Enhancing the selective extracellular location of a recombinant E. coli domain antibody by management of fermentation conditions

The preparation of a recombinant protein using Escherichia coli often involves a challenging primary recovery sequence. This is due to the inability to secrete the protein to the extracellular space without a significant degree of cell lysis. This results in the release of nucleic acids, leading to a high viscosity, difficulty to clarify, broth and also to contamination with cell materials such as lipopolysaccharides and host cell proteins. In this paper, we present different fermentation strategies to facilitate the recovery of a V H domain antibody (13.1 kDa) by directing it selectively to the extracellular space and changing the balance between domain antibody to nucleic acid release. The manipulation of the cell growth rate in order to increase the outer cell membrane permeability gave a small ~1.5-fold improvement in released domain antibody to nucleic acid ratio without overall loss of yield. The introduction during fermentation of release agents such as EDTA gave no improvement in the ratio of released domain antibody to nucleic acid and a loss of overall productivity. The use of polyethyleneimine (PEI) during fermentation was with the aim to (a) permeabilise the outer bacterial membrane to release selectively domain antibody and (b) remove selectively by precipitation nucleic acids released during cell lysis. This strategy resulted in up to ~4-fold increase in the ratio of domain antibody to soluble nucleic acid with no reduction in domain antibody overall titre. In addition, a reduction in host cell protein contamination was achieved and there was no increase in endotoxin levels. Similar results were demonstrated with a range of other antibody products prepared in E. coli

Violent Behavior During Psychiatric Inpatient Treatment in a German Prison Hospital

Violent behavior in correctional facilities is common and differs substantially in type, target, implication, and trigger. Research on frequency and characteristics of violent behavior in correctional facilities and psychiatric hospitals is limited. Results from recent research suggest that comorbidity of severe mental disorder, personality disorder, and diagnosis of substance abuse is related to a higher risk of violent behavior. In the Berlin prison hospital, a database was created to collect data from all violent incidences (n=210) between 1997 and 2006 and between 2010 and 2016. In a retrospective, case-control study, we analyzed specific socioeconomic data and psychiatric diagnosis and compared the group of prisoners with violent behavior with randomly selected prisoners of the same department without violent behavior (n = 210). Diagnosis of schizophrenia, non-German nationality, no use of an interpreter, no children, and no previous sentence remained significantly associated with the dependent variable violent behavior. There were no significant differences regarding age and legal statuses. Practical implications for clinical work are discussed

Evaluation of options for harvest of a recombinant E. coli fermentation producing a domain antibody using ultra scale-down techniques and pilot-scale verification

Ultra scale-down (USD) methods operating at the millilitre scale were used to characterise full-scale processing of E. coli fermentation broths autolysed to different extents for release of a domain antibody. The focus was on the primary clarification stages involving continuous centrifugation followed by depth filtration. The performance of this sequence was predicted by USD studies to decrease significantly with increased extents of cell lysis. The use of polyethyleneimine (PEI) reagent was studied to treat the lysed cell broth by precipitation of soluble contaminants such as DNA and flocculation of cell debris material. The USD studies were used to predict the impact of this treatment on the performance and here it was found that the fermentation could be run to maximum productivity using an acceptable clarification process (e.g a centrifugation stage operating at 0.11 L per m(2) equivalent gravity settling area per h followed by a resultant required depth filter area of 0.07 m(2) per L supernatant). A range of USD predictions was verified at the pilot scale for centrifugation followed by depth filtration. This article is protected by copyright. All rights reserved

Hybrid Dissemination: Adding Determinism to Probabilistic Multicasting in Large-Scale P2P Systems

Abstract. Epidemic protocols have demonstrated remarkable scalability and robustness in disseminating information on internet-scale, dynamic P2P systems. However, popular instances of such protocols suffer from a number of significant drawbacks, such as increased message overhead in push-based systems, or low dissemination speed in pull-based ones. In this paper we study push-based epidemic dissemination algorithms, in terms of hit ratio, communication overhead, dissemination speed, and resilience to failures and node churn. We devise a hybrid push-based dissemination algorithm, combining probabilistic with deterministic properties, which limits message overhead to an order of magnitude lower than that of the purely probabilistic dissemination model, while retaining strong probabilistic guarantees for complete dissemination of messages. Our extensive experimentation shows that our proposed algorithm outperforms that model both in static and dynamic network scenarios, as well as in the face of large-scale catastrophic failures. Moreover, the proposed algorithm distributes the dissemination load uniformly on all participating nodes. Keywords: Epidemic/Gossip protocols, Information Dissemination, Peer-to-Peer

