Efficiency of delivery of DNA to cells by bovine papillomavirus type-1 L1/L2 pseudovirions

To investigate the efficiency of encapsidation of plasmid by papillomavirus virus-like particles (PV VLPs), and the infectivity of the resultant PV pseudovirions, Cos-1 cells were transfected with an 8-kb plasmid incorporating a green fluorescent protein (GFP) reporter gene (pGSV), and infected with bovine PV (BPV-1) L1/L2 recombinant vaccinia virus to produce BPV1 pseudovirions. Approximately 1 in 1.5x10(4) of dense (1.35 g/ml) PV pseudovirions and 0.3 in 10(4) Of less-dense (1.29 g/ml) pseudovirions packaged an intact pGSV plasmid. The majority (>75%) of packaged plasmids contained deletions, and the deletions affected all tested genes. After exposure of Cos-1 cells to BPV-1 pseudovirions at an MOI of 40,000:1, 6% of cells expressed GFP giving a calculated efficiency of delivery of the pGSV plasmid, by pseudovirions which had packaged an intact plasmid, of approximately 5%. Plasmid delivery was not effected by purified pGSV plasmid, was blocked by antiserum against BPV-1, and was not blocked by DNase treatment of pseudovirions, confirming that delivery was mediated by DNA within the pseudovirion. We conclude that a major limitation to the use of PV pseudovirions as a gene delivery system is that intact plasmid DNA is not efficiently selected for packaging by VLPs in cell-based pseudovirions production systems.

Parenteral and mucosal delivery of a novel multi-epitope M protein-based group A streptococcal vaccine construct: investigation of immunogenicity in mice

Primary vaccine strategies against group A streptococci (GAS) have focused on the M protein-the target of opsonic antibodies important for protective immunity. We have previously reported protection of mice against GAS infection following parenteral delivery of a multi-epitope vaccine construct, referred to as a heteropolymer. This current report has assessed mucosal (intranasal (i.n.) and oral) delivery of the heteropolymer in mice with regard to the induction and specificity of mucosal and systemic antibody responses, and compared this to parenteral delivery. GAS-specific IgA responses were detected in saliva and gut upon i.n. and oral delivery of the heteropolymer co-administered with cholera toxin B subunit, respectively. High titre serum IgG responses were elicited to the heteropolymer following all routes of delivery when administered with adjuvant. Moreover, as with parenteral delivery, serum IgG antibodies were detected to the individual heteropolymer peptides following i.n. but not oral delivery. These data support the potential of the i.n. route in the mucosal delivery of a GAS vaccine. (C) 2002 Elsevier Science Ltd. All rights reserved.

Enhancement of drug affinity for cell membranes by conjugation with lipoamino acids II. Experimental and computational evidence using biomembrane models

Lipoamino acids (LAAs) are promoieties able to enhance the amphiphilicity of drugs, facilitating their interaction with cell membranes. Experimental and computational studies were carried out on two series of lipophilic amide conjugates between a model drug (tranylcypromine, TCP) and LAA or alkanoic acids containing a short, medium or long alkyl side chain (C-4 to C-16). The effects of these compounds were evaluated by monolayer surface tension analysis and differential scanning calorimetry using dimyristoylphosphatidylcholine nnonolayers and liposomes as biomembrane models. The experimental results were related to independent calculations to determine partition coefficient and blood-brain partitioning. The comparison of TCP-LAA conjugates with the related series of TCP alkanoyl amides confirmed that the ability to interact with the biomembrane models is not due to the mere increase of lipophilicity, but mainly to the amphipatic nature and the kind of LAA residue. (C) 2005 Elsevier B.V. All rights reserved.

ESEM observations of SCC initiation for 4340 high strength steel in distilled water

The stress corrosion cracking (SCC) initiation process for 4340 high strength steel in distilled water at room temperature was studied using a new kind of instrument: an environmental scanning electron microscope (ESEM). It was found that the applied stress accelerated oxide film formation which has an important influence on the subsequent SCC initiation. SCC was observed to initiate in the following circumstances: (1) cracking of a thick oxide film leading to SCC initiation along metal grain boundaries, (2) the initiation of pits initiating SCC in the metal and (3) SCC initiating from the edge of the specimen. All these three SCC initiation circumstances are consistent with the following model which couples SCC initiation with cracking of a surface protective oxide. There is a dynamic interaction between oxide formation, the applied stress, oxide cracking, pitting and the initiation of SCC. An aspect of the dynamic interaction is cracks forming in a protective surface oxide because of the applied stress, exposing to the water bare metal at the oxide crack tip, and oxidation of the bare metal causing crack healing. Oxide crack healing would be competing with the initiation of intergranular SCC if an oxide crack meets the metal surface at a grain boundary. If the intergranular SCC penetration is sufficiently fast along the metal grain boundary, then the crack yaws open preventing healing of the oxide crack. If intergranular SCC penetration is not sufficiently fast, then the oxidation process could produce sufficient oxide to fill both the stress corrosion crack and the oxide crack; in this case there would be initiation of SCC but only limited propagation of SCC. Stress-induced cracks in very thin oxide can induce pits which initiate SCC, and under some conditions such stress induced cracks in a thin oxide can directly initiate SCC.

