Regulation of cardiolipin biosynthesis by fatty acid transport protein-1 IN HEK 293 cells

AbstractCardiolipin (CL) is a major phospholipid involved in energy metabolism mammalian mitochondria and fatty acid transport protein-1 (FATP-1) is a fatty acid transport protein that may regulate the intracellular level of fatty acyl-Coenzyme A's. Since fatty acids are required for oxidative phosphorylation via mitochondrial oxidation, we examined the effect of altering FATP-1 levels on CL biosynthesis. HEK-293 mock- and FATP-1 siRNA transfected cells or mock and FATP-1 expressing cells were incubated for 24 h with 0.1 mM oleic acid bound to albumin (1:1 molar ratio) then incubated for 24 h with 0.1 mM [1,3-3H]glycerol and radioactivity incorporated into CL determined. FATP-1 siRNA transfected cells exhibited reduced FATP-1 mRNA and increased incorporation of [1,3-3H]glycerol into CL (2-fold, p<0.05) compared to controls indicating elevation in de novo CL biosynthesis. The reason for this was an increase in [1,3-3H]glycerol uptake and increase in activity and mRNA expression of the CL biosynthetic enzymes. In contrast, expression of FATP-1 resulted a reduction in incorporation of [1,3-3H]glycerol into CL (65%, p<0.05) indicating reduced CL synthesis. [1,3-3H]Glycerol uptake was unaltered whereas activity of cytidine-5′-diphosphate-1,2-diacyl-sn-glycerol synthetase (CDS) and CDS-2 mRNA expression were reduced in FATP-1 expressing cells compared to control. In addition, in vitro CDS activity was reduced by exogenous addition of oleoyl-Coenzyme A. The data indicate that CL de novo biosynthesis may be regulated by FATP-1 through CDS-2 expression in HEK 293 cells

Contributors to the May Issue/Notes

Notes by William B. Lawless, Timothy M. Green, Thomas J. Mitchell, John D. Ryan, Charles Boynton, John R. Baty, and Theodore P. Frericks

Morphological and organic spectroscopic studies of a 44-million-year-old leaf beetle (Coleoptera: Chrysomelidae) in amber with endogenous remains of chitin

This study details the quality of preservation of amber deposits in the Eocene. Through Baltic amber crack-out studies using Synchrotron Micro-Computed Tomography and Scanning Electron Microscopy it was found that the cuticle of a specimen of leaf beetle (Crepidodera tertiotertiaria (Alticini: Galerucinae: Chrysomelidae)) is exceptionally well preserved. Spectroscopic analysis using Synchrotron Fourier Transform Infrared Spectroscopy suggests presence of degraded α -chitin in multiple areas of the cuticle, and Energy Dispersive Spectroscopy supports the presence of organic preservation. This remarkable preservation is likely the result of several factors such as the favourable antimicrobial and physical shielding properties of Baltic amber as compared to other depositional media, coupled to rapid dehydration of the beetle early in its taphonomic process. We provide evidence that crack-out studies of amber inclusions, although inherently destructive of fossils, are an underutilised method for probing exceptional preservation in deep time

Surface effects on the orbital order in the single layered manganite La0.5Sr1.5MnO4

We report the first observation of `orbital truncation rods' -- the scattering arising from the termination of bulk orbital order at the surface of a crystal. The x-ray measurements, performed on a cleaved, single-layered perovskite, La0.5Sr1.5MnO4, reveal that while the crystallographic surface is atomically smooth, the orbital `surface' is much rougher, with an r.m.s. deviation from the average `surface' of ~0.7nm. The temperature dependence of this scattering shows evidence of a surface-induced second order transition.Comment: 13 pages, 4 figure

Influence of HFE variants and cellular iron on monocyte chemoattractant protein-1

<p>Abstract</p> <p>Background</p> <p>Polymorphisms in the <it>MHC class 1-like </it>gene known as <it>HFE </it>have been proposed as genetic modifiers of neurodegenerative diseases that include neuroinflammation as part of the disease process. Variants of <it>HFE </it>are relatively common in the general population and are most commonly associated with iron overload, but can promote subclinical cellular iron loading even in the absence of clinically identified disease. The effects of the variants as well as the resulting cellular iron dyshomeostasis potentially impact a number of disease-associated pathways. We tested the hypothesis that the two most common HFE variants, H63D and C282Y, would affect cellular secretion of cytokines and trophic factors.</p> <p>Methods</p> <p>We screened a panel of cytokines and trophic factors using a multiplexed immunoassay in human neuroblastoma SH-SY5Y cells expressing different variants of HFE. The influence of cellular iron secretion on the potent chemokine monocyte chemoattractant protein-1 (MCP-1) was assessed using ferric ammonium citrate and the iron chelator, desferroxamine. Additionally, an antioxidant, Trolox, and an anti-inflammatory, minocycline, were tested for their effects on MCP-1 secretion in the presence of HFE variants.</p> <p>Results</p> <p>Expression of the HFE variants altered the labile iron pool in SH-SY5Y cells. Of the panel of cytokines and trophic factors analyzed, only the release of MCP-1 was affected by the HFE variants. We further examined the relationship between iron and MCP-1 and found MCP-1 secretion tightly associated with intracellular iron status. A potential direct effect of HFE is considered because, despite having similar levels of intracellular iron, the association between HFE genotype and MCP-1 expression was different for the H63D and C282Y HFE variants. Moreover, HFE genotype was a factor in the effect of minocycline, a multifaceted antibiotic used in treating a number of neurologic conditions associated with inflammation, on MCP-1 secretion.</p> <p>Conclusion</p> <p>Our results demonstrate that HFE polymorphisms influence the synthesis and release of MCP-1. The mechanism of action involves cellular iron status but it appears there could be additional influences such as ER stress. Finally, these data demonstrate a pharmacogenetic effect of HFE polymorphisms on the ability of minocycline to inhibit MCP-1 secretion.</p

