Recombinant Listeria monocytogenes as a live vaccine vehicle for the induction of protective anti-viral cell-mediated immunity.

Listeria monocytogenes (LM) is a Gram-positive bacterium that is able to enter host cells, escape from the endocytic vesicle, multiply within the cytoplasm, and spread directly from cell to cell without encountering the extracellular milieu. The ability of LM to gain access to the host cell cytosol allows proteins secreted by the bacterium to efficiently enter the pathway for major histocompatibility complex class I antigen processing and presentation. We have established a genetic system for expression and secretion of foreign antigens by recombinant strains, based on stable site-specific integration of expression cassettes into the LM genome. The ability of LM recombinants to induce protective immunity against a heterologous pathogen was demonstrated with lymphocytic choriomeningitis virus (LCMV). LM strains expressing the entire LCMV nucleoprotein or an H-2Ld-restricted nucleoprotein epitope (aa 118-126) were constructed. Immunization of mice with LM vaccine strains conferred protection against challenge with virulent strains of LCMV that otherwise establish chronic infection in naive adult mice. In vivo depletion of CD8+ T cells from vaccinated mice abrogated their ability to clear viral infection, showing that protective anti-viral immunity was due to CD8+ T cells.

At work or play : a comparison of private property vehicle crashes with those occurring on public roads in north Queensland.

Objective: To define characteristics of vehicle crashes occurring on rural private property in north Queensland with an exploration of associated risk factors. Design: Descriptive analysis of private property crash data collected by the Rural and Remote Road Safety Study. Setting: Rural and remote north Queensland. Participants: A total of 305 vehicle controllers aged 16 years or over hospitalised at Atherton, Cairns, Mount Isa or Townsville for at least 24 hours as a result of a vehicle crash. Main outcome measure: A structured questionnaire completed by participants covering crash details, lifestyle and demographic characteristics, driving history, medical history, alcohol and drug use and attitudes to road use. Results: Overall, 27.9% of interviewees crashed on private property, with the highest proportion of private road crashes occurring in the North West Statistical Division (45%). Risk factors shown to be associated with private property crashes included male sex, riding off-road motorcycle or all-terrain vehicle, first-time driving at that site, lack of licence for vehicle type, recreational use and not wearing a helmet or seatbelt. Conclusions: Considerable trauma results from vehicle crashes on rural private property. These crashes are not included in most crash data sets, which are limited to public road crashes. Legislation and regulations applicable to private property vehicle use are largely focused on workplace health and safety, yet work-related crashes represent a minority of private property crashes in north Queensland.

The File Marked "Rita". Selections from the Letters of Herbert Mueller and Rita Klein (1939-1945) 1939-1946

Exchange of letters between Herbert Mueller and Rita Klein after Mueller's emigration to England in 1939; Rita Klein's attempt to obtain divorce; suicide attempt by her husband Leo Klein; correspondence through intermediaries after outbreak of World War II; notice of Rita Klein's deportation to Auschwitz; (translation from original German)

Anomalous reaction-diffusion as a model of nonexponential DNA escape kinetics

We show that data from recent experiments carried out on the kinetics of DNA escape from alpha-hemolysin nanopores [M. Wiggin, C. Tropini, C. T. Cossa, N. N. Jetha, and A. Marziali, Biophys. J. 95, 5317 (2008)] may be rationalized by a model of chain dynamics based on the anomalous diffusion of a particle moving in a harmonic well in the presence of a delta function sink. The experiments of Wiggin found, among other things, that the occasional occurrence of unusually long escape times in the distribution of chain trapping events led to nonexponential decays in the survival probability, S(t), of the DNA molecules within the nanopore. Wiggin ascribed this nonexponentiality to the existence of a distribution of trapping potentials, which they suggested was theresult of stochastic interactions between the bases of the DNA and the amino acids located on the surface of the nanopore. Based on this idea, they showed that the experimentally determined S(t) could be well fit in both the short and long time regimes by a function of the form (1+t/tau)(-alpha) (the so called Becquerel function). In our model, S(t) is found to be given by a Mittag-Leffler function at short times and by a generalized Mittag-Leffler function at long times. By suitable choice of certain parameter values, these functions are found to fit the experimental S(t) even better than the Becquerel function. Anomalous diffusion of DNA within the trap prior to escape over a barrier of fixed height may therefore provide a second, plausible explanation of the data, and may offer fresh perspectives on similar trapping and escape problems.

