A Multimetric Assessment of Stream Condition in the Northern Lakes and Forests Ecoregion Using Spatially Explicit Statistical Modeling and Regional Normalization

We sampled fish communities, water temperature, water chemistry, physical habitat, and catchment characteristics for 94 stream sites selected randomly throughout the Northern Lakes and Forests ecoregion and used those data to explicitly model reference conditions and assess ecological stream condition at each site via a regional normalization framework. The streams we sampled were first order through fourth order, and the catchments ranged from 0.9 to 458 km2. We developed multiple linear regression (MLR) models that predicted fish community metrics, water chemistry characteristics, and local physical habitat from catchment characteristics; we used these models to compare existing conditions with the conditions that would be expected based on the regression models. Our results indicated that the fish communities were relatively unimpaired because the catchment variables associated with human‐induced land use change were important in only 1 of the 10 fish metric models. Agricultural land use was a significant variable in the MLR equation for species of Lepomis (sunfish). Agricultural land use and urban land use were both significant variables in all of the MLR models predicting water chemistry variables; urban land use was a significant variable in the MLR model predicting the percent coverage of all instream cover types. Regional normalization indicated that none of the sites were impaired based on fish community attributes. However, our analysis based on water chemistry metrics indicated that 22– 35% of the sites were impaired and that, based on physical habitat, 6–14% of the sites were impaired. A comparison with other published studies of the ecoregion suggested that the regional normalization process correctly characterized stream condition.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141590/1/tafs0697.pd

A Landscapeâ Based Classification of Fish Assemblages in Sampled and Unsampled Lakes

We related fish species patterns and landscapeâ scale environmental data from 216 Michigan lakes to identify repeatable types of fish assemblages, identify environmental factors related to assemblage types, and classify fish assemblages in unsampled lakes. Multivariate regression tree modeling of fish species abundances identified six assemblage types that were explained by degreeâ days during the iceâ free period, lake surface area, and mean lake surface temperature. Warmwater species dominated southern lakes, while coolwater and coldwater species had higher abundances in northern lakes. Coolwater species were present in large southern lakes, whereas warmwater species were excluded from northern lakes that had low mean surface temperatures or low degreeâ days. These results suggest that patterns of lake fish assemblages are shaped by differences in climate as well as lakeâ specific differences in surface temperature regimes and in vertical availability of coldwater and coolwater habitats. Because we related fish patterns to readily available landscapeâ level data, our approach can be used to characterize fish assemblages in all lakes across broad geographic extents.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/142256/1/tafs0414.pd

The Great Lakes Hydrography Dataset: Consistent, Binational Watersheds for the Laurentian Great Lakes Basin

Ecosystem‐based management of the Laurentian Great Lakes, which spans both the United States and Canada, is hampered by the lack of consistent binational watersheds for the entire Basin. Using comparable data sources and consistent methods, we developed spatially equivalent watershed boundaries for the binational extent of the Basin to create the Great Lakes Hydrography Dataset (GLHD). The GLHD consists of 5,589 watersheds for the entire Basin, covering a total area of approximately 547,967 km2, or about twice the 247,003 km2 surface water area of the Great Lakes. The GLHD improves upon existing watershed efforts by delineating watersheds for the entire Basin using consistent methods; enhancing the precision of watershed delineation using recently developed flow direction grids that have been hydrologically enforced and vetted by provincial and federal water resource agencies; and increasing the accuracy of watershed boundaries by enforcing embayments, delineating watersheds on islands, and delineating watersheds for all tributaries draining to connecting channels. In addition, the GLHD is packaged in a publically available geodatabase that includes synthetic stream networks, reach catchments, watershed boundaries, a broad set of attribute data for each tributary, and metadata documenting methodology. The GLHD provides a common set of watersheds and associated hydrography data for the Basin that will enhance binational efforts to protect and restore the Great Lakes.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/134077/1/jawr12435_am.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/134077/2/jawr12435.pd

