Eimeria tenella protein trafficking: differential regulation of secretion versus surface tethering during the life cycle

Eimeria spp. are intracellular parasites that have a major impact on poultry. Effective live vaccines are available and the development of reverse genetic technologies has raised the prospect of using Eimeria spp. as recombinant vectors to express additional immunoprotective antigens. To study the ability of Eimeria to secrete foreign antigens or display them on the surface of the sporozoite, transiently transfected populations of E. tenella expressing the fluorescent protein mCherry, linked to endogenous signal peptide (SP) and glycophosphatidylinositol-anchor (GPI) sequences, were examined. The SP from microneme protein EtMIC2 (SP2) allowed efficient trafficking of mCherry to cytoplasmic vesicles and following the C-terminal addition of a GPI-anchor (from surface antigen EtSAG1) mCherry was expressed on the sporozoite surface. In stable transgenic populations, mCherry fused to SP2 was secreted into the sporocyst cavity of the oocysts and after excystation, secretion was detected in culture supernatants but not into the parasitophorous vacuole after invasion. When the GPI was incorporated, mCherry was observed on the sporozites surface and in the supernatant of invading sporozoites. The proven secretion and surface exposure of mCherry suggests that antigen fusions with SP2 and GPI of EtSAG1 may be promising candidates to examine induction of protective immunity against heterologous pathogens

On the geometry of Riemannian cubic polynomials

AbstractWe continue the work of Crouch and Silva Leite on the geometry of cubic polynomials on Riemannian manifolds. In particular, we generalize the theory of Jacobi fields and conjugate points and present necessary and sufficient optimality condition

Vapor phase growth of group 3, 4, and 5 compounds by HCl transport of elements

Technique has been devised for vapor-phase epitaxial growth of group 3, 4, and 5 binary, ternary, or quaternary compounds by HCl transport of the constituent elements or dopants. Technique uses all the constituents of the alloy system in their elemental form. Transport of these elements by an HCl + H2 carrier gas facilitates their transport as subchlorides

Viral proteins expressed in the protozoan parasite Eimeria tenella are detected by the chicken immune system

BACKGROUND: Eimeria species are parasitic protozoa that cause coccidiosis, an intestinal disease commonly characterised by malabsorption, diarrhoea and haemorrhage that is particularly important in chickens. Vaccination against chicken coccidiosis is effective using wild-type or attenuated live parasite lines. The development of protocols to express foreign proteins in Eimeria species has opened up the possibility of using Eimeria live vaccines to deliver heterologous antigens and function as multivalent vaccine vectors that could protect chickens against a range of pathogens. RESULTS: In this study, genetic complementation was used to express immunoprotective virus antigens in Eimeria tenella. Infectious bursal disease virus (IBDV) causes Gumboro, an immunosuppressive disease that affects productivity and can interfere with the efficacy of poultry vaccination programmes. Infectious laryngotracheitis virus (ILTV) causes a highly transmissible respiratory disease for which strong cellular immunity and antibody responses are required for effective vaccination. Genes encoding the VP2 protein from a very virulent strain of IBDV (vvVP2) and glycoprotein I from ILTV (gI) were cloned downstream of 5’Et-Actin or 5’Et-TIF promoter regions in plasmids that also contained a mCitrine fluorescent reporter cassette under control of the 5’Et-MIC1 promoter. The plasmids were introduced by nucleofection into E. tenella sporozoites, which were then used to infect chickens. Progeny oocysts were sorted by FACS and passaged several times in vivo until the proportion of fluorescent parasites in each transgenic population reached ~20 % and the number of transgene copies per parasite genome decreased to < 10. All populations were found to transcribe and express the transgene and induced the generation of low titre, transgene-specific antibodies when used to immunise chickens. CONCLUSIONS: E. tenella can express antigens of other poultry pathogens that are successfully recognised by the chicken immune system. Nonetheless, further work has to be done in order to improve the levels of expression for its future use as a multivalent vaccine vector. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13071-016-1756-2) contains supplementary material, which is available to authorized users

Existence of solutions for a higher order non-local equation appearing in crack dynamics

In this paper, we prove the existence of non-negative solutions for a non-local higher order degenerate parabolic equation arising in the modeling of hydraulic fractures. The equation is similar to the well-known thin film equation, but the Laplace operator is replaced by a Dirichlet-to-Neumann operator, corresponding to the square root of the Laplace operator on a bounded domain with Neumann boundary conditions (which can also be defined using the periodic Hilbert transform). In our study, we have to deal with the usual difficulty associated to higher order equations (e.g. lack of maximum principle). However, there are important differences with, for instance, the thin film equation: First, our equation is nonlocal; Also the natural energy estimate is not as good as in the case of the thin film equation, and does not yields, for instance, boundedness and continuity of the solutions (our case is critical in dimension $1$ in that respect)

Comparison Of Structure And Properties Of Femtosecond And Nanosecond Laser-Structured Silicon

We compare the optical properties,chemical composition, and crystallinity of siliconmicrostructures formed in the presence of SF6 by femtosecond laserirradiation and by nanosecond laser irradiation. In spite of very different morphology and crystallinity, the optical properties and chemical composition of the two types of microstructures are very similar. The structures formed with femtosecond (fs) pulses are covered with a disordered nanocrystalline surface layer less than 1 μm thick, while those formed with nanosecond (ns) pulses have very little disorder. Both ns-laser-formed and fs-laser-formed structures absorb near-infrared (1.1–2.5 μm) radiation strongly and have roughly 0.5% sulfur impurities

Comparison of Structure and Properties of Femtosecond and Nanosecond Laser-Structured Silicon

We compare the optical properties, chemical composition, and crystallinity of silicon microstructures formed in the presence of SF6 by femtosecond laser irradiation and by nanosecond laser irradiation. In spite of very different morphology and crystallinity, the optical properties and chemical composition of the two types of microstructures are very similar. The structures formed with femtosecond (fs) pulses are covered with a disordered nanocrystalline surface layer less than 1 um thick, while those formed with nanosecond (ns) pulses have very little disorder. Both ns-laser-formed and fs-laser-formed structures absorb near-infrared (1.1 – 2.5 um) radiation strongly and have roughly 0.5% sulfur impurities.Engineering and Applied Science

Tunneling Conductance and Coulomb Blockade Peak Splitting of Two Quantum Dots Connected by a Quantum Point Contact

By using bosonization method and unitary transformation, we give a general relation between the dimensionless tunneling conductance and the fractional Coulomb blockade conductance peak splitting which is valid both for weak and strong transmission between two quantum dots, and show that the tunneling conductance has a linear temperature dependence in the low energy and low temperature limit.Comment: 12 pages, Revtex, no figures, to appear in Phys. Rev.

Large-Area Liquid Scintillation Detector Slab

A low-cost detector 18' x 2' x 5" has been developed for an underground cosmic ray neutrino experiment. The liquid employed is a high-clarity mineral oil-based mixture, and light is guided to the ends of the detector by total internal reflection at the surface of the Lucite container. Signals from 2 five-inch photomultipliers at each end give energy and event location for single penetrating particles, with relatively good discrimination against natural radioactivity by virtue of the substantial thickness. Data are presented on the response function of the tank, energy resolution, rates and thresholds. A number of modifications that have been tried are also described