Claire Evans

Viral infection of Emiliania huxleyi (Prymnesiophyceae) leads to elevated production of reactive oxygen species

The effect of viral infection of Emiliania huxleyi (Lohman) Hay and Mohler on the concentration of intracellular reactive oxygen species (ROS), hydrogen peroxide (H2O2) excretion and cell photosynthetic capacity (CPC) was examined. During the crash of an E. huxleyi culture induced by viruses intracellular ROS concentrations were generally elevated and reached levels of approximately double those observed in non‐infected control cultures. H2O2 concentrations also increased in the media of the infected cultures from background levels of around 130 nM to approximately 580 nM while levels in the controls decreased. These data suggest that oxidative stress is elevated in infected cells. Although the precise mechanism for ROS production was not identified, a traditional defense related oxidative burst was ruled out, as no evidence of a rapid intracellular accumulation of ROS following addition of the virus was found. CPC declined substantially in the infected culture from a healthy 0.6–0 arbitrary units. Clearly infection disrupted normal photosynthetic processes, which could lead to the production of ROS via interruption of the electron transport chain at the PSII level. Alternatively, ROS may also be a necessary requirement for viral replication in E. huxleyi, possibly due to a link with viral‐induced cell death or associated with general death processes.

Regional Variation in Lytic and Lysogenic Viral Infection in the Southern Ocean and Its Contribution to Biogeochemical Cycling

ABSTRACT Lytic and lysogenic viral infection was investigated throughout the Southern Ocean at sites spanning the sub-Antarctic zone, the Antarctic Circumpolar Current, and an Antarctic continental sea. Higher lytic virus activity was recorded in the more productive sub-Antarctic zone than in the iron-limited waters of the Antarctic Circumpolar Current during two transects. Reduced lytic viral activity in the Antarctic Circumpolar Current was combined with a shift toward lysogenic infection, probably resulting from the lower concentration of potential prokaryotic hosts. Superimposed on this variation, lytic viral production was lower in a transect completed in the Drake Passage in autumn (1.8 × 108 to 1.5 × 109 liter−1 day−1) than over the Greenwich Meridian during summer (5.1 × 108 to 2.0 × 1010 cells liter−1 day−1), indicating that viral activity is linked to the overall seasonal fluctuations in biotic activity. Interestingly, while prokaryotic abundance was lowest in the coastal Weddell Sea, levels of bacterial and lytic viral production (4.3 × 108 to 1.7 × 1010 cells liter−1 day−1) in this area were similar to those of the other zones. This may explain the weak relationship between the distribution of prokaryotes and chlorophyll in the Weddell Sea, as a high turnover of prokaryotic biomass may have been stimulated by the availability of substrates in the form of viral lysate. With estimated carbon and iron releases of 0.02 to 7.5 μg liter−1 day−1 and 1.5 to 175.7 pg liter−1 day−1, respectively, viral activity in the Southern Ocean is shown to be a major contributor to satisfying the elemental requirements of microbes, notably prokaryotes in the Weddell Sea and phytoplankton in the sub-Antarctic zone.

Viral‐mediated lysis of microbes and carbon release in the sub‐Antarctic and Polar Frontal zones of the Australian Southern Ocean

Viral production was determined in the sub-Antarctic zone (SAZ) to the southwest and southeast of Tasmania and in the Polar Frontal zone (PFZ) of the Australian sector of the Southern Ocean during Austral summer (January-February 2007). Concentrations of viruses were the lowest (6.6 x 10(9) particles l(-1)) in the PFZ and the highest (2.1 x 10(10) particles l(-1)) in the eastern SAZ where nutrient input from the East Australian Current (EAC) sustained higher concentrations of bacteria and bacterial production relative to the west. Rates of viral production in the PFZ (1.8 x 10(10) viruses l(-1) day(-1)) were lower than those in the western SAZ (2.5 x 10(10) viruses l(-1) day(-1)). Viral production in the eastern SAZ (2.2 x 10(11) viruses l(-1) day(-1)) was the highest recorded and was approximately one order of magnitude higher than at the other sites. In the western SAZ and PFZ, the percentage of available bacterial biomass lysed by viruses was similar (23.5% and 23% respectively) equating to the release of 3.3 and 2.3 microg carbon l(-1) day(-1) respectively (assuming a burst size of 50 viruses host(-1)). In the eastern SAZ the potential bacterial biomass lysed was higher (on average 40%) and corresponded to the release of 26.5 microg carbon l(-1) day(-1). These findings suggest the importance of the viral shunt in carbon cycling within these regions.

