Regulation of cytokine receptors by Golgi N-glycan processing and endocytosis

The Golgi enzyme beta1,6 N-acetylglucosaminyltransferase V (Mgat5) is up-regulated in carcinomas and promotes the substitution of N-glycan with poly N-acetyllactosamine, the preferred ligand for galectin-3 (Gal-3). Here, we report that expression of Mgat5 sensitized mouse cells to multiple cytokines. Gal-3 cross-linked Mgat5-modified N-glycans on epidermal growth factor and transforming growth factor-beta receptors at the cell surface and delayed their removal by constitutive endocytosis. Mgat5 expression in mammary carcinoma was rate limiting for cytokine signaling and consequently for epithelial-mesenchymal transition, cell motility, and tumor metastasis. Mgat5 also promoted cytokine-mediated leukocyte signaling, phagocytosis, and extravasation in vivo. Thus, conditional regulation of N-glycan processing drives synchronous modification of cytokine receptors, which balances their surface retention against loss via endocytosis.     

Trans-endocytosis of CD80 and CD86: a molecular basis for the cell-extrinsic function of CTLA-4

Cytotoxic T lymphocyte antigen 4 (CTLA-4) is an essential negative regulator of T cell immune responses whose mechanism of action is the subject of debate. CTLA-4 shares two ligands (CD80 and CD86) with a stimulatory receptor, CD28. Here, we show that CTLA-4 can capture its ligands from opposing cells by a process of trans-endocytosis. After removal, these costimulatory ligands are degraded inside CTLA-4-expressing cells, resulting in impaired costimulation via CD28. Acquisition of CD86 from antigen-presenting cells is stimulated by T cell receptor engagement and observed in vitro and in vivo. These data reveal a mechanism of immune regulation in which CTLA-4 acts as an effector molecule to inhibit CD28 costimulation by the cell-extrinsic depletion of ligands, accounting for many of the known features of the CD28-CTLA-4 system.     

Evolution of the ratio of strontium-87 to strontium-86 in seawater from cretaceous to present

A detailed record of the strontium-87 to strontium-86 ratio in seawater during the last 100 million years was determined by measuring this ratio in 137 well-preserved and well-dated fossil foraminifera samples. Sample preservation was evaluated from scanning electron microscopy studies, measured strontium-calcium ratios, and pore water strontium isotope ratios. The evolution of the strontium isotopic ratio in seawater offers a means to evaluate long-term changes in the global strontium isotope mass balance. Results show that the marine strontium isotope composition can be used for correlating and dating well-preserved authigenic marine sediments throughout much of the Cenozoic to a precision of +/-1 million years. The strontium-87 to strontium-86 ratio in seawater increased sharply across the Cretaceous/Tertiary boundary, but this feature is not readily explained as strontium input from a bolide impact on land.     

Using Participatory Workshops to Integrate State-and-Transition Models Created With Local Knowledge and Ecological Data

State-and-transition models (STMs) depict current understanding of vegetation dynamics and are being created for most ecological sites in the United States. Model creation is challenging due to inadequate long-term data, and most STMs rely on expert knowledge. There has been little systematic documentation of how different types of knowledge have been integrated in STMs, or what these distinct knowledge sources offer. We report on a series of participatory workshops where stakeholders helped to integrate STMs developed for the same region using local knowledge and ecological field data. With this exploratory project, we seek to understand what kinds of information local knowledge and ecological field data can provide to STMs, assess workshops as a method of integrating knowledge and evaluate how different stakeholders perceive models created with different types of knowledge. Our analysis is based on meeting notes, comments on draft models, and workshop evaluation questionnaires. We conclude that local knowledge and ecological data can complement one another, providing different types of information at different spatial and temporal scales. Participants reported that the workshop increased their knowledge of STMs and vegetation dynamics, suggesting that engaging potential model users in developing STMs is an effective outreach and education approach. Agency representatives and ranchers expressed the value of both the local knowledge and data-driven models. Agency participants were likely to critique or add components based on monitoring data or prior research, and ranchers were more likely to add states and transitions based on personal experience. As STM development continues, it is critical that range professionals think systematically about what different forms of data might contribute to model development, how we can best integrate existing knowledge and data to create credible and useful models, and how to validate the resulting STMs.     

