Erythrocyte enrichment in hematopoietic progenitor cell cultures based on magnetic susceptibility of the hemoglobin

Using novel media formulations, it has been demonstrated that human placenta and umbilical cord blood-derived CD34+ cells can be expanded and differentiated into erythroid cells with high efficiency. However, obtaining mature and functional erythrocytes from the immature cell cultures with high purity and in an efficient manner remains a significant challenge. A distinguishing feature of a reticulocyte and maturing erythrocyte is the increasing concentration of hemoglobin and decreasing cell volume that results in increased cell magnetophoretic mobility (MM) when exposed to high magnetic fields and gradients, under anoxic conditions. Taking advantage of these initial observations, we studied a noninvasive (label-free) magnetic separation and analysis process to enrich and identify cultured functional erythrocytes. In addition to the magnetic cell separation and cell motion analysis in the magnetic field, the cell cultures were characterized for cell sedimentation rate, cell volume distributions using differential interference microscopy, immunophenotyping (glycophorin A), hemoglobin concentration and shear-induced deformability (elongation index, EI, by ektacytometry) to test for mature erythrocyte attributes. A commercial, packed column high-gradient magnetic separator (HGMS) was used for magnetic separation. The magnetically enriched fraction comprised 80% of the maturing cells (predominantly reticulocytes) that showed near 70% overlap of EI with the reference cord blood-derived RBC and over 50% overlap with the adult donor RBCs. The results demonstrate feasibility of label-free magnetic enrichment of erythrocyte fraction of CD34+ progenitor-derived cultures based on the presence of paramagnetic hemoglobin in the maturing erythrocytes. © 2012 Jin et al

(G)hosting television: Ghostwatch and its medium

This article’s subject is Ghostwatch (BBC, 1992), a drama broadcast on Halloween night of 1992 which adopted the rhetoric of live non-fiction programming, and attracted controversy and ultimately censure from the Broadcasting Standards Council. In what follows, we argue that Ghostwatch must be understood as a televisually-specific artwork and artefact. We discuss the programme’s ludic relationship with some key features of television during what Ellis (2000) has termed its era of ‘availability’, principally liveness, mass simultaneous viewing, and the flow of the television super-text. We trace the programme’s television-specific historicity whilst acknowledging its allusions and debts to other media (most notably film and radio). We explore the sophisticated ways in which Ghostwatch’s visual grammar and vocabulary and deployment of ‘broadcast talk’ (Scannell 1991) variously ape, comment upon and subvert the rhetoric of factual programming, and the ends to which these strategies are put. We hope that these arguments collectively demonstrate the aesthetic and historical significance of Ghostwatch and identify its relationship to its medium and that medium’s history. We offer the programme as an historically-reflexive artefact, and as an exemplary instance of the work of art in television’s age of broadcasting, liveness and co-presence

Recovery of magnetic catalysts: advanced design for process intensification

The design of microdevices in which components with magnetic character must be separated and recovered from reactive media benefits from the advantages of microfluidics and meets the criteria for process intensification; however, there are open questions, such as the design of the most appropriate magnet arrangement, that need further research in order to increase the magnetic gradient exerted on the particles. Herein, we focus on the continuous recovery of magnetic microparticles, that can be used as support to facilitate the recovery of biocatalysts (magnetic microcatalysts, MMCs) from biological fluids. We analyze and compare the performance of two typical magnetophoretic microdevices for addressing bead recovery: (i) annular channels with a quadrupole orientation of the permanent magnets (quadrupole magnetic sorter, QMS) and (ii) the standard design, which consists of rectangular channels with a single permanent magnet to generate the magnetic field. To this end, an experimentally validated computational fluid dynamics (CFD) numerical model has been employed. Our results reveal that for devices with the same width and length, the micro QMS, in comparison to a rectangular channel, could accomplish the complete particle retrieval while (i) processing more than 4 times higher fluid velocities, treating more than 360 times higher flow rates or (ii) working with smaller particles, thus reducing by 55% the particle mass. Additionally, the parallel performance of +/-300 micro-QMSs fulfills the processing of flow rates as high as 200 L·h-1 while entirely capturing the magnetic beads. Thereby, this work shows the potential of the QMS advanced design in the intensification of the recovery of catalysts supports of magnetic character.Financial support from the Spanish Ministry of Science, Innovation and Universities under the project RTI2018- 093310-B-I00 is gratefully acknowledged. Cristina González-Fernández acknowledges the FPU (FPU18/03525) postgraduate research grants. We also wish to thank the United States National Institutes of Health (1R01HL131720-01A1, CA62349) and the United States Defense Advanced Research Projects Agency (BAA07-21) for financial assistance

Continuous Magnetophoretic Separation of Blood Cells from Plasma at the Microscale

We present a method for the direct and continuous separation of red and white blood cells from plasma at the microscale. The method is implemented in a microfluidic system with magnetic functionality. The fluidic structure within the microsystem consists of an inlet and a single microfluidic channel with multiple outlets. The magnetic functionality is provided by an array of integrated soft-magnetic elements that are embedded transverse and adjacent to the microchannel. The elements are magnetized using an external field, and once magnetized they produce a magnetic force on blood cells as they flow through the microchannel. In whole blood, white blood cells (WBCs) behave as diamagnetic microparticles, while red blood cells (RBCs) exhibit diamagnetic or paramagnetic behavior depending on the oxygenation of their hemoglobin. We study the motion of blood cells through the microchannel using a mathematical model that takes into account the magnetic, fluidic and gravitational forces on the cells. We use the model to study blood cell separation, and our analysis indicates that the microsystem is capable of separating WBC-rich plasma, deoxygenated RBC-rich plasma and cell-depleted plasma into respective outlets.Comment: Submitted to Journal of Applied Physic

An amphitropic cAMP-binding protein in yeast mitochondria

ABSTRACT: We describe the first example of a mitochondrial protein with a covalently attached phos-phatidylinositol moiety acting as a membrane anchor. The protein can be metabolically labeled with both stearic acid and inositol. The stearic acid label is removed by phospholipase D whereupon the protein with the retained inositol label is released from the membrane. This protein is a cAMP receptor of the yeast Saccharomyces cereuisiae and tightly associated with the inner mitochondrial membrane. However, it is converted into a soluble form during incubation of isolated mitochondria with Ca2+ and phospholipid (or lipid derivatives). This transition requires the action of a proteinaceous, N-ethylmaleimide-sensitive component of the intermembrane space and is accompanied by a decrease in the lipophilicity of the cAMP receptor. We propose that the component of the intermembrane space triggers the amphitropic behavior of the mitochondrial lipid-modified CAMP-binding protein through a phospholipase activity. Only in recent years specific fatty acids have been recog-nized to play important roles in the association of proteins with membranes. Both noncovalent and covalent interactions be-tween fatty acids and proteins have been reported. Among the latter are GTP-binding proteins (Molenaar et al., 1988)