C/EBPβ-1 promotes transformation and chemoresistance in Ewing sarcoma cells.

CEBPB copy number gain in Ewing sarcoma was previously shown to be associated with worse clinical outcome compared to tumors with normal CEBPB copy number, although the mechanism was not characterized. We employed gene knockdown and rescue assays to explore the consequences of altered CEBPB gene expression in Ewing sarcoma cell lines. Knockdown of EWS-FLI1 expression led to a decrease in expression of all three C/EBPβ isoforms while re-expression of EWS-FLI1 rescued C/EBPβ expression. Overexpression of C/EBPβ-1, the largest of the three C/EBPβ isoforms, led to a significant increase in colony formation when cells were grown in soft agar compared to empty vector transduced cells. In addition, depletion of C/EBPβ decreased colony formation, and re-expression of either C/EBPβ-1 or C/EBPβ-2 rescued the phenotype. We identified the cancer stem cell marker ALDH1A1 as a target of C/EBPβ in Ewing sarcoma. Furthermore, increased expression of C/EBPβ led to resistance to chemotherapeutic agents. In summary, we have identified CEBPB as an oncogene in Ewing sarcoma. Overexpression of C/EBPβ-1 increases transformation, upregulates expression of the cancer stem cell marker ALDH1A1, and leads to chemoresistance

Chemical–Genetic Profiling of Imidazo[1,2-a]pyridines and -Pyrimidines Reveals Target Pathways Conserved between Yeast and Human Cells

Small molecules have been shown to be potent and selective probes to understand cell physiology. Here, we show that imidazo[1,2-a]pyridines and imidazo[1,2-a]pyrimidines compose a class of compounds that target essential, conserved cellular processes. Using validated chemogenomic assays in Saccharomyces cerevisiae, we discovered that two closely related compounds, an imidazo[1,2-a]pyridine and -pyrimidine that differ by a single atom, have distinctly different mechanisms of action in vivo. 2-phenyl-3-nitroso-imidazo[1,2-a]pyridine was toxic to yeast strains with defects in electron transport and mitochondrial functions and caused mitochondrial fragmentation, suggesting that compound 13 acts by disrupting mitochondria. By contrast, 2-phenyl-3-nitroso-imidazo[1,2-a]pyrimidine acted as a DNA poison, causing damage to the nuclear DNA and inducing mutagenesis. We compared compound 15 to known chemotherapeutics and found resistance required intact DNA repair pathways. Thus, subtle changes in the structure of imidazo-pyridines and -pyrimidines dramatically alter both the intracellular targeting of these compounds and their effects in vivo. Of particular interest, these different modes of action were evident in experiments on human cells, suggesting that chemical–genetic profiles obtained in yeast are recapitulated in cultured cells, indicating that our observations in yeast can: (1) be leveraged to determine mechanism of action in mammalian cells and (2) suggest novel structure–activity relationships

Evaluation of effects of FMVSS 213 seat back's flexibility on booster seat responses. Final report.

Mode of access: Internet.Author corporate affiliation: National Highway Traffic Safety Administration, Vehicle Research and Test Center, East Liberty, OhioSubject code: DGEORFBSubject code: DGESSubject code: JLTSubject code: OAISubject code: XMC*E

Slightly and Moderately Lame Cows in Tie Stalls Behave Differently From Non-lame Controls. A Matched Case-Control Study.

