Communication between distinct subunit interfaces of the cohesin complex promotes its topological entrapment of DNA.

Cohesin mediates higher order chromosome structure. Its biological activities require topological entrapment of DNA within a lumen(s) formed by cohesin subunits. The reversible dissociation of cohesins Smc3p and Mcd1p subunits is postulated to form a regulated gate that allows DNA entry and exit into the lumen. We assessed gate-independent functions of this interface in yeast using a fusion protein that joins Smc3p to Mcd1p. We show that in vivo all the regulators of cohesin promote DNA binding of cohesin by mechanisms independent of opening this gate. Furthermore, we show that this interface has a gate-independent activity essential for cohesin to bind chromosomes. We propose that this interface regulates DNA entrapment by controlling the opening and closing of one or more distal interfaces formed by cohesin subunits, likely by inducing a conformation change in cohesin. Furthermore, cohesin regulators modulate the interface to control both DNA entrapment and cohesin functions after DNA binding

Evaluation of photografted charged sites within polymer monoliths in capillary columns using contactless conductivity detection

Capacitively coupled contactless conductivity detection (C4D) is presented as a novel and versatile means of visualising discrete zones of charged functional groups grafted onto polymer based monoliths. Monoliths were formed within 100 μm UV transparent fused silica capillaries and photografting methods were subsequently used to graft a charged functional monomer, 2-acrylamido-2-methyl-1-propanesulfonic acid (AMPS) onto discrete regions of the “generic” monolith using a photomask. Post-modification monolith evaluation involves scanning the C4D detector along the length of the monolith to obtain a profile of the exact spatial location of grafted charged functionalities with millimetre accuracy. The methodology was extended to the visualisation of several zones of immobilised protein (bovine serum albumin) using photografted azlactone groups to enable covalent attachment of the protein to the monolith at precise locations along its length. In addition, the extent of non-specific binding of protein to the ungrafted regions of the monolith due to hydrophobic interactions could be monitored as an increase in background conductivity of the stationary phase. Finally, the technique was cross-validated using fluorescence microscopy by immobilising green fluorescent protein (GFP) in discrete zones and comparing the profiles obtained using both complementary techniques

The interaction of reading, spelling and handwriting difficulties with writing development

Editorial

О методике испытаний вентиляторов местного проветривания

In this article we examine how consumer knowledge and two aspects of email ad design (copy type and testimonial type) influence attitudes and purchase intentions. Results from a\ud field experiment reveal differences between experts and novices in their responses to email advertising. Specifically, experts report more favorable evaluations for email advertising than novices. Experts also demonstrate a preference for expert testimonials, when exposed to\ud attribute copy. Yet when benefits-only ad copy was used, experts are most influenced by novice testimonials. In contrast, novice consumers show no copy-testimonial preference. Expert testimonials are also more effective than novice testimonials for expert and novice consumers. We discuss the results with respect to theoretical contributions and managerial implications

Development and application of an empirical probability distribution for the prediction error of re-entry body maximum dynamic pressure

The relationship between actual and predicted re-entry maximum dynamic pressure is characterized using a probability density function and a cumulative distribution function derived from sounding rocket flight data. This paper explores the properties of this distribution and demonstrates applications of this data with observed sounding rocket re-entry body damage characteristics to assess probabilities of sustaining various levels of heating damage. The results from this paper effectively bridge the gap existing in sounding rocket reentry analysis between the known damage level/flight environment relationships and the predicted flight environment

I am your trans patient

For the first time we are publishing a What Your Patient is Thinking article by a group of people. These transgender authors share their experiences of healthcare and the important messages they would like doctors to know

Precision, Reliability, and Responsiveness of a Novel Automated Quantification Tool for Cartilage Thickness: Data from the Osteoarthritis Initiative