Rhesus TRIM5α disrupts the HIV-1 capsid at the inter-hexamer interfaces

TRIM proteins play important roles in the innate immune defense against retroviral infection, including human immunodeficiency virus type-1 (HIV-1). Rhesus macaque TRIM5α (TRIM5αrh) targets the HIV-1 capsid and blocks infection at an early post-entry stage, prior to reverse transcription. Studies have shown that binding of TRIM5α to the assembled capsid is essential for restriction and requires the coiled-coil and B30.2/SPRY domains, but the molecular mechanism of restriction is not fully understood. In this study, we investigated, by cryoEM combined with mutagenesis and chemical cross-linking, the direct interactions between HIV-1 capsid protein (CA) assemblies and purified TRIM5αrh containing coiled-coil and SPRY domains (CC-SPRYrh). Concentration-dependent binding of CC-SPRYrh to CA assemblies was observed, while under equivalent conditions the human protein did not bind. Importantly, CC-SPRYrh, but not its human counterpart, disrupted CA tubes in a non-random fashion, releasing fragments of protofilaments consisting of CA hexamers without dissociation into monomers. Furthermore, such structural destruction was prevented by inter-hexamer crosslinking using P207C/T216C mutant CA with disulfide bonds at the CTD-CTD trimer interface of capsid assemblies, but not by intra-hexamer crosslinking via A14C/E45C at the NTD-NTD interface. The same disruption effect by TRIM5αrh on the inter-hexamer interfaces also occurred with purified intact HIV-1 cores. These results provide insights concerning how TRIM5α disrupts the virion core and demonstrate that structural damage of the viral capsid by TRIM5α is likely one of the important components of the mechanism of TRIM5α-mediated HIV-1 restriction. © 2011 Zhao et al

Maturation-Induced Cloaking of Neutralization Epitopes on HIV-1 Particles

To become infectious, HIV-1 particles undergo a maturation process involving proteolytic cleavage of the Gag and Gag-Pol polyproteins. Immature particles contain a highly stable spherical Gag lattice and are impaired for fusion with target cells. The fusion impairment is relieved by truncation of the gp41 cytoplasmic tail (CT), indicating that an interaction between the immature viral core and gp41 within the particle represses HIV-1 fusion by an unknown mechanism. We hypothesized that the conformation of Env on the viral surface is regulated allosterically by interactions with the HIV-1 core during particle maturation. To test this, we quantified the binding of a panel of monoclonal antibodies to mature and immature HIV-1 particles by immunofluorescence imaging. Surprisingly, immature particles exhibited markedly enhanced binding of several gp41-specific antibodies, including two that recognize the membrane proximal external region (MPER) and neutralize diverse HIV-1 strains. Several of the differences in epitope exposure on mature and immature particles were abolished by truncation of the gp41 CT, thus linking the immature HIV-1 fusion defect with altered Env conformation. Our results suggest that perturbation of fusion-dependent Env conformational changes contributes to the impaired fusion of immature particles. Masking of neutralization-sensitive epitopes during particle maturation may contribute to HIV-1 immune evasion and has practical implications for vaccine strategies targeting the gp41 MPER

Developing User‐Friendly Habitat Suitability Tools from Regional Stream Fish Survey Data

We developed user‐friendly fish habitat suitability tools (plots) for fishery managers in Michigan; these tools are based on driving habitat variables and fish population estimates for several hundred stream sites throughout the state. We generated contour plots to show patterns in fish biomass for over 60 common species (and for 120 species grouped at the family level) in relation to axes of catchment area and low‐flow yield (90% exceedance flow divided by catchment area) and also in relation to axes of mean and weekly range of July temperatures. The plots showed distinct patterns in fish habitat suitability at each level of biological organization studied and were useful for quantitatively comparing river sites. We demonstrate how these plots can be used to support stream management, and we provide examples pertaining to resource assessment, trout stocking, angling regulations, chemical reclamation of marginal trout streams, indicator species, instream flow protection, and habitat restoration. These straightforward and effective tools are electronically available so that managers can easily access and incorporate them into decision protocols and presentations.Received April 9, 2010; accepted November 8, 2010Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/141005/1/nafm0041.pd

The Interferon Response Inhibits HIV Particle Production by Induction of TRIM22

Treatment of human cells with Type 1 interferons restricts HIV replication. Here we report that the tripartite motif protein TRIM22 is a key mediator. We used transcriptional profiling to identify cellular genes that were induced by interferon treatment and identified TRIM22 as one of the most strongly up-regulated genes. We confirmed, as in previous studies, that TRIM22 over-expression inhibited HIV replication. To assess the role of TRIM22 expressed under natural inducing conditions, we compared the effects of interferon in cells depleted for TRIM22 using RNAi and found that HIV particle release was significantly increased in the knockdown, implying that TRIM22 acts as a natural antiviral effector. Further studies showed that TRIM22 inhibited budding of virus-like particles containing Gag only, indicating that Gag was the target of TRIM22. TRIM22 did not block the release of MLV or EIAV Gag particles. Inhibition was associated with diffuse cytoplasmic staining of HIV Gag rather than accumulation at the plasma membrane, suggesting TRIM22 disrupts proper trafficking. Mutational analyses of TRIM22 showed that the catalytic amino acids Cys15 and Cys18 of the RING domain are required for TRIM22 antiviral activity. These data disclose a pathway by which Type 1 interferons obstruct HIV replication

