Comparative effects of trichothecene mycotoxins on bovine platelet function: Acetyl T-2 toxin,a more potent inhibitor than T-2 toxin

The effects of the trichothecene mycotoxins (acetyl T-2 toxin, T-2 toxin, HT-2 toxin, palmityl T-2 toxin, diacetoxyscirpenol (DAS), deoxynivalenol (DON), and T-2 tetraol) on bovine platelet function were examined in homologous plasma stimulated with platelet activating factor (PAF). The mycotoxins inhibited platelet function with the following order of potency: acetyl T-2 toxin > palmityl T-2 toxin = DAS > HT-2 toxin = T-2 toxin. While T-2 tetraol was completely ineffective as an inhibitor, DON exhibited minimal inhibitory activity at concentrations above 10×10^?4M. The stability of the platelet aggregates formed was significantly reduced in all mycotoxin treated platelets compared to that of the untreated PAF controls. It is suggested that the increased sensitivity of PAF stimulated bovine platelets to the more lipophilic mycotoxins may be related to their more efficient partitioning into the platelet membrane compared to the more hydrophilic compounds.     

Correlation of greenhouse bioassay with field response to fertilizer by paper birch

Summary Bulk soil samples were collected from the top 15 cm of untreated areas adjacent to field fertilizer trials at 2 locations. Amounts of N, P, K, and lime equivalent to the field treatments were mixed with the soil in 15-cm diameter pots. Paper birch (Betula papyrifera March.) trees were grown from seed for a greenhouse bioassay. Height and dry weight of the bioassay seedlings were significantly correlated with 3-year volume growth of 10-year-old paper birch seedlings in the field. Correlation coefficients were 0.88 for height growth and 0.91 for dry weight growth on one site, and 0.72 and 0.63 on the other. With further refinements and observations on a larger number of sites, this bioassay technique should be a valuable tool for estimating potential response to fertilizer by young paper birch in the field, and for ranking the relative productivity of different soils.     

Monoclonal antibodies to rabbit liver cytochrome P-448

Cytochrome P-448 (mol wt 55,000 Daltons) from rabbit liver was purified to a specific content of 16.6 nmol/mg. Mice were immunised with this preparation, their spleens removed and dissociated lymphocytes hybridised with myeloma cells. Four monoclonal antibodies against cytochrome P-448 were raised and partially characterised. All four antibodies interacted with cytochrome P-448 in intact microsomal fractions and selectively immunoadsorbed cytochrome P-448 from solubilised microsomal preparations. One of the antibodies inhibited benzo[a] pyrene hydroxylase activity in a reconstituted system, one had no effect on activity and two increased activity. The possible applications of such antibodies are discussed.     

Regulated expression of the rat medium chain hydrolase gene in transgenic rape seed

Medium chain hydrolase (MCH) is an enzyme which regulates the chain length of fatty acid synthesis specifically in the mammary gland of the rat. During lactation, MCH interacts with fatty acid synthase (FAS) to cause premature release of acyl chains, thus providing medium chain fatty acids for synthesis of milk fat. In this study we have investigated the ability of rat MCH to interact with the phylogenetically more distant FAS structure present in plant systems and to cause a perturbation of fatty acid synthesis. Inin vitro experiments, addition of purified MCH to rapeseed homogenates was found to cause a significant perturbation of fatty acid synthesis towards medium chain length products. The rat MCH gene was expressed in transgenic oilseed rape using a seed specific rape acyl carrier protein (ACP) promoter and a rape ACP plastid targeting sequence. Western analysis showed MCH protein to be present in transgenic seed and for its expression to be developmentally regulated in concert with storage lipid synthesis. The chimaeric preprotein was correctly processed and immunogold labelling studies confirmed MCH to be localized within plastid organelles. However, fatty acid analysis of oil from MCH-expressing rape seed showed no significant differences to that from control seed.     

3, 3′-diindolylmethane Enhances the Effectiveness of Herceptin against HER-2/Neu-Expressing Breast Cancer Cells

Herceptin failure is a major clinical problem in breast cancer. A subset of breast cancer patients with high HER-2/neu levels eventually experience metastatic disease progression when treated with Herceptin as a single agent. Mechanistic details of development of this aggressive disease are not clear. Therefore, there is a dire need to better understand the mechanisms by which drug resistance develops and to design new combined treatments that benefit patients with aggressive breast cancer and have minimal toxicity. We hypothesized that 3, 3′-diindolylmethane (DIM), a non-toxic agent can be combined with Herceptin to treat breast cancers with high levels of HER-2/neu. Here, we evaluated the effects of Herceptin alone and in combination with DIM on cell viability, apoptosis and clonogenic assays in SKBR3 (HER-2/neu-expressing) and MDA-MB-468 (HER-2/neu negative) breast cancer cells. We found that DIM could enhance the effectiveness of Herceptin by significantly reducing cell viability, which was associated with apoptosis-induction and significant inhibition of colony formation, compared with single agent treatment. These results were consistent with the down-regulation of Akt and NF-kB p65. Mechanistic investigations revealed a significant upregulation of miR-200 and reduction of FoxM1 expression in DIM and Herceptin-treated breast cancer cells. We, therefore, transfected cells with pre-miR-200 or silenced FoxM1 in these cells for understanding the molecular mechanism involved. These results provide experimental evidence, for the first time, that DIM plus Herceptin therapy could be translated to the clinic as a therapeutic modality to improve treatment outcome of patients with breast cancer, particularly for the patients whose tumors express high levels of HER-2/neu.     

Quantitative Evidence for Increasing Forest Fire Severity in the Sierra Nevada and Southern Cascade Mountains, California and Nevada, USA

Recent research has concluded that forest wildfires in the western United States are becoming larger and more frequent. A more significant question may be whether the ecosystem impacts of wildfire are also increasing. We show that a large area (approximately 120000 km²) of California and western Nevada experienced a notable increase in the extent of forest stand-replacing (“high severity”) fire between 1984 and 2006. High severity forest fire is closely linked to forest fragmentation, wildlife habitat availability, erosion rates and sedimentation, post-fire seedling recruitment, carbon sequestration, and various other ecosystem properties and processes. Mean and maximum fire size, and the area burned annually have also all risen substantially since the beginning of the 1980s, and are now at or above values from the decades preceding the 1940s, when fire suppression became national policy. These trends are occurring in concert with a regional rise in temperature and a long-term increase in annual precipitation. A close examination of the climate–fire relationship and other evidence suggests that forest fuels are no longer limiting fire occurrence and behavior across much of the study region. We conclude that current trends in forest fire severity necessitate a re-examination of the implications of all-out fire suppression and its ecological impacts. Author Contributions: Jay Miller designed the study, performed research, analyzed data, and wrote the article. Hugh Safford performed research, analyzed data, and wrote the article. Michael Crimmins performed research and analyzed data. Andi Thode designed the study and performed research.