Characterizing the LIR Domain of ABIN1 and Identifying its Role as a Regulator of Mitophagy

A20 Inhibitor of NF-κB (ABIN1/TNIP1) is a known regulator of TNFα signaling induced cell death and inflammation. The regulatory activity has been attributed to ABIN1’s recruitment of the ubiquitin editing enzyme TNF-α-induced protein 3 (TNFAIP3/A20) to Receptor-interacting serine/threonine-protein kinase 1 (RIPK1). The regulation of RIPK1 by ABIN1 and A20 relies on a third player, linear ubiquitin chain assembly complex (LUBAC), which is involved in the recruitment of ABIN1 to RIPK1. Loss of LUBAC or ABIN1 is embryonic lethal, but loss of A20 is not embryonic lethal. The embryonic lethality due to loss of ABIN1, paired with the lack of lethality with loss of A20, provides evidence that ABIN1 plays a role in cellular regulation beyond its role in recruiting A20 for RIPK1 regulation. We established that ABIN1 binds LC3 and is degraded in an autophagy-dependent manner. We also found that ABIN1 is a binding partner of mitochondria and plays a role in general housekeeping of mitochondria. ABIN1’s regulation of mitophagy is an effect of its regulation of AMP-activated protein kinase (AMPK) phosphorylation, an upstream regulator of mitophagy. Loss of ABIN1 and its disruption of mitophagy appears to influence the release of mitochondrial DNA (mtDNA) from the mitochondria. The presence of mtDNA in the cytosol presents a potential explanation for Lupus in ABIN1 mutant patients

ANAEROBIC AND AEROBIC BIODEGRADATION OF THE OIL DISPERSANT COMPONENTS 1,2-PROPANEDIOL AND 2-BUTOXYETHANOL IN SEAWATER

Oil spills are a recurring issue associated with fossil fuel consumption. The largest accidental oil spill in the history of the petroleum industry was the Deepwater Horizon explosion and seafloor well blowout, where the Deepwater Horizon oil rig exploded and sank, allowing the well to gush uncontrolled from April 20, 2010, until it was capped on July 15, 2010, releasing an estimated 210,000,000 gallons of oil. Oil dispersants were used in unprecedented quantities during the cleanup response to the spill with a total of 1,840,000 gallons of the dispersant COREXIT being applied. The goal of this research was to evaluate the biodegradation of the oil dispersant components 1,2-propanediol (1,2-PD) and 2-butoxyethanol (2-BE) in seawater and under anaerobic conditions, such as those found in near-shore estuarine environments of the Gulf of Mexico. Relatively little is known about the biodegradability of these two compounds in seawater. The specific research objectives were: 1) to evaluate the terminal electron acceptor (TEA) conditions under which 2-BE was biodegraded in microcosms and enrichment cultures based on the stoichiometry of electron acceptor and 2-BE utilization; and 2) to develop enrichment cultures capable of using 2-BE as a growth substrate under anaerobic and aerobic conditions and evaluate the role of sulfate-reducing bacteria through the use of molybdate as an inhibitor. The work conducted and described herein showed that: 1) Analytical methods were successfully developed for quantification of 1,2-PD, 2-BE, nitrate, ferrous iron, and sulfide in the high ionic strength matrix of seawater; 2) 1,2-PD biodegraded in seawater under both aerobic and anaerobic conditions in microcosms constructed with seawater and sediment from Bay Jimmy, Louisiana, which appears to be the first report of 1,2-PD biodegradation in a high salt environment. Because of the high level of biodegradable organic matter in the microcosms, it was not possible to associate the anaerobic consumption of 1,2-PD to a specific terminal electron acceptor; 3) 2-BE biodegraded in seawater under both aerobic and anaerobic conditions in microcosms constructed with seawater and sediment from Bay Jimmy, Louisiana, which appears to be the first report of 2-BE biodegradation in a high salt environment. Because of the high level of biodegradable organic matter in the microcosms, it was not possible to associate the anaerobic consumption of 2-BE to a specific terminal electron acceptor; and 4) Biodegradation of 2-BE was sustained in aerobic and anaerobic enrichment cultures, through two transfers (1% v/v). Nitrate, Fe(III) and sulfate did not appear to be used as the TEAs under anaerobic conditions. Molybdate slowed but did not stop consumption of 2-BE. Sulfide was detected, but at levels well below the amount expected if complete mineralization occurred with sulfate as the TEA. Based on these results, the presumptive pathway for anaerobic biodegradation of 2-BE in the study was via fermentation. Products were not measured, although 2-butoxy acetic acid and hydrogen are likely intermediates. The results of the study definitively show that 1,2-PD and 2-BE biodegraded in microcosms constructed using Bay Jimmy sediment and transfer enrichment cultures. Analytical methods for the measurement of 1,2-PD, 2-BE, Fe(II), nitrate, and sulfide were developed, which has laid the foundation from which future students will investigate the biodegradation of 1,2-PD, 2-BE, and other COREXIT compounds in seawater and in the presence of petroleum hydrocarbons

