Diminished Reovirus Capsid Stability Alters Disease Pathogenesis and Littermate Transmission

Reovirus is a nonenveloped mammalian virus that provides a useful model system for studies of viral infections in the young. Following internalization into host cells, the outermost capsid of reovirus virions is removed by endosomal cathepsin proteases. Determinants of capsid disassembly kinetics reside in the viral σ3 protein. However, the contribution of capsid stability to reovirus-induced disease is unknown. In this study, we found that mice inoculated intramuscularly with a serotype 3 reovirus containing σ3-Y354H, a mutation that reduces viral capsid stability, succumbed at a higher rate than those infected with wild-type virus. At early times after inoculation, σ3-Y354H virus reached higher titers than wild-type virus at several sites within the host. Animals inoculated perorally with a serotype 1 reassortant reovirus containing σ3-Y354H developed exaggerated myocarditis accompanied by elaboration of pro-inflammatory cytokines. Surprisingly, unchallenged littermates of mice infected with σ3-Y354H virus displayed higher titers in the intestine, heart, and brain than littermates of mice inoculated with wild-type virus. Together, these findings suggest that diminished capsid stability enhances reovirus replication, dissemination, lethality, and host-to-host spread, establishing a new virulence determinant for nonenveloped viruses.     

Effects of ambient temperature on avian incubation behavior

Ambient temperature is commonly thought to influence avian incubation behavior. However, results of empirical studies examining correlationsbetween ambient temperature and bout duration are equivocal.We propose that these equivocal results can be partly explainedby developing a conceptual understanding of how we should expecttemperature to influence incubation. We demonstrate why linearcorrelation analyses across a wide range of temperatures canbe inappropriate based on development of an incubation model for small birds that incorporates how ambient temperature influencesboth embryonic development and adult metabolism. We found supportfor predictions of the model using incubation data from orange-crownedwarblers (Vermivora celata) in Arizona. Both off- and on-boutduration were positively correlated with ambient temperaturebetween 9° and 26°C, but unrelated to ambient temperature<9° and 26-40°C. Bout durations declined as ambienttemperature approached or exceeded 40°C. Incubating orange-crowned warblers appeared to avoid bouts off the nest <7 min andbouts on the nest <20 min. Time of day, duration of theprevious bout, and variation among nests all explained variationin both on- and off-bout duration. Although we found supportfor the general shape of the incubation model, temperature still explained only a small portion of the overall variation in on-and off-bout duration. Results of previous studies were generallyconsistent with the model for off-bout duration; most studiesin colder environments reported positive correlations withtemperature, and the one negative correlation reported was from a hot environment. However, the relationships between on-boutduration and temperature reported in previous studies wereless consistent with our model and our data. Although somediscrepancies could be explained by considering our model,some studies reported negative correlations in cold environments.The effect of ambient temperature on duration of on-bouts probablydiffers among species based on the amount of fat reserves females typically carry during incubation and the extent of male incubationfeeding. Additional studies of the effects of temperature onavian incubation will help improve the general model and ultimatelyaid our understanding of energetic and ecological constraintson avian incubation.     

Evaluation of past and potential phosphorus uptake at the Orlando Easterly Wetland

The Orlando Easterly Wetland (OEW), located near Christmas, Florida, USA, is among the longer-lived treatment wetlands in the United States. It was established in the late 1980s to reduce nitrogen and phosphorus concentrations from tertiary treated wastewater bound for the St. Johns River. A goal of 0.07 mg/l total phosphorus concentration has been set by the regulating agency (St. Johns River Water Management District). In order to understand and define the operating conditions for which this target could be met, a systematic study of historic phosphorus uptake was performed using a traditional first-order model. Phosphorus uptake performance is shown to correlate well with hydraulic performance for two parallel upstream cells. The first-order model is enhanced with predictive capabilities that acknowledge the correlation between the phosphorus uptake rate constant and the hydraulic loading rate observed in the system. Inherent limitations with the first-order modeling approach are addressed and uncertainty in model performance is used to bound predictions.     

