High-throughput Automated Muropeptide Analysis (HAMA) Reveals Peptidoglycan Composition of Gut Microbial Cell Walls

Peptidoglycan (PGN), a net-like polymer constituted by muropeptides, provides protection for microorganisms and has been a major target for antibiotics for decades. Researchers have explored host-microbiome interactions through PGN recognition systems and discovered key muropeptides modulating host responses. However, most common characterization techniques for muropeptides are labor-intensive and require manual analysis of mass spectra due to the complex cross-linked PGN structures. Each species has unique moiety modifications and inter-/intra-bridges, which further complicates the structural analysis of PGN. Here, we developed a high-throughput automated muropeptide analysis (HAMA) platform leveraging tandem mass spectrometry and in silico muropeptide MS/MS fragmentation matching to comprehensively identify muropeptide structures, quantify their abundance, and infer PGN cross-linking types. We demonstrated the effectiveness of HAMA platform using well-characterized PGNs from E. coli and S. aureus and further applied it to common gut bacteria including Bifidobacterium, Bacteroides, Lactobacillus, Enterococcus, and Akkermansia muciiniphila. Specifically, we found that the stiffness and strength of the cell envelopes may correspond to the lengths and compositions of interpeptide bridges within Bifidobacterium species. In summary, the HAMA framework exhibits an automated, intuitive, and accurate analysis of PGN compositions, which may serve as a potential tool to investigate the post-synthetic modifications of saccharides, the variation in interpeptide bridges, and the types of cross-linking within bacterial PGNs.</p

InGaN-based light-emitting diodes with an embedded conical air-voids structure

The conical air-void structure of an InGaN light-emitting diode (LEDs) was formed at the GaN/sapphire interface to increase the light extraction efficiency. The fabrication process of the conical air-void structure consisted of a dry process and a crystallographic wet etching process on an undoped GaN layer, followed by a re-growth process for the InGaN LED structure. A higher light output power (1.54 times) and a small divergent angle (120o) were observed, at a 20mA operation current, on the treated LED structure when compared to a standard LED without the conical air-void structure. In this electroluminescence spectrum, the emission intensity and the peak wavelength varied periodically by corresponding to the conical air-void patterns that were measured through a 100nm-optical-aperture fiber probe. The conical air-void structure reduced the compressed strain at the GaN/sapphire interface by inducing the wavelength blueshift phenomenon and the higher internal quantum efficiency of the photoluminescence spectra for the treated LED structure

The microtubule-associated protein, EB1, links AIM2 inflammasomes with autophagy-dependent secretion

Inflammasomes are multi-protein complexes that regulate chronic inflammation-associated diseases by inducing interleukin-1 β (IL-1β) secretion. Numerous components involved in inflammasome activation have been identified, but the mechanisms of inflammasome-mediated IL-1β secretion have not yet been fully explored. Here, we demonstrate that end-binding protein 1 (EB1), which is required for activation of AIM2 inflammasome complex, links the AIM2 inflammasome to autophagy-dependent secretion. Imaging studies revealed that AIM2 inflammasomes colocalize with microtubule organizing centers and autophagosomes. Biochemical analyses showed that poly(dA-dT)-activated AIM2 inflammasomes induce autophagy and IL-1β secretion in an LC3-dependent fashion. Furthermore, depletion of EB1 decreases autophagic shedding and intracellular trafficking. Finally, we found that the 5′-AMP activated protein kinase may regulate this EB1-mediated autophagy-based inflammasome-induced secretion of IL-1β. These findings reveal a novel EB1-mediated pathway for the secretion of IL-1β

