Molecular Envelope Research Articles

SOFIA/GREAT observations of OD and OH rotational lines towards high-mass star forming regions

Context. Only recently, OD, the deuterated isotopolog of hydroxyl, OH, has become accessible in the interstellar medium; spectral lines from both species have been observed in the supra-Terahertz and far infrared regime. Studying variations of the OD/OH abundance amongst different types of sources can deliver key information on the formation of water, H2O. Aims. With observations of rotational lines of OD and OH towards 13 Galactic high-mass star forming regions, we aim to constrain the OD abundance and infer the deuterium fractionation of OH in their molecular envelopes. For the best studied source in our sample, G34.26+0.15, we were able to perform detailed radiative transfer modelling to investigate the OD abundance profile in its inner envelope. Methods. We used the Stratospheric Observatory for Infrared Astronomy (SOFIA) to observe the 2Π3∕2 J = 5∕2−3∕2 ground-state transition of OD at 1.3 THz (215 μm) and the rotationally excited OH line at 1.84 THz (163 μm). We also used published high-spectral-resolution SOFIA data of the OH ground-state transition at 2.51 THz (119.3 μm). Results. Absorption from the 2Π3∕2 OD J = 5∕2−3∕2 ground-state transition is prevalent in the dense clumps surrounding active sites of high-mass star formation. Our modelling suggests that part of the absorption arises from the denser inner parts, while the bulk of it as seen with SOFIA originates in the outer, cold layers of the envelope for which our constraints on the molecular abundance suggest a strong enhancement in deuterium fractionation. We find a weak negative correlation between the OD abundance and the bolometric luminosity to mass ratio, an evolutionary indicator, suggesting a slow decrease of OD abundance with time. A comparison with HDO shows a similarly high deuterium fractionation for the two species in the cold envelopes, which is of the order of 0.48% for the best studied source, G34.26+0.15. Conclusions. Our results are consistent with chemical models that favour rapid exchange reactions to form OD in the dense cold gas. Constraints on the OD/OH ratio in the inner envelope could further elucidate the water and oxygen chemistry near young high-mass stars.

Topological Effects of Dendronized Polymers on Their Thermoresponsiveness and Microconfinement

Topological structures can provide dendritic polymers with featured properties through cooperative effects beyond their molecular structures. Herein, we report the synthesis of a series of oligoethylene glycol (OEG)-based dendronized polymers, which carry the first generation of 2-fold, 4-fold, or 6-fold dendritic OEG pendants with methoxyl or ethoxyl terminals. Depending on the OEG branching density, these polymers show different molecular thicknesses and exhibit characteristic thermoresponsiveness with phase transition temperatures in the range of 16.6–67.2 °C. Furthermore, the higher the OEG branching density, the higher the shielding to the interior parts from the dendritic OEGs cooperatively to exhibit “molecular envelope” effects. This molecular microconfinement formed from the crowded OEG dendrons was further investigated through covalently linked or physically mixed dye. Topological structures of the polymers are discovered to play a key role in encapsulation of the dye guest and can modulate its transition within the aggregates through the thermally induced phase transitions.

Amphotericin B Polymer Nanoparticles Show Efficacy against Candida Species Biofilms.

Purpose: Chronic infections of Candida albicans are characterised by the embedding of budding and entwined filamentous fungal cells into biofilms. The biofilms are refractory to many drugs and Candida biofilms are associated with ocular fungal infections. The objective was to test the activity of nanoparticulate amphotericin B (AmB) against Candida biofilms. Methods: AmB was encapsulated in the Molecular Envelope Technology (MET, N-palmitoyl-N-monomethyl-N,N-dimethyl-N,N,N-trimethyl-6-O-glycolchitosan) nanoparticles and tested against Candida biofilms in vitro. Confocal laser scanning microscopy (CLSM) imaging of MET nanoparticles’ penetration into experimental biofilms was carried out and a MET-AmB eye drop formulation was tested for its stability. Results: MET-AmB formulations demonstrated superior activity towards C. albicans biofilms in vitro with the EC50 being ~30 times lower than AmB alone (EC50 MET-AmB = 1.176 μg mL−1, EC50 AmB alone = 29.09 μg mL−1). A similar superior activity was found for Candida glabrata biofilms, where the EC50 was ~10× lower than AmB alone (EC50 MET-AmB = 0.0253 μg mL−1, EC50 AmB alone = 0.289 μg mL−1). CLSM imaging revealed that MET nanoparticles penetrated through the C. albicans biofilm matrix and bound to fungal cells. The activity of MET-AmB was no different from the activity of AmB alone against C. albicans cells in suspension (MET-AmB MIC90 = 0.125 μg mL−1, AmB alone MIC90 = 0.250 μg mL−1). MET-AmB eye drops were stable at room temperature for at least 28 days. Conclusions: These biofilm activity findings raise the possibility that MET-loaded nanoparticles may be used to tackle Candida biofilm infections, such as refractory ocular fungal infections.

