Multivariate Normality of Cartesian-Framed Covariances: Evaluation and Operational Significance

Collision avoidance relies on representative Cartesian uncertainty volumes in order to calculate probabilities of collision. Among the potential shortcomings of a covariance matrix representation of state errors, the most worrisome is the coordinate mismatch between the Cartesian framework in which these matrices are distributed and the curvilinear path that satellite orbits actually follow. The present study compares curvilinear-based and Cartesian covariance representations for ~50,000 conjunctions to determine the frequency in which significant deviations from Gaussianity are observed, then compares the 2-D Pc result from the Cartesian covariance to a Monte Carlo Pc conducted in element space to assess operational significance

Assessment and Validation of Collision "Consequence" Method of Assessing Orbital Regime Risk Posed by Potential Satellite Conjunctions

Collision risk management theory requires a thorough assessment of both the likelihood and consequence of potential collision events. Satellite conjunction risk assessment has produced a highly-developed theory for assessing the likelihood of collision but neglects to account for the consequences of a given collision. While any collision may compromise the operational survival of a spacecraft, the amount of debris produced by the potential collision, and therefore the degree to which the orbital corridor may be compromised, can vary greatly among satellite conjunctions. Previous studies leveraged work on satellite collision modeling to develop a method to estimate whether a particular collision is likely to produce a relatively large or relatively small amount of resultant debris. The approximation of the number of debris pieces is dependent on a mass estimation process for the secondary objects utilizing the radar cross section of said object. This study examines the validity of the mass estimation process and establishes uncertainty bounds on the secondary object mass, which will be used to best approximate the possible consequences of a potential collision. This process is then applied to a large set of historical conjunctions to assess the frequency at which possible collisions may significantly augment the orbital debris environment in operational spacecraft

Multivariate Normality of Cartesian Frame Covariances: Evaluation and Operational Significance

Collision avoidance relies on representative Cartesian uncertainty volumes in order to calculate probabilities of collision. Among the potential shortcomings of a covariance matrix representation of state errors, the most worrisome is the coordinate mismatch between the Cartesian framework in which these matrices are distributed and the curvilinear path that satellite orbits actually follow. The present study compares curvilinear-based and Cartesian covariance representations for ~50,000 conjunctions to determine the frequency in which significant deviations from Gaussianity are observed, then compares the 2-D Pc result from the Cartesian covariance to a Monte Carlo Pc conducted in element space to assess operational significance

Using a CO2 Surgical Laser for Piglet Castration to Reduce Pain and Inflammation, and to Improve Wound Healing

The objectives of this preliminary study were to determine the ability of a CO2 surgical laser to 1) reduce pain, 2) reduce inflammation, and 3) improve wound healing of piglets undergoing surgical castration. Two-day old male Yorkshire × Landrace piglets were used and randomly assigned to one of three treatments (n = 10 piglets/treatment group): surgical castration with the CO2 laser, surgical castration with a scalpel, or sham (uncastrated control). Piglets were video recorded in their pens for 1 h pre-procedure and from 0-2, 6-8, and at 24 h post-procedure for behavior scoring. Surgical site images were collected at baseline, 0, 8, 24, 48, 72, 96, 120, 144, and 168 h post-castration for wound healing assessment. Infrared thermography (IRT) images of the surgical site were also taken at baseline, 0, 0.5, 8, and 24 h post-procedure to assess inflammation. Finally, blood was collected from each piglet at baseline and 0.5 h post-castration to assess cortisol levels, prostaglandin E metabolite (PGEM), and pig-major acute phase protein (pig-MAP) concentration. Laser-castrated piglets displayed more pain behav­iors across the observation period than scalpel-castrated piglets (P = 0.049). Laser-castrated piglets also displayed significantly more agonistic behavior than both scalpel-castrated and sham piglets (P = 0.005 and P = 0.036, respectively); yet, laser-castrated piglets had significantly lower temperatures at the site of incision compared to scalpel-castrated piglets (P = 0.0211). There was no significant difference in wound healing or any of the blood parameters assessed between laser-castrated and scalpel-castrated piglets. There was evidence of thermal tissue damage on the scrotum of piglets that were castrated using the CO2 laser. This may have resulted in the unremarkable healing time and the increased pain behavior observed in this study. The surgical laser technique should be refined before conclusions can be made regarding the utility of a CO2 laser for piglet castration

Counter-current chromatography for the separation of terpenoids: A comprehensive review with respect to the solvent systems employed

Copyright @ 2014 The Authors.This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.Natural products extracts are commonly highly complex mixtures of active compounds and consequently their purification becomes a particularly challenging task. The development of a purification protocol to extract a single active component from the many hundreds that are often present in the mixture is something that can take months or even years to achieve, thus it is important for the natural product chemist to have, at their disposal, a broad range of diverse purification techniques. Counter-current chromatography (CCC) is one such separation technique utilising two immiscible phases, one as the stationary phase (retained in a spinning coil by centrifugal forces) and the second as the mobile phase. The method benefits from a number of advantages when compared with the more traditional liquid-solid separation methods, such as no irreversible adsorption, total recovery of the injected sample, minimal tailing of peaks, low risk of sample denaturation, the ability to accept particulates, and a low solvent consumption. The selection of an appropriate two-phase solvent system is critical to the running of CCC since this is both the mobile and the stationary phase of the system. However, this is also by far the most time consuming aspect of the technique and the one that most inhibits its general take-up. In recent years, numerous natural product purifications have been published using CCC from almost every country across the globe. Many of these papers are devoted to terpenoids-one of the most diverse groups. Naturally occurring terpenoids provide opportunities to discover new drugs but many of them are available at very low levels in nature and a huge number of them still remain unexplored. The collective knowledge on performing successful CCC separations of terpenoids has been gathered and reviewed by the authors, in order to create a comprehensive document that will be of great assistance in performing future purifications. © 2014 The Author(s)

