Structure of the Pseudorabies Virus Capsid: Comparison with Herpes Simplex Virus Type 1 and Differential Binding of Essential Minor Proteins

The structure of pseudorabies virus (PRV) capsids isolated from the nucleus of infected cells and from PRV virions was determined by cryo-electron microscopy (cryo-EM) and compared to herpes simplex virus type 1 (HSV-1) capsids. PRV capsid structures closely resemble those of HSV-1, including distribution of the capsid vertex specific component (CVSC) of HSV-1, which is a heterodimer of the pUL17 and pUL25 proteins. Occupancy of CVSC on all PRV capsids is near 100%, compared to ~ 50% reported for HSV-1 C-capsids and 25% or less that we measure for HSV-1 A- and B-capsids. A PRV mutant lacking pUL25 does not produce C-capsids and lacks visible CVSC density in the cryo-EM-based reconstruction. A reconstruction of PRV capsids in which green fluorescent protein was fused within the N-terminus of pUL25 confirmed previous studies with a similar HSV-1 capsid mutant localizing pUL25 to the CVSC density region that is distal to the penton. However, comparison of the CVSC density in a 9-Å-resolution PRV C-capsid map with the available crystal structure of HSV-1 pUL25 failed to find a satisfactory fit, suggesting either a different fold for PRV pUL25 or a capsid-bound conformation for pUL25 that does not match the X-ray model determined from protein crystallized in solution. The PRV capsid imaged within virions closely resembles C-capsids with the addition of weak but significant density shrouding the pentons that we attribute to tegument proteins. Our results demonstrate significant structure conservation between the PRV and HSV capsids.     

New approaches to the study of Antarctic lithobiontic microorganisms and their inorganic traces,and their application in the detection of life in Martian rocks

Microbial life in the harsh conditions of Antarctica's cold desert may be considered an analogue of potential life on early Mars. In order to explore the development and survival of this epilithic and endolithic form of microbial life, our most sophisticated, state-of-the-art visualization technologies have to be used to their full potential. The study of any ecosystem requires a knowledge of its components and the processes that take place within it. If we are to understand the structure and function of each component of the microecosystems that inhabit lithic substrates, we need to be able to quantify and identify the microorganisms present in each lithobiontic ecological niche and to accurately characterize the mineralogical features of these hidden microhabitats. Once we have established the techniques that will allow us to observe and identify these microorganisms and mineral substrates in situ, and have confirmed the presence of water, the following questions can be addressed: How are the microorganisms organized in the fissures or cavities? Which microorganisms are present and how many are there? Additional questions that logically follow include: What are the existing water relationships in the microhabitat and what effects do the microorganisms have on the mineral composition? Mechanical and chemical changes in minerals and mineralization of microbial cells can give rise to physical and/or chemical traces (biomarkers) and to microbial fossil formation. In this report, we describe the detection of chains of magnetite within the Martian meteorite ALH84001, as an example of the potential use of SEM-BSE in the search for plausible traces of life on early Mars. Electronic Publication     

Microstructural Characterization of Cyanobacterial Mats from the McMurdo Ice Shelf, Antarctica

The three-dimensional structures of two types of cyanobacterium-dominated microbial mats from meltwater ponds on the McMurdo Ice Shelf were as determined by using a broad suite of complementary techniques, including optical and fluorescence microscopy, confocal scanning laser microscopy, scanning electron microscopy with back-scattered electron-imaging mode, low-temperature scanning electron microscopy, and microanalyitical X-ray energy dispersive spectroscopy. By using a combination of the different in situ microscopic techniques, the Antarctic microbial mats were found to be structures with vertical stratification of groups of cyanobacteria and mineral sediments, high contents of extracellular polymeric substances, and large void spaces occupied by water. In cyanobacterium-rich layers, heterocystous nostocalean and nonheterocystous oscillatorialean taxa were the most abundant taxa and appeared to be intermixed with fine-size deposits of epicellular silica and calcium carbonate. Most of the cyanobacterial filaments had similar orientations in zones without sediment particles, but thin filaments were tangled among thicker filaments. The combination of the microscopic techniques used showed the relative positions of biological and mineral entities within the microbial mats and enabled some speculation about their interactions.     

Detection of significant disease risks using a spatial conditional autoregressive model

SUMMARY: The conditional autoregressive (CAR) model is widely used to describe the geographical distribution of a specific disease risk in lattice mapping. Successful developments based on frequentist and Bayesian procedures have been extensively applied to obtain two-stage disease risk predictions at the subregional level. Bayesian procedures are preferred for making inferences, as the posterior standard errors (SE) of the two-stage prediction account for the variability in the variance component estimates; however, some recent work based on frequentist procedures and the use of bootstrap adjustments for the SE has been undertaken. In this article we investigate the suitability of an analytical adjustment for disease risk inference that provides accurate interval predictions by using the penalized quasilikelihood (PQL) technique to obtain model parameter estimates. The method is a first-order approximation of the naive SE based on a Taylor expansion and is interpreted as a conditional measure of variability providing conditional calibrated prediction intervals, given the data. We conduct a simulation study to demonstrate how the method can be used to estimate the specific subregion risk by interval. We evaluate the proposed methodology by analyzing the commonly used example data set of lip cancer incidence in the 56 counties of Scotland for the period 1975-1980. This evaluation reveals a close similarity between the solutions provided by the method proposed here and those of its fully Bayesian counterpart.     