Traits of Expanded Gender in Hemingway\u27s Lady Brett Ashley

Ernest Hemingway was a male chauvinist pig. This ad hominem attack against him and his writing is an often-repeated, stereotypical polemic that hardly needs scholarly citation. And yet Hemingway paradoxically also created strong female characters who were tough, witty, and supremely independent. Highlighted in this poster is one such character from his 1926 novel, The Sun Also Rises. Lady Brett Ashley is Hemingway\u27s shrewd heroine whose sexual behavior, physical appearance, and personal philosophy helped expand modern sociological gender norms

Brain Regeneration in Drosophila Involves Comparison of Neuronal Fitness

SummaryDarwinian-like cell selection has been studied during development and cancer [1–11]. Cell selection is often mediated by direct intercellular comparison of cell fitness, using “fitness fingerprints” [12–14]. In Drosophila, cells compare their fitness via several isoforms of the transmembrane protein Flower [12, 13]. Here, we studied the role of intercellular fitness comparisons during regeneration. Regeneration-competent organisms are traditionally injured by amputation [15, 16], whereas in clinically relevant injuries such as local ischemia or traumatic injury, damaged tissue remains within the organ [17–19]. We reasoned that “Darwinian” interactions between old and newly formed tissues may be important in the elimination of damaged cells. We used a model of adult brain regeneration in Drosophila in which mechanical puncture activates regenerative neurogenesis based on damage-responsive stem cells [20]. We found that apoptosis after brain injury occurs in damage-exposed tissue located adjacent to zones of de novo neurogenesis. Injury-affected neurons start to express isoforms of the Flower cell fitness indicator protein not found on intact neurons. We show that this change in the neuronal fitness fingerprint is required to recognize and eliminate such neurons. Moreover, apoptosis is inhibited if all neurons express “low-fitness” markers, showing that the availability of new and healthy cells drives tissue replacement. In summary, we found that elimination of impaired tissue during brain regeneration requires comparison of neuronal fitness and that tissue replacement after brain damage is coordinated by injury-modulated fitness fingerprints. Intercellular fitness comparisons between old and newly formed tissues could be a general mechanism of regenerative tissue replacement

Restrictions in the ankle sagittal- and frontal-plane range of movement during simulated walking with different types of orthoses