An early transition to magnetic supercriticality in star formation

Magnetic fields have an important role in the evolution of interstellar medium and star formation1,2. As the only direct probe of interstellar field strength, credible Zeeman measurements remain sparse owing to the lack of suitable Zeeman probes, particularly for cold, molecular gas3. Here we report the detection of a magnetic field of +3.8 ± 0.3 microgauss through the H I narrow self-absorption (HINSA)4,5 towards L15446,7—a well-studied prototypical prestellar core in an early transition between starless and protostellar phases8–10 characterized by a high central number density11 and a low central temperature12. A combined analysis of the Zeeman measurements of quasar H I absorption, H I emission, OH emission and HINSA reveals a coherent magnetic field from the atomic cold neutral medium (CNM) to the molecular envelope. The molecular envelope traced by the HINSA is found to be magnetically supercritical, with a field strength comparable to that of the surrounding diffuse, magnetically subcritical CNM despite a large increase in density. The reduction of the magnetic flux relative to the mass, which is necessary for star formation, thus seems to have already happened during the transition from the diffuse CNM to the molecular gas traced by the HINSA. This is earlier than envisioned in the classical picture where magnetically supercritical cores capable of collapsing into stars form out of magnetically subcritical envelopes13,14.

Deriving RNA topological structure from SAXS.

Structures of well-folded RNA molecules can be determined with atomic resolution by either X-ray crystallography, cryo-EM, or NMR spectroscopy, but those of conformationally-flexible RNAs often are difficult to study with these methods. However, flexible RNAs have biological relevance and likely represent the majority of the RNA conformational space. Due to the high electron density of the phosphate-sugar backbone, RNA is very sensitive to small-angle X-ray scattering (SAXS), and SAXS data can be recorded with sub-μM concentrations and under near-physiological solution conditions without the need for labeling. For these reasons, SAXS has significant advantages over other techniques for obtaining global structural information of flexible RNAs in the form of molecular envelopes or low-resolution topological structural models. The SAXS-derived information is extremely valuable for bridging secondary structure data, often determined by other techniques, with a three-dimensional structure description. In this chapter, we present a detailed account of the principle, algorithms, and experimental and computational protocols for topological structure determination of RNA molecules in solution. To illustrate the applications of the methodology, we provide several case studies that cover a broad spectrum of the RNA conformational landscape.

A Revised Description of the Cosmic Ray Induced Desorption of Interstellar Ices

Abstract Nonthermal desorption of ices on interstellar grains is required to explain observations of molecules that are not synthesized efficiently in the gas phase in cold dense clouds. Perhaps the most important nonthermal desorption mechanism is one induced by cosmic rays (CRs), which, when passing through a grain, heat it transiently to a high temperature—the grain cools back to its original equilibrium temperature via the (partial) sublimation of the ice. Current cosmic ray induced desorption (CRD) models assume a fixed grain cooling time. In this work, we present a revised description of CRD in which the desorption efficiency depends dynamically on the ice content. We apply the revised desorption scheme to two-phase and three-phase chemical models in physical conditions corresponding to starless and prestellar cores, and to molecular cloud envelopes. We find that, inside starless and prestellar cores, introducing dynamic CRD can decrease gas-phase abundances by up to an order of magnitude in two-phase chemical models. In three-phase chemical models, our model produces results very similar to those of the static cooling scheme—when only one monolayer of ice is considered active. Ice abundances are generally insensitive to variations in the grain cooling time. Further improved CRD models need to take into account additional effects in the transient heating of the grains—introduced, for example, by the adoption of a spectrum of CR energies.

