Influence of Substitutions in the Binding Motif of Proline-Rich Antimicrobial Peptide ARV-1502 on 70S Ribosome Binding and Antimicrobial Activity

Proline-rich antimicrobial peptides (PrAMPs) are promising candidates to treat bacterial infections. The designer peptide ARV-1502 exhibits strong antimicrobial effects against Enterobacteriaceae both in vitro and in vivo. Since the inhibitory effects of ARV-1502 reported for the 70 kDa heat-shock protein DnaK do not fully explain the antimicrobial activity of its 176 substituted analogs, we further studied their effect on the bacterial 70S ribosome of Escherichia coli, a known target of PrAMPs. ARV-1502 analogues, substituted in positions 3, 4, and 8 to 12 (underlined) of the binding motif D3KPRPYLPRP12 with aspartic acid, lysine, serine, phenylalanine or leucine, were tested in a competitive fluorescence polarization (FP) binding screening assay using 5(6)-carboxyfluoresceinlabeled (Cf-) ARV-1502 and the 70S ribosome isolated from E. coli BW25113. While their effect on ribosomal protein expression was studied for green fluorescent protein (GFP) in a cell-free expression system (in vitro translation), the importance of known PrAMP transporters SbmA and MdtM was investigated using E. coli BW25113 and the corresponding knockout mutants. The dissociation constant (Kd) of 201 16 nmol/L obtained for Cf-ARV-1502 suggests strong binding to the E. coli 70S ribosome. An inhibitory binding assay indicated that the binding site overlaps with those of other PrAMPs including Onc112 and pyrrhocoricin as well as the non-peptidic antibiotics erythromycin and chloramphenicol. All these drugs and drug candidates bind to the exit-tunnel of the 70S ribosome. Substitutions of the C-terminal fragment of the binding motif YLPRP reduced binding. At the same time, inhibition of GFP expression increased with net peptide charge. Interestingly, the MIC values of wild-type and DsbmA and DmdtM knockout mutants indicated that substitutions in the ribosomal binding motif altered also the bacterial uptake, which was generally improved by incorporation of hydrophobic residues. In conclusion, most substituted ARV-1502 analogs bound weaker to the 70S ribosome than ARV-1502 underlining the importance of the YLPRP binding motif. The weaker ribosomal binding correlated well with decreased antimicrobial activity in vitro. Substituted ARV-1502 analogs with a higher level of hydrophobicity or positive net charge improved the ribosome binding, inhibition of translation, and bacterial uptake

Safety and Pharmacokinetics of a Four Monoclonal Antibody Combination Against Botulinum C and D Neurotoxins.

Botulism is caused by botulinum neurotoxin (BoNT), the most poisonous substance known. BoNTs are also classified as Tier 1 biothreat agents due to their high potency and lethality. The existence of seven BoNT serotypes (A-G), which differ between 35% to 68% in amino acid sequence, necessitates the development of serotype specific countermeasures. We present results of a Phase 1 clinical study of an anti-toxin to BoNT serotypes C and D, NTM-1634, which consists of an equimolar mixture of four fully human IgG1 monoclonal antibodies (mAbs), each binding to non-overlapping epitopes on BoNT serotypes C and D resulting in potent toxin neutralization in rodents. This first-in-human study evaluated the safety and pharmacokinetics of escalating doses of NTM-1634 administered intravenously to healthy adults (NCT03046550). Three cohorts of eight healthy subjects received a single intravenous dose of NTM-1634 or placebo at 0.33 mg/kg, 0.66 mg/kg or 1 mg/kg. Follow-up examinations and pharmacokinetic evaluations were continued up to 121 days post-infusion. Subjects were monitored using physical examinations, hematology and chemistry blood tests, and electrocardiograms. Pharmacokinetic parameters were estimated using noncompartmental methods. The results demonstrated that the materials were safe and well-tolerated with the expected half-lives for human mAbs and with minimal anti-drug antibodies detected over the dose ranges and duration of the study

A Triple Protostar System Formed via Fragmentation of a Gravitationally Unstable Disk