Aerodynamics of aero-engine installation

This paper describes current progress in the development of methods to assess aero-engine airframe installation effects. The aerodynamic characteristics of isolated intakes, a typical transonic transport aircraft as well as a combination of a through-flow nacelle and aircraft configuration have been evaluated. The validation task for an isolated engine nacelle is carried out with concern for the accuracy in the assessment of intake performance descriptors such as mass flow capture ratio and drag rise Mach number. The necessary mesh and modelling requirements to simulate the nacelle aerodynamics are determined. Furthermore, the validation of the numerical model for the aircraft is performed as an extension of work that has been carried out under previous drag prediction research programmes. The validation of the aircraft model has been extended to include the geometry with through flow nacelles. Finally, the assessment of the mutual impact of the through flow nacelle and aircraft aerodynamics was performed. The drag and lift coefficient breakdown has been presented in order to identify the component sources of the drag associated with the engine installation. The paper concludes with an assessment of installation drag for through-flow nacelles and the determination of aerodynamic interference between the nacelle and the aircraft

A Test of a Strong Ground Motion Prediction Methodology for the 7 September 1999, Mw=6.0 Athens Earthquake

We test a methodology to predict the range of ground-motion hazard for a fixed magnitude earthquake along a specific fault or within a specific source volume, and we demonstrate how to incorporate this into probabilistic seismic hazard analyses (PSHA). We modeled ground motion with empirical Green's functions. We tested our methodology with the 7 September 1999, Mw=6.0 Athens earthquake, we: (1) developed constraints on rupture parameters based on prior knowledge of earthquake rupture processes and sources in the region; (2) generated impulsive point shear source empirical Green's functions by deconvolving out the source contribution of M < 4.0 aftershocks; (3) used aftershocks that occurred throughout the area and not necessarily along the fault to be modeled; (4) ran a sufficient number of scenario earthquakes to span the full variability of ground motion possible; (5) found that our distribution of synthesized ground motions span what actually occurred and their distribution is realistically narrow; (6) determined that one of our source models generates records that match observed time histories well; (7) found that certain combinations of rupture parameters produced ''extreme'' ground motions at some stations; (8) identified that the ''best fitting'' rupture models occurred in the vicinity of 38.05{sup o} N 23.60{sup o} W with center of rupture near 12 km, and near unilateral rupture towards the areas of high damage, and this is consistent with independent investigations; and (9) synthesized strong motion records in high damage areas for which records from the earthquake were not recorded. We then developed a demonstration PSHA for a source region near Athens utilizing synthesized ground motion rather that traditional attenuation. We synthesized 500 earthquakes distributed throughout the source zone likely to have Mw=6.0 earthquakes near Athens. We assumed an average return period of 1000 years for this magnitude earthquake in the particular source zone, thereby having simulated a catalog of ground motion for a period of 500,000 years. The distribution of traditional ground motion parameters of peak acceleration or spectral ordinates then becomes the synthesized record from which we develop hazard curves in the form of the annual probability of exceedance. This approach replaces the aleatory uncertainty that current PSHA studies estimate by regression of empirical parameters from the worldwide database with epistemic uncertainty on what specific sources actually do at specific sites. This is a fundamental change for PSHA and eliminates the need to extrapolate current empirical data that was gathered over about 50 years to represent values for 10{sup -3} annual probability of exceedance or less. This difference becomes especially significant for very sensitive structures that require estimates for 10{sup -5} or less exceedance

First Results from The GlueX Experiment

The GlueX experiment at Jefferson Lab ran with its first commissioning beam in late 2014 and the spring of 2015. Data were collected on both plastic and liquid hydrogen targets, and much of the detector has been commissioned. All of the detector systems are now performing at or near design specifications and events are being fully reconstructed, including exclusive production of $\pi^{0}$, $\eta$ and $\omega$ mesons. Linearly-polarized photons were successfully produced through coherent bremsstrahlung and polarization transfer to the $\rho$ has been observed.Comment: 8 pages, 6 figures, Invited contribution to the Hadron 2015 Conference, Newport News VA, September 201