SAXS Analysis of the Intrinsic Tenase Complex Bound to Lipid Nanodisc Highlights Intermolecular Contacts between Factors VIIIa/IXa

The intrinsic tenase (Xase) complex is formed by activated clotting factors VIII (fVIIIa) and IX (fIXa) on activated platelet surfaces and catalyzes the activation of factor X to promote blood coagulation. The structural organization of the membrane-bound Xase complex remains largely unknown, hindering a mechanistic understanding of how mutations to fVIIIa or fIXa lead to thrombotic disorders. In the present study, we aimed to structurally characterize the Xase complex bound to a lipid nanodisc using biolayer interferometry and small-angle X-ray scattering. By immobilizing lipid nanodiscs, we measured binding rates and nanomolar affinities for fVIIIa, fIXa, and the Xase complex. An ab initio molecular envelope of the nanodisc-bound Xase complex allowed us to computationally model fVIIIa and fIXa docked onto a flexible lipid membrane and identify protein-protein interactions. Our results suggest that interactions between the Gla and catalytic domains of fIXa and the C2 and A2 domains of fVIIIa, respectively, mediate Xase complex assembly. Additionally, our computational modelling highlighted a novel interaction between fIXa residue K80 and an acidic patch formed at the fVIIIa A1/A3 domain interface. Our model of the Xase complex sheds light on how hemophilia A/B-related mutations with varying severities disrupt Xase complex assembly. We also speculate on how gain-of-function mutations to fIXa residue R338, such as the Padua (R338L) and Shanghai (R338Q) variants, bind to the fVIIIa A2 domain and stabilize the Xase complex, leading to excessive blood clotting. Together, our results support the use of SAXS as an emergent tool to investigate the membrane-bound Xase complex and illustrate how mutations at the fVIIIa/fIXa dimer interface may disrupt or stabilize the activated enzyme complex. DisclosuresLollar: Expression Therapeutics: Divested equity in a private or publicly-traded company in the past 24 months, Patents & Royalties. Spencer: Expression Therapeutics: Divested equity in a private or publicly-traded company in the past 24 months. Doering: Expression Therapeutics: Divested equity in a private or publicly-traded company in the past 24 months.

Analysis of Protein-Glycosaminoglycan Interactions Using Traveling Wave Ion-Mobility Mass Spectrometry.

Traveling wave ion-mobility mass spectrometry (TWIMS) combined with native mass spectrometry (MS) has emerged as a powerful tool for analyzing biomolecules, including complexes of protein and heparan sulfate (HS). This technique allows determination of the stoichiometry of the protein-HS interaction and information on the overall 3D molecular envelope.

The Specific Elongation Factor to Selenocysteine Incorporation in Escherichia coli: Unique tRNASec Recognition and its Interactions

Several molecular mechanisms are involved in the genetic code interpretation during translation, as codon degeneration for the incorporation of rare amino acids. One mechanism that stands out is selenocysteine (Sec), which requires a specific biosynthesis and incorporation pathway. In Bacteria, the Sec biosynthesis pathway has unique features compared with the eukaryote pathway as Ser to Sec conversion mechanism is accomplished by a homodecameric enzyme (selenocysteine synthase, SelA) followed by the action of an elongation factor (SelB) responsible for delivering the mature Sec-tRNASec into the ribosome by the interaction with the Selenocysteine Insertion Sequence (SECIS). Besides this mechanism being already described, the sequential events for Sec-tRNASec and SECIS specific recognition remain unclear. In this study, we determined the order of events of the interactions between the proteins and RNAs involved in Sec incorporation. Dissociation constants between SelB and the native as well as unacylated-tRNASec variants demonstrated that the acceptor stem and variable arm are essential for SelB recognition. Moreover, our data support the sequence of molecular events where GTP-activated SelB strongly interacts with SelA.tRNASec. Subsequently, SelB.GTP.tRNASec recognizes the mRNA SECIS to deliver the tRNASec to the ribosome. SelB in complex with its specific RNAs were examined using Hydrogen/Deuterium exchange mapping that allowed the determination of the molecular envelopes and its secondary structural variations during the complex assembly. Our results demonstrate the ordering of events in Sec incorporation and contribute to the full comprehension of the tRNASec role in the Sec amino acid biosynthesis, as well as extending the knowledge of synthetic biology and the expansion of the genetic code.