Binary and multiple star systems are a frequent outcome of the star formation process, and as a result, almost half of all sun-like stars have at least one companion star. Theoretical studies indicate that there are two main pathways that can operate concurrently to form binary/multiple star systems: large scale fragmentation of turbulent gas cores and filaments or smaller scale fragmentation of a massive protostellar disk due to gravitational instability. Observational evidence for turbulent fragmentation on scales of $>$1000~AU has recently emerged. Previous evidence for disk fragmentation was limited to inferences based on the separations of more-evolved pre-main sequence and protostellar multiple systems. The triple protostar system L1448 IRS3B is an ideal candidate to search for evidence of disk fragmentation. L1448 IRS3B is in an early phase of the star formation process, likely less than 150,000 years in age, and all protostars in the system are separated by $<$200~AU. Here we report observations of dust and molecular gas emission that reveal a disk with spiral structure surrounding the three protostars. Two protostars near the center of the disk are separated by 61 AU, and a tertiary protostar is coincident with a spiral arm in the outer disk at a 183 AU separation. The inferred mass of the central pair of protostellar objects is $\sim$1 M$_{sun}$, while the disk surrounding the three protostars has a total mass of $\sim$0.30 M_{\sun}. The tertiary protostar itself has a minimum mass of $\sim$0.085 M$_{sun}$. We demonstrate that the disk around L1448 IRS3B appears susceptible to disk fragmentation at radii between 150~AU and 320~AU, overlapping with the location of the tertiary protostar. This is consistent with models for a protostellar disk that has recently undergone gravitational instability, spawning one or two companion stars.Comment: Published in Nature on Oct. 27th. 24 pages, 8 figure

TESS Hunt for Young and Maturing Exoplanets (THYME). X. A Two-planet System in the 210 Myr MELANGE-5 Association

Young (&lt;500 Myr) planets are critical to studying how planets form and evolve. Among these young planetary systems, multiplanet configurations are particularly useful, as they provide a means to control for variables within a system. Here, we report the discovery and characterization of a young planetary system, TOI-1224. We show that the planet host resides within a young population we denote as MELANGE-5. By employing a range of age-dating methods—isochrone fitting, lithium abundance analysis, gyrochronology, and Gaia excess variability—we estimate the age of MELANGE-5 to be 210 ± 27 Myr. MELANGE-5 is situated in close proximity to previously identified younger (80–110 Myr) associations, Crius 221 and Theia 424/Volans-Carina, motivating further work to map out the group boundaries. In addition to a planet candidate detected by the TESS pipeline and alerted as a TESS object of interest, TOI-1224 b, we identify a second planet, TOI-1224 c, using custom search tools optimized for young stars (Notch and LOCoR). We find that the planets are 2.10 ± 0.09 R⊕ and 2.88 ± 0.10 R⊕ and orbit their host star every 4.18 and 17.95 days, respectively. With their bright (K = 9.1 mag), small (R* = 0.44 R⊙), and cool (Teff = 3326 K) host star, these planets represent excellent candidates for atmospheric characterization with JWST

TESS Hunt for Young and Maturing Exoplanets (THYME). X. A Two-planet System in the 210 Myr MELANGE-5 Association

Young (&lt;500 Myr) planets are critical to studying how planets form and evolve. Among these young planetary systems, multiplanet configurations are particularly useful, as they provide a means to control for variables within a system. Here, we report the discovery and characterization of a young planetary system, TOI-1224. We show that the planet host resides within a young population we denote as MELANGE-5. By employing a range of age-dating methods—isochrone fitting, lithium abundance analysis, gyrochronology, and Gaia excess variability—we estimate the age of MELANGE-5 to be 210 ± 27 Myr. MELANGE-5 is situated in close proximity to previously identified younger (80–110 Myr) associations, Crius 221 and Theia 424/Volans-Carina, motivating further work to map out the group boundaries. In addition to a planet candidate detected by the TESS pipeline and alerted as a TESS object of interest, TOI-1224 b, we identify a second planet, TOI-1224 c, using custom search tools optimized for young stars (Notch and LOCoR). We find that the planets are 2.10 ± 0.09 R⊕ and 2.88 ± 0.10 R⊕ and orbit their host star every 4.18 and 17.95 days, respectively. With their bright (K = 9.1 mag), small (R* = 0.44 R⊙), and cool (Teff = 3326 K) host star, these planets represent excellent candidates for atmospheric characterization with JWST

TESS Hunt for Young and Maturing Exoplanets (THYME) IX: a 27 Myr extended population of Lower-Centaurus Crux with a transiting two-planet system