Pilot Visual Detection of Small Unmanned Aircraft Systems (sUAS) Equipped with Strobe Lighting

When operating under Visual Flight Rules, pilots primarily rely on visual scanning to avoid other aircraft and airborne collision threats. Records from the Federal Aviation Administration indicate that near encounters with unmanned aircraft are on the rise, reaching 1,761 reported unmanned aircraft system (UAS) sightings or near-misses in 2016. This study sought to assess the effectiveness of pilot visual detection of UAS platforms that were equipped with strobe lighting. A sample of 10 pilots flew a general aviation aircraft on a scripted series of five intercepts with a small UAS (sUAS) that was equipped with strobe lighting. Participants were asked to indicate when they visually detected the unmanned aircraft. Geolocation information for both the aircraft and sUAS platform was compared to assess visibility distance. Findings were used to evaluate the efficacy of daytime strobe lighting as a method to enhance pilot sUAS detection, visibility, and collision avoidance. Participants detected the unmanned aircraft during 7.7% of the intercepts. Due to a lack of data points, the authors were unable to conclusively determine if strobe lighting improved UAS visual detection. The authors recommend further research to explore the effectiveness of using sUAS-mounted strobe lights for nighttime visual detection

Detecting and Assessing Collision Potential of Aircraft and Small Unmanned Aircraft Systems (sUAS) by Visual Observers

Visual observers are used to assist the Remote Pilot with maintaining sight of the unmanned aircraft as well as scanning the surrounding airspace for potential collision hazards. The purpose of this study was to examine the effectiveness of visual observers in detecting an intruding general aviation aircraft approaching the small unmanned aircraft system (sUAS) operations area. The study sought to determine the effectiveness of sUAS visual observers in detecting a general aviation aircraft collision hazard with a sUAS. Ten participants were asked to perform visual observer duties in support of a sUAS operation. Participants were asked to indicate when they were able to hear and see an aircraft that conducted a scripted series of close intercepts with a sUAS. Additionally, researchers assessed each visual observer’s ability to accurately judge the closure rate of the aircraft, by estimating the duration from initial sighting until the aircraft would intercept the airborne sUAS platform. Geolocation data from both the aircraft and sUAS were time correlated and compared to determine estimation accuracy. Findings were used to formulate operational recommendations to improve visual observer performance in detecting and assessing intruder aircraft collision potential

Pilot Visual Detection of Small Unmanned Aircraft on Final Approach during Nighttime Conditions

In December 2020, the Federal Aviation Administration (FAA) announced the release of a new final rule, permitting operators of small unmanned aircraft systems (sUAS) to perform routine night operations. Public comments to the Notice of Proposed Rulemaking indicated potential safety concerns regarding a pilot’s ability to spot a low-altitude sUAS during nighttime conditions. Leveraging data from the FAA’s UAS Sighting Report Database, the research team evaluated the significance of aircraft encounters with UAS at night. Researchers conducted an inflight experiment in which 10 pilots performed an instrument approach to airport during nighttime conditions in which a multi-rotor sUAS presented a potential collision hazard. The sUAS was equipped with lighting visible for 3 miles with a sufficient flash rate to avoid a collision, as specified by the new regulation. Participants performed five approaches, with the sUAS flying different scripted encounter profiles. Participants were asked to indicate when they visually spotted the sUAS, with sighting data recorded via an onboard observer. Geolocation information from both the aircraft and sUAS were compared at the time of each reported sighting to assess visibility distance and orientation. The sUAS was successfully spotted during 30 percent (n = 12) of the testing passes. Hovering sUAS were spotted at the same rate as moving sUAS, however, sUAS in motion were spotted at a much greater range. Researchers noted disproportionately higher spotting rates occurred when the sUAS was oriented on the starboard side of the aircraft vs. the port side. It is believed that airport lighting systems may have obscured or otherwise camouflaged portside sUAS encounters. When asked to estimate distance to an encountered sUAS, most participants underestimated, perceiving the sUAS to be much closer than reality. Additionally, the researchers assessed the potential for the participants to initiate evasive maneuvers, based on the distance and closure rate of the aircraft and sUAS at the time of sighting. Based on the FAA’s Aircraft Identification and Reaction Time Chart, collision avoidance would only have been successful during 15 percent of encounters (n = 6). The research team recommends Remote Pilots employ vigilant traffic awareness during nighttime operations, and leverage use of ADS-B (In) technology and monitor Common Traffic Advisory Frequencies to maintain situational awareness—particularly when operating in proximity to airports