Polymeric Micelles for the Enhanced Deposition of Hydrophobic Drugs into Ocular Tissues, without Plasma Exposure.

Commercial topical ocular formulations for hydrophobic actives rely on the use of suspensions or oil in water emulsions and neither of these formulation modalities adequately promote drug penetration into ocular tissues. Using the ocular relevant hydrophobic drug, cyclosporine A (CsA), a non-irritant ocular penetration enhancer is showcased, which may be used for the formulation of hydrophobic actives. The activity of this penetration enhancer is demonstrated in a healthy rabbit model. The Molecular Envelope Technology (MET) polymer (N-palmitoyl-N-monomethyl-N,N-dimethyl-N,N,N-trimethyl-6-O-glycolchitosan), a self-assembling, micelle-forming polymer, was used to formulate CsA into sterile filtered nanoparticulate eye drop formulations and the stability of the formulation tested. Healthy rabbits were dosed with a single dose of a MET–CsA (NM133) 0.05% formulation and ocular tissues analyzed. Optically clear NM133 formulations were prepared containing between 0.01–0.1% w/v CsA and 0.375–0.75% w/v MET polymer. NM133 0.01%, NM133 0.02% and NM133 0.05% were stable for 28 days when stored at refrigeration temperature (5–6 °C) and room temperature (16–23 °C), but there was evidence of evaporation of the formulation at 40 °C. There was no change in drug content when NM133 0.05% was stored for 387 days at 4 °C. On topical dosing to rabbits, corneal, conjunctival and scleral AUC0–24 levels were 25,780 ng.h g−1, 12,046 ng.h g−1 and 5879 ng.h g−1, respectively, with NM133 0.05%. Meanwhile, a similar dose of Restasis 0.05% yielded lower values of 4726 ng.h/g, 4813 ng.h/g and 1729 ng.h/g for the drug corneal, conjunctival and scleral levels, respectively. NM133 thus delivered up to five times more CsA to the ocular surface tissues when compared to Restasis. The MET polymer was non-irritant up to a concentration of 4% w/v. The MET polymer is a non-irritant ocular penetration enhancer that may be used to deliver hydrophobic drugs in optically clear topical ocular formulations.

Stable stratification promotes multiple zonal jets in a turbulent jovian dynamo model

The ongoing NASA's Juno mission puts new constraints on the internal dynamics of Jupiter. Data gathered by its onboard magnetometer reveal a dipole-dominated surface magnetic field accompanied by strong localised magnetic flux patches. The gravity measurements indicate that the fierce surface zonal jets extend several thousands of kilometers below the cloud level before rapidly decaying below 0.94 − 0.96RJ, RJ being the mean Jovian radius at the one bar level. Jupiter's internal structure can be grossly decomposed in two parts: (i) an outer layer filled with a mixture of molecular hydrogen and helium where the zonal flows are thought to be driven; (ii) an inner region where hydrogen becomes metallic and dynamo action is expected to sustain the magnetic field. Several internal models however suggest a more intricate structure with a thin intermediate region in which helium would segregate from hydrogen, forming a compositionally-stratified layer. Here, we develop the first global Jovian dynamo which incorporates an intermediate stably-stratified layer between 0.82RJ and 0.86RJ. Using much lower diffusivities than previous models enables us to more clearly separate the dynamics of the metallic core and the molecular envelope. Analysing the energy balance reveals that magnetic energy is almost one order of magnitude larger than kinetic energy in the metallic region, while most of the kinetic energy is pumped into zonal motions in the molecular envelope. Those result from the different underlying force hierarchy with a triple balance between Lorentz, Archimedean and ageostrophic Coriolis forces in the metallic core and inertia, buoyancy and ageostrophic Coriolis forces controlling the external layers. The simulation presented here is the first to demonstrate that multiple zonal jets and dipole-dominated dynamo action can be consolidated in a global simulation. The inclusion of a stable layer is a necessary ingredient that allows zonal jets to develop in the outer envelope without contributing to the dynamo action in the deeper metallic region. Stable stratification however also smooths out the small-scale features of the magnetic field by skin effect, yielding a too-dipolar surface field as compared to the observations. These constraints suggest that possible stable layers in Jupiter should be located much closer to the surface (0.9 − 0.95RJ).