We report the discovery and characterization of a nearby (~ 85 pc), older (27 +/- 3 Myr), distributed stellar population near Lower-Centaurus-Crux (LCC), initially identified by searching for stars co-moving with a candidate transiting planet from TESS (HD 109833; TOI 1097). We determine the association membership using Gaia kinematics, color-magnitude information, and rotation periods of candidate members. We measure it's age using isochrones, gyrochronology, and Li depletion. While the association is near known populations of LCC, we find that it is older than any previously found LCC sub-group (10-16 Myr), and distinct in both position and velocity. In addition to the candidate planets around HD 109833 the association contains four directly-imaged planetary-mass companions around 3 stars, YSES-1, YSES-2, and HD 95086, all of which were previously assigned membership in the younger LCC. Using the Notch pipeline, we identify a second candidate transiting planet around HD 109833. We use a suite of ground-based follow-up observations to validate the two transit signals as planetary in nature. HD 109833 b and c join the small but growing population of <100 Myr transiting planets from TESS. HD 109833 has a rotation period and Li abundance indicative of a young age (< 100 Myr), but a position and velocity on the outskirts of the new population, lower Li levels than similar members, and a CMD position below model predictions for 27 Myr. So, we cannot reject the possibility that HD 109833 is a young field star coincidentally nearby the population.Comment: 23 pages, 15 figures, Accepted for publication in A

<i>TESS</i> Spots a Compact System of Super-Earths around the Naked-eye Star HR 858

Transiting Exoplanet Survey Satellite (TESS) observations have revealed a compact multiplanet system around the sixth-magnitude star HR 858 (TIC 178155732, TOI 396), located 32 pc away. Three planets, each about twice the size of Earth, transit this slightly evolved, late F-type star, which is also a member of a visual binary. Two of the planets may be in mean motion resonance. We analyze the TESS observations, using novel methods to model and remove instrumental systematic errors, and combine these data with follow-up observations taken from a suite of ground-based telescopes to characterize the planetary system. The HR 858 planets are enticing targets for precise radial velocity observations, secondary eclipse spectroscopy, and measurements of the Rossiter–McLaughlin effect

Association of Resistance Exercise With the Incidence of Hypercholesterolemia in Men.

OBJECTIVE: To examine the associations of resistance exercise, independent of and combined with aerobic exercise, with the risk of development of hypercholesterolemia in men. PATIENTS AND METHODS: This study used data from the Aerobics Center Longitudinal Study, which is a cohort examining the associations of clinical and lifestyle factors with the development of chronic diseases and mortality. Participants received extensive preventive medical examinations at the Cooper Clinic in Dallas, Texas, between January 1, 1987, and December 31, 2006. A total of 7317 men aged 18 to 83 years (mean age, 46 years) without hypercholesterolemia at baseline were included. Frequency (times per week) and total amount (min/wk) of resistance and aerobic exercise were determined by self-report. Hypercholesterolemia was defined as a total cholesterol level of 240 mg/dL or higher or physician diagnosis. RESULTS: During a median (interquartile range) follow-up of 4 (2 to 7) years, hypercholesterolemia developed in 1430 of the 7317 men (20%). Individuals meeting the resistance exercise guidelines (≥2 d/wk) had a 13% lower risk of development of hypercholesterolemia (hazard ratio [HR], 0.87; 95% CI, 0.76-0.99; P=.04) after adjustment for general characteristics, lifestyle factors, and aerobic exercise. In addition, less than 1 h/wk and 2 sessions per week of resistance exercise were associated with 32% and 31% lower risks of hypercholesterolemia (HR, 0.68; 95% CI, 0.54-0.86; P=.001; and HR, 0.69; 95% CI, 0.54-0.88; P=.003), respectively, compared with no resistance exercise. Higher levels of resistance exercise did not provide benefits. Meeting both resistance and aerobic exercise guidelines (≥500 metabolic equivalent task min/wk) lowered the risk of development of hypercholesterolemia by 21% (HR, 0.79; 95% CI, 0.68-0.91; P=.002). compared with meeting none of the guidelines. CONCLUSION: Compared with no resistance exercise, less than 1 h/wk of resistance exercise, independent of aerobic exercise, is associated with a significantly lower risk of development of hypercholesterolemia in men (P=.001). However, the lowest risk of hypercholesterolemia was found at 58 min/wk of resistance exercise. This finding suggests that resistance exercise should be encouraged to prevent hypercholesterolemia in men. However, future studies with a more rigorous analysis including major potential confounders (eg, diet, medications) are warranted