RBR ligase–mediated ubiquitin transfer: a tale with many twists and turns

RBR ligases are an enigmatic class of E3 ubiquitin ligases that combine properties of RING and HECT-type E3s and undergo multilevel regulation through autoinhibition, post-translational modifications, multimerization and interaction with binding partners. Here, we summarize recent progress in RBR structures and function, which has uncovered commonalities in the mechanisms by which different family members transfer ubiquitin through a multistep process. However, these studies have also highlighted clear differences in the activity of different family members, suggesting that each RBR ligase has evolved specific properties to fit the biological process it regulates

Structural Studies of a Four-MBT Repeat Protein MBTD1

The Polycomb group (PcG) of proteins is a family of important developmental regulators. The respective members function as large protein complexes involved in establishment and maintenance of transcriptional repression of developmental control genes. MBTD1, Malignant Brain Tumor domain-containing protein 1, is one such PcG protein. MBTD1 contains four MBT repeats.We have determined the crystal structure of MBTD1 (residues 130-566aa covering the 4 MBT repeats) at 2.5 A resolution by X-ray crystallography. The crystal structure of MBTD1 reveals its similarity to another four-MBT-repeat protein L3MBTL2, which binds lower methylated lysine histones. Fluorescence polarization experiments confirmed that MBTD1 preferentially binds mono- and di-methyllysine histone peptides, like L3MBTL1 and L3MBTL2. All known MBT-peptide complex structures characterized to date do not exhibit strong histone peptide sequence selectivity, and use a "cavity insertion recognition mode" to recognize the methylated lysine with the deeply buried methyl-lysine forming extensive interactions with the protein while the peptide residues flanking methyl-lysine forming very few contacts [1]. Nevertheless, our mutagenesis data based on L3MBTL1 suggested that the histone peptides could not bind to MBT repeats in any orientation.The four MBT repeats in MBTD1 exhibits an asymmetric rhomboid architecture. Like other MBT repeat proteins characterized so far, MBTD1 binds mono- or dimethylated lysine histones through one of its four MBT repeats utilizing a semi-aromatic cage.This article can also be viewed as an enhanced version in which the text of the article is integrated with interactive 3D representations and animated transitions. Please note that a web plugin is required to access this enhanced functionality. Instructions for the installation and use of the web plugin are available in Text S1

Activity-based E3 ligase profiling uncovers an E3 ligase with esterification activity

Ubiquitination is initiated by transfer of ubiquitin (Ub) from a ubiquitin-activating enzyme (E1) to a ubiquitin-conjugating enzyme (E2), producing a covalently linked intermediate (E2-Ub)(1). Ubiquitin ligases (E3s) of the 'really interesting new gene' (RING) class recruit E2-Ub via their RING domain and then mediate direct transfer of ubiquitin to substrates(2). By contrast, 'homologous to E6-AP carboxy terminus' (HECT) E3 ligases undergo a catalytic cysteine-dependent transthiolation reaction with E2-Ub, forming a covalent E3-Ub intermediate(3,4). Additionally, RING-between-RING (RBR) E3 ligases have a canonical RING domain that is linked to an ancillary domain. This ancillary domain contains a catalytic cysteine that enables a hybrid RING-HECT mechanism(5). Ubiquitination is typically considered a post-translational modification of lysine residues, as there are no known human E3 ligases with non-lysine activity. Here we perform activity-based protein profiling of HECT or RBR-like E3 ligases and identify the neuron-associated E3 ligase MYCBP2 (also known as PHR1) as the apparent single member of a class of RING-linked E3 ligase with esterification activity and intrinsic selectivity for threonine over serine. MYCBP2 contains two essential catalytic cysteine residues that relay ubiquitin to its substrate via thioester intermediates. Crystallographic characterization of this class of E3 ligase, which we designate RING-Cys-relay (RCR), provides insights into its mechanism and threonine selectivity. These findings implicate non-lysine ubiquitination in cellular regulation of higher eukaryotes and suggest that E3 enzymes have an unappreciated mechanistic diversity

The serine protease domain of MASP-3: enzymatic properties and crystal structure in complex with ecotin.

International audienceMannan-binding lectin (MBL), ficolins and collectin-11 are known to associate with three homologous modular proteases, the MBL-Associated Serine Proteases (MASPs). The crystal structures of the catalytic domains of MASP-1 and MASP-2 have been solved, but the structure of the corresponding domain of MASP-3 remains unknown. A link between mutations in the MASP1/3 gene and the rare autosomal recessive 3MC (Mingarelli, Malpuech, Michels and Carnevale,) syndrome, characterized by various developmental disorders, was discovered recently, revealing an unexpected important role of MASP-3 in early developmental processes. To gain a first insight into the enzymatic and structural properties of MASP-3, a recombinant form of its serine protease (SP) domain was produced and characterized. The amidolytic activity of this domain on fluorescent peptidyl-aminomethylcoumarin substrates was shown to be considerably lower than that of other members of the C1r/C1s/MASP family. The E. coli protease inhibitor ecotin bound to the SP domains of MASP-3 and MASP-2, whereas no significant interaction was detected with MASP-1, C1r and C1s. A tetrameric complex comprising an ecotin dimer and two MASP-3 SP domains was isolated and its crystal structure was solved and refined to 3.2 Å. Analysis of the ecotin/MASP-3 interfaces allows a better understanding of the differential reactivity of the C1r/C1s/MASP protease family members towards ecotin, and comparison of the MASP-3 SP domain structure with those of other trypsin-like proteases yields novel hypotheses accounting for its zymogen-like properties in vitro