Dust Growth in Molecular Cloud Envelopes: A Numerical Approach

Abstract Variations in the grain size distribution are to be expected in the interstellar medium (ISM) owing to grain growth and destruction. In this work, we present a dust collision model to be implemented inside a magnetohydrodynamic (MHD) code that takes into account grain growth and shattering of charged dust grains of a given composition (silicate or graphite). We integrate this model in the MHD code Athena and build on a previous implementation of the dynamics of charged dust grains in the same code. To demonstrate the performance of this coagulation model, we study the variations in the grain size distribution of a single-sized population of dust with radius 0.05 μm inside several dust filaments formed during a 2D MHD simulation. We also consider a realistic dust distribution with sizes ranging from 50 Å to 0.25 μm and analyze the variations in both the size distribution for graphite and silicates and the far-ultraviolet extinction curve. From the obtained results, we conclude that the methodology here presented, based on the MHD evolution of the equation of motion for a charged particle, is optimal for studying the coagulation of charged dust grains in a diffuse regime such as a molecular cloud envelope. Observationally, these variations in the dust size distribution are translated into variations in the far-ultraviolet extinction curve, and they are mainly caused by small graphite dust grains.

Challenging the electrostatic σ-hole picture of halogen bonding using minimal models and the interacting quantum atoms approach.

Among the different noncovalent interactions, halogen bonds have captured wide attention in the last years. Their stability has been rationalized in electrostatic terms by appealing to the σ-hole concept, a charge-depleted region that is able to interact favorably with electron rich moieties. This interpretation has been questioned, and in this work a set of anionic halogen model systems are used to shed some light on this issue. We use the interacting quantum atoms method, which provides an orbital invariant energy decomposition in which pure electrostatic terms are well isolated, and we complement our insights with the analysis of electrostatic potentials (ESPs) as well as with traditional descriptors of charge accumulation like the Laplacian of the electron density. The total electrostatic interaction between the interacting species is surprisingly destabilizing in many of the systems examined, demonstrating that although σ-holes might be qualitatively helpful, much care has to be taken in ascribing the stability of these systems to electrostatics. It is clearly shown that electron delocalization is essential to understand the stability of the complexes. The evolution of atomic charges as the aggregates forms reveals a charge transfer picture in which the central, σ-hole bearing halogen acts as a mere spectator. These systems may then be not far from engaging in a classical 3c-4e interaction. Since the presence of a σ-hole as characterized by the ESP mapped on a suitable molecular envelope isosurface does not guarantee attractive electrostatic interactions, we encourage to employ a wider perspective that takes into account the full charge distribution.

A polymeric aqueous tacrolimus formulation for topical ocular delivery

Tacrolimus (TAC) suspension is used to treat moderate to severe atopic keratoconjunctivitis (AKC) and vernal keratoconjunctivitis (VKC). The objectives of this study were to formulate the hydrophobic compound TAC (TAC) in an aqueous eye drop formulation and study its ocular biodistribution on topical ocular application to a healthy rabbit model, with the overall aim of using the formulation to treat AKC and VKC. A thin-film hydration method was used to encapsulate TAC within the chitosan-based amphiphile: N-palmitoyl-N-monomethyl-N,N-dimethyl-N,N,N-trimethyl-6-O-glycolchitosan (Molecular Envelope Technology – MET) in an aqueous formulation. The formulation was characterized, and its stability studied under three storage conditions for one month. The ocular distribution of the formulation was studied in healthy rabbits and the ocular tissues and the whole blood analyzed by LC-MS/MS. A 200 nm nanoparticle formulation (MET-TAC) containing 0.1 ± 0.002% w/v TAC was produced with viscosity, osmolarity and pH within the ocular comfort range, and the formulation was stable on refrigeration for one month. On topical application, the TAC concentrations in rabbit cornea and conjunctiva one hour after dosing were 4452 ± 2289 and 516 ± 180 ng/g of tissue, respectively. A topical ocular aqueous TAC eye drop formulation has been prepared with the ability to deliver sufficient drug to the relevant ocular surface tissues.

