Functional Evaluation of Plasmodium Export Signals in Plasmodium berghei Suggests Multiple Modes of Protein Export

The erythrocytic stage development of malaria parasites occurs within the parasitophorous vacuole inside the infected-erythrocytes, and requires transport of several parasite-encoded proteins across the parasitophorous vacuole to several locations, including the cytosol and membrane of the infected cell. These proteins are called exported proteins; and a large number of such proteins have been predicted for Plasmodium falciparum based on the presence of an N-terminal motif known as the Plasmodium export element (PEXEL) or vacuolar transport signal (VTS), which has been shown to mediate export. The majority of exported proteins contain one or more transmembrane domains at the C-terminus and one of three types of N-terminus domain architectures. (1) The majority, including the knob-associated histidine rich protein (KAHRP), contain a signal/hydrophobic sequence preceding the PEXEL/VTS motif. (2) Other exported proteins, including the P. berghei variant antigen family bir and the P. falciparum skeleton binding protein-1, do not appear to contain a PEXEL/VTS motif. (3) The P. falciparum erythrocyte membrane protein-1 (PfEMP1) family lacks a signal/hydrophobic sequence before the motif. These different domain architectures suggest the presence of multiple export pathways in malaria parasites. To determine if export pathways are conserved in plasmodia and to develop an experimental system for studying these processes, we investigated export of GFP fused with N- and C-terminus putative export domains in the rodent malaria parasite P. berghei. Export was dependent on specific N- and C-terminal domains. Constructs with a KAHRP-like or bir N-terminus, but not the PfEMP1 N-terminus, exported GFP into the erythrocyte. The C-terminus of a P. falciparum variant antigen rifin prevented GFP export by the KAHRP-like N-terminus. In contrast, GFP chimeras containing KAHRP-like N-termini and the PfEMP1 C-terminus were exported to the surface of erythrocytes. Taken together, these results suggest that proteins with KAHRP-like architecture follow a common export pathway, but that PfEMP1s utilize an alternative pathway. Functional validation of common putative export domains of malaria parasites in P. berghei provides an alternative and simpler system to investigate export mechanisms

Remnant lipoproteins inhibit malaria sporozoite invasion of hepatocytes

Remnants of lipoproteins, intestinal chylomicrons, and very low density lipoprotein (VLDL), are rapidly cleared from plasma and enter hepatocytes. It has been suggested that remnant lipoproteins are initially captured in the space of Disse by heparan sulfate proteoglycans (HSPGs), and that their subsequent internalization into hepatocytes is mediated by members of the LDL-receptor gene family. Similarly to lipoprotein remnants, malaria sporozoites are removed from the blood circulation by the liver within minutes after injection by Anopheles mosquitoes. The sporozoite's surface is covered by the circumsporozoite protein (CS), and its region II-plus has been implicated in the binding of the parasites to glycosaminoglycan chains of hepatocyte HSPGs. Lactoferrin, a protein with antibacterial properties found in breast milk and neutrophil granules, is also rapidly cleared from the circulation by hepatocytes, and can inhibit the hepatic uptake of lipoprotein remnants. Here we provide evidence that sporozoites, lactoferrin, and remnant lipoproteins are cleared from the blood by similar mechanisms. CS, lactoferrin, and remnant lipoproteins compete in vitro and in vivo for binding sites on liver cells. The relevance of this binding event for sporozoite infectivity is highlighted by our demonstration that apoliprotein E-enriched beta-VLDI and lactoferrin inhibit sporozoite invasion of HepG2 cells. In addition, malaria sporozoites are less infective in LDL-receptor knockout (LDLR -/-) mice maintained on a high fat diet, as compared with littermates maintained on a normal diet. We conclude that the clearance of lipoprotein remnants and sporozoites from the blood is mediated by the same set of highly sulfated HSPGs on the hepatocyte plasma membrane

Evidence for an axion-like particle from PKS 1222+216?

The surprising discovery by MAGIC of an intense, rapidly varying emission in the energy range 70 - 400 GeV from the flat spectrum radio quasar PKS 1222+216 represents a challenge for all interpretative scenarios. Indeed, in order to avoid absorption of \gamma rays in the dense ultraviolet radiation field of the broad line region (BLR), one is forced to invoke some unconventional astrophysical picture, like for instance the existence of a very compact (r\sim 10^{14} cm) emitting blob at a large distance (R \sim10^{18} cm) from the jet base. We offer the investigation of a scenario based on the standard blazar model for PKS 1222+216 where \gamma rays are produced close to the central engine, but we add the new assumption that inside the source photons can oscillate into axion-like particles (ALPs), which are a generic prediction of several extensions of the Standard Model of elementary particle interactions. As a result, a considerable fraction of very-high-energy photons can escape absorption from the BLR through the mechanism of photon-ALP oscillations much in the same way as they largely avoid absorption from extragalactic background light when propagating over cosmic distances in the presence of large-scale magnetic fields in the nG range. In addition we show that the above MAGIC observations and the simultaneous Fermi/LAT observations in the energy range 0.3 - 3 GeV can both be explained by a standard spectral energy distribution for experimentally allowed values of the model parameters. In particular, we need a very light ALP just like in the case of photon-ALP oscillations in cosmic space. Moreover, we find it quite tantalizing that the most favorable value of the photon-ALP coupling happens to be the same in both situations. Although our ALPs cannot contribute to the cold dark matter, they are a viable candidate for the quintessential dark energy. [abridged]Comment: 32 pages, 10 figures, accepted for publication in Physical Review

