Protein crystals in adenovirus type 5-infected cells: requirements for intranuclear crystallogenesis, structural and functional analysis

Intranuclear crystalline inclusions have been observed in the nucleus of epithelial cells infected with Adenovirus serotype 5 (Ad5) at late steps of the virus life cycle. Using immuno-electron microscopy and confocal microscopy of cells infected with various Ad5 recombinants modified in their penton base or fiber domains, we found that these inclusions represented crystals of penton capsomers, the heteromeric capsid protein formed of penton base and fiber subunits. The occurrence of protein crystals within the nucleus of infected cells required the integrity of the fiber knob and part of the shaft domain. In the knob domain, the region overlapping residues 489–492 in the FG loop was found to be essential for crystal formation. In the shaft, a large deletion of repeats 4 to 16 had no detrimental effect on crystal inclusions, whereas deletion of repeats 8 to 21 abolished crystal formation without altering the level of fiber protein expression. This suggested a crucial role of the five penultimate repeats in the crystallisation process. Chimeric pentons made of Ad5 penton base and fiber domains from different serotypes were analyzed with respect to crystal formation. No crystal was found when fiber consisted of shaft (S) from Ad5 and knob (K) from Ad3 (heterotypic S5-K3 fiber), but occurred with homotypic S3K3 fiber. However, less regular crystals were observed with homotypic S35-K35 fiber. TB5, a monoclonal antibody directed against the Ad5 fiber knob was found by immunofluorescence microscopy to react with high efficiency with the intranuclear protein crystals in situ. Data obtained with Ad fiber mutants indicated that the absence of crystalline inclusions correlated with a lower infectivity and/or lower yields of virus progeny, suggesting that the protein crystals might be involved in virion assembly. Thus, we propose that TB5 staining of Ad-infected 293 cells can be used as a prognostic assay for the viability and productivity of fiber-modified Ad5 vectors

TOI-3235 b: A Transiting Giant Planet around an M4 Dwarf Star

We present the discovery of TOI-3235 b, a short-period Jupiter orbiting an M dwarf with a stellar mass close to the critical mass at which stars transition from partially to fully convective. TOI-3235 b was first identified as a candidate from TESS photometry and confirmed with radial velocities from ESPRESSO and ground-based photometry from HATSouth, MEarth-South, TRAPPIST-South, LCOGT, and ExTrA. We find that the planet has a mass of 0.665 ± 0.025 M J and a radius of 1.017 ± 0.044 R J. It orbits close to its host star, with an orbital period of 2.5926 days but has an equilibrium temperature of ≈ 604 K, well below the expected threshold for radius inflation of hot Jupiters. The host star has a mass of 0.3939 ± 0.0030 M ☉, a radius of 0.3697 ± 0.0018 R ☉, an effective temperature of 3389 K, and a J-band magnitude of 11.706 ± 0.025. Current planet formation models do not predict the existence of gas giants such as TOI-3235 b around such low-mass stars. With a high transmission spectroscopy metric, TOI-3235 b is one of the best-suited giants orbiting M dwarfs for atmospheric characterization

The deuteron: structure and form factors

A brief review of the history of the discovery of the deuteron in provided. The current status of both experiment and theory for the elastic electron scattering is then presented.Comment: 80 pages, 33 figures, submited to Advances in Nuclear Physic

Spider Silk Constructs Enhance Axonal Regeneration and Remyelination in Long Nerve Defects in Sheep

BACKGROUND: Surgical reapposition of peripheral nerve results in some axonal regeneration and functional recovery, but the clinical outcome in long distance nerve defects is disappointing and research continues to utilize further interventional approaches to optimize functional recovery. We describe the use of nerve constructs consisting of decellularized vein grafts filled with spider silk fibers as a guiding material to bridge a 6.0 cm tibial nerve defect in adult sheep. METHODOLOGY/PRINCIPAL FINDINGS: The nerve constructs were compared to autologous nerve grafts. Regeneration was evaluated for clinical, electrophysiological and histological outcome. Electrophysiological recordings were obtained at 6 months and 10 months post surgery in each group. Ten months later, the nerves were removed and prepared for immunostaining, electrophysiological and electron microscopy. Immunostaining for sodium channel (NaV 1.6) was used to define nodes of Ranvier on regenerated axons in combination with anti-S100 and neurofilament. Anti-S100 was used to identify Schwann cells. Axons regenerated through the constructs and were myelinated indicating migration of Schwann cells into the constructs. Nodes of Ranvier between myelin segments were observed and identified by intense sodium channel (NaV 1.6) staining on the regenerated axons. There was no significant difference in electrophysiological results between control autologous experimental and construct implantation indicating that our construct are an effective alternative to autologous nerve transplantation. CONCLUSIONS/SIGNIFICANCE: This study demonstrates that spider silk enhances Schwann cell migration, axonal regrowth and remyelination including electrophysiological recovery in a long-distance peripheral nerve gap model resulting in functional recovery. This improvement in nerve regeneration could have significant clinical implications for reconstructive nerve surgery

HATS-47b, HATS-48Ab, HATS-49b and HATS-72b: Four Warm Giant Planets Transiting K Dwarfs