Assessment of plant-available phosphorus in a fine textured sodic substrate

The objective of this study was to determine how common soil P extractants compared when assessed on revegetated bauxite residue. Samples from contrasting residue preparation, amended with gypsum and/or organic matter, were analyzed for phosphorus using CaCl₂, Morgans, Olsen and Colwell extractants. Herbage samples were also taken from the residue and phosphorus content and biomass determined. Phosphorus adsorption in the different treatments was also assessed.Amounts determined by CaCl₂ were below limits of detection and Morgans extractant overestimated amounts of plant available P. P levels determined by Olsen extractant were within the deficiency range. Colwell P, although overestimated plant available P, did correlate with vegetation performance (biomass production) and amount of P added to the substrate. Plant indices (P-content and biomass) were correlated with organic and Ca content of the substrate.Fine particle size of the substrate resulted in increased rate of P adsorption and Ca-content further increased the rate. However, overall Ca content of the residue improved P-uptake and nutrition.     

Human Amnion Epithelial Cells Induced to Express Functional Cystic Fibrosis Transmembrane Conductance Regulator

Cystic fibrosis, an autosomal recessive disorder caused by a mutation in a gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR), remains a leading cause of childhood respiratory morbidity and mortality. The respiratory consequences of cystic fibrosis include the generation of thick, tenacious mucus that impairs lung clearance, predisposing the individual to repeated and persistent infections, progressive lung damage and shortened lifespan. Currently there is no cure for cystic fibrosis. With this in mind, we investigated the ability of human amnion epithelial cells (hAECs) to express functional CFTR. We found that hAECs formed 3-dimensional structures and expressed the CFTR gene and protein after culture in Small Airway Growth Medium (SAGM). We also observed a polarized CFTR distribution on the membrane of hAECs cultured in SAGM, similar to that observed in polarized airway cells in vivo. Further, hAECs induced to express CFTR possessed functional iodide/chloride (I^−/Cl⁻) ion channels that were inhibited by the CFTR-inhibitor CFTR-172, indicating the presence of functional CFTR ion channels. These data suggest that hAECs may be a promising source for the development of a cellular therapy for cystic fibrosis.     

An Evaluation of Revegetation Success on Bauxite Residue

The current study was undertaken to evaluate the success of a revegetation program on three sites within the Bauxite Residue Disposal Area at the Aughinish Alumina Ltd. refinery. This was achieved by determining botanical diversity, substrate conditions, and plant uptake. Two sites revegetated in 1999, with and without the use of gypsum, were assessed and compared to a site revegetated in 1997. Compared to an initial 6 species used in seeding, a total of 47 species were recorded growing on the three sites with greatest diversity on the 1997 site. There was limited variation in the residue properties of the three treatments indicating that diversity was most influenced by succession and not substrate conditions. Limited available manganese was found in all treatments and significantly lower exchangeable magnesium in the gypsum-amended treatment. Exchangeable sodium, aluminum, and pH in the substrate were not at levels of concern. Appreciable nitrogen, phosphorus, and potassium were found as a result of a fertilizing program. Dominant species in the 1999 treatments, Holcus lanatus and Trifolium pratense , were analyzed for elemental composition. Compared to previous studies, foliar nitrogen, phosphorus, potassium, and calcium were adequate and sodium levels were low. Manganese and magnesium levels were low, and availability should be assessed as part of the monitoring program. Furthermore, the effect of a fertilizing regime on plant uptake and substrate conditions needs to be assessed.     

Defining the limits: Protein aggregation and toxicity in vivo

Abstract

The proper folding of proteins to their functional forms is essential to cellular homeostasis. Perhaps not surprisingly, cells have evolved multiple pathways, some overlapping and others complementary, to resolve mis-folded proteins when they arise, ranging from refolding through the action of molecular chaperones to elimination through regulated proteolytic mechanisms. These protein quality control pathways are sufficient, under normal conditions, to maintain a functioning proteome, but in response to diverse environmental, genetic and/or stochastic events, protein mis-folding exceeds the corrective capacity of these pathways, leading to the accumulation of aggregates and ultimately toxicity. Particularly devastating examples of these effects include certain neurodegenerative diseases, such as Huntington’s Disease, which are associated with the expansion of polyglutamine tracks in proteins. In these cases, protein mis-folding and aggregation are clear contributors to pathogenesis, but uncovering the precise mechanistic links between the two events remains an area of active research. Studies in the yeast Saccharomyces cerevisiae and other model systems have uncovered previously unanticipated complexity in aggregation pathways, the contributions of protein quality control processes to them and the cellular perturbations that result from them. Together these studies suggest that aggregate interactions and localization, rather than their size, are the crucial considerations in understanding the molecular basis of toxicity.