. IV. Variability and Modeling of the 900 ks CHANDRA Spectrum

We present a detailed spectral analysis of the data obtained from NGC 3783 during the period 2000-2001 using Chandra. This analysis leads us to the following results. 1) NGC 3783 fluctuated in luminosity by a factor ~1.5 during individual observations (~170 ks duration). These fluctuations were not associated with significant spectral variations. 2) On a longer time scale (20-120 days), we found the source to exhibit two very different spectral shapes. The main difference between these can be well-described by the appearance and disappearance of a spectral component that dominates the underlying continuum at the longest wavelengths. The spectral variations are not related to the brightening or the fading of the continuum at short wavelengths in any simple way. 3) The appearance of the soft continuum component is consistent with being the only spectral variation, and there is no need to invoke changes in the opacity of the absorbers. 4) Photoionization modeling indicates that a combination of three ionized absorbers, each split into two kinematic components, can explain the strengths of almost all the absorption lines and bound-free edges. All three components are thermally stable and seem to have the same gas pressure. 5) The only real discrepancy between our model and the observations concerns the range of wavelengths absorbed by the iron M-shell UTA feature. This most likely arises as the result of our underestimation of the poorly-known dielectronic recombination rates appropriate for these ions. 6) The lower limit on the distance of the absorbing gas in NGC 3783 is between 0.2 and 3.2 pc. The assumption of pressure equilibrium imposes an upper limit of about 25 pc on the distance of the least-ionized component from the central source. (abridged)Comment: 16 pages, 12 figures (9 in color), emulateapj5, accepted for publication in The Astrophysical Journa

First-in-Human Clinical Trial of Oral ONC201 in Patients with Refractory Solid Tumors

Purpose: ONC201 is a small-molecule selective antagonist of the G protein–coupled receptor DRD2 that is the founding member of the imipridone class of compounds. A first-in-human phase I study of ONC201 was conducted to determine its recommended phase II dose (RP2D). Experimental Design: This open-label study treated 10 patients during dose escalation with histologically confirmed advanced solid tumors. Patients received ONC201 orally once every 3 weeks, defined as one cycle, at doses from 125 to 625 mg using an accelerated titration design. An additional 18 patients were treated at the RP2D in an expansion phase to collect additional safety, pharmacokinetic, and pharmacodynamic information. Results: No grade \u3e 1 drug-related adverse events occurred, and the RP2D was defined as 625 mg. Pharmacokinetic analysis revealed a Cmax of 1.5 to 7.5 μg/mL (∼3.9–19.4 μmol/L), mean half-life of 11.3 hours, and mean AUC of 37.7 h·μg/L. Pharmacodynamic assays demonstrated induction of caspase-cleaved keratin 18 and prolactin as serum biomarkers of apoptosis and DRD2 antagonism, respectively. No objective responses by RECIST were achieved; however, radiographic regression of several individual metastatic lesions was observed along with prolonged stable disease (\u3e 9 cycles) in prostate and endometrial cancer patients. Conclusions: ONC201 is a selective DRD2 antagonist that is well tolerated, achieves micromolar plasma concentrations, and is biologically active in advanced cancer patients when orally administered at 625 mg every 3 weeks

The Ionized Gas and Nuclear Environment in NGC 3783 II. Averaged HST/STIS and FUSE Spectra

We present observations of the intrinsic absorption in the Seyfert 1 galaxy NGC 3783 obtained with the STIS/HST and FUSE. We have coadded multiple STIS and FUSE observations to obtain a high S/N averaged spectrum spanning 905-1730 A. The averaged spectrum reveals absorption in O VI, N V, C IV, N III, C III and the Lyman lines up to LyE in the three blueshifted kinematic components previously detected in the STIS spectrum (at radial velocities of -1320, -724, and -548 km/s). The highest velocity component exhibits absorption in Si IV. We also detect metastable C III* in this component, indicating a high density in this absorber. We separate the individual covering factors of the continuum and emission-line sources as a function of velocity in each kinematic component using the LyA and LyB lines. Additionally, we find that the continuum covering factor varies with velocity within the individual kinematic components, decreasing smoothly in the wings of the absorption by at least 60%. The covering factor of Si IV is found to be less than half that of H I and N V in the high velocity component. Additionally, the FWHM of N III and Si IV are narrower than the higher ionization lines in this component. These results indicate there is substructure within this absorber. We derive a lower limit on the total column (N_H>=10^{19}cm^{-2}) and ionization parameter (U>=0.005) in the low ionization subcomponent of this absorber. The metastable-to-total C III column density ratio implies n_e~10^9 cm^{-3} and an upper limit on the distance of the absorber from the ionizing continuum of R<=8x10^{17} cm.Comment: 29 pages, 8 figures (Figures 1-3 are in color), Accepted for pulication in the Astrophysical Journa

Distinct disease mutations in DNMT3A result in a spectrum of behavioral, epigenetic, and transcriptional deficits

Phenotypic heterogeneity in monogenic neurodevelopmental disorders can arise from differential severity of variants underlying disease, but how distinct alleles drive variable disease presentation is not well understood. Here, we investigate missense mutations in DNA methyltransferase 3A (DNMT3A), a DNA methyltransferase associated with overgrowth, intellectual disability, and autism, to uncover molecular correlates of phenotypic heterogeneity. We generate a Dnmt3

The structural basis for partitioning of the XRCC1/DNA ligase III-α BRCT-mediated dimer complexes

The ultimate step common to almost all DNA repair pathways is the ligation of the nicked intermediate to form contiguous double-stranded DNA. In the mammalian nucleotide and base excision repair pathways, the ligation step is carried out by ligase III-α. For efficient ligation, ligase III-α is constitutively bound to the scaffolding protein XRCC1 through interactions between the C-terminal BRCT domains of each protein. Although structural data for the individual domains has been available, no structure of the complex has been determined and several alternative proposals for this interaction have been advanced. Interpretation of the models is complicated by the formation of homodimers that, depending on the model, may either contribute to, or compete with heterodimer formation. We report here the structures of both homodimer complexes as well as the heterodimer complex. Structural characterization of the heterodimer formed from a longer XRCC1 BRCT domain construct, including residues comprising the interdomain linker region, revealed an expanded heterodimer interface with the ligase III-α BRCT domain. This enhanced linker-mediated binding interface plays a significant role in the determination of heterodimer/homodimer selectivity. These data provide fundamental insights into the structural basis of BRCT-mediated dimerization, and resolve questions related to the organization of this important repair complex