Nitric oxide (NO), an important effector molecule of the innate immune system, can also regulate adaptive immunity. In this study, the molecular effects of NO on the toll-like receptor signaling pathway were determined using interleukin-12 (IL-12) as an immunologically relevant target gene. The principal conclusion of these experiments is that NO inhibits IL-1 receptor-associated kinase (IRAK) activity and attenuates the molecular interaction between tumor necrosis factor receptor-associated factor-6 and IRAK. As a consequence, the NO donor S-nitroso-N-acetylpenicillamine (SNAP) inhibits lipopolysaccharide (LPS)-induced IL-12 p40 mRNA expression, protein production, and promoter activity in murine macrophages, dendritic cells, and the murine macrophage cell line RAW 264.7. Splenocytes from inducible nitric-oxide synthase-deficient mice demonstrate markedly increased IL-12 p40 protein and mRNA expression compared with wild type splenocytes. The inhibitory action of NO on IL-12 p40 is independent of the cytokine IL-10. The effects of NO can be directly attributed to inhibition of NF-κB activation through IRAK-dependent pathways. Accordingly, SNAP strongly reduces LPS-induced NF-κB DNA binding to the p40 promoter and inhibits LPS-induced IκB phosphorylation. Similarly, NO attenuates IL-1β-induced NF-κB activation. These experiments provide another example of how an innate immune molecule may have a profound effect on adaptive immunity.

The protein arginine methyltransferases PRMT7 and PRMT5, respectively, monomethylate and symmetrically dimethylate arginine side-chains of proteins involved in diverse cellular mechanisms, including chromatin-mediated control of gene transcription, splicing, and the RAS to ERK transduction cascade. It is believed that PRMT5 and PRMT7 act in conjunction to methylate their substrates, and genetic deletions support the notion that these enzymes derepress cell proliferation and migration in cancer. Using available structures of PRMT5, we designed DS-437, a PRMT5 inhibitor with an IC₅₀ value of 6 μM against both PRMT5 and PRMT7 that is inactive against 29 other human protein-, DNA-, and RNA-methyltransferases and inhibits symmetrical dimethylation of PRMT5 substrates in cells. This compound behaves as a cofactor competitor and represents a valid scaffold to interrogate the potential of the PRMT5–PRMT7 axis as a target for therapy.

Brassica species represent several important crops including canola ( Brassica napus ). Understanding of genetic elements that contribute to seed-associated functions will impact future improvements in the canola crop. Brassica species share a very close taxonomic and molecular relationship with Arabidopsis thaliana. However, there are several subtle but distinct seed-associated agronomic characteristics that differ among the oil seed crop species. To address these, we have generated 67 535 ESTs predominately from Brassica seeds, analyzed these sequences, and identified 10 642 unigenes for the preparation of a targeted seed cDNA array. A set of 10 642 PCR primer pairs was designed and corresponding amplicons were produced for spotting, along with relevant controls. Critical quality control tests produced satisfactory results for use of this microarray in biological experiments. The microarray was also tested with specific RNA targets from embryos, germinating seeds, and leaf tissues. The hybridizations, signal intensities, and overall quality of these slides were consistent and reproducible. Additionally, there are 429 ESTs represented on the array that show no homology with any A. thaliana annotated gene or any gene in the Brassica genome databases or other plant databases; however, all of these probes hybridized to B. napus transcripts, indicating that the array also will be useful in defining expression patterns for genes so far unique to Brassica species.

Grazing removal rate of grasses needs to be determined for various climate conditions to address eco-environmental concerns (e.g., desertification) related to steppe grassland degradation. The conventional approach, which requires survey data on animal species and heads as well as grass consumption per individual animal, is too costly and time-consuming to be applied at a watershed scale. The objective of this study was to present a new approach that can be used to estimate grazing removal rate with no requirement of animal-related data. The application of this new approach was demonstrated in a Eurasian semiarid typical-steppe watershed for an analysis period of 2000 to 2010. The results indicate that the removal rate tended to become larger, but its temporal variation tended to become smaller, from the upstream to downstream. Averaged across the watershed, the removal rate ranged from 63.9 to 401.0 g DM m⁻² (or 22.4 to 60.9%) during the analysis period. As expected, the removal rate in an atmospherically wetter year was higher than that in an atmospherically drier year. Nevertheless, none of the eleven analysis years had a removal rate higher than the threshold value of 65%, above which the risk of grassland degradation would become much greater.