Lameness affects dairy cows worldwide and is usually associated with pain. Behavioral differences in lame compared to non-lame tie-stall-housed dairy cows might be less pronounced than in free-stall-housed, since the principle demands to a cow's locomotor system and thus the impact of lameness on behavior seem to be lower in tie stalls. Behavioral differences between lame and non-lame cows might be used to estimate the impact of lameness on the well-being of tied dairy cows. In the current study, lame cows were categorized as locomotion scoring between 2.25 and 3.25 on a 1-5 scale. The aim was to compare the eating, rumination and lying behavior of lame cows against non-lame tied dairy cows, in order to draw conclusions on the association of lameness, behavior and well-being in tied dairy cows. The eating and rumination behavior of 26, the lying behavior of 30, and the relative upright and lying activities of 25 matched case-control pairs were analyzed, considering the matching criteria farm, breed-type, and parity-group. Lame cows had fewer [mean of the pairwise differences (case-control) (meandiff) = -2.6 bouts, CI95% (-3.8--1.4) bouts, p = 0.001], but longer lying bouts [meandiff = 26.7 min per bout, CI95% (10.1-43.4) min per bout, p = 0.006]. The lying time was shorter [meandiff = -64.7 min, CI95% (-104.4--24.9) min, p = 0.006] in lame cows compared to their non-lame controls. Lame cows had a shorter eating time [meandiff = -27.7 min, CI95% (-51.5--4.0) min, p = 0.042] and spent a larger proportion of their upright time ruminating [meandiff = 7.2%, CI95% (3.2-11.1)%, p = 0.001] instead of eating. The results of the current study indicate that the eating, rumination, and lying behavior of lame tied dairy cows is altered. These findings indicate that slight and moderate lameness (locomotion score between 2.25 and 3.25 on a 1-5 scale) are likely to be associated with an impaired well-being in affected tied dairy cows. This underlines the need to continuously reduce the lameness prevalence and severity in tied dairy herds

In vitro selective cytotoxicity of the dietary chalcone cardamonin (CD) on melanoma compared to healthy cells is mediated by apoptosis.

Malignant melanoma is an aggressive type of cancer and the deadliest form of skin cancer. Even though enormous efforts have been undertaken, in particular the treatment options against the metastasizing form are challenging and the prognosis is generally poor. A novel therapeutical approach is the application of secondary plant constituents occurring in food and food products. Herein, the effect of the dietary chalcone cardamonin, inter alia found in Alpinia species, was tested using human malignant melanoma cells. These data were compared to cardamonin treated normal melanocytes and dermal fibroblasts representing healthy cells. To investigate the impact of cardamonin on tumor and normal cells, it was added to monolayer cell cultures and cytotoxicity, proliferation, tumor invasion, and apoptosis were studied with appropriate cell biological and biochemical methods. Cardamonin treatment resulted in an apoptosis-mediated increase in cytotoxicity towards tumor cells, a decrease in their proliferation rate, and a lowered invasive capacity, whereas the viability of melanocytes and fibroblasts was hardly affected at such concentrations. A selective cytotoxic effect of cardamonin on melanoma cells compared to normal (healthy) cells was shown in vitro. This study along with others highlights that dietary chalcones may be a valuable tool in anticancer therapies which has to be proven in the future in vivo

CNP mediated selective toxicity on melanoma cells is accompanied by mitochondrial dysfunction.

Cerium (Ce) oxide nanoparticles (CNP; nanoceria) are reported to have cytotoxic effects on certain cancerous cell lines, while at the same concentration they show no cytotoxicity on normal (healthy) cells. Redox-active CNP exhibit both selective prooxidative as well as antioxidative properties. The former is proposed to be responsible for impairment of tumor growth and invasion and the latter for rescuing normal cells from reactive oxygen species (ROS)-induced damage. Here we address possible underlying mechanisms of prooxidative effects of CNP in a metastatic human melanoma cell line. Malignant melanoma is the most aggressive form of skin cancer, and once it becomes metastatic the prognosis is very poor. We have shown earlier that CNP selectively kill A375 melanoma cells by increasing intracellular ROS levels, whose basic amount is significantly higher than in the normal (healthy) counterpart, the melanocytes. Here we show that CNP initiate a mitochondrial increase of ROS levels accompanied by an increase in mitochondrial thiol oxidation. Furthermore, we observed CNP-induced changes in mitochondrial bioenergetics, dynamics, and cristae morphology demonstrating mitochondrial dysfunction which finally led to tumor cell death. CNP-induced cell death is abolished by administration of PEG-conjugated catalase. Overall, we propose that cerium oxide nanoparticles mediate cell death via hydrogen peroxide production linked to mitochondrial dysfunction