Indirect monitoring of TORC1 signalling pathway reveals molecular diversity among different yeast strains.

Saccharomyces cerevisiae is the main species responsible for the alcoholic fermentation in wine production. One of the main problems in this process is the deficiency of nitrogen sources in the grape must, which can lead to stuck or sluggish fermentations. Currently, yeast nitrogen consumption and metabolism are under active inquiry, with emphasis on the study of the TORC1 signalling pathway, given its central role responding to nitrogen availability and influencing growth and cell metabolism. However, the mechanism by which different nitrogen sources activates TORC1 is not completely understood. Existing methods to evaluate TORC1 activation by nitrogen sources are time-consuming, making difficult the analyses of large numbers of strains. In this work, a new indirect method for monitoring TORC1 pathway was developed on the basis of the luciferase reporter gene controlled by the promoter region of RPL26A gene, a gene known to be expressed upon TORC1 activation. The method was tested in strains representative of the clean lineages described so far in S. cerevisiae. The activation of the TORC1 pathway by a proline-to-glutamine upshift was indirectly evaluated using our system and the traditional direct methods based on immunoblot (Sch9 and Rps6 phosphorylation). Regardless of the different molecular readouts obtained with both methodologies, the general results showed a wide phenotypic variation between the representative strains analysed. Altogether, this easy-to-use assay opens the possibility to study the molecular basis for the differential TORC1 pathway activation, allowing to interrogate a larger number of strains in the context of nitrogen metabolism phenotypic differences.

Advances in multimodal treatment for stage IIIA-N2 non-small cell lung cancer.

BACKGROUND AND AIM: In Stage IIIA-N2 non-small cell lung cancer (NSCLC), the accuracy of combined positron-emission tomography/computed tomography imaging (PET-CT), together with mediastinal staging techniques, has led to a wide range of challenging clinical scenarios in terms of therapeutic management. Concurrent chemoradiotherapy followed by consolidation immunotherapy remains the standard of care. In patients with potentially-resectable disease, surgery plays an important role in multimodal therapy. The introduction of targeted therapies and immune-checkpoint inhibitors has revolutionized multimodal treatment. In the present article, we review current treatment options and future trends in stage IIIA-N2 NSCLC. RELEVANCE FOR PATIENTS: This article provides insight into the current status of multimodal treatment for NSCLC to support decision-making in routine clinical practice.

Factors Affecting Thermal Insulation of Songbird Nests as Measured Using Temperature Loggers.

AbstractBird nests represent an extended phenotype that is variable among and within species in terms of nest location and construction materials. It has also been suggested that nests indicate niche construction, although empirical evidence to support this is lacking. The nest wall is often considered to confer some insulation because this would help minimize the energetic expenditure by the incubating adult. However, it has been previously suggested that nests are constructed primarily for their structural role rather than for insulation, but to date, studies have used a variety of techniques to investigate the insulation of nest walls but only for relatively few species. This study used temperature loggers to determine insulatory values of nests for 16 previously undescribed species and to replicate values for eight previously described species. In addition, data for nest wall insulation that were determined using temperature loggers have been collated for a total of 32 different passerine species. The effects of nest dimensions, mass, and composition on these values were examined. Base thickness, but not wall thickness, and nest mass significantly positively affected insulatory values. This study found that the proportions of feathers and moss in the nest wall significantly positively correlated with insulatory values. This suggests that there is a key role for nest materials in determining insulation, which provides empirical evidence that nest building constitutes niche construction. The data will also help us interpret data from future studies of the thermal properties of nests. Ultimately, we need to develop our understanding of the role of insulatory values in the evolution of nest function.