We report the discovery of four transiting giant planets around K dwarfs. The planets HATS-47b, HATS-48Ab, HATS49b, and HATS-72b have masses of 0.369+ 0.0210.031MJ, 0.243+ 0.0300.022 MJ, 0.353+ 0.0270.038 MJ, and 0.1254. 0.0039 MJ, respectively, and radii of 1.117. 0.014 RJ, 0.800. 0.015 RJ, 0.765. 0.013 RJ, and 0.7224. 0.0032 RJ, respectively. The planets orbit close to their host stars with orbital periods of 3.9228 days, 3.1317 days, 4.1480 days, and 7.3279 days, respectively. The hosts are main-sequence K dwarfs with masses of 0.674+ 0.0120.016.M, 0.7279. 0.0066.M, 0.7133. 0.0075.M, and 0.7311. 0.0028, and with V-band magnitudes of V = 14.829. 0.010, 14.35. 0.11, 14.998. 0.040 and 12.469. 0.010. The super-Neptune HATS-72b (a.k.a. WASP-191b and TOI 294.01) was independently identified as a transiting planet candidate by the HATSouth, WASP, and TESS surveys, and we present a combined analysis of all of the data gathered by each of these projects (and their follow-up programs). An exceptionally precise mass is measured for HATS-72b thanks to high-precision radial velocity (RV) measurements obtained with VLT/ESPRESSO, FEROS, HARPS, and Magellan/PFS. We also incorporate TESS observations of the warm Saturn-hosting systems HATS-47 (a.k.a. TOI.1073.01), HATS-48A, and HATS-49. HATS-47 was independently identified as a candidate by the TESS team, while the other two systems were not previously identified from the TESS data. The RV orbital variations are measured for these systems using Magellan/PFS. HATS-48A has a resolved 5.. 4 neighbor in Gaia.DR2, which is a common-proper-motion binary star companion to HATS-48A with a mass of 0.22.M and a current projected physical separation of similar to 1400 au

PAX2 Regulates ADAM10 Expression and Mediates Anchorage-Independent Cell Growth of Melanoma Cells

PAX transcription factors play an important role during development and carcinogenesis. In this study, we investigated PAX2 protein levels in melanocytes and melanoma cells by Western Blot and immunofluorescence analysis and characterized the role of PAX2 in the pathogenesis of melanoma. In vitro we found weak PAX2 protein expression in keratinocytes and melanocytes. Compared to melanocytes increased PAX2 protein levels were detectable in melanoma cell lines. Interestingly, in tissue sections of melanoma patients nuclear PAX2 expression strongly correlated with nuclear atypia and the degree of prominent nucleoli, indicating an association of PAX2 with a more atypical cellular phenotype. In addition, with chromatin immunoprecipitation assay, PAX2 overexpression and PAX2 siRNA we present compelling evidence that PAX2 can regulate ADAM10 expression, a metalloproteinase known to play important roles in melanoma metastasis. In human tissue samples we found co-expression of PAX2 and ADAM10 in melanocytes of benign nevi and in melanoma cells of patients with malignant melanoma. Importantly, the downregulation of PAX2 by specific siRNA inhibited the anchorage independent cell growth and decreased the migratory and invasive capacity of melanoma cells. Furthermore, the downregulation of PAX2 abrogated the chemoresistance of melanoma cells against cisplatin, indicating that PAX2 expression mediates cell survival and plays important roles during melanoma progression

Genome-Wide Scan on Total Serum IgE Levels Identifies FCER1A as Novel Susceptibility Locus

High levels of serum IgE are considered markers of parasite and helminth exposure. In addition, they are associated with allergic disorders, play a key role in anti-tumoral defence, and are crucial mediators of autoimmune diseases. Total IgE is a strongly heritable trait. In a genome-wide association study (GWAS), we tested 353,569 SNPs for association with serum IgE levels in 1,530 individuals from the population-based KORA S3/F3 study. Replication was performed in four independent population-based study samples (total n = 9,769 individuals). Functional variants in the gene encoding the alpha chain of the high affinity receptor for IgE (FCER1A) on chromosome 1q23 (rs2251746 and rs2427837) were strongly associated with total IgE levels in all cohorts with P values of 1.85×10−20 and 7.08×10−19 in a combined analysis, and in a post-hoc analysis showed additional associations with allergic sensitization (P = 7.78×10−4 and P = 1.95×10−3). The “top” SNP significantly influenced the cell surface expression of FCER1A on basophils, and genome-wide expression profiles indicated an interesting novel regulatory mechanism of FCER1A expression via GATA-2. Polymorphisms within the RAD50 gene on chromosome 5q31 were consistently associated with IgE levels (P values 6.28×10−7−4.46×10−8) and increased the risk for atopic eczema and asthma. Furthermore, STAT6 was confirmed as susceptibility locus modulating IgE levels. In this first GWAS on total IgE FCER1A was identified and replicated as new susceptibility locus at which common genetic variation influences serum IgE levels. In addition, variants within the RAD50 gene might represent additional factors within cytokine gene cluster on chromosome 5q31, emphasizing the need for further investigations in this intriguing region. Our data furthermore confirm association of STAT6 variation with serum IgE levels