Epitope-based DNA vaccine for Alzheimer's disease: Translational study in macaques

Clinical trials with passive and active Alzheimers disease (AD) vaccines suggest that early interventions are needed for improvement of cognitive and/or functional performance in patients, providing impetus for the development of safe and immunologically potent active vaccines targeting amyloid β (Aβ). The AN‐1792 trial has indicated that Aβ‐specific T cells may be unsafe for humans; therefore, other vaccines based on small Aβ epitopes are undergoing preclinical and clinical testing.

Delivery of a DNA vaccine for Alzheimer's disease by electroporation versus gene gun generates potent and similar immune responses.

Background: Induction of a humoral response against amyloid-β peptide may be beneficial for Alzheimer’s disease (AD) patients and may alleviate the onset and progression of AD. DNA-based vaccination provides a unique alternative method of immunization for treatment and prevention of AD. Currently, the two major delivery methods used for enhancing DNA uptake and immune responses to DNA vaccines in humans are electroporation (EP) and gene gun (GG). Objective: The goal of this translational study was to evaluate the efficacy of an AD DNA epitope vaccine (DepVac) delivered intramuscularly by EP or intradermally by GG. Methods: Humoral and cellular immune responses to immunization with DepVac were evaluated by ELISA and ELISPOT, respectively. Functional activity of the antibodies was also assessed. Results: EP- and GG-mediated immunizations with DepVac induced similar anti-amyloid-β (Aβ) antibody and T cell responses. Anti-Aβ antibodies bound to amyloid plaques in AD brain tissue and to toxic forms of Aβ42 peptide. Conclusion: Both delivery methods are effective at promoting potent antibodies specific for Aβ.

Substrate kinetics of DMSP-lyases in axenic cultures and mesocosm populations of Emiliania huxleyi

Abstract.Most oceanic dimethyl sulphide (DMS) is produced through the enzymatic cleavage of dimethylsulphoniopropionate (DMSP), a process governed by the activity of DMSP-lyases in algae and bacteria. Laboratory and field experiments with the DMS-producing coccolithophorid Emiliania huxleyi have advanced our understanding of algal DMSP lyases. However, despite their central role in the global biogeochemical cycle of sulphur, little is known about the physiological and biochemical properties of this group of isozymes. Here we investigate the apparent substrate kinetic properties of two axenic E. huxleyi cultures (CCMP 373 and CCMP 379) under two light conditions and a mesocosm assemblage dominated by this species. Assays of in vivo and in vitro DMSP-lyase activity (DLA) were used to estimate the kinetic parameters Km and Vmax. Whereas in vivo DLA was not detected in CCMP 379, both laboratory cultures showed high DLA during in vitro tests. The mesocosm population was initially characterised by low substrate affinity (low Vmax and high Km), but shifted to a high affinity community when more dissolved DMSP became available after the collapse of the E. huxleyi bloom. At natural DMSP concentrations, DMS production is probably more affected by changes in Km than Vmax and future studies should include further investigations of the kinetic properties of DMSP-lyases.

In vivo electroporation improves therapeutic potency of a DNA vaccine targeting hepadnaviral proteins.

