Characterization of Ribeye Subunits in Zebrafish Hair Cells Reveals That Exogenous Ribeye B-Domain and CtBP1 Localize to the Basal Ends of Synaptic Ribbons

Synaptic ribbons are presynaptic structures formed by the self-association of RIBEYE–the main structural component of ribbon synapses. RIBEYE consists of two domains: a unique N-terminal A-domain and a C-terminal B-domain that is identical to the transcription co-repressor C-terminal binding protein 2 (CtBP2). Previous studies in cell lines have shown that RIBEYE A-domain alone is sufficient to form ribbon-like aggregates and that both A- and B- domains form homo-and heterotypic interactions. As these interactions are likely the basis for synaptic-ribbon assembly and structural plasticity, we wanted to examine how zebrafish Ribeye A- and B- domains interact with synaptic ribbons in vivo. To that end, we characterized the localization of exogenously expressed Ribeye A- and B- domains and the closely related protein, CtBP1, in the hair cells of transgenic zebrafish larvae. Unexpectedly, exogenously expressed Ribeye A-domain showed variable patterns of localization in hair cells; one zebrafish paralog of A-domain failed to self-associate or localize to synaptic ribbons, while the other self-assembled but sometimes failed to localize to synaptic ribbons. By contrast, Ribeye B-domain/CtBP2 was robustly localized to synaptic ribbons. Moreover, both exogenously expressed B-domain/CtBP2 and CtBP1 were preferentially localized to the basal end of ribbons adjacent to the postsynaptic density. Overexpression of B-domain/CtBP2 also appeared to affect synaptic-ribbon composition; endogenous levels of ribbon-localized Ribeye were significantly reduced as hair cells matured in B-domain/CtBP2 transgenic larvae compared to wild-type. These results reveal how exogenously expressed Ribeye domains interact with synaptic ribbons, and suggest a potential organization of elements within the ribbon body

The Quest for the Missing Dust: II -- Two Orders of Magnitude of Evolution in the Dust-to-Gas Ratio Resolved Within Local Group Galaxies

We explore evolution in the dust-to-gas ratio with density within four well-resolved Local Group galaxies - the LMC, SMC, M31, and M33. We do this using new ${\it Herschel}$ maps, which restore extended emission that was missed by previous ${\it Herschel}$ reductions. This improved data allows us to probe the dust-to-gas ratio across 2.5 orders of magnitude in ISM surface density. We find significant evolution in the dust-to-gas ratio, with dust-to-gas varying with density within each galaxy by up to a factor 22.4. We explore several possible reasons for this, and our favored explanation is dust grain growth in denser regions of ISM. We find that the evolution of the dust-to-gas ratio with ISM surface density is very similar between M31 and M33, despite their large differences in mass, metallicity, and star formation rate; conversely, we find M33 and the LMC to have very different dust-to-gas evolution profiles, despite their close similarity in those properties. Our dust-to-gas ratios address previous disagreement between UV- and FIR-based dust-to-gas estimates for the Magellanic Clouds, removing the disagreement for the LMC, and considerably reducing it for the SMC - with our new dust-to-gas measurements being factors of 2.4 and 2.0 greater than the previous far-infrared estimates, respectively. We also observe that the dust-to-gas ratio appears to fall at the highest densities for the LMC, M31, and M33; this is unlikely to be an actual physical phenomenon, and we posit that it may be due to a combined effect of dark gas, and changing dust mass opacity.Comment: Accepted for publication in the Astrophysical Journa

A multi-institutional study evaluating and describing atypical parathyroid tumors discovered after parathyroidectomy

