Reversible or irreversible modification of [3H]PAF binding on rabbit platelet membranes differentiates various PAF receptor antagonists

[³H]Platelet-activating factor (PAF) binding to rabbit platelet membranes was examined before and after 20 min preincubation at 25°C in the presence of PAF, lysoPAF, or of five different PAF receptor antagonists (L 652731, BN 52021, WEB 2086, BN 52111 and BN 52115). When platelet membranes were not washed after preincubation with PAF or PAF antagonists, no significant specific binding of [³H]PAF was observed, which suggests full occupancy of the binding sites. When membranes were extensively washed, full recovery of specific [³H]PAF binding was attained with L 652731 and partial recoveries (60%, 55% and 30%) were reached with BN52021, WEB 2086 and PAF, respectively; no recovery was seen with the dioxolanes BN 52111 and BN 52115. Scatchard analysis of the binding data indicated that no significant change in the dissociation constant (K_d) and maximum number of binding sites (B_max) occurred after preincubation of platelet membrane with L 652731, whereas a reduction of B_max was observed when PAF and BN 52021 were measured. When platelet membranes were preincubated with WEB 2086, B_max and K_d significantly increased. The data suggest differing binding properties for PAF and the PAF antagonists. Some of the PAF antagonists may tightly bind to the PAF receptor site(s) and/or irreversibly modify or downregulate PAF recognition sites. Our results also suggest that the interaction of PAF receptor antagonists with PAF receptor can be divided into at least two components, namely a reversible component and an irreversible one. Based on a paper presented at the Third International Conference on Platelet-Activating Factor and Structurally Related Alkyl Ether Lipids, Tokyo, Japan, May 1989.     

Purification of the human anionic polypeptide fraction of the apo-bile lipoprotein complex by zonal ultracentrifugation

The two main proteic constituents of the human Apo-bile lipoprotein complex (BLC), i.e., the anionic polypeptide fraction (APF) and the IgA fragments, were separated by preparative zonal ultracentrifugation using a sucrose gradient containing 1.5 mM glycodesoxycholate. The purification of the APF was verified by sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis and immunology, and its amino acid composition then was determined. This procedure was used to obtain a polyclonal antiserum directed solely against the APF.     

Improving Neural Networks for Mechanism Kinematic Chain Isomorphism Identification

Detection of isomorphism among kinematic chains is essential in mechanical design, but difficult and computationally expensive. It has been shown that both traditional methods and previously presented neural networks still have a lot to be desired in aspects such as simplifying procedure of identification and adapting automatic computation. Therefore, a new algorithm based on a competitive Hopfield network is developed for automatic computation in the kinematic chain isomorphism problem. The neural approach provides directly interpretable solutions and does not demand tuning of parameters. We have tested the algorithm by solving problems reported in the recent mechanical literature. Simulation results show the effectiveness of the network that rapidly identifies isomorphic kinematic chains.     

A variable rate multipulse speech coder for CDMA cellular systems

Variable Rate (VR) speech coders are classified into: source-controlled VR coders where the rate is selected depending on the local character of the speech, and network-controlled VR coders where an external control signal selects the coding rate. The first category benefits from the variable rate channels used by Code Division Multiple Access (CDMA) mobile communications. The second category is indispensable for the right behaviour of the CDMA systems under conditions as high traffic levels. The VR speech coder presented in this communication exhibits both types of control. The source control is achieved by means of a Voice Activity Detector (VAD) and a phonetic classifier. The network control acts on the selection procedure of the multipulse excitation sequence to the synthesis filter. This is the main advantage of our VR MultiPulse speech coder because by means of an external signal the bit rate can be changed only every 4 msec, without transitions or distortions. Considering one-way communication, six different operating rates can be externally selected ranging from 4.8 to 9.1 kbps for the active frames; an average bit rate of 380 bps is required for the noise frames.This work has been partly funded by the Spanish Research National Plan under grant no. TIC92-0800-C05-02 and by Northern Telecom.     

Techniques for the mechanical characterisation of civil structures

Broadly speaking, structural defects include all those changes affecting a structure so that its behaviour is no longer in keeping with its intended use characteristics or differs from the behaviour it was originally designed for. In recent years, damage recognition and location methods, relying on the measurement of parameters that are indicative of a structure's mechanical behaviour, have benefitted from the introduction of innovative approaches, which make it possible to process and correlate an unprecedented wealth of measurble data. The aim of this work is to illustrate the basic problems associated with the different methods used for structural identification (parametric and non-parametric, linear and non-linear, with known and unknown inputs, etc.) and to report on some interesting applications for civil structural diagnosis.     

A panoramic view and swot analysis of artificial intelligence for achieving the sustainable development goals by 2030: progress and prospects

Applied Intelligence - The17 Sustainable Development Goals (SDGs) established by the United Nations Agenda 2030 constitute a global blueprint agenda and instrument for peace and prosperity...     

Is implementation of continuous improvement possible? An evolutionary model of enablers and inhibitors

The purpose of this article is to present an evolutionary model of the enablers and inhibitors affecting the implementation and sustainability of continuous improvement. With this aim, a qualitative study using the grounded theory was carried out. Interviews were conducted with managers responsible for continuous improvement in first‐tier suppliers in the automotive sector. As a result, two existing lines of research in the continuous improvement literature are synthesized: on one hand, the literature concerning the enablers and barriers associated with continuous improvement sustainability; and on the other, the literature concerning the evolutionary model. © 2011 Wiley Periodicals, Inc.     

The GAUGAA Motif Is Responsible for the Binding between circSMARCA5 and SRSF1 and Related Downstream Effects on Glioblastoma Multiforme Cell Migration and Angiogenic Potential

Circular RNAs (circRNAs) are a large class of RNAs with regulatory functions within cells. We recently showed that circSMARCA5 is a tumor suppressor in glioblastoma multiforme (GBM) and acts as a decoy for Serine and Arginine Rich Splicing Factor 1 (SRSF1) through six predicted binding sites (BSs). Here we characterized RNA motifs functionally involved in the interaction between circSMARCA5 and SRSF1. Three different circSMARCA5 molecules (Mut1, Mut2, Mut3), each mutated in two predicted SRSF1 BSs at once, were obtained through PCR-based replacement of wild-type (WT) BS sequences and cloned in three independent pcDNA3 vectors. Mut1 significantly decreased its capability to interact with SRSF1 as compared to WT, based on the RNA immunoprecipitation assay. In silico analysis through the “Find Individual Motif Occurrences” (FIMO) algorithm showed GAUGAA as an experimentally validated SRSF1 binding motif significantly overrepresented within both predicted SRSF1 BSs mutated in Mut1 (q-value = 0.0011). U87MG and CAS-1, transfected with Mut1, significantly increased their migration with respect to controls transfected with WT, as revealed by the cell exclusion zone assay. Immortalized human brain microvascular endothelial cells (IM-HBMEC) exposed to conditioned medium (CM) harvested from U87MG and CAS-1 transfected with Mut1 significantly sprouted more than those treated with CM harvested from U87MG and CAS-1 transfected with WT, as shown by the tube formation assay. qRT-PCR showed that the intracellular pro- to anti-angiogenic Vascular Endothelial Growth Factor A (VEGFA) mRNA isoform ratio and the amount of total VEGFA mRNA secreted in CM significantly increased in Mut1-transfected CAS-1 as compared to controls transfected with WT. Our data suggest that GAUGAA is the RNA motif responsible for the interaction between circSMARCA5 and SRSF1 as well as for the circSMARCA5-mediated control of GBM cell migration and angiogenic potential.