Fractionation of human red blood cells based on intrinsic magnetization

Red blood cell (RBC) transfusion is clinically used to treat hemodynamic instability and O2 carrying deficits in patients with acute blood loss, and patients with chronic anemia caused by bone marrow failure/suppression. Currently, cold storage of human RBCs (hRBCs) can preserve hRBCs for a maximum of six weeks (i.e. 42 days), set by the United States Food and Drug Administration (US FDA). However, as stored RBCs age, they undergo biochemical and biophysical changes that are often referred to as the storage lesion, which decreases the efficacy of transfusion while increasing the risk for transfusion-associated adverse effects. It is well known that upon transfusion of stored RBCs, there is a population of RBCs (i.e. healthy RBCs) that circulate for more than 24 hours, and another smaller population (i.e. damaged RBCs) that are cleared within 24 hours post transfusion. This population of cells destined to be cleared quickly can be higher than 25% in units stored for a mean of 30 days. The objective of our current project is to remove aged RBCs based on hemoglobin content. Under the influence of ultra-high magnetic fields and gradients, we have demonstrated that it is possible to fractionate RBCs into multiple factions based solely on difference in the intrinsic magnetization of the deoxygenated form of hemoglobin inside the RBCs (i.e. labeless separation). We hypothesize for our currently funded National Institute of Heart Lung and Blood project that healthy RBCs with higher Hb content correlate with longer half lives in transfused animal models than unhealthy RBCs which have lost some of their hemoglobin. In addition, material balances are being performed to track the hemoglobin molecules that are lost during the extended periods of storage. This work will reveal the mechanism behind the lost hemoglobin during RBC storage, deepen the knowledge about aged RBCs and RBC-associated exosomes, and facilitate bulk separation of RBCs without labeling the cells. Therefore, it could be clinically beneficial if the damaged RBCs in any unit of RBCs could be separated leaving a population of only healthy RBCs behind for transfusion. When a recipient is transfused with a dose of RBCs that overwhelms their circulatory system’s ability to compensate for the increased intravascular volume, heart failure can ensue. This condition is known as Transfusion Associated Circulatory Overload (TACO). It is the second leading cause of death related to transfusion reported to the FDA

A subpopulation of monocytes in normal human blood has significant magnetic susceptibility : quantification and potential implications

The presence of iron in circulating monocytes is well known as they play essential roles in iron recycling. Also, the storage of this metal as well as its incorrect uptake and/or release are important data to diagnose different pathologies. It has been demonstrated that iron storage in human blood cells can be measured through their magnetic behavior with high accuracy; however, the magnetic characteristics of monocytes have not been reported so far to the best of our knowledge. Therefore, in this work, we report, for the first time, the physical and magnetic properties of human monocytes, along with plasma platelets, oxyhemoglobin red blood cells (oxyHb‐RBCs), and methemoglobin red blood cells (metHb‐RBCs). The different cell populations were separated by Ficoll‐density gradient centrifugation, followed by a flow sorting step to isolate monocytes from peripheral blood mononuclear cells. The different fractions were analyzed by Coulter Counter (for determining the size distribution and concentration) and the sorted monocytes were qualitatively analyzed on ImageStream, a state‐of‐the‐art imaging cytometer. The analysis of the Coulter Counter and ImageStream data suggests that although there exists contamination in the monocyte fraction, the integrity of the sorted monocytes appears to be intact and the concentration was high enough to precisely measure their magnetic velocity by Cell Tracking Velocimetry. Surprisingly, monocytes reported the highest magnetic mobility from the four fractions under analysis, with an average magnetic velocity 7.8 times higher than MetHb‐RBCs, which is the only type of cells with positive magnetic velocities. This value is equivalent to a susceptibility 2.5 times higher than the value reported by fresh MetHb‐RBCs. It should be noted that this is the first study that reports that a subpopulation of human monocytes is much more magnetic than MetHb‐RBCs, opening the door to the possible isolation of human monocytes by label‐free magnetic techniques. Further, it is suggested that these magnetic monocytes could “contaminate” positively selected, immunomagnetically labeled blood cells (i.e., during a process using magnetically conjugated antibodies targeting cells, such as CD34 positive cells). Conversely, these magnetic monocytes could be inadvertently removed from a desired blood population when one is using a negative magnetic isolation technique to target cells for removal.The National Heart, Lung, and Blood Institute (1R01HL131720-01A1) and DARPA (BAA07-21).https://onlinelibrary.wiley.com/journal/155249302020-05-01hj2019BiochemistryGeneticsMicrobiology and Plant PathologyPlant Production and Soil Scienc