Hemagglutinin sequence conservation guided stem immunogen design from influenza A H3 subtype

Seasonal epidemics caused by influenza A (H1 and H3 subtypes) and B viruses are a major global health threat. The traditional, trivalent influenza vaccines have limited efficacy because of rapid antigenic evolution of the circulating viruses. This antigenic variability mediates viral escape from the host immune responses, necessitating annual vaccine updates. Influenza vaccines elicit a protective antibody response, primarily targeting the viral surface glycoprotein hemagglutinin (HA). However, the predominant humoral response is against the hypervariable head domain of HA, thereby restricting the breadth of protection. In contrast, the conserved, subdominant stem domain of HA is a potential ``universal'' vaccine candidate. We designed an HA stem-fragment immunogen from the 1968 pandemic H3N2 strain (A/Hong Kong/1/68) guided by a comprehensive H3 HA sequence conservation analysis. The biophysical properties of the designed immunogen were further improved by C-terminal fusion of a trimerization motif, ``isoleucine-zipper'', or ``foldon''. These immunogens elicited cross-reactive, antiviral antibodies and conferred partial protection against a lethal, homologous HK68 virus challenge in vivo. Furthermore, bacterial expression of these immunogens is economical and facilitates rapid scale-up.

Influence of recombination on acquisition and reversion of immune escape and compensatory mutations in HIV-1

Following transmission, HIV-1 adapts in the new host by acquiring mutations that allow it to escape from the host immune response at multiple epitopes. It also reverts mutations associated with epitopes targeted in the transmitting host but not in the new host. Moreover, escape mutations are often associated with additional compensatory mutations that partially recover fitness costs. It is unclear whether recombination expedites this process of multi-locus adaptation. To elucidate the role of recombination, we constructed a detailed population dynamics model that integrates viral dynamics, host immune response at multiple epitopes through cytotoxic T lymphocytes, and viral evolution driven by mutation, recombination, and selection. Using this model, we compute the expected waiting time until the emergence of the strain that has gained escape and compensatory mutations against the new host's immune response, and reverted these mutations at epitopes no longer targeted. We find that depending on the underlying fitness landscape, shaped by both costs and benefits of mutations, adaptation proceeds via distinct dominant pathways with different effects of recombination, in particular distinguishing escape and reversion. When adaptation at a single epitope is involved, recombination can substantially accelerate immune escape but minimally affects reversion. When multiple epitopes are involved, recombination can accelerate or inhibit adaptation depending on the fitness landscape. Specifically, recombination tends to delay adaptation when a purely uphill fitness landscape is accessible at each epitope, and accelerate it when a fitness valley is associated with each epitope. Our study points to the importance of recombination in shaping the adaptation of HIV-1 following its transmission to new hosts, a process central to T cell-based vaccine strategies. (C) 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license.

Subcellular localization and characterization of G protein-coupled receptor homolog from lymphocystis disease virus isolated in China