Formation of Dust Filaments in the Diffuse Envelopes of Molecular Clouds

Abstract The path to understanding star formation processes begins with the study of the formation of molecular clouds. The outskirts of these clouds are characterized by low column densities that allow the penetration of ultraviolet radiation, resulting in a nonnegligible ionization fraction and the charging of the small dust grains that are mixed with the gas; this diffuse phase is then coupled to the ambient magnetic field. Despite the general assumption that dust and gas are tightly correlated, several observational and theoretical studies have reported variations in the dust-to-gas ratio toward diffuse and cold clouds. In this work, we present the implementation of a new charged particles module for analyzing the dust dynamics in molecular cloud envelopes. We study the evolution of a single population of small charged grains (0.05 μm) in the turbulent, magnetized molecular cloud envelope using this module. We show that variations in the dust-to-gas ratio arise due to the coupling of the grains with the magnetic field, forming elongated dust structures decoupled from the gas. This emphasizes the importance of considering the dynamics of charged dust when simulating the different phases of the interstellar medium, especially for star formation studies.

CHEOPS observations of the HD 108236 planetary system: a fifth planet, improved ephemerides, and planetary radii

Context. The detection of a super-Earth and three mini-Neptunes transiting the bright (V = 9.2 mag) star HD 108236 (also known as TOI-1233) was recently reported on the basis of TESS and ground-based light curves. Aims. We perform a first characterisation of the HD 108236 planetary system through high-precision CHEOPS photometry and improve the transit ephemerides and system parameters. Methods. We characterise the host star through spectroscopic analysis and derive the radius with the infrared flux method. We constrain the stellar mass and age by combining the results obtained from two sets of stellar evolutionary tracks. We analyse the available TESS light curves and one CHEOPS transit light curve for each known planet in the system. Results. We find that HD 108236 is a Sun-like star with R⋆ = 0.877 ± 0.008 R⊙, M⋆ = 0.869−0.048+0.050 M⊙, and an age of 6.7−5.1+4.0 Gyr. We report the serendipitous detection of an additional planet, HD 108236 f, in one of the CHEOPS light curves. For this planet, the combined analysis of the TESS and CHEOPS light curves leads to a tentative orbital period of about 29.5 days. From the light curve analysis, we obtain radii of 1.615 ± 0.051, 2.071 ± 0.052, 2.539−0.065+0.062, 3.083 ± 0.052, and 2.017−0.057+0.052 R⊕ for planets HD 108236 b to HD 108236 f, respectively. These values are in agreement with previous TESS-based estimates, but with an improved precision of about a factor of two. We perform a stability analysis of the system, concluding that the planetary orbits most likely have eccentricities smaller than 0.1. We also employ a planetary atmospheric evolution framework to constrain the masses of the five planets, concluding that HD 108236 b and HD 108236 c should have an Earth-like density, while the outer planets should host a low mean molecular weight envelope. Conclusions. The detection of the fifth planet makes HD 108236 the third system brighter than V = 10 mag to host more than four transiting planets. The longer time span enables us to significantly improve the orbital ephemerides such that the uncertainty on the transit times will be of the order of minutes for the years to come. A comparison of the results obtained from the TESS and CHEOPS light curves indicates that for a V ~ 9 mag solar-like star and a transit signal of ~500 ppm, one CHEOPS transit light curve ensures the same level of photometric precision as eight TESS transits combined, although this conclusion depends on the length and position of the gaps in the light curve.

Macromolecular phasing using diffraction from multiple crystal forms.

A phasing algorithm for macromolecular crystallography is proposed that utilizes diffraction data from multiple crystal forms - crystals of the same molecule with different unit-cell packings (different unit-cell parameters or space-group symmetries). The approach is based on the method of iterated projections, starting with no initial phase information. The practicality of the method is demonstrated by simulation using known structures that exist in multiple crystal forms, assuming some information on the molecular envelope and positional relationships between the molecules in the different unit cells. With incorporation of new or existing methods for determination of these parameters, the approach has potential as a method for ab initio phasing.

