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Biological and chemical measurements of sediment are useful parameters when establishing long-term assessment and monitoring tools for designated areas of concern (AOCs) in the Great Lakes. An intensified Environmental Monitoring and Assessment Program (EMAP) sampling grid in the St. Louis River estuary of western Lake Superior was used to assess the relationship between surfi-cial sediment characteristics andbenthic community structure. Ninety sites within two habitat classes(< 5.5 m and > 5.5 m depth) were randomly sampled. Sediment for chemical analysis was collected with a cylindrical drop core while benthic macroinvertebrate abundance and composition were determined from petite Ponar grab samples. Taxa richness was variable (1 to 25 taxa) among sites in the St. Louis River AOC. Oligochaeta were the most abundant taxa while Chironomidae larvae provided a majority of the taxa richness with 43 genera. Results from multivariate redundancy analysis (RDA) on 13 environmental parameters revealed that the majority of variation in benthic community structure was attributed to water depth and site distance from the headwaters. Although physical habitat alterations occur over large spatial scales and are more subtle than those conditions associated with chemically impacted sites, only a small portion of the variability in benthic community structure was explained by sediment chemistry variables. Variability in benthic community structure during this survey was best explained by physical habitat features and must first be quantified prior to understanding benthic response to contaminated sediments. [ABSTRACT FROM AUTHOR]

Abstract: We developed ecological indicators of stream macroinvertebrates in two regions of the Midwestern USA dominated by row-crop agriculture. Indicators were identified in a hierarchical fashion. Reach-scale variables related to macroinvertebrate attributes were first identified, and then catchment-scale variables related to those reach-scale variables were identified. Reach-scale indicators common to both regions were % fine sediments, number of habitats, and width:depth ratio. SD of elevation and % commercial land use were selected as catchment-scale indicators in both regions. Our analyses revealed a multi-scale mechanistic relationship between macroinvertebrate attributes associated with degraded conditions (i.e., fewer taxa of Plecoptera and Trichoptera, and a higher proportion of chironomids, burrowers, and depositional taxa) and % fine sediments in stream reaches, which, in turn, was negatively related to catchment characteristics (i.e., SD of elevation) in one region. Understanding how natural variables such as topography influence channel shape and within-channel structure can help guide management options and expectations for different regions. We suggest that developing multi-scale indicators in a mechanistic fashion will be more effective than developing indicators at only one spatial scale for protecting and restoring stream structure and function. [Copyright &y& Elsevier]

The synergistic effect of IL-18/IL-15/IL-12 stimulation potently activates NK cells, inducing high levels of IFN-γ production. As a result of this potent stimulatory effect, NK cell pre-activation with IL-18/IL-15/IL-12 is being developed as a cancer immunotherapy. Ex vivo expansion of NK cells enables the efficient generation of large numbers of NK cells for wide-scale and repeated therapeutic use, and is thus an important source of NK cells for clinical application. However, the effects of IL-18/IL-15/IL-12 stimulation on ex vivo expanded NK cells have not yet been assessed. Thus, the present study assessed the effects of IL-18/IL-15/IL-12 stimulation on NK cells expanded ex vivo using K562-based artificial antigen presenting cells expressing membrane-bound IL-21. We report that ex vivo expanded NK cells stimulated with IL-18/IL-15/IL-12 produce high levels of IFN-γ and TNFα, have potent cytotoxicity, and maintain prolonged IFN-γ production following removal of stimulation. IL-18/IL-15/IL-12 stimulation induces a phenotypically unique IFN-γ-producing population with reduced CD16 expression and greater CD25 expression as compared to stimulated IFN-γ- NK cells and unstimulated NK cells. We elucidate that the mechanism of synergy for induction and maintenance of IFN-γ production is not due to a further enhancement of STAT4 activation compared to stimulation with IL-12 alone. Furthermore, we demonstrate that the synergistic increase in IFN-γ is not solely under translational regulation, as elevated levels of IFN-γ mRNA contribute to the synergistic increase in IFN-γ. Overall, this study characterizes the response of ex vivo expanded NK cells to IL-18/IL-15/IL-12 stimulation and supports the use of ex vivo expanded NK cells as a feasible and efficient source of IL-18/IL-15/IL-12 pre-activated NK cells for adoptive transfer in cancer immunotherapies. [ABSTRACT FROM AUTHOR]