High-contrast Polarimetry Observation of the T Tau Circumstellar Environment

We conducted high-contrast polarimetry observations of T Tau in the H-band, using the HiCIAO instrument mounted on the Subaru Telescope, revealing structures as near as 0.\arcsec1 from the stars T Tau N and T Tau S. The whole T Tau system is found to be surrounded by nebula-like envelopes, and several outflow-related structures are detected in these envelopes. We analyzed the detailed polarization patterns of the circumstellar structures near each component of this triple young star system and determined constraints on the circumstellar disks and outflow structures. We suggest that the nearly face-on circumstellar disk of T Tau N is no larger than 0.\arcsec8, or 117 AU, in the northwest, based on the existence of a hole in this direction, and no larger than 0.\arcsec27, or 40 AU, in the south. A new structure "N5" extends to about 0.\arcsec42, or 59 AU, on the southwest of the star, believed to be part of the disk. We suggest that T Tau S is surrounded by a highly inclined circumbinary disk with a radius of about 0.\arcsec3, or 44 AU, with a position angle of about 30$^\circ$, that is misaligned with the orbit of the T Tau S binary. After analyzing the positions and polarization vector patterns of the outflow-related structures, we suggest that T Tau S should trigger the well-known E-W outflow, and is also likely to be responsible for a southwest precessing outflow "coil" and a possible south outflow

High Affinity Antibodies to Plasmodium falciparum Merozoite Antigens Are Associated with Protection from Malaria

Malaria kills almost 1 million people every year, but the mechanisms behind protective immunity against the disease are still largely unknown. In this study, surface plasmon resonance technology was used to evaluate the affinity (measured as k(d)) of naturally acquired antibodies to the Plasmodium falciparum antigens MSP2 and AMA1. Antibodies in serum samples from residents in endemic areas bound with higher affinities to AMA1 than to MSP2, and with higher affinities to the 3D7 allele of MSP2-3D7 than to the FC27 allele. The affinities against AMA1 and MSP2-3D7 increased with age, and were usually within similar range as the affinities for the monoclonal antibodies also examined in this study. The finding of MSP2-3D7 type parasites in the blood was associated with a tendency for higher affinity antibodies to both forms of MSP2 and AMA1, but this was significant only when analyzing antibodies against MSP2-FC27, and individuals infected with both allelic forms of MSP2 at the same time showed the highest affinities. Individuals with the highest antibody affinities for MSP2-3D7 at baseline had a prolonged time to clinical malaria during 40 weeks of follow-up, and among individuals who were parasite positive at baseline higher antibody affinities to all antigens were seen in the individuals that did not experience febrile malaria during follow up. This study contributes important information for understanding how immunity against malaria arises. The findings suggest that antibody affinity plays an important role in protection against disease, and differs between antigens. In light of this information, antibody affinity measurements would be a key assessment in future evaluation of malaria vaccine formulations

The Multiplanet System TOI-421*: A Warm Neptune and a Super Puffy Mini-Neptune Transiting a G9 V Star in a Visual Binary*

We report the discovery of a warm Neptune and a hot sub-Neptune transiting TOI-421 (BD-14 1137, TIC 94986319), a bright (V = 9.9) G9 dwarf star in a visual binary system observed by the Transiting Exoplanet Survey Satellite (TESS) space mission in Sectors 5 and 6. We performed ground-based follow-up observations—comprised of Las Cumbres Observatory Global Telescope transit photometry, NIRC2 adaptive optics imaging, and FIbre-fed Echellé Spectrograph, CORALIE, High Accuracy Radial velocity Planet Searcher, High Resolution Échelle Spectrometer, and Planet Finder Spectrograph high-precision Doppler measurements—and confirmed the planetary nature of the 16 day transiting candidate announced by the TESS team. We discovered an additional radial velocity signal with a period of five days induced by the presence of a second planet in the system, which we also found to transit its host star. We found that the inner mini-Neptune, TOI-421 b, has an orbital period of Pb = 5.19672 ± 0.00049 days, a mass of Mb = 7.17 ± 0.66 M⊕, and a radius of Rb = ${2.68}_{-0.18}^{+0.19}$ R⊕, whereas the outer warm Neptune, TOI-421 c, has a period of Pc = 16.06819 ± 0.00035 days, a mass of Mc = ${16.42}_{-1.04}^{+1.06}$ M⊕, a radius of Rc = ${5.09}_{-0.15}^{+0.16}$ R⊕, and a density of ρc = ${0.685}_{-0.072}^{+0.080}$ g cm−3. With its characteristics, the outer planet (ρc = ${0.685}_{-0.072}^{+0.080}$ g cm−3) is placed in the intriguing class of the super-puffy mini-Neptunes. TOI-421 b and TOI-421 c are found to be well-suited for atmospheric characterization. Our atmospheric simulations predict significant Lyα transit absorption, due to strong hydrogen escape in both planets, as well as the presence of detectable CH4 in the atmosphere of TOI-421 c if equilibrium chemistry is assumed