Macrophage Replication Screen Identifies a Novel Francisella Hydroperoxide Resistance Protein Involved in Virulence

Francisella tularensis is a Gram-negative facultative intracellular pathogen and the causative agent of tularemia. Recently, genome-wide screens have identified Francisella genes required for virulence in mice. However, the mechanisms by which most of the corresponding proteins contribute to pathogenesis are still largely unknown. To further elucidate the roles of these virulence determinants in Francisella pathogenesis, we tested whether each gene was required for replication of the model pathogen F. novicida within macrophages, an important virulence trait. Fifty-three of the 224 genes tested were involved in intracellular replication, including many of those within the Francisella pathogenicity island (FPI), validating our results. Interestingly, over one third of the genes identified are annotated as hypothetical, indicating that F. novicida likely utilizes novel virulence factors for intracellular replication. To further characterize these virulence determinants, we selected two hypothetical genes to study in more detail. As predicted by our screen, deletion mutants of FTN_0096 and FTN_1133 were attenuated for replication in macrophages. The mutants displayed differing levels of attenuation in vivo, with the FTN_1133 mutant being the most attenuated. FTN_1133 has sequence similarity to the organic hydroperoxide resistance protein Ohr, an enzyme involved in the bacterial response to oxidative stress. We show that FTN_1133 is required for F. novicida resistance to, and degradation of, organic hydroperoxides as well as resistance to the action of the NADPH oxidase both in macrophages and mice. Furthermore, we demonstrate that F. holarctica LVS, a strain derived from a highly virulent human pathogenic species of Francisella, also requires this protein for organic hydroperoxide resistance as well as replication in macrophages and mice. This study expands our knowledge of Francisella's largely uncharacterized intracellular lifecycle and demonstrates that FTN_1133 is an important novel mediator of oxidative stress resistance

Host-Adaptation of Francisella tularensis Alters the Bacterium's Surface-Carbohydrates to Hinder Effectors of Innate and Adaptive Immunity

The gram-negative bacterium Francisella tularensis survives in arthropods, fresh water amoeba, and mammals with both intracellular and extracellular phases and could reasonably be expected to express distinct phenotypes in these environments. The presence of a capsule on this bacterium has been controversial with some groups finding such a structure while other groups report that no capsule could be identified. Previously we reported in vitro culture conditions for this bacterium which, in contrast to typical methods, yielded a bacterial phenotype that mimics that of the bacterium's mammalian, extracellular phase.SDS-PAGE and carbohydrate analysis of differentially-cultivated F. tularensis LVS revealed that bacteria displaying the host-adapted phenotype produce both longer polymers of LPS O-antigen (OAg) and additional HMW carbohydrates/glycoproteins that are reduced/absent in non-host-adapted bacteria. Analysis of wildtype and OAg-mutant bacteria indicated that the induced changes in surface carbohydrates involved both OAg and non-OAg species. To assess the impact of these HMW carbohydrates on the access of outer membrane constituents to antibody we used differentially-cultivated bacteria in vitro to immunoprecipitate antibodies directed against outer membrane moieties. We observed that the surface-carbohydrates induced during host-adaptation shield many outer membrane antigens from binding by antibody. Similar assays with normal mouse serum indicate that the induced HMW carbohydrates also impede complement deposition. Using an in vitro macrophage infection assay, we find that the bacterial HMW carbohydrate impedes TLR2-dependent, pro-inflammatory cytokine production by macrophages. Lastly we show that upon host-adaptation, the human-virulent strain, F. tularensis SchuS4 also induces capsule production with the effect of reducing macrophage-activation and accelerating tularemia pathogenesis in mice.F. tularensis undergoes host-adaptation which includes production of multiple capsular materials. These capsules impede recognition of bacterial outer membrane constituents by antibody, complement, and Toll-Like Receptor 2. These changes in the host-pathogen interface have profound implications for pathogenesis and vaccine development