Objective: To describe common intraoperative and pathologic findings of atypical parathyroid tumors (APTs) and evaluate clinical outcomes in patients undergoing parathyroidectomy. Methods: In this multi-institutional retrospective case series, data were collected from patients who underwent parathyroidectomy from 2000 to 2018 from three tertiary care institutions. APTs were defined according to the AJCC eighth edition guidelines and retrospective chart review was performed to evaluate the incidence of recurrent laryngeal nerve injury, recurrence of disease, and disease-specific mortality. Results: Twenty-eight patients were identified with a histopathologic diagnosis of atypical tumor. Mean age was 56 years (range, 23-83) and 68% (19/28) were female. All patients had an initial diagnosis of primary hyperparathyroidism with 21% (6/28) exhibiting clinical loss of bone density and 32% (9/28) presenting with nephrolithiasis or renal dysfunction. Intraoperatively, 29% (8/28) required thyroid lobectomy, 29% (8/28) had gross adherence to adjacent structures and 46% (13/28) had RLN adherence. The most common pathologic finding was fibrosis 46% (13/28). Postoperative complications include RLN paresis/paralysis in 14% (4/28) and hungry bone syndrome in 7% (2/28). No patients with a diagnosis of atypical tumor developed recurrent disease, however there was one patient that had persistent disease and hypercalcemia that is being observed. There were 96% (27/28) patients alive at last follow-up, with one death unrelated to disease. Conclusion: Despite the new AJCC categorization of atypical tumors staged as Tis, we observed no recurrence of disease after resection and no disease-specific mortality. However, patients with atypical tumors may be at increased risk for recurrent laryngeal nerve injury and incomplete resection

An Advanced Trajectory-Based Operations Prototype Tool and Focus Group Evaluation

Trajectory-based operations (TBO) is a key concept in the Next Generation Air Transportation System transformation of the National Airspace System (NAS) that will increase the predictability and stability of traffic flows, support a common operational picture through the use of digital data sharing, facilitate more effective collaborative decision making between airspace users and air navigation service providers, and enable increased levels of integrated automation across the NAS. The National Aeronautics and Space Administration (NASA) has been developing trajectory-based systems to improve the efficiency of the NAS during specific phases of flight and is now also exploring Advanced 4-Dimensional Trajectory (4DT) operational concepts that will integrate these technologies and incorporate new technology where needed to create both automation and procedures to support gate-to-gate TBO. A TBO Prototype simulation toolkit has been developed that demonstrates initial functionality that may reside in an Advanced 4DT TBO concept. Pilot and controller subject matter experts (SMEs) were brought to the Air Traffic Operations Laboratory at NASA Langley Research Center for discussions on an Advanced 4DT operational concept and were provided an interactive demonstration of the TBO Prototype using four example scenarios. The SMEs provided feedback on potential operational, technological, and procedural opportunities and concerns. After viewing the interactive demonstration scenarios, the SMEs felt the operational capabilities demonstrated would be useful for performing TBO while maintaining situation awareness and low mental workload. The TBO concept demonstrated produced defined routings around weather which resulted in a more organized, consistent flow of traffic where it was clear to both the controller and pilot what route the aircraft was to follow. In general, the controller SMEs felt that traffic flow management should be responsible for generating and negotiating the operational constraints demonstrated, in cooperation with the Air Traffic Control System Command Center, while air traffic control should be responsible for the implementation of those constraints. The SMEs also indicated that digital data communications would be very beneficial for TBO operations and would result in less workload due to reduced communications, would eliminate issues due to language barriers and frequency problems, and would make receiving, loading, accepting, and executing clearances easier, less ambiguous, and more expeditious. This paper describes an Advanced 4DT operational concept, the TBO Prototype, the demonstration scenarios and methods used, and the feedback obtained from the pilot and controller SMEs in this focus group evaluation

Effects of habitat composition and landscape structure on worker foraging distances of five bumblebee species