This preclinical study investigated the therapeutic efficacy of electroporation (EP)-based delivery of plasmid DNA (pDNA) encoding viral proteins (envelope, core) and IFN-γ in the duck model of chronic hepatitis B virus (DHBV) infection. Importantly, only DNA EP-therapy resulted in a significant decrease in mean viremia titers and in intrahepatic covalently closed circular DNA (cccDNA) levels in chronic DHBV-carrier animals, compared with standard needle pDNA injection (SI). In addition, DNA EP-therapy stimulated in all virus-carriers a humoral response to DHBV preS protein, recognizing a broader range of major antigenic regions, including neutralizing epitopes, compared with SI. DNA EP-therapy led also to significant higher intrahepatic IFN-γ RNA levels in DHBV-carriers compared to other groups, in the absence of adverse effects. We provide the first evidence on DNA EP-therapy benefit in terms of hepadnaviral infection clearance and break of immune tolerance in virus-carriers, supporting its clinical application for chronic hepatitis B.

How many Coccolithovirus genotypes does it take to terminate an Emiliania huxleyi bloom

Giant viruses are known to be significant mortality agents of phytoplankton, often being implicated in the terminations of large Emiliania huxleyi blooms. We have previously shown the high temporal variability of E. huxleyi-infecting coccolithoviruses (EhVs) within a Norwegian fjord mesocosm. In the current study we investigated EhV dynamics within a naturally-occurring E. huxleyi bloom in the Western English Channel. Using denaturing gradient gel electrophoresis and marker gene sequencing, we uncovered a spatially highly dynamic Coccolithovirus population that was associated with a genetically stable E. huxleyi population as revealed by the major capsid protein gene (mcp) and coccolith morphology motif (CMM), respectively. Coccolithoviruses within the bloom were found to be variable with depth and unique virus populations were detected at different stations sampled indicating a complex network of EhV-host infections. This ultimately will have significant implications to the internal structure and longevity of ecologically important E. huxleyi blooms.

Enhanced magnitude and breadth of neutralizing humoral response to a DNA vaccine targeting the DHBV envelope protein delivered by in vivo electroporation.

We explored in the duck hepatitis B virus (DHBV) model the impact of electroporation (EP)-mediated DNA vaccine delivery on the neutralizing humoral response to viral preS/S large envelope protein. EP enhanced the kinetics and magnitude of anti-preS response compared to the standard needle DNA injection (SI). Importantly, EP dramatically enhanced the neutralizing potency of the humoral response, since antibodies induced by low DNA dose (10 μg) were able to highly neutralize DHBV and to recognize ten antigenic regions, including four neutralization epitopes. Whereas, SI-induced antibodies by the same low DNA dose were not neutralizing and the epitope pattern was extremely narrow, since it was limited to only one epitope. Thus, EP-based delivery was able to improve the dose efficiency of DNA vaccine and to maintain a highly neutralizing, multi-specific B-cell response, suggesting that it may be an effective approach for chronic hepatitis B therapy at clinically feasible DNA dose.

Microbial biogeography of the North Sea during summer

Micro-organisms are vital for the functioning of all food webs and are the major drivers of the global biogeochemical cycles. The microbial community compositions and physicochemical conditions of the different water masses in the North Sea, a biologically productive sea on the northwestern European continental shelf, were studied during two summer cruises, in order to provide detailed baseline data for this region and examine its microbial biogeography. For each cruise the stations were clustered according to their physicochemical characteristics and their microbial community composition. The largest cluster, which covered most of the central and northern North Sea, consisted of stations that were characterized by a thermally stratified water column and had low chlorophyll a autofluorescence and generally low microbial abundances. The second main cluster contained stations that were dominated by picoeukaryotes and showed the influence of influxes of North Atlantic water via the English Channel and south of the Shetland Islands. The third main cluster was formed by stations that were dominated by cyanobacteria and nanoeukaryotes in the reduced salinity Norwegian Coastal and Skagerrak waters, while the fourth cluster represented the German Bight, a region with strong riverine input, high nutrient concentrations, and consequently high heterotrophic bacterial and viral abundances. Despite the complex and dynamic hydrographic nature of the North Sea, the consistent distinctions in microbiology between these different hydrographic regions during both cruises illustrate the strong links between the microbial community and its environment, as well as the possibility to use microorganisms for long-term monitoring of environmental change.