Cumulative Risk of Bovine Mastitis Treatments in Denmark,Finland, Norway and Sweden

Data from the national dairy cow recording systems during 1997 were used to calculate lactation-specific cumulative risk of mastitis treatments and cumulative risk of removal from the herds in Denmark, Finland Norway and Sweden. Sweden had the lowest risk of recorded mastitis treatments during 305 days of lactation and Norway had the highest risk. The incidence risk of recorded mastitis treatments during 305 days of lactation in Denmark, Finland, Norway and Sweden was 0.177, 0.139, 0.215 and 0.127 for first parity cows and 0.228, 0.215, 0.358 and 0.204 for parities higher than three, respectively. The risk of a first parity cow being treated for mastitis was almost 3 times higher at calving in Norway than in Sweden. The period with the highest risk for mastitis treatments was from 2 days before calving until 14 days after calving and the highest risk for removal was from calving to 10 days after calving in all countries.The study clearly demonstrated differences in bovine mastitis treatment patterns among the Nordic countries. The most important findings were the differences in treatment risks during different lactations within each country, as well as differences in strategies with respect to the time during lactation mastitis was treated.     

Acid microenvironments in microbial biofilms of antarctic endolithic microecosystems

Antarctic endolithic microecosystems harbour distinct biofilms. The lithic substrate and the microorganisms comprising these films are intimately linked, leading to complex mineral-microbe interactions. Hence, the microhabitats and microenvironments of these microecosystems are not only determined by the physicochemical features of the lithic substrate, but are also conditioned by the biological components of these biofilms. The Antarctic biofilms analysed in this study are characterized by the presence of extracellular polymer substances and acid microenvironments in the proximity of the cells; cyanobacteria appearing as key components. On ultrastructural analysis, these endolithic cyanobacteria showed differences in sheath organization, probably related to their spatial position in the lithic substrate. It is proposed that in this type of ecosystem, biofilm structure could favour the formation of microsites with specific physicochemical conditions appropriate for the survival of microbial communities in this extreme environment.     

Heterologous expression in <Emphasis Type="Italic">Tritrichomonas foetus</Emphasis> of functional <Emphasis Type="Italic">Trichomonas vaginalis</Emphasis> AP65 adhesin

Background  

Trichomonosis, caused by Trichomonas vaginalis, is the number one, nonviral sexually transmitted infection that has adverse consequences for the health of women and children. The interaction of T. vaginalis with vaginal epithelial cells (VECs), a step preparatory to infection, is mediated in part by the prominent surface protein AP65. The bovine trichomonad, Tritrichomonas foetus, adheres poorly to human VECs. Thus, we established a transfection system for heterologous expression of the T. vaginalis AP65 in T. foetus, as an alternative approach to confirm adhesin function for this virulence factor.     

Microhabitats and Chemical Microenvironments under Saxicolous Lichens Growing on Granite

Lasallia hispanica, Parmelia omphalodes, and Cornicularia normoerica, saxicolous thalli growing on granite, show a close relationship with other lichens and microorganisms living in the lithic substrate beneath them. The lithobiontic community is an accumulation of microorganisms at an interface forming a biofilm, which interacts with the lithic substrate both geophysically and geochemically. Because of their fruticose and foliose morphology, the saxicolous species examined here are mainly involved in geophysical processes, but in the proximity of their attachment structures, geochemical processes may also be observed. On the other hand, fungi, algae and cyanobacteria forming crustose lichens, as well as free-living lithobiontic microorganisms, are known to show combined geophysical and geochemical action, mainly on laminar minerals. The substrate zone where the saxicolous lichens are attached is most affected by weathering reactions and shows the highest co-occurrence of lithobiontic microorganisms. The physical and chemical properties of the substrate, along with lichen and microorganism activity, determine different microenvironments and microhabitats. The ecological functioning of these lithobiontic communities is not yet fully understood, and research efforts similar to the present are needed to confirm that their development is influenced by interrelations between different community members and the substrate, as suggested here.     

Study of lichens with different state of hydration by the combination of low temperature scanning electron and confocal laser scanning microscopies.

The use of techniques such as low temperature scanning electron microscopy (LTSEM) and confocal laser scanning microscopy (CLSM) allows the study of lichen thalli in different states of hydration and also near the natural state. The spatial organization of desiccated thalli, with reduced, very compact algal layers, is different from that of hydrated ones. Sometimes, the observation with transmission electron microscopy (TEM) of photobiont pyrenoids from desiccated thalli reports pyrenoids with a central part of a weak stained matrix lacking pyrenoglobuli, named "empty zones". "Empty zones" are not distinguishable with LTSEM and do not present immunolabelling with rubisco antibody in TEM. These zones could be originated by an expansion process during rehydration produced in chemical fixation.