Complement activation and protein adsorption by carbon nanotubes

As a first step to validate the use of carbon nanotubes as novel vaccine or drug delivery devices, their interaction with a part of the human immune system, complement, has been explored. Haemolytic assays were conducted to investigate the activation of the human serum complement system via the classical and alternative pathways. Western blot and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) techniques were used to elucidate the mechanism of activation of complement via the classical pathway, and to analyse the interaction of complement and other plasma proteins with carbon nanotubes. We report for the first time that carbon nanotubes activate human complement via both classical and alternative pathways. We conclude that complement activation by nanotubes is consistent with reported adjuvant effects, and might also in various circumstances promote damaging effects of excessive complement activation, such as inflammation and granuloma formation. C1q binds directly to carbon nanotubes. Protein binding to carbon nanotubes is highly selective, since out of the many different proteins in plasma, very few bind to the carbon nanotubes. Fibrinogen and apolipoproteins (AI, AIV and CIII) were the proteins that bound to carbon nanotubes in greatest quantit

Detection and characterisation of Complement protein activity in bovine milk by bactericidal sequestration assay

Susan Maye is in receipt of a Teagasc Walsh Fellowship. Financial support by the Department of Agriculture, Food and the Marine is gratefully acknowledged.Copyright © Proprietors of Journal of Dairy Research 2015 (Institute of Food Research and the Hannah Research Institute)peer-reviewedWhile the Complement protein system in human milk is well characterised, there is little information on its presence and activity in bovine milk. Complement forms part of the innate immune system, hence the importance of its contribution during milk ingestion to the overall defences of the neonate. A bactericidal sequestration assay, featuring a Complement sensitive strain, Escherichia coli 0111, originally used to characterise Complement activity in human milk was successfully applied to freshly drawn bovine milk samples, thus, providing an opportunity to compare Complement activities in both human and bovine milks. Although not identical in response, the levels of Complement activity in bovine milk were found to be closely comparable with that of human milk. Differential counts of Esch. coli 0111 after 2 h incubation were 6·20 and 6·06 log CFU/ml, for raw bovine and human milks, respectively – the lower value representing a stronger Complement response. Exposing bovine milk to a range of thermal treatments e.g. 42, 45, 65, 72, 85 or 95 °C for 10 min, progressively inhibited Complement activity by increasing temperature, thus confirming the heat labile nature of this immune protein system. Low level Complement activity was found, however, in 65 and 72 °C heat treated samples and in retailed pasteurised milk which highlights the outer limit to which high temperature, short time (HTST) industrial thermal processes should be applied if retention of activity is a priority. Concentration of Complement in the fat phase was evident following cream separation, and this was also reflected in the further loss of activity recorded in low fat variants of retailed pasteurised milk. Laboratory-based churning of the cream during simulated buttermaking generated an aqueous (buttermilk) phase with higher levels of Complement activity than the fat phase, thus pointing to a likely association with the milk fat globule membrane (MFGM) layer.Department of Agriculture, Food and the Marin

Stratified Morse Theory in Arrangements

This paper is a survey of our work based on the stratified Morse theory of Goresky and MacPherson. First we discuss the Morse theory of Euclidean space stratified by an arrangement. This is used to show that the complement of a complex hyperplane arrangement admits a minimal cell decomposition. Next we review the construction of a cochain complex whose cohomology computes the local system cohomology of the complement of a complex hyperplane arrangement. Then we present results on the Gauss-Manin connection for the moduli space of arrangements of a fixed combinatorial type in rank one local system cohomology.Comment: For Robert MacPherson on the occasion of his sixtieth birthda

On the Modulation Equations and Stability of Periodic GKdV Waves via Bloch Decompositions

In this paper, we complement recent results of Bronski and Johnson and of Johnson and Zumbrun concerning the modulational stability of spatially periodic traveling wave solutions of the generalized Korteweg-de Vries equation. In this previous work it was shown by rigorous Evans function calculations that the formal slow modulation approximation resulting in the Whitham system accurately describes the spectral stability to long wavelength perturbations. Here, we reproduce this result without reference to the Evans function by using direct Bloch-expansion methods and spectral perturbation analysis. This approach has the advantage of applying also in the more general multi-periodic setting where no conveniently computable Evans function is yet devised. In particular, we complement the picture of modulational stability described by Bronski and Johnson by analyzing the projectors onto the total eigenspace bifurcating from the origin in a neighborhood of the origin and zero Floquet parameter. We show the resulting linear system is equivalent, to leading order and up to conjugation, to the Whitham system and that, consequently, the characteristic polynomial of this system agrees (to leading order) with the linearized dispersion relation derived through Evans function calculation.Comment: 19 pages

Coherent states and global entanglement in an N qubit system

We consider an $N$ qubit system and show that in the symmetric subspace, $\mathbb{S}$ a state is not globally entangled, iff it is a coherent state. It is also proven that in the orthogonal complement $\mathbb{S}_{\bot}$ all states are globally entangled

Sublytic Terminal Complement Components Induce Eryptosis in Autoimmune Haemolytic Anaemia Related to IgM Autoantibodies

BACKGROUND/AIMS: Eryptosis, the suicidal death of red blood cells (RBCs), is characterized by phosphatidylserine (PS) exposure at the cell surface. It can be catalysed by a variety of abnormal conditions and diseases. Until now, the many questions surrounding the physiology and pathophysiology of eryptosis have not been sufficiently answered. Recently, we demonstrated IgM and IgA autoantibodies (aab) to induce PS exposure on circulating RBCs of patients with autoimmune haemolytic anaemia (AIHA). However, it remained unclear how these aab lead to eryptosis. METHODS: Serum and plasma samples from patients with clinically relevant AIHA of cold type were used to induce eryptosis in O RBCs. Serum containing fresh complement from healthy donors, antibodies to complement component, and complement factor depleted sera were added to examine the influence of the complement on PS-exposure. RBC bound annexin V PE were analysed by flow cytometry. RESULTS: Eryptosis related to IgM aab was found to be dependent on complement activation and could be effectively inhibited by EDTA, serum heat inactivation and anti-C5. PS exposure increased with sequential activation of the sublytic terminal complement components C5b6, C5b-7 and was most significant at the C5b-8 stage. A decrease was observed following the formation of the lytic membrane attack complex C5b-9, either because of lysis of eryptotic RBCs or because of inhibition of eryptosis by C9. CONCLUSION: Our findings reflect new aspects on RBC destruction in AIHA as well the impact of the terminal complement complexes on the RBC membrane. The striking differences to nucleated cell apoptosis may even have physiological meaning of RBC acting as a buffer of the complement system