Microbiota accompanying different arbuscular mycorrhizal fungal isolates influence soil aggregation

Arbuscular mycorrhizal fungi (AMF) are key components of ecosystems through their influence on plant communities and ecosystem processes. A major source of information regarding the importance of AMF species richness on process rates are mesocosm experiments using different levels of diversity of AMF as provided by single-species cultures of AMF. Since AMF inocula are generally made available in the form of non-sterile pot culture material, it is possible that AMF symbiosis-associated microbiota are at least partially responsible for some effects hitherto directly attributed to the AMF mycelium. Here, we provide evidence that microbiota associated with single-species cultures of AMF (after long-term pot culture enrichment of 7–8 years) can strongly affect the ecosystem process of soil aggregation. This effect occurred in an AMF isolate specific manner, but in the absence of live and active AMF mycelium. We additionally documented large differences in microbiota communities associated with the different AMF inocula (using PLFA analyses), suggesting that these differences were at least partly responsible for the observed changes in soil aggregation. This result points to AMF–microbiota interactions as a largely unexplored mechanism underlying soil aggregation (and potentially other ecosystem processes). We suggest that a reinterpretation of previous experiments using greenhouse-derived AMF cultures may be necessary, and the need to consider AMF symbiosis-associated microbiota in mechanistic studies of AMF and mycorrhizae in general is emphasized.     

Microbial activity and bacterial composition of H2-treated soils with net CO2 fixation

The effects of H₂ gas treatment of an agricultural soil cultivated previously with a mixture of clover (Trifolium pratense) and alfalfa (Medicago sativa) on CO₂ dynamics and microbial activity and composition were analyzed. The H₂ emission rate of 250 nmol H₂ g⁻¹ soil h⁻¹ was similar to the upper limit of estimated H₂ amounts emitted from N₂ fixing nodules into the surrounding soil ([Dong, Z., Layzell, D.B., 2001. H₂ oxidation, O₂ uptake and CO₂ fixation in hydrogen treated soil. Plant and Soil 229, 1-12.]). After 1 week of H₂ supply to soil samples simultaneously with H₂ uptake net CO₂ production declined continuously and this finally led to a net CO₂ fixation rate in the H₂-treated soil of 8 nmol CO₂ g⁻¹ soil h⁻¹. The time course of H₂ uptake and CO₂ fixation in the soils corresponded with an increase in microbial activity and biomass of the H₂-treated soil determined by microcalorimetric measurements, fluorescence in situ hybridization analysis (FISH) and DNA staining (DAPI). Shifts in the bacterial community structure caused by the supply of H₂ were recorded. While the H₂ treatment stimulated β-and γ-subclasses of Proteobacteria, it had no significant effect on α-Proteobacteria. In addition, FISH-detectable bacteria of the Cytophaga-Flavobacterium-Bacteroides phylum increased in numbers.     

Comparison of the odds of isolation, genotypes, and in vivo production of major toxins by Clostridium perfringens obtained from the gastrointestinal tract of dairy cows with hemorrhagic bowel syndrome or left-displaced abomasum

OBJECTIVE: To compare the frequency of isolation, genotypes, and in vivo production of major lethal toxins of Clostridium perfringens in adult dairy cows affected with hemorrhagic bowel syndrome (HBS) versus left-displaced abomasum (LDA). DESIGN: Case-control study. ANIMALS: 10 adult dairy cattle with HBS (cases) and 10 adult dairy cattle with LDA matched with cases by herd of origin (controls). PROCEDURE: Samples of gastrointestinal contents were obtained from multiple sites during surgery or necropsy examination. Each sample underwent testing for anaerobic bacteria by use of 3 culture methods. The genotype of isolates of C. perfringens was determined via multiplex polymerase chain reaction assay. Major lethal toxins were detected by use of an ELISA. Data were analyzed with multivariable logistic regression and chi2 analysis. RESULTS: C. perfringens type A and type A with the beta2 gene (A + beta2) were the only genotypes isolated. Isolation of C. perfringens type A and type A + beta2 was 6.56 and 3.3 times as likely, respectively, to occur in samples from cattle with HBS than in cattle with LDA. Alpha toxin was detected in 7 of 36 samples from cases and in 0 of 32 samples from controls. Beta2 toxin was detected in 9 of 36 samples from cases and 0 of 36 samples from controls. CONCLUSIONS AND CLINICAL RELEVANCE: C. perfringens type A and type A + beta2 can be isolated from the gastrointestinal tract with significantly greater odds in cattle with HBS than in herdmates with LDA. Alpha and beta2 toxins were detected in samples from cows with HBS but not from cows with LDA.