Bumblebees (Bombus spp.) are important pollinators of both crops and wild flowers. Their contribution to this essential ecosystem service has been threatened over recent decades by changes in land use, which have led to declines in their populations. In order to design effective conservation measures it is important to understand the effects of variation in landscape composition and structure on the foraging activities of worker bumblebees. This is because the viability of individual colonies is likely to be affected by the trade-off between the energetic costs of foraging over greater distances and the potential gains from access to additional resources. We used field surveys, molecular genetics and fine resolution remote sensing to estimate the locations of wild bumblebee nests and to infer foraging distances across a 20 km2 agricultural landscape in southern England. We investigated five species, including the rare B. ruderatus and ecologically similar but widespread B. hortorum. We compared worker foraging distances between species and examined how variation in landscape composition and structure affected foraging distances at the colony level. Mean worker foraging distances differed significantly between species. Bombus terrestris, B. lapidarius and B. ruderatus exhibited significantly greater mean foraging distances (551 m, 536 m, 501 m, respectively) than B. hortorum and B. pascuorum (336 m, 272 m, respectively). There was wide variation in worker foraging distances between colonies of the same species, which was in turn strongly influenced by the amount and spatial configuration of available foraging habitats. Shorter foraging distances were found for colonies where the local landscape had high coverage and low fragmentation of semi-natural vegetation, including managed agri-environmental field margins. The strength of relationships between different landscape variables and foraging distance varied between species, for example the strongest relationship for B. ruderatus being with floral cover of preferred forage plants. Our findings suggest that favourable landscape composition and configuration has the potential to minimise foraging distances across a range of bumblebee species. There is thus potential for improvements in the design and implementation of landscape management options, such as agri-environment schemes, aimed at providing foraging habitat for bumblebees and enhancing crop pollination services

Luminal Plasma Treatment for Small Diameter Polyvinyl Alcohol Tubular Scaffolds

Plasma-based surface modification is recognized as an effective way to activate biomaterial surfaces, and modulate their interactions with cells, extracellular matrix proteins, and other materials. However, treatment of a luminal surface of a tubular scaffold remains non-trivial to perform in small diameter tubes. Polyvinyl alcohol (PVA) hydrogel, which has been widely used for medical applications, lacks functional groups to mediate cell attachment. This poses an issue for vascular applications, as endothelialization in a vascular graft lumen is crucial to maintain long term graft patency. In this study, a Radio Frequency Glow Discharges (RFGD) treatment in the presence of NH3 was used to modify the luminal surface of 3-mm diameter dehydrated PVA vascular grafts. The grafted nitrogen containing functional groups demonstrated stability, and in vitro endothelialization was successfully maintained for at least 30 days. The plasma-modified PVA displayed a higher percentage of carbonyl groups over the untreated PVA control. Plasma treatment on PVA patterned with microtopographies was also studied, with only the concave microlenses topography demonstrating a significant increase in platelet adhesion. Thus, the study has shown the possibility of modifying a small diameter hydrogel tubular scaffold with the RFGD plasma treatment technique and demonstrated stability in ambient storage conditions for up to 30 days

HDAC9 is implicated in atherosclerotic aortic calcification and affects vascular smooth muscle cell phenotype.

Aortic calcification is an important independent predictor of future cardiovascular events. We performed a genome-wide association meta-analysis to determine SNPs associated with the extent of abdominal aortic calcification (n = 9,417) or descending thoracic aortic calcification (n = 8,422). Two genetic loci, HDAC9 and RAP1GAP, were associated with abdominal aortic calcification at a genome-wide level (P < 5.0 × 10-8). No SNPs were associated with thoracic aortic calcification at the genome-wide threshold. Increased expression of HDAC9 in human aortic smooth muscle cells promoted calcification and reduced contractility, while inhibition of HDAC9 in human aortic smooth muscle cells inhibited calcification and enhanced cell contractility. In matrix Gla protein-deficient mice, a model of human vascular calcification, mice lacking HDAC9 had a 40% reduction in aortic calcification and improved survival. This translational genomic study identifies the first genetic risk locus associated with calcification of the abdominal aorta and describes a previously unknown role for HDAC9 in the development of vascular calcification

Discovery of Inhibitors of Burkholderia pseudomallei Methionine Aminopeptidase with Antibacterial Activity

Evaluation of a series of MetAP inhibitors in an in vitro enzyme activity assay led to the first identification of potent molecules that show significant growth inhibition against Burkholderia pseudomallei. Nitroxoline analogues show excellent inhibition potency in the BpMetAP1 enzyme activity assay with the lowest IC50 of 30 nM and inhibit the growth of B. pseudomallei and B. thailandensis at concentrations ≥31 μM