Physical Characterization and Cellular Toxicity Studies of Commercial NiO Nanoparticles

Nickel oxide (NiO) nanoparticles from several manufacturers with different reported sizes and surface coatings were characterized prior to assessing their cellular toxicity. The physical characterization of these particles revealed that sizes often varied from those reported by the supplier, and that particles were heavily agglomerated when dispersed in water, resulting in a smaller surface area and larger hydrodynamic diameter upon dispersion. Cytotoxicity testing of these materials showed differences between samples; however, correlation of these differences with the physical properties of the materials was not conclusive. Generally, particles with higher surface area and smaller hydrodynamic diameter were more cytotoxic. While all samples produced an increase in reactive oxygen species (ROS), there was no correlation between the magnitude of the increase in ROS and the difference in cytotoxicity between different materials

Characterization of Engineered Cerium Oxide Nanoparticles and Their Effects on Lung and Macrophage Cells

Cerium oxide nanoparticles are promising materials as novel nanoscale therapeutics and are commonly used materials in industrial processes. Most cytotoxicity studies on cerium oxide nanoparticles are made from in-lab prepared materials making comparison between studies challenging, especially when performed on unique cell lines under non-standard conditions. Using commercially available nanoparticles we show that particle stability/agglomeration may be critical in determining the cytotoxicity in some cell lines, while in other cell lines, larger sized primary particles are linked to higher cytotoxicity, contrasting what has been reported in the literature for smaller cerium nanoparticles. To accelerate the development of cerium oxide enabled commercial processes and biomedical innovations, a clearer understanding of the interactions between cerium oxide nanoparticles and cells is needed to better understand their fate in and impact on biological systems

Structure and Mineralogy of Hydrophilic and Biwettable Sub-2 µm Clay Aggregates in Oil Sands Bitumen Froth

A primary concern of commercial mined oil sands operations is the extent to which one can minimize the content of water and solids contaminants in the solvent-diluted bitumen products resulting from the bitumen production processes. During bitumen production, particles of about 2 µm or less may be responsible for the stabilization of water-in-bitumen emulsions that form during aqueous extraction of bitumen and purification of bitumen froth subsequently during the froth treatment processes, thus leading to the presence of those contaminants in solvent-diluted bitumen products. In this study, we separate and analyze sub-2 µm clay solids isolated from typical bitumen froth fed to a froth treatment plant at a commercial mined oil sands operation. Analytical transmission electron microscopy (TEM) with spatially-resolved energy-dispersive X-ray spectroscopy (EDX) and electron energy-loss spectroscopy (EELS) demonstrate key differences in morphology and composition between sub-2 µm clay aggregates with two distinct wettability characteristics: hydrophilic vs. biwettable particle surfaces. In particular, clay platelets with <200 nm lateral dimensions and thicknesses of a few atomic layers, which are intermixed within coarser sub-2 µm clay aggregates, are found to confer clear differences in morphological characteristics and wettability behaviors to the sub-2 µm clay aggregates. The <200 nm clay platelets found within sub-2 µm biwettable clays tend to arrange themselves with random orientations, whereas <200 nm clay platelets within sub-2 µm hydrophilic clays typically form well-ordered face-to-face stacks. Moreover, in biwettable sub-2 µm clay aggregates, <200 nm clay platelets often cover the surfaces of ~1–2 µm sized mineral particles, whereas similarly sized mineral particles in hydrophilic sub-2 µm clay aggregates, in contrast, generally have exposed surfaces without clay platelet coverage. These biwettable vs. hydrophilic behaviors are attributed to a difference in the surface characteristics of the <200 nm clay platelets caused by toluene-unextractable organic carbon coatings. Nanometer-scale carbon mapping reveals an inhomogeneous toluene-unextractable organic carbon coating on the surfaces of <200 nm platelets in biwettable clays. In contrast, hydrophilic clays have a significantly lower amount of toluene-unextractable organic carbon, which tends to be concentrated at steps or near metal oxide nanoparticles on clay particle surfaces. Mixing surface-active organic species, such as asphaltene, resin, or carboxylic organic acids of various types with inorganic solids can lead to a dramatically enhanced emulsion stability. Consequently, understanding the origin and characteristics of sub-2 µm clay solids in bitumen froth is important to (i) clarify their potential role in the formation of stable water-in-oil emulsions during bitumen production and (ii) improve froth treatment process performance to further reduce contaminant solids in solvent-diluted bitumen products. We discuss the implications of our results from these two perspectives

