Mass loss is a key property to understand stellar evolution and in particular for low-metallicity environments. Our knowledge has improved dramatically over the last decades both for single and binary evolutionary models. However, episodic mass loss although definitely present observationally, is not included in the models, while its role is currently undetermined. A major hindrance is the lack of large enough samples of classified stars. We attempted to address this by applying an ensemble machine-learning approach using color indices (from IR/Spitzer and optical/Pan-STARRS photometry) as features and combining the probabilities from three different algorithms. We trained on M31 and M33 sources with known spectral classification, which we grouped into Blue/Yellow/Red/B[e] Supergiants, Luminous Blue Variables, classical Wolf-Rayet and background galaxies/AGNs. We then applied the classifier to about one million Spitzer point sources from 25 nearby galaxies, spanning a range of metallicites (). Equipped with spectral classifications we investigated the occurrence of these populations with metallicity.

The Orai1-STIM1 constructed store-operated Ca2+ channels (SOCs) have been found to exert several essential Ca2+ entry/signaling cascades, e.g., the generation of immune response in T lymphocytes. Although biochemical and novel imaging evidence appear to indicate that Orai1 and STIM1 interact with each other to achieve store-operated Ca2+ entry (SOCE), the detailed mechanism of functional SOCE in situ has yet to be fully understood. In this study, green fluorescence protein (EGFP as donor) targeted to either the N- or C-terminal of Orai1 (wild type or ▵1-90+▵267-301 double deletion type) and mOrange (as acceptor) tagged STIM1 were used to comprise a fluorescence resonance energy transfer (FRET) pair within living PC12 cells. The fluorescence lifetime map and histogram/distribution of each single cell, determined by one-photon excitation fluorescence lifetime imaging microscopy (FLIM), was used to visualize FRET and show the Orai1 homodimer and Orai1-STIM1 binding. Both the color-coded lifetime map and the distribution of EGFP-tagged Orai1 significantly changed after the administration of thapsigargin, the SOCE stimulating agent. The FRET efficiency from each experimental set was also calculated and compared using double exponential analysis. In summary, we show the detailed interactions Orai1-Orai1 and Orai1-STIM1 within intact living cells by using the FLIM-FRET technique.

The high sensitivity and spatial resolution enabled by two-photon excitation fluorescence lifetime imaging microscopy/fluorescence resonance energy transfer (2PE-FLIM/FRET) provide an effective approach that reveals protein-protein interactions in a single cell during stimulated exocytosis. Enhanced green fluorescence protein (EGFP)–labeled synaptosomal associated protein of 25 kDa (SNAP25A) and red fluorescence protein (mRFP)–labeled Rabphillin 3A (RPH3A) were co-expressed in PC12 cells as the FRET donor and acceptor, respectively. The FLIM images of EGFP-SNAP25A suggested that SNAP25A/RPH3A interaction was increased during exocytosis. In addition, the multidimensional (three-dimensional with time) nature of the 2PE-FLIM image datasets can also resolve the protein interactions in the z direction, and we have compared several image analysis methods to extract more accurate and detailed information from the FLIM images. Fluorescence lifetime was fitted by using one and two component analysis. The lifetime FRET efficiency was calculated by the peak lifetime (τpeak) and the left side of the half-peak width (τ1/2), respectively. The results show that FRET efficiency increased at cell surface, which suggests that SNAP25A/RPH3A interactions take place at cell surface during stimulated exocytosis. In summary, we have demonstrated that the 2PE-FLIM/FRET technique is a powerful tool to reveal dynamic SNAP25A/RPH3A interactions in single neuroendocrine cells.

The Pr55–xAl12+xFe33–yCuy (0 ≤ x ≤ 5, 0 ≤ y ≤ 8) bulk metallic glasses (BMGs) 5 mm in diameter and 100 mm in length were prepared by copper mold suction casting. Hysteresis loops of the Pr55–xAl12+xFe33–yCuy BMGs and the corresponding Pr55Al12Fe30Cu3 crystallized alloy were measured, and the results showed that the Pr55–xAl12+xFe33–yCuy BMGs are hard magnetic, while the completely crystallized Pr55Al12Fe30Cu3 alloy is paramagnetic at room temperature. The thermal behavior and crystallization of the Pr55Al12Fe30Cu3 BMG were studied by differential scanning calorimetry, and the results indicated that the Pr-based BMG has obvious glass transition and a wide supercooled liquid region up to 75 K. The crystallization activation energy for the Pr55Al12Fe30Cu3 BMG is much smaller than that of Zr–Ti–Cu–Ni–Be BMG.