A simplified immunoprecipitation method for quantitatively measuring antibody responses in clinical sera samples by using mammalian-produced Renilla luciferase-antigen fusion proteins

BACKGROUND: Assays detecting human antigen-specific antibodies are medically useful. However, the usefulness of existing simple immunoassay formats is limited by technical considerations such as sera antibodies to contaminants in insufficiently pure antigen, a problem likely exacerbated when antigen panels are screened to obtain clinically useful data. RESULTS: We developed a novel and simple immunoprecipitation technology for identifying clinical sera containing antigen-specific antibodies and for generating quantitative antibody response profiles. This method is based on fusing protein antigens to an enzyme reporter, Renilla luciferase (Ruc), and expressing these fusions in mammalian cells, where mammalian-specific post-translational modifications can be added. After mixing crude extracts, sera and protein A/G beads together and incubating, during which the Ruc-antigen fusion become immobilized on the A/G beads, antigen-specific antibody is quantitated by washing the beads and adding coelenterazine substrate and measuring light production. We have characterized this technology with sera from patients having three different types of cancers. We show that 20–85% of these sera contain significant titers of antibodies against at least one of five frequently mutated and/or overexpressed tumor-associated proteins. Five of six colon cancer sera tested gave responses that were statistically significantly greater than the average plus three standard deviations of 10 control sera. The results of competition experiments, preincubating positive sera with unmodified E. coli-produced antigens, varied dramatically. CONCLUSION: This technology has several advantages over current quantitative immunoassays including its relative simplicity, its avoidance of problems associated with E. coli-produced antigens and its use of antigens that can carry mammalian or disease-specific post-translational modifications. This assay should be generally useful for analyzing sera for antibodies recognizing any protein or its post-translational modifications

Debris cover and surface melt at a temperate maritime alpine glacier: Franz Josef Glacier, New Zealand

Melt rates on glaciers are strongly influenced by the presence of supraglacial debris, which can either enhance or reduce ablation relative to bare ice. Most recently, Franz Josef Glacier has entered into a phase of strong retreat and downwasting, with the increasing emergence of debris on the surface in the ablation zone. Previously at Franz Josef Glacier, melt has only been measured on bare ice. During February 2012, a network of 11 ablation stakes was drilled into locations of varying supraglacial debris thickness on the lower glacier. Mean ablation rates over 9 days varied over the range 1.2–10.1 cm d−1, and were closely related to debris thickness. Concomitant observations of air temperature allowed the application of a degree-day approach to the calculation of melt rates, with air temperature providing a strong indicator of melt. Degree-day factors (d f) varied over the range 1.1–8.1 mm d−1 °C−1 (mean of 4.4 mm d−1 °C−1), comparable with rates reported in other studies. Mapping of the current debris cover revealed 0.7 km2 of the 4.9 km2 ablation zone surface was debris-covered, with thicknesses ranging 1–50 cm. Based on measured debris thicknesses and d f, ablation on debris-covered areas of the glacier is reduced by a total of 41% which equates to a 6% reduction in melt overall across the entire ablation zone. This study highlights the usefulness of a short-term survey to gather representative ablation data, consistent with numerous overseas ablation studies on debris-covered glaciers

Orbital operations study. Appendix B: Operational procedures

Operational procedures for each alternate approach for each interfacing activity of the orbital operations study are presented. The applicability of the procedures to interfacing element pairs is identified

Non-Newtonian fluid flow through three-dimensional disordered porous media

We investigate the flow of various non-Newtonian fluids through three-dimensional disordered porous media by direct numerical simulation of momentum transport and continuity equations. Remarkably, our results for power-law (PL) fluids indicate that the flow, when quantified in terms of a properly modified permeability-like index and Reynolds number, can be successfully described by a single (universal) curve over a broad range of Reynolds conditions and power-law exponents. We also study the flow behavior of Bingham fluids described in terms of the Herschel-Bulkley model. In this case, our simulations reveal that the interplay of ({\it i}) the disordered geometry of the pore space, ({\it ii}) the fluid rheological properties, and ({\it iii}) the inertial effects on the flow is responsible for a substantial enhancement of the macroscopic hydraulic conductance of the system at intermediate Reynolds conditions. This anomalous condition of ``enhanced transport'' represents a novel feature for flow in porous materials.Comment: 5 pages, 5 figures. This article appears also in Physical Review Letters 103 194502 (2009

