RESUMEN
Mutations in the estrogen receptor (ESR1) gene are common in ER-positive breast cancer patients who progress on endocrine therapies. Most mutations localise to just three residues at, or near, the C-terminal helix 12 of the hormone binding domain, at leucine-536, tyrosine-537 and aspartate-538. To investigate these mutations, we have used CRISPR-Cas9 mediated genome engineering to generate a comprehensive set of isogenic mutant breast cancer cell lines. Our results confirm that L536R, Y537C, Y537N, Y537S and D538G mutations confer estrogen-independent growth in breast cancer cells. Growth assays show mutation-specific reductions in sensitivities to drugs representing three classes of clinical anti-estrogens. These differential mutation- and drug-selectivity profiles have implications for treatment choices following clinical emergence of ER mutations. Our results further suggest that mutant expression levels may be determinants of the degree of resistance to some anti-estrogens. Differential gene expression analysis demonstrates up-regulation of estrogen-responsive genes, as expected, but also reveals that enrichment for interferon-regulated gene expression is a common feature of all mutations. Finally, a new gene signature developed from the gene expression profiles in ER mutant cells predicts clinical response in breast cancer patients with ER mutations.
Asunto(s)
Neoplasias de la Mama , Receptores de Estrógenos , Humanos , Femenino , Receptores de Estrógenos/genética , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Receptor alfa de Estrógeno/genética , Receptor alfa de Estrógeno/metabolismo , Pronóstico , Antagonistas de Estrógenos/uso terapéutico , Mutación , Estrógenos/farmacologíaRESUMEN
Plant-derived molluscicides have been indicated as biodegradable and low-cost strategies for control of Biomphalaria spp., intermediate host for the Schistosoma. This study evaluated whether the crude ethanolic extract of the Persea americana stem bark has molluscicidal activity against embryos, newly-hatched and adults of Biomphalaria glabrata. The extract was obtained, characterized and its toxicity analyzed by snail embryotoxicity test (144 h) and acute toxicity test with newly-hatching and adult snails (96 h). Results showed the presence of flavonoids, anthraquinone heterosides, coumarins and tannins in the crude ethanolic extract, which showed molluscicidal activity against all life cycle stages of B. glabrata. The LC50 for embryos, newly-hatched and adults were 27.06, 30.60 and 55.55 ppm, respectively. Embryos exposed to the extract at 50 ppm showed hatching inhibition and at 6.2 and 25 ppm had the highest rates of morphological alterations, such as shell malformations and coagulation of the perivitelline substance. Adult snails exposed to the extract at 75 ppm showed a peak of behavioral changes, such as lethargy and shell reclusion, in addition to answers like hemolymph release in most concentrations. Further studies are required, prioritizing toxicity testing on non-target organisms and further elucidation of the active molecules.
Asunto(s)
Biomphalaria , Lauraceae , Moluscocidas , Persea , Animales , Moluscocidas/toxicidad , Corteza de la Planta , Extractos Vegetales/farmacología , CaracolesRESUMEN
Controlling nitrogen (N) losses from pig slurry (PS) is a challenge under no-till because amendments are left on the soil surface. We investigated the potential of shallow injection of PS, with and without addition of the nitrification inhibitor dicyandiamide (DCD), to abate gaseous ammonia (NH) and nitrous oxide (NO) emissions in winter crops in subtropical soils. Injection was compared with surface broadcasting of PS, with and without DCD. The significance of winter season on annual NO emissions was assessed. Injecting PS reduced NH volatilization compared with surface application. However, this reduction was partly offset because NO emissions increased by 77% (+1.53 kg NO-N ha) when PS was injected. Adding DCD to injected PS reduced NO emission below levels of surface-broadcast PS without the inhibitor, indicating that DCD may be a management option when injecting PS. Compared with a reference urea treatment, PS without DCD increased cumulative NO emissions 5.7-fold (from 613 to 3515 g NO-N ha) when injected, and 3.2-fold (from 613 to 1980 g NO-N ha) when surface applied. Adding DCD significantly reduced emissions with injected PS, whereas reduction was not always significant with surface-applied PS. Nitrous oxide emissions during the winter cropping season contributed 30 to 44% of annual emissions, indicating that controlling gaseous N losses in that season is required to reduce the environmental footprint of the whole cropping system. Overall, combining PS injection with DCD was an efficient practice for reducing winter-season gaseous N losses from no-till soils under subtropical climate.