2,989 research outputs found
Complement’s C1 complex, Factor H and the X factor: A personal tribute to Prof. Robert B. Sim
No full text© 2021 by the author. It is with great sadness that I am writing this obituary for my mentor, colleague and friend, Bob, i.e., Prof. Robert B. Sim, who passed away on 6 February 2021
Surface-bound myeloperoxidase is a ligand for recognition of late apoptotic neutrophils by human lung surfactant proteins A and D
No full textSurfactant proteins A (SP-A) and D (SP-D), both members of the collectin family, play a well established role in apoptotic cell recognition and clearance. Recent in vitro data show that SP-A and SP-D interact with apoptotic neutrophils in a distinct manner. SP-A and SP-D bind in a Ca2+-dependent manner to viable and early apoptotic neutrophils whereas the much greater interaction with late apoptotic neutrophils is Ca2+-independent. Cell surface molecules on the apoptotic target cells responsible for these interactions had not been identified and this study was done to find candidate target molecules. Myeloperoxidase (MPO), a specific intracellular defense molecule of neutrophils that becomes exposed on the outside of the cell upon apoptosis, was identified by affinity purification, mass-spectrometry and western blotting as a novel binding molecule for SP-A and SP-D. To confirm its role in recognition, it was shown that purified immobilised MPO binds SP-A and SP-D, and that MPO is surface-exposed on late apoptotic neutrophils. SP-A and SP-D inhibit binding of an anti-MPO monoclonal Ab to late apoptotic cells. Fluorescence microscopy confirmed that anti-MPO mAb and SP-A/SP-D colocalise on late apoptotic neutrophils. Desmoplakin was identified as a further potential ligand for SP-A, and neutrophil defensin as a target for both proteins. <br/
The human lung surfactant proteins A (SP-A) and D (SP-D) interact with apoptotic target cells by different binding mechanisms
No full textThe role of the lung surfactant proteins SP-A and SP-D in immune defence is well established. They bind to foreign organisms that invade the lungs and target them for phagocytic clearance by resident alveolar macrophages. SP-A and SP-D also bind to various apoptotic cells and facilitate their phagocytic uptake. To date, the molecular mechanisms by which the lung surfactant proteins interact with apoptotic cells and phagocytes are poorly understood.The aims of this study were to investigate further the interactions between SP-A and SP-D and apoptotic cells using human neutrophils and Jurkat cells as model systems.Specifically the binding behaviour of SP-A and SP-D with viable, early apoptotic and late apoptotic cells was investigated and compared. SP-A and SP-D show very distinct binding to the various cell types. SP-A bound to viable and early apoptotic cells in a predominantly Ca2+-dependent manner but the interaction with late apoptotic cells was Ca2+-independent, suggesting involvement of other than the lectin- or Ca2+-binding sites. This was consistent for neutrophils and Jurkat cells.SP-D in contrast, did not interact with viable and early apoptotic Jurkat cells but strongly and in a Ca2+-independent manner with late apoptotic Jurkat cells. SP-D-binding to viable and early apoptotic neutrophils was inhibited by maltose and ethylene-diamin-tetra-acetate (EDTA), suggesting lectin-binding site involvement whereas the binding to late apoptotic neutrophils was predominantly Ca2+-independent.These results represent a detailed study of the binding behaviour of SP-A and SP-D with different cell types and stages of viability. The mechanisms of these interactions appear to involve preferential recognition of different ligands on the apoptotic cell surface, which may include nucleic acid, phospholipid, protein and glycan structure
A Tribute to Robert (Bob) Sim—Personal Memories of Working in Bob’s Lab
No full textThis article is intended as a tribute to Robert B. Sim through the sharing of personal memories and anecdotes from two of Bob’s lab members who worked in his lab between 1989 to 1994
A Personal Tribute to Robert B. Sim with Reflections on Our Work Together on Factor H
No full textRobert (Bob) Sim had a profound effect on almost every aspect of my approach to scientific research, acting as a mentor and moral compass through the many different stages of my career [...
Similarities between 2D and 3D convection for large Prandtl number
No full textUsing direct numerical simulations of Rayleigh-B\'enard convection (RBC), we perform a comparative study of the spectra and fluxes of energy and entropy for large and infinite Prandtl numbers in two (2D) and three (3D) dimensions. We observe close similarities between the 2D and 3D RBC, in particular the kinetic energy spectrum , and the entropy spectrum exhibits a dual branch with a dominant spectrum. We showed that the dominant Fourier modes in the 2D and 3D flows are very close
Mannan-binding lectin in human serum, cerebrospinal fluid and brain tissue and its role in Alzheimer's disease
No full textMannan-binding lectin (MBL) is a serum lectin which can activate the classical complement pathway. Complement proteins of the classical pathway have been found in the brains of patients with Alzheimer's disease (AD) in association with AD brain pathology. To investigate the role for MBL in AD we have looked for its presence in the brain by immunohistochemistry and determined the levels of MBL in paired samples of cerebrospinal fluid and serum from AD patients and controls. MBL was detected in association with blood vessels in the brain tissue of both AD patients and control subjects. There was no apparent difference in the distribution of MBL in the brain tissue between the two groups. The mean concentration of MBL in the CSF was 44% lower in AD patients than in controls (AD 154 ± 35 pg/ml, n = 19; non- AD 276 ± 50 pg/ml, n = 15, p < 0.05). The levels of MBL in serum were not significantly different in the two groups. Thus, this study shows that MBL is associated with blood vessels in the brains of both AD and control subjects. Moreover, CSF levels of MBL appear to be lower in AD patients than in control subjects which may indicate a higher degree of MBL consumption connected with complement activation in the AD patients.</p
"Square in a Circle" by Ellen Braaten. Collegiate Legacy: Emeritus Faculty Exhibition
No full textExhibition of works by College of Architecture and Urban Studies emeritus faculty, celebrating the 50th anniversary of the college. Curated by Truman Capone and Deb Sim. Moss Arts Center, Virginia Tech.ELLEN BRAATEN. Square in a Circle. 1994. Clay. Collection of Robert Turner
Chemical labelling of active serum thioester proteins for quantification
No full textAbstractThe complement serum proteins C3 and C4 and the protease inhibitor α-2 macroglobulin are all members of the C3/α-2M thioester protein family, an evolutionarily ancient and conserved family that contains an intrachain thioester bond. The chemistry of the thioester bond is a key to the function of the thioester proteins. All these proteins function by covalently linking to their target by acyl transfer of the protein via the thioester moiety. We show that the signature thioester bond can be targeted with nucleophiles linked to a bioreporter molecule, site-specifically modifying the whole, intact thioester protein. Conditions were optimised to label selectively and efficiently pull-down unprocessed thioester-containing proteins from serum. We demonstrated pull-down of full-length C3, α-2M and C4 from sera in high salt, using a biotinylated nucleophile and streptavidin-coated resin, confirmed by MALDI-TOF MS identification of the gel bands. The potential for the development of a quantitative method for measuring active C3 in serum was investigated in patient sera pre and post operation. Quantifying active C3 in clinical assays using current methods is difficult. Methods based on antibody detection (e.g. nephelometry) do not distinguish between active C3 and inactive breakdown products. C3-specific haemolytic assays can be used, but these require use of relatively unstable reagents. The current work represents a promising robust, enzyme- and antibody-free chemical method for detecting active thioester proteins in blood, plasma or serum
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