Off-Equilibrium Dynamics in Finite-Dimensional Spin Glass Models

The low temperature dynamics of the two- and three-dimensional Ising spin glass model with Gaussian couplings is investigated via extensive Monte Carlo simulations. We find an algebraic decay of the remanent magnetization. For the autocorrelation function $C(t,t_w)=[]_{av}$ a typical aging scenario with a $t/t_w$ scaling is established. Investigating spatial correlations we find an algebraic growth law $\xi(t_w)\sim t_w^{\alpha(T)}$ of the average domain size. The spatial correlation function $G(r,t_w)=[< S_i(t_w)S_{i+r}(t_w)>^2]_{av}$ scales with $r/\xi(t_w)$. The sensitivity of the correlations in the spin glass phase with respect to temperature changes is examined by calculating a time dependent overlap length. In the two dimensional model we examine domain growth with a new method: First we determine the exact ground states of the various samples (of system sizes up to $100\times 100$) and then we calculate the correlations between this state and the states generated during a Monte Carlo simulation.Comment: 38 pages, RevTeX, 14 postscript figure

Creating and Sharing Digital Instructional Activities: A Practical Tutorial

BCBAs may encounter situations, such as the current COVID-19 pandemic, that preclude them from providing traditional in-person ABA services to clients. When conditions prevent BCBAs and behavior technicians from working directly with clients, digital instructional activities designed by BCBAs and delivered via a computer or tablet may be a viable substitute. Google applications, including Google Slides, Google Forms, and Google Classroom, can be particularly useful for creating and sharing digital instructional activities. In the current paper, we provide task analyses for utilizing basic Google Slides functions, developing independent instructional activities, developing caregiver-supported instructional activities, and sharing activities with clients and caregivers. We also provide practical recommendations for implementing digital instructional activities with clients and caregivers

An Intact Kidney Slice Model to Investigate Vasa Recta Properties and Function in situ

Background: Medullary blood flow is via vasa recta capillaries, which possess contractile pericytes. In vitro studies using isolated descending vasa recta show that pericytes can constrict/dilate descending vasa recta when vasoactive substances are present. We describe a live kidney slice model in which pericyte-mediated vasa recta constriction/dilation can be visualized in situ. Methods: Confocal microscopy was used to image calcein, propidium iodide and Hoechst labelling in ‘live’ kidney slices, to determine tubular and vascular cell viability and morphology. DIC video-imaging of live kidney slices was employed to investigate pericyte-mediated real-time changes in vasa recta diameter. Results: Pericytes were identified on vasa recta and their morphology and density were characterized in the medulla. Pericyte-mediated changes in vasa recta diameter (10–30%) were evoked in response to bath application of vasoactive agents (norepinephrine, endothelin-1, angiotensin-II and prostaglandin E2) or by manipulating endogenous vasoactive signalling pathways (using tyramine, L-NAME, a cyclo-oxygenase (COX-1) inhibitor indomethacin, and ATP release). Conclusions: The live kidney slice model is a valid complementary technique for investigating vasa recta function in situ and the role of pericytes as regulators of vasa recta diameter. This technique may also be useful in exploring the role of tubulovascular crosstalk in regulation of medullary blood flow

Publication and patent analysis of European researchers in the field of production technology and manufacturing systems

This paper develops a structured comparison among a sample of European researchers in the field of Production Technology and Manufacturing Systems, on the basis of scientific publications and patents. Researchers are evaluated and compared by a variegated set of indicators concerning (1) the output of individual researchers and (2) that of groups of researchers from the same country. While not claiming to be exhaustive, the results of this preliminary study provide a rough indication of the publishing and patenting activity of researchers in the field of interest, identifying (dis)similarities between different countries. Of particular interest is a proposal for aggregating analysis results by means of maps based on publication and patent indicators. A large amount of empirical data are presented and discusse

Multimodal perioperative pain protocol for Gynecologic Oncology laparotomy reduces length of hospital stay

Our primary objective was to evaluate the impact of a multimodal perioperative pain regimen on length of hospital stay for patients undergoing laparotomy with a gynecologic oncologist