Planning the Future of U.S. Particle Physics (Snowmass 2013): Chapter 4: Cosmic Frontier

These reports present the results of the 2013 Community Summer Study of the APS Division of Particles and Fields ("Snowmass 2013") on the future program of particle physics in the U.S. Chapter 4, on the Cosmic Frontier, discusses the program of research relevant to cosmology and the early universe. This area includes the study of dark matter and the search for its particle nature, the study of dark energy and inflation, and cosmic probes of fundamental symmetries.Comment: 61 page

Alpha-v-containing integrins are host receptors for the Plasmodium falciparum sporozoite surface protein, TRAP.

Malaria-causing Plasmodium sporozoites are deposited in the dermis by the bite of an infected mosquito and move by gliding motility to the liver where they invade and develop within host hepatocytes. Although extracellular interactions between Plasmodium sporozoite ligands and host receptors provide important guidance cues for productive infection and are good vaccine targets, these interactions remain largely uncharacterized. Thrombospondin-related anonymous protein (TRAP) is a parasite cell surface ligand that is essential for both gliding motility and invasion because it couples the extracellular binding of host receptors to the parasite cytoplasmic actinomyosin motor; however, the molecular nature of the host TRAP receptors is poorly defined. Here, we use a systematic extracellular protein interaction screening approach to identify the integrin αvβ3 as a directly interacting host receptor for Plasmodium falciparum TRAP. Biochemical characterization of the interaction suggests a two-site binding model, requiring contributions from both the von Willebrand factor A domain and the RGD motif of TRAP for integrin binding. We show that TRAP binding to cells is promoted in the presence of integrin-activating proadhesive Mn2+ ions, and that cells genetically targeted so that they lack cell surface expression of the integrin αv-subunit are no longer able to bind TRAP. P. falciparum sporozoites moved with greater speed in the dermis of Itgb3-deficient mice, suggesting that the interaction has a role in sporozoite migration. The identification of the integrin αvβ3 as the host receptor for TRAP provides an important demonstration of a sporozoite surface ligand that directly interacts with host receptors. Copyright © 2018 the Author(s). Published by PNAS

The Highest Energy Neutrinos

Measurements of the arrival directions of cosmic rays have not revealed their sources. High energy neutrino telescopes attempt to resolve the problem by detecting neutrinos whose directions are not scrambled by magnetic fields. The key issue is whether the neutrino flux produced in cosmic ray accelerators is detectable. It is believed that the answer is affirmative, both for the galactic and extragalactic sources, provided the detector has kilometer-scale dimensions. We revisit the case for kilometer-scale neutrino detectors in a model-independent way by focussing on the energetics of the sources. The real breakthrough though has not been on the theory but on the technology front: the considerable technical hurdles to build such detectors have been overcome. Where extragalactic cosmic rays are concerned an alternative method to probe the accelerators consists in studying the arrival directions of neutrinos produced in interactions with the microwave background near the source, i.e. within a GZK radius. Their flux is calculable within large ambiguities but, in any case, low. It is therefore likely that detectors that are larger yet by several orders of magnitudes are required. These exploit novel techniques, such as detecting the secondary radiation at radio wavelengths emitted by neutrino induced showers.Comment: 16 pages, pdflatex, 7 jpg figures, ICRC style files included. Highlight talk presented at the 30th International Cosmic Ray Conference, Merida, Mexico, 200

ADMX-Orpheus First Search for 70 μ\mueV Dark Photon Dark Matter: Detailed Design, Operations, and Analysis

Dark matter makes up 85% of the matter in the universe and 27% of its energy density, but we don't know what comprises dark matter. It is possible that dark matter may be composed of either axions or dark photons, both of which can be detected using an ultra-sensitive microwave cavity known as a haloscope. The haloscope employed by ADMX consists of a cylindrical cavity operating at the TM$_{010}$ mode and is sensitive to the QCD axion with masses of few $\mu$eV. However, this haloscope design becomes challenging to implement for higher masses. This is because higher masses require smaller-diameter cavities, consequently reducing the detection volume which diminishes the detected signal power. ADMX-Orpheus mitigates this issue by operating a tunable, dielectrically-loaded cavity at a higher-order mode, allowing the detection volume to remain large. This paper describes the design, operation, analysis, and results of the inaugural ADMX-Orpheus dark photon search between 65.5 $\mu$eV (15.8 GHz) and 69.3 $\mu$eV (16.8 GHz), as well as future directions for axion searches and for exploring more parameter space.Comment: 21 pages, 29 figures. To be submitted to Physical Review D. arXiv admin note: substantial text overlap with arXiv:2112.0454