Metrology Camera System of Prime Focus Spectrograph for Subaru Telescope

The Prime Focus Spectrograph (PFS) is a new optical/near-infrared multi-fiber spectrograph designed for the prime focus of the 8.2m Subaru telescope. PFS will cover a 1.3 degree diameter field with 2394 fibers to complement the imaging capabilities of Hyper SuprimeCam. To retain high throughput, the final positioning accuracy between the fibers and observing targets of PFS is required to be less than 10um. The metrology camera system (MCS) serves as the optical encoder of the fiber motors for the configuring of fibers. MCS provides the fiber positions within a 5um error over the 45 cm focal plane. The information from MCS will be fed into the fiber positioner control system for the closed loop control. MCS will be located at the Cassegrain focus of Subaru telescope in order to to cover the whole focal plane with one 50M pixel Canon CMOS camera. It is a 380mm Schmidt type telescope which generates a uniform spot size with a 10 micron FWHM across the field for reasonable sampling of PSF. Carbon fiber tubes are used to provide a stable structure over the operating conditions without focus adjustments. The CMOS sensor can be read in 0.8s to reduce the overhead for the fiber configuration. The positions of all fibers can be obtained within 0.5s after the readout of the frame. This enables the overall fiber configuration to be less than 2 minutes. MCS will be installed inside a standard Subaru Cassgrain Box. All components that generate heat are located inside a glycol cooled cabinet to reduce the possible image motion due to heat. The optics and camera for MCS have been delivered and tested. The mechanical parts and supporting structure are ready as of spring 2016. The integration of MCS will start in the summer of 2016.Comment: 11 pages, 15 figures. SPIE proceeding. arXiv admin note: text overlap with arXiv:1408.287

Metrology Camera System of Prime Focus Spectrograph for Subaru Telescope

The Prime Focus Spectrograph (PFS) is a new optical/near-infrared multi-fiber spectrograph designed for the prime focus of the 8.2m Subaru telescope. The metrology camera system of PFS serves as the optical encoder of the COBRA fiber motors for the configuring of fibers. The 380mm diameter aperture metrology camera will locate at the Cassegrain focus of Subaru telescope to cover the whole focal plane with one 50M pixel Canon CMOS sensor. The metrology camera is designed to provide the fiber position information within 5{\mu}m error over the 45cm focal plane. The positions of all fibers can be obtained within 1s after the exposure is finished. This enables the overall fiber configuration to be less than 2 minutes.Comment: 10 pages, 12 figures, SPIE Astronomical Telescopes and Instrumentation 201

Prime Focus Spectrograph (PFS): the metrology camera system

The Prime Focus Spectrograph (PFS) is a new optical/near-infrared multi-fiber spectrograph designed for the prime focus of the 8.2m Subaru telescope. PFS will cover a 1.3 degree diameter field with 2394 fibers to complement the imaging capabilities of Hyper SuprimeCam. To retain high throughput, the final positioning accuracy between the fibers and observing targets of PFS is required to be less than 10 µ m. The metrology camera system (MCS) serves as the optical encoder of the fiber positioners for configuring of fibers. The MCS locates at the Cassegrain focus of the Subaru telescope to cover the whole focal plane with one 50M pixel CMOS sensor. The information from MCS will be fed into the fiber positioner control system for closed loop control. The MCS was delivered to Subaru Observatory in Apr. 2018 and it had two engineering runs in Oct. 2018 and Aug. 2019. The 1st engineering run concluded that the original mirror supports need to be improved to provide better image quality. The newly designed mirror supports were installed before the 2nd engineering run. The 2nd engineering run result shows that the MCS overall position accuracy is better than 4μm and the image processing time is less than 4 seconds. The MCS is ready for the system integration with other PFS components

Neutrophilic Inflammation in the Immune Responses of Chronic Obstructive Pulmonary Disease: Lessons from Animal Models

Chronic obstructive pulmonary disease (COPD) is a major cause of mortality worldwide, which is characterized by chronic bronchitis, destruction of small airways, and enlargement/disorganization of alveoli. It is generally accepted that the neutrophilic airway inflammation observed in the lungs of COPD patients is intrinsically linked to the tissue destruction and alveolar airspace enlargement, leading to disease progression. Animal models play an important role in studying the underlying mechanisms of COPD as they address questions involving integrated whole body responses. This review aims to summarize the current animal models of COPD, focusing on their advantages and disadvantages on immune responses and neutrophilic inflammation. Also, we propose a potential new animal model of COPD, which may mimic the most characteristics of human COPD pathogenesis, including persistent moderate-to-high levels of neutrophilic inflammation