Sequential Analysis of Trans-SNARE Formation in Intracellular Membrane Fusion

SM proteins stabilize cis-SNARE complexes leading to a specific preferred topology for trans-SNARE formation

Validation of first chemistry mode retrieval results from the new limb-imaging FTS GLORIA with correlative MIPAS-STR observations

We report first chemistry mode retrieval results from the new airborne limb-imaging infrared FTS (Fourier transform spectrometer) GLORIA (Gimballed Limb Observer for Radiance Imaging of the Atmosphere) and comparisons with observations by the conventional airborne limb-scanning infrared FTS MIPAS-STR (Michelson Interferometer for Passive Atmospheric Sounding – STRatospheric aircraft). For GLORIA, the flights aboard the high-altitude research aircraft M55 Geophysica during the ESSenCe campaign (ESa Sounder Campaign 2011) were the very first in field deployment after several years of development. The simultaneous observations of GLORIA and MIPAS-STR during the flight on 16 December 2011 inside the polar vortex and under conditions of optically partially transparent polar stratospheric clouds (PSCs) provided us the first opportunity to compare the observations by two different infrared FTS generations directly. We validate the GLORIA results with MIPAS-STR based on the lower vertical resolution of MIPAS-STR and compare the vertical resolutions of the instruments derived from their averaging kernels. The retrieval results of temperature, HNO3, O3, H2O, CFC-11 and CFC-12 show reasonable agreement of GLORIA with MIPAS-STR and collocated in situ observations. For the horizontally binned hyperspectral limb images, the GLORIA sampling outnumbered the horizontal cross-track sampling of MIPAS-STR by up to 1 order of magnitude. Depending on the target parameter, typical vertical resolutions of 0.5 to 2.0 km were obtained for GLORIA and are typically a factor of 2 to 4 better compared to MIPAS-STR. While the improvement of the performance, characterization and data processing of GLORIA are the subject of ongoing work, the presented first results already demonstrate the considerable gain in sampling and vertical resolution achieved with GLORIA

Validation of first chemistry mode retrieval results from new limb-imaging FTS GLORIA with correlative MIPAS-STR observations

We report first chemistry mode retrieval results from the new airborne limb-imaging infrared FTS (Fourier transform spectrometer) GLORIA (Gimballed Limb Observer for Radiance Imaging of the Atmosphere) and comparisons with observations by the conventional airborne limb-scanning infrared FTS MIPAS-STR (Michelson Interferometer for Passive Atmospheric Sounding - STRatospheric aircraft). For GLORIA, the flights aboard the high-altitude research aircraft M55 Geophysica during the ESSenCe campaign (ESa Sounder Campaign 2011) were the very first in field deployment after several years of development. The simultaneous observations of GLORIA and MIPAS-STR during the flight on 16 December 2011 inside the polar vortex and under conditions of optically partially transparent polar stratospheric clouds (PSCs) provided us the first opportunity to compare the observations by two different infrared FTS generations directly. We validate the GLORIA results with MIPAS-STR based on the lower vertical resolution of MIPAS-STR and compare the vertical resolutions of the instruments derived from their averaging kernels. The retrieval results of temperature, HNO3, O3, H2O, CFC-11 and CFC-12 show reasonable agreement of GLORIA with MIPAS-STR and collocated in situ observations. For the horizontally binned hyperspectral limb images, the GLORIA sampling outnumbered the horizontal cross-track sampling of MIPAS-STR by up to 1 order of magnitude. Depending on the target parameter, typical vertical resolutions of 0.5 to 2.0 km were obtained for GLORIA and are typically a factor of 2 to 4 better compared to MIPAS-STR. While the improvement of the performance, characterization and data processing of GLORIA are the subject of ongoing work, the presented first results already demonstrate the considerable gain in sampling and vertical resolution achieved with GLORIA

Validation of first chemistry mode retrieval results from new limb-imaging FTS GLORIA with correlative MIPAS-STR observations