A Subpopulation of Monocytes in Normal Human Blood Has Significant Magnetic Susceptibility: Quantification and Potential Implications

The presence of iron in circulating monocytes is well known as they play essential roles in iron recycling. Also, the storage of this metal as well as its incorrect uptake and/or release are important data to diagnose different pathologies. It has been demonstrated that iron storage in human blood cells can be measured through their magnetic behavior with high accuracy; however, the magnetic characteristics of monocytes have not been reported so far to the best of our knowledge. Therefore, in this work, we report, for the first time, the physical and magnetic properties of human monocytes, along with plasma platelets, oxyhemoglobin red blood cells (oxyHb‐RBCs), and methemoglobin red blood cells (metHb‐RBCs). The different cell populations were separated by Ficoll‐density gradient centrifugation, followed by a flow sorting step to isolate monocytes from peripheral blood mononuclear cells. The different fractions were analyzed by Coulter Counter (for determining the size distribution and concentration) and the sorted monocytes were qualitatively analyzed on ImageStream, a state‐of‐the‐art imaging cytometer. The analysis of the Coulter Counter and ImageStream data suggests that although there exists contamination in the monocyte fraction, the integrity of the sorted monocytes appears to be intact and the concentration was high enough to precisely measure their magnetic velocity by Cell Tracking Velocimetry. Surprisingly, monocytes reported the highest magnetic mobility from the four fractions under analysis, with an average magnetic velocity 7.8 times higher than MetHb‐RBCs, which is the only type of cells with positive magnetic velocities. This value is equivalent to a susceptibility 2.5 times higher than the value reported by fresh MetHb‐RBCs. It should be noted that this is the first study that reports that a subpopulation of human monocytes is much more magnetic than MetHb‐RBCs, opening the door to the possible isolation of human monocytes by label‐free magnetic techniques. Further, it is suggested that these magnetic monocytes could “contaminate” positively selected, immunomagnetically labeled blood cells (i.e., during a process using magnetically conjugated antibodies targeting cells, such as CD34 positive cells). Conversely, these magnetic monocytes could be inadvertently removed from a desired blood population when one is using a negative magnetic isolation technique to target cells for removal.The National Heart, Lung, and Blood Institute (1R01HL131720-01A1) and DARPA (BAA07-21).https://onlinelibrary.wiley.com/journal/155249302020-05-01hj2019BiochemistryGeneticsMicrobiology and Plant PathologyPlant Production and Soil Scienc

Characterization of solids in froths, diluted bitumen products, tailings, and rag layers from naphtha based froth treatment processes

NRC publication: Ye

Stratified analyses refine association between TLR7 rare variants and severe COVID-19

Summary: Despite extensive global research into genetic predisposition for severe COVID-19, knowledge on the role of rare host genetic variants and their relation to other risk factors remains limited. Here, 52 genes with prior etiological evidence were sequenced in 1,772 severe COVID-19 cases and 5,347 population-based controls from Spain/Italy. Rare deleterious TLR7 variants were present in 2.4% of young (<60 years) cases with no reported clinical risk factors (n = 378), compared to 0.24% of controls (odds ratio [OR] = 12.3, p = 1.27 × 10−10). Incorporation of the results of either functional assays or protein modeling led to a pronounced increase in effect size (ORmax = 46.5, p = 1.74 × 10−15). Association signals for the X-chromosomal gene TLR7 were also detected in the female-only subgroup, suggesting the existence of additional mechanisms beyond X-linked recessive inheritance in males. Additionally, supporting evidence was generated for a contribution to severe COVID-19 of the previously implicated genes IFNAR2, IFIH1, and TBK1. Our results refine the genetic contribution of rare TLR7 variants to severe COVID-19 and strengthen evidence for the etiological relevance of genes in the interferon signaling pathway