Abstract: Oncostatin M (OSM) is an IL-6/LIF cytokine family member whose role has been identified in a range of biological activities in vitro, including upregulation of inflammatory gene expression and regulation of connective tissue metabolism. Previously, we identified murine OSM (mOSM) as an inducer of the eosinophil-chemoattractant protein eotaxin-1, both in vitro using fibroblast cell lines and in vivo from mouse lung tissues. Using the NIH 3T3 cell line, we demonstrate the requirement of PI3′K activation for mOSM induction of eotaxin-1 through inhibition of both mOSM-stimulated mRNA and protein expression using the PI3′K antagonist LY294002. By assessment of phosphorylation of the downstream mediator Akt, we show mOSM to be differentially capable of activating PI3′K relative to the related gp130-utilizing cytokine IL-6. Assessment of eotaxin-1 gene expression utilizing PKB/Akt mutant-transfected NIH 3T3 cell lines demonstrated Akt is not involved in upstream regulation of eotaxin-1 through mOSM, indicating an alternate kinase pathway downstream of PI3′K may be involved. We demonstrate that mOSM stimulation of expression of eotaxin-1 is reduced by PD98059, a MAPK kinase inhibitor selective for MEK1. Both LY294002 and PD98059 attenuated mOSM-induced phosphorylation of ERK1/2 MAP kinase and also reduced binding of an AP-1 responsive promoter element, a transcriptional complex known to be MAPK-sensitive. Further, LY294002 pretreatment reduced mOSM-stimulated expression of the downstream AP-1 co-factor JunB, while PD98059 reduced levels of JunB as well as c-Fos. These results provide evidence for a previously unidentified signaling mechanism utilized by mOSM for the induction of eotaxin-1. [Copyright &y& Elsevier]

Oncostatin M (OSM) regulates expression of various genes in connective tissue (CT) cells, including tissue inhibitor of metalloproteinases-1 (TIMP-1). In mouse fibroblast cell lines MLg, NIH 3T3 and primary mouse lung fibroblasts (MLF), murine OSM (muOSM) stimulated high TIMP-1 mRNA expression in comparison to leukemia inhibitory factor (LIF), epidermal growth factor (EGF), interleukin (IL)-1β and transforming growth factor (TGF)β. In cell signaling, muOSM induced strong phosphorylation of extracellular-signal regulated protein kinase (Erk) 1/2, p38 and Akt in addition to phosphorylation of signal transducer and activator of transcription (STAT) 1, STAT3 and STAT5 within 15 min. LIF and TGFβ had no such effects. EGF stimulated comparable or lower Erk1/2, p38 and Akt phosphorylation while IL-1β induced p38 phosphorylation in the fibroblast cell lines. The Erk1/2 inhibitor PD98059 and the p38 inhibitor SB203580 inhibited TIMP-1 mRNA response to muOSM, whereas the phosphoinositide 3-kinase (PI3K) inhibitor LY294002 enhanced the TIMP-1 mRNA response in NIH 3T3 and MLg cells. PD98059 and SB203580, but not LY294002, also inhibited fold induction of a chloramphenicol acetyltransferase (CAT) reporter gene driven by a minimal TIMP-1 promoter that contained a proximal activator protein-1 (AP-1) site. Co-transfection with JunB or c-Jun expression vector in NIH 3T3 cells caused marked transactivation of the TIMP-1 promoter/CAT reporter gene. muOSM caused a rapid increase of JunB and c-Jun protein in NIH 3T3 cells. PD98059 partially inhibited the increase of JunB, but not c-Jun, whereas SB203580 did not induce detectable changes in expression of either AP-1 factor in response to muOSM. These results demonstrate that Erk1/2 and p38 contribute to the elevation of muOSM induced TIMP-1 expression, but PI3K does not, and suggest that Erk1/2 does so by enhancing JunB expression. [Copyright &y& Elsevier]