The diversity of the Digenea of Australian animals

The Digenea is one of five major helminth assemblages represented in Australian animals. History of the study of digeneans in Australia is reviewed briefly to show that it has never been subjected to the kind of sustained study needed to reach an understanding of it. The Australian vertebrate fauna comprises over 5500 species. These have so far been shown to harbour just over 70 families, about 306 genera and 566 species of digeneans. Digeneans occur in all classes of vertebrates in Australia but are distributed very unevenly; aquatic hosts are generally most heavily infected, but many terrestrial species are also infected. Particular weaknesses in knowledge of the fauna concern the bats, cetaceans and teleosts. Another weakness is in knowledge of life-cycles; representative life-cycles are known for only about 20 of the 70 families known in Australia. Estimates of the overall size of the fauna are dependent on an understanding of sampling strategies, the heterogeneity of distribution of the fauna, and the nature of host-specificity. These subjects are reviewed briefly and an estimate of the total fauna is made. There may be as many as 6000 species of digeneans in Australia. (C) 1998 Australian Society for Parasitology. Published by Elsevier Science Ltd.

Determination of pore size distributions by regularization and finite element collocation

The problem of extracting pore size distributions from characterization data is solved here with particular reference to adsorption. The technique developed is based on a finite element collocation discretization of the adsorption integral, with fitting of the isotherm data by least squares using regularization. A rapid and simple technique for ensuring non-negativity of the solutions is also developed which modifies the original solution having some negativity. The technique yields stable and converged solutions, and is implemented in a package RIDFEC. The package is demonstrated to be robust, yielding results which are less sensitive to experimental error than conventional methods, with fitting errors matching the known data error. It is shown that the choice of relative or absolute error norm in the least-squares analysis is best based on the kind of error in the data. (C) 1998 Elsevier Science Ltd. All rights reserved.

Finite element modelling of temperature gradient driven rock alteration and mineralization in porous rock masses

We present finite element simulations of temperature gradient driven rock alteration and mineralization in fluid saturated porous rock masses. In particular, we explore the significance of production/annihilation terms in the mass balance equations and the dependence of the spatial patterns of rock alteration upon the ratio of the roll over time of large scale convection cells to the relaxation time of the chemical reactions. Special concepts such as the gradient reaction criterion or rock alteration index (RAI) are discussed in light of the present, more general theory. In order to validate the finite element simulation, we derive an analytical solution for the rock alteration index of a benchmark problem on a two-dimensional rectangular domain. Since the geometry and boundary conditions of the benchmark problem can be easily and exactly modelled, the analytical solution is also useful for validating other numerical methods, such as the finite difference method and the boundary element method, when they are used to dear with this kind of problem. Finally, the potential of the theory is illustrated by means of finite element studies related to coupled flow problems in materially homogeneous and inhomogeneous porous rock masses. (C) 1998 Elsevier Science S.A. All rights reserved.

Numerical modelling of double diffusion driven reactive flow transport in deformable fluid-saturated porous media with particular consideration of temperature-dependent chemical reaction rates

Numerical methods ave used to solve double diffusion driven reactive flow transport problems in deformable fluid-saturated porous media. in particular, thp temperature dependent reaction rate in the non-equilibrium chemical reactions is considered. A general numerical solution method, which is a combination of the finite difference method in FLAG and the finite element method in FIDAP, to solve the fully coupled problem involving material deformation, pore-fluid flow, heat transfer and species transport/chemical reactions in deformable fluid-saturated porous media has been developed The coupled problem is divided into two subproblems which are solved interactively until the convergence requirement is met. Owing to the approximate nature of the numerical method, if is essential to justify the numerical solutions through some kind of theoretical analysis. This has been highlighted in this paper The related numerical results, which are justified by the theoretical analysis, have demonstrated that the proposed solution method is useful for and applicable to a wide range of fully coupled problems in the field of science and engineering.

Finite element modelling of heat transfer through permeable cracks in hydrothermal systems with upward throughflow

We use the finite element method to model the heat transfer phenomenon through permeable cracks in hydrothermal systems with upward throughflow. Since the finite element method is an approximate numerical method, the method must be validated before it is used to soh,e any new, kind of problem. However, the analytical solution, which can be used to validate the finite element method and other numerical methods, is rather limited in the literature, especially, for the problem considered here. Keeping this in mind, we have derived analytical solutions for the temperature distribution along the vertical axis of a crack in a fluid-saturated porous layer. After the finite element method is validated by comparing the numerical solution with the analytical solution for the same benchmark problem, it is used to investigate the pore-fluid flow and heat transfer in layered hydrothermal systems with vertical permeable cracks. The related analytical and numerical results have demonstrated that vertical cracks are effective and efficient members to transfer heat energy from the bottom section to the top section in hydrothermal systems with upward throughflow.