G protein-coupled receptors (GPCRs) constitute a large superfamily involved in various types of signal transduction pathways, and play an important role in coordinating the activation and migration of leukocytes to sites of infection and inflammation. Viral GPCRs, on the other hand, can help the virus to escape from host immune surveillance and contribute to viral pathogenesis. Lymphocystis disease virus isolated in China (LCDV-C) contains a putative homolog of cellular GPCRs, LCDV-C GPCR. In this paper, LCDV-C GPCR was cloned, and the subcellular localization and characterization of GPCR protein were investigated in fish cells. LCDV-C GPCR encoded a 325-amino acid peptide, containing a typical seven-transmembrane domain characteristic of the chemokine receptors and a conserved DRY motif that is usually essential for receptor activation. Transient transfection of GPCR-EGFP in fathead minnow (FHM) cells and epithelioma papulosum cyprini (EPC) cells indicated that LCDV-C GPCR was expressed abundantly in both the cytoplasm and nucleoplasm. Transient overexpression of GPCR in these two cells cannot induce obvious apoptosis. FHM cells stably expressing GPCR showed enhanced cell proliferation and significant anchorage-independent growth. The effects of GPCR protein on external apoptotic stimuli were examined. Few apoptotic bodies were observed in cells expressing GPCR treated with actinomycin D (ActD). Quantitative analysis of apoptotic cells indicated that a considerable decrease in the apoptotic fraction of cells expressing GPCR, compared with. the control cells, was detected after exposure to ActD and cycloheximide. These data suggest that LCDV-C GPCR may inhibit apoptosis as part of its potential mechanism in mediating cellular transformation.

Inhibition of progesterone receptor activity in recombinant yeast by soot from fossil fuel combustion emissions and air particulate materials

Numerous environmental pollutants have been detected for estrogenic activity by interacting with the estrogen receptor, but little information is available about their interactions with the progesterone receptor. In this study, emission samples generated by fossil fuel combustion (FFC) and air particulate material (APM) collected from an urban location near a traffic line in a big city of China were evaluated to interact with the human progesterone receptor (hPR) signaling pathway by examining their ability to interact with the activity of hPR expressed in yeast. The results showed that the soot of a petroleum-fired vehicle possessed the most potent anti-progesteronic activity, that of coal-fired stove and diesel fired agrimotor emissions took the second place, and soot samples of coal-fired heating work and electric power station had lesser progesterone inhibition activity. The anti-progesteronic activity of APM was between that of soot from petroleum-fired vehicle and soot from coal-fired establishments and diesel fired agrimotor. Since there was no other large pollution source near the APM sampling sites, the endocrine disrupters were most likely from vehicle emissions, tire attrition and house heating sources. The correlation analysis showed that a strong relationship existed between estrogenic activity and anti-progesteronic activity in emissions of fossil fuel combustion. The discoveries that some environmental pollutants with estrogenic activity can also inhibit OR activity indicate that further studies are required to investigate potential mechanisms for the reported estrogenic activities of these pollutants. (c) 2005 Elsevier B.V. All rights reserved.

Escaping From The Past? The Falkland Islands in the Twenty-First Century

This paper discusses the competing claims for the sovereignty of the Falkland Islands in the South Atlantic, which led to a war in 1982 between the United Kingdom and Argentina, which given that it was over competing claims for sovereignty over a non-independent territory seemed to be redolent of the nineteenth rather than the late-twentieth century. Post-war developments are outlined whilst the paper considers whether or not the Falkland Islands can ever escape from the Conflict, now more than thirty years ago.

The Impact of Vehicle-to-Grid on Electric Power Systems: A Review

Environmental concerns and fossil fuel shortage put pressure on both power and transportation systems. Electric vehicles (EVs) are thought to be a good solution to these problems. With EV adoption, energy flow is two way: from grid to vehicle and from vehicle to grid, which is known as vehicle-to-grid (V2G) today. This paper considers electric power systems and provides a review of the impact of V2G on power system stability. The concept and basics of V2G technology are introduced at first, followed by a description of EV application in the world. Several technical issues are detailed in V2G modeling and capacity forecasting, steady-state analysis and stability analysis. Research trends of such topics are declared at last.