We report first chemistry mode retrieval results from the new airborne limb-imaging infrared FTS (Fourier transform spectrometer) GLORIA (Gimballed Limb Observer for Radiance Imaging of the Atmosphere) and comparisons with observations by the conventional airborne limb-scanning infrared FTS MIPAS-STR (Michelson Interferometer for Passive Atmospheric Sounding - STRatospheric aircraft). For GLORIA, the flights aboard the high-altitude research aircraft M55 Geophysica during the ESSenCe campaign (ESa Sounder Campaign 2011) were the very first in field deployment after several years of development. The simultaneous observations of GLORIA and MIPAS-STR during the flight on 16 December 2011 inside the polar vortex and under conditions of optically partially transparent polar stratospheric clouds (PSCs) provided us the first opportunity to compare the observations by two different infrared FTS generations directly. We validate the GLORIA results with MIPAS-STR based on the lower vertical resolution of MIPAS-STR and compare the vertical resolutions of the instruments derived from their averaging kernels. The retrieval results of temperature, HNO3, O3, H2O, CFC-11 and CFC-12 show reasonable agreement of GLORIA with MIPAS-STR and collocated in situ observations. For the horizontally binned hyperspectral limb images, the GLORIA sampling outnumbered the horizontal cross-track sampling of MIPAS-STR by up to 1 order of magnitude. Depending on the target parameter, typical vertical resolutions of 0.5 to 2.0 km were obtained for GLORIA and are typically a factor of 2 to 4 better compared to MIPAS-STR. While the improvement of the performance, characterization and data processing of GLORIA are the subject of ongoing work, the presented first results already demonstrate the considerable gain in sampling and vertical resolution achieved with GLORIA

CAIRT - The changing-atmosphere infra-red tomography explorer

Optics InfoBase Conference Papers2021 Article number FTh4G.6. Fourier Transform Spectroscopy, FTS 2021 - Part of OSA Optical Sensors and Sensing Congress 2021Virtual, Online19 July 2021 through 23 July 2021Code 174136CAIRT, a candidate for ESA’s Earth Explorer 11 mission, will observe the Earth’s limb with an imaging Fourier-transform spectrometer. It will provide global observations of ozone, temperature, water vapour and key halogen and nitrogen compounds.With funding from the Spanish government through the Severo Ochoa Centre of Excellence accreditation SEV-2017-0709Peer reviewe

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

A Role for Glutamate Transporters in the Regulation of Insulin Secretion

In the brain, glutamate is an extracellular transmitter that mediates cell-to-cell communication. Prior to synaptic release it is pumped into vesicles by vesicular glutamate transporters (VGLUTs). To inactivate glutamate receptor responses after release, glutamate is taken up into glial cells or neurons by excitatory amino acid transporters (EAATs). In the pancreatic islets of Langerhans, glutamate is proposed to act as an intracellular messenger, regulating insulin secretion from beta-cells, but the mechanisms involved are unknown. By immunogold cytochemistry we show that insulin containing secretory granules express VGLUT3. Despite the fact that they have a VGLUT, the levels of glutamate in these granules are low, indicating the presence of a protein that can transport glutamate out of the granules. Surprisingly, in beta-cells the glutamate transporter EAAT2 is located, not in the plasma membrane as it is in brain cells, but exclusively in insulin-containing secretory granules, together with VGLUT3. In EAAT2 knock out mice, the content of glutamate in secretory granules is higher than in wild type mice. These data imply a glutamate cycle in which glutamate is carried into the granules by VGLUT3 and carried out by EAAT2. Perturbing this cycle by knocking down EAAT2 expression with a small interfering RNA, or by over-expressing EAAT2 or a VGLUT in insulin granules, significantly reduced the rate of granule exocytosis. Simulations of granule energetics suggest that VGLUT3 and EAAT2 may regulate the pH and membrane potential of the granules and thereby regulate insulin secretion. These data suggest that insulin secretion from beta-cells is modulated by the flux of glutamate through the secretory granules