ALMA Observations and Modeling of the Rotating Outflow in Orion Source I

Abstract We present 29SiO(J = 8–7) ν = 0, SiS (J = 19–18) ν = 0, and 28SiO (J = 8–7) ν = 1 molecular line archive observations made with the Atacama Large Millimeter/Submillimeter Array (ALMA) of the molecular outflow associated with Orion Source I. The observations show velocity asymmetries about the flow axis that are interpreted as outflow rotation. We find that the rotation velocity (∼4–8 km s−1) decreases with the vertical distance to the disk. In contrast, the cylindrical radius (∼100–300 au), the expansion velocity (∼2–15 km s−1), and the axial velocity v z (∼−1–10 km s−1) increase with the vertical distance. The mass estimated of the molecular outflow M outflow ∼ 0.66–1.3 M ⊙. Given a kinematic time ∼130 yr, this implies a mass-loss rate M ⊙ yr−1. This massive outflow sets important constraints on disk wind models. We compare the observations with a model of a shell produced by the interaction between an anisotropic stellar wind and an Ulrich accretion flow that corresponds to a rotating molecular envelope in collapse. We find that the model cylindrical radii are consistent with the 29SiO(J = 8–7) ν = 0 data. The expansion velocities and the axial velocities of the model are similar to the observed values, except for the expansion velocity close to the disk (z ∼ ±150 au). Nevertheless, the rotation velocities of the model are a factor ∼3–10 lower than the observed values. We conclude that the Ulrich flow alone cannot explain the rotation observed and other possibilities should be explored, like the inclusion of the angular momentum of a disk wind.

Structural Characterization of Non-structural Protein 9 Complexed With Specific Nanobody Pinpoints Two Important Residues Involved in Porcine Reproductive and Respiratory Syndrome Virus Replication.

Porcine reproductive and respiratory syndrome (PRRS), caused by PRRS virus (PRRSV), is a widespread viral disease that has led to huge economic losses for the global swine industry. Non-structural protein 9 (Nsp9) of PRRSV possesses essential RNA-dependent RNA polymerase (RdRp) activity for viral RNA replication. Our previous report showed that Nsp9-specific nanobody, Nb6, was able to inhibit PRRSV replication. In this study, recombinant Nsp9 and Nsp9-Nb6 complex were prepared then characterized using bio-layer interferometry (BLI) and dynamic light scattering (DLS) analyses that demonstrated high-affinity binding of Nb6 to Nsp9 to form a homogeneous complex. Small-angle X-ray scattering (SAXS) characterization analyses revealed that spatial interactions differed between Nsp9 and Nsp9-Nb6 complex molecular envelopes. Enzyme-linked immunosorbent assays (ELISAs) revealed key involvement of Nsp9 residues Ile588, Asp590, and Leu643 and Nb6 residues Tyr62, Trp105, and Pro107 in the Nsp9-Nb6 interaction. After reverse genetics-based techniques were employed to generate recombinant Nsp9 mutant viruses, virus replication efficiencies were assessed in MARC-145 cells. The results revealed impaired viral replication of recombinant viruses bearing I588A and L643A mutations as compared with replication of wild type virus, as evidenced by reduced negative-strand genomic RNA [(−) gRNA] synthesis and attenuated viral infection. Moreover, the isoleucine at position 588 of Nsp9 was conserved across PRRSV genotypes. In conclusion, structural analysis of the Nsp9-Nb6 complex revealed novel amino acid interactions involved in viral RNA replication that will be useful for guiding development of structure-based anti-PRRSV agents.

Exclusion of Cosmic Rays from Molecular Clouds by Self-generated Electric Fields

Abstract It was recently discovered that in some regions of the Galaxy, the cosmic-ray (CR) abundance is several orders of magnitude higher than previously thought. Additionally, there is evidence that in molecular cloud envelopes, the CR ionization may be dominated by electrons. We show that for regions with high, electron-dominated ionization, the penetration of CR electrons into molecular clouds is modulated by the electric field that develops as a result of the charge they deposit. We evaluate the significance of this novel mechanism of self-modulation and show that the CR penetration can be reduced by a factor of a few to a few hundred in high-ionization environments, such as those found near the Galactic center.