Dynamic Axle Force and Road Profile Identification Using a Moving Vehicle

In the interaction between vehicles, pavements and bridges, it is essential to aim towards a reduction of vehicle axle forces to promote longer pavement life spans and to prevent bridges loads becoming too high. Moreover, as the road surface roughness affects the vehicle dynamic forces, an efficient monitoring of pavement condition is also necessary to achieve this aim. This paper uses a novel algorithm to identify the dynamic interaction forces and pavement roughness from vehicle accelerations in both theoretical simulations and a laboratory experiment; moving force identification theory is applied to a vehicle model for this purpose. Theoretical simulations are employed to evaluate the ability of the algorithm to predict forces over a range of bridge spans and to evaluate the influence of road roughness level on the accuracy of the results. Finally, in addressing the challenge for the real-world problem, the effects of vehicle configuration and speed on the predicted road roughness are also investigated in a laboratory experiment.

Identification of Road Irregularities via Vehicle Accelerations

A periodic monitoring of the pavement condition facilitates a cost-effective distribution of the resources available for maintenance of the road infrastructure network. The task can be accurately carried out using profilometers, but such an approach is generally expensive. This paper presents a method to collect information on the road profile via accelerometers mounted in a fleet of non-specialist vehicles, such as police cars, that are in use for other purposes. It proposes an optimisation algorithm, based on Cross Entropy theory, to predict road irregularities. The Cross Entropy algorithm estimates the height of the road irregularities from vehicle accelerations at each point in time. To test the algorithm, the crossing of a half-car roll model is simulated over a range of road profiles to obtain accelerations of the vehicle sprung and unsprung masses. Then, the simulated vehicle accelerations are used as input in an iterative procedure that searches for the best solution to the inverse problem of finding road irregularities. In each iteration, a sample of road profiles is generated and an objective function defined as the sum of squares of differences between the ‘measured’ and predicted accelerations is minimized until convergence is reached. The reconstructed profile is classified according to ISO and IRI recommendations and compared to its original class. Results demonstrate that the approach is feasible and that a good estimate of the short-wavelength features of the road profile can be detected, despite the variability between the vehicles used to collect the data.

Do Children still need to Escape Childhood? A Reassessment of John Holt and his Vision for Children's Rights

The work of children’s liberationists have been long been critiqued for pushing the parameters of rights discourse too far; specifically, by suggesting that there are no significant differences between children and adults, including their ability for self-determination. John Holt’s 1974 text Escape from Childhood is one such work which was deemed highly controversial for its time. This article uses Holt’s Escape from Childhood as an overarching framework against which to examine the current state of play on children’s rights as explicated through the UN Convention on the Rights of the Child. It suggests that whilst Holt has often been critiqued for being too radical, in the context of current children’s rights discourse Holt’s visioning is not as radical as it might first appear.

Escape and connection : a phenomenological investigation into the meaning of an after-school program for adolescent boys

The purpos e of this phenomenological research is to explore the meaning of a YMCA-sponsored after-school recreation program in the lives of four adolescent boys. Listening to youth voice is impor t ant to the ability of othe r s to design, implement and evaluate high-quality programs tha t facilitate learning opportunities tha t a r e meaningful to participants. Within the context of interviews, task-based activities we r e used to ga the r data. Guided by Creswell's analytic spiral (1998), data wa s analyzed according to van Manen's (1990) thematic analysis and Caeilli's (2000) creative narrative analysis. It wa s found tha t this after-school progr am provided the s e adolescents with the opportunity to escape from the i r monotonous after-school activities and the instability of the i r home and school environments. Also, they we r e connected wi th positive peers, caring adults and the wide r community, opportunities tha t we r e limited in othe r aspects of the i r lives. Methodological issues a r e also discussed.

Model of single-electron decay from a strongly isolated quantum dot

Recent measurements of electron escape from a nonequilibrium charged quantum dot are interpreted within a two-dimensional (2D) separable model. The confining potential is derived from 3D self-consistent Poisson-Thomas-Fermi calculations. It is found that the sequence of decay lifetimes provides a sensitive test of the confining potential and its dependence on electron occupation