Objective; Accurate automated segmentation of cartilage should provide rapid reliable outcomes for both epidemiological studies and clinical trials. We aimed to assess the precision and responsiveness of cartilage thickness measured with careful manual segmentation or a novel automated technique. Methods; Agreement of automated segmentation was assessed against two manual segmentation datasets: 379 MR images manually segmented in-house (Training set), and 582 from the OAI with data available at 0, 1, and 2 years (Biomarkers set). Agreement of mean thickness was assessed using Bland-Altman plots, change with pairwise Students t-test, in the central medial femur and tibia regions (cMF, cMT). Repeatability was assessed on a set of 19 knees imaged twice on the same day. Responsiveness was assessed using standardised response means (SRMs). Results; Agreement of manual vs automated methods was excellent with no meaningful systematic bias (Training set cMF bias 0.1mm 95%CI ±0.35, Biomarkers set bias 0.1mm ±0.4). The smallest detectable difference (SDD) for cMF was 0.13mm, coefficient of variation (CoV) 3.1%; cMT 0.16 mm, 2.65%. Reported change using manual segmentations in the cMF region at 1 year was -0.031mm, confidence limit (-0.022, -0.039), p<10-4, SRM -0.31 (-0.23,-0.38); at 2 years was -0.071 (-0.058,-0.085), p<10-4, SRM -0.43(-0.36,-0.49). Reported change using automated segmentations in the cMF at 1 year was - 0.059 (-0.047, -0.071), p<10-4, SRM -0.41(-0.34,-0.48) ; 2 years: -0.14 (-0.123,-0.157), p<10-4, SRM -0.67 (-0.6,-0.72). Conclusion; A novel cartilage segmentation method provides highly accurate and repeatable measures with comparable cartilage thickness measurements to careful manual segmentation, but with improved responsiveness

Informing Management of Henrys Lake, Idaho, using an Integrated Catch-at-Age Model

Henrys Lake, Idaho, supports a popular fishery for Yellowstone Cutthroat Trout Oncorhynchus clarkii bouvieri and Yellowstone Cutthroat Trout × Rainbow Trout O. mykiss hybrids. A majority of the adult population of fish in Henrys Lake are of hatchery origin that were stocked as fingerlings. The fishery is closed to angling during the late winter and spring months, but fisheries managers are considering opening the fishery year-round with catch-and-release- only regulations or with a two-fish bag limit during the extended season. However, there is concern that the proposed management actions may negatively affect the current fishery. Therefore, we developed an integrated catch-atage model to estimate population parameters for trout in Henrys Lake and used a simulation model to evaluate alternative management actions. Results of this study suggest that catch and release of both Yellowstone Cutthroat Trout and hybrids would increase and that abundance of trout in the spring (i.e., the start of the traditional season) would decrease under both proposed bag limits. Losses in abundance can be mitigated by stocking additional fish as long as no more than approximately 1,520,000 Yellowstone Cutthroat Trout are stocked annually. If catch-and-release-only regulations are implemented during the newly proposed season, total harvest is expected to decrease compared to the current fishery due to additional catch-and-release mortality. Ultimately, managers will need to prioritize harvest or catch-and-release opportunity, both of which provide additional utility to anglers, when choosing how to proceed with bag limit regulations

Overcoming scale-up challenges for a First-In-Human (FIH) antibody production process at the 2000L scale: Successful optimization of bioreactor equipment and harvest conditions to improve process performance and product yield

During the development and scale-up of a FIH fed-batch cell culture process, we have encountered two major challenges that impacted culture performance and harvest process yield significantly. The first challenge relates to the oxygen requirement for this particular process, where a higher oxygen demand was observed compared to our platform processes. The high oxygen demand is met by increasing agitation and sparging of air and oxygen in bioreactors, but also negatively impacted cell health due to higher gas entrance velocities (GEV) from the higher sparging rate. The observation is exemplified in the manufacturing scale which significantly impacted cell culture performance and antibody production. Further exacerbating this issue, many sparger holes were later found to be plugged leading to a theoretical GEV as high as 300 m/s. Extensive troubleshooting studies were carried out at small scale to confirm the impact of high GEV on this antibody production process and led to the re-design of the sparger for the manufacturing scale. Once implemented, the new sparger successfully mitigated the issue. The second challenge was relating to harvest operation, where the final membrane filter was clogged at the manufacturing scale. Studies were performed at the pilot scale to evaluate different solutions including increasing depth filter area, testing different types of the final filter, and changing the chase buffer at the manufacturing scale. In the end, successful implementation of the new sparger design and optimized harvest conditions led to comparable process performance between the manufacturing scale and the pilot scale. The final process resulted not only in an increase in the overall product yield, but also prevented the need of filter change-outs during the harvest operation, therefore, significantly improving manufacturing ease