Different types of orthoses are available to clinicians for non-surgical treatment of acute ankle sprains. The goal of this study was to scientifically compare the movement restrictions in the sagittal and frontal plane during simulated walking between one adaptable semi-rigid brace (OrthoTri-PhaseTM), four non-adaptable semi-rigid braces (OrthoStandardTM, MalleoLocTM, MalleoSprintTM, VACOankleTM), and one rigid cast. Predefined time sequences of rotational moments and axial loading during gait were applied via an ankle joint simulator, with the pneumatic pressure inside the orthoses kept constant to ensure the same condition for different trials and orthoses. The peak ranges of motion (RoMs) in the frontal and sagittal plane during gait were analyzed for statistically significant differences using single-factorial ANOVA with post-hoc Bonferroni analysis. Significant differences in peak plantar-/dorsiflexion and in-/eversion RoM during gait were found between different types of orthoses. In the sagittal plane, the rigid cast most significantly restricted overall RoM followed by the Ortho Tri-PhaseTM in Phase 1 and the Ortho StandardTM. The peak restriction in-/eversion RoM of the VACOankleTM came closest to the rigid cast, with a shift towards inversion. The VACOankleTM allowed for significantly larger dorsiflexion movement compared to all other orthoses. The present results may help clinicians in the decision-making process of finding the optimal orthosis for individual patients

Darwin's multicellularity: from neurotrophic theories and cell competition to fitness fingerprints

Metazoans have evolved ways to engage only the most appropriate cells for long-term tissue development and homeostasis. In many cases, competitive interactions have been shown to guide such cell selection events. In Drosophila, a process termed cell competition eliminates slow proliferating cells from growing epithelia. Recent studies show that cell competition is conserved in mammals with crucial functions like the elimination of suboptimal stem cells from the early embryo and the replacement of old T-cell progenitors in the thymus to prevent tumor formation. Moreover, new data in Drosophila has revealed that fitness indicator proteins, required for cell competition, are also involved in the culling of retinal neurons suggesting that 'fitness fingerprints' may play a general role in cell selection

Targeting eosinophils by active vaccination against interleukin-5 reduces basophil counts in horses with insect bite hypersensitivity in the 2nd year of vaccination

Previously, virus-like particle (VLP)-based self-vaccinations targeting interleukin (IL)-5 or IL-31 have been suggested to treat equine insect bite hypersensitivity (IBH), a seasonal recurrent allergic dermatitis in horses. The IL-5-targeting equine vaccine significantly reduced blood eosinophil counts in horses, similar to human monoclonal antibodies targeting IL-5 or the IL-5 receptor alpha (IL-5Rα). Previous studies in humans have also reported an additional effect on reduction of basophil counts. The aim of the present study was to evaluate whether an equine anti-IL-5 vaccine affected blood basophil counts. Horses with IBH were followed in a 3-year trial consisting of a placebo administered in the 1st year, followed by vaccination using an equine (e)IL-5-VLP vaccine in the 2nd and 3rd years. There was a strong reduction in circulating eosinophil counts after vaccination against IL-5. Additionally, there were reduced basophil counts, but only in the 3rd year of the study, suggesting a bystander effect of the anti-IL-5 vaccine on basophil counts

Editing graphs for maximum effect

The paper contains over eighty rules for editing graphs, arranged under nine major headings in a logical sequence for editing all the graphs in a manuscript. It is excerpted from a monograph used at the Lawrence Livermore National Laboratory to train beginning technical editors in editing graphs; a corresponding Hypercard stack is also used in this training. 6 refs., 4 figs

Flower Forms an Extracellular Code that Reveals the Fitness of a Cell to its Neighbors in Drosophila

SummaryCell competition promotes the elimination of weaker cells from a growing population. Here we investigate how cells of Drosophila wing imaginal discs distinguish “winners” from “losers” during cell competition. Using genomic and functional assays, we have identified several factors implicated in the process, including Flower (Fwe), a cell membrane protein conserved in multicellular animals. Our results suggest that Fwe is a component of the cell competition response that is required and sufficient to label cells as “winners” or “losers.” In Drosophila, the fwe locus produces three isoforms, fweubi, fweLose-A, and fweLose-B. Basal levels of fweubi are constantly produced. During competition, the fweLose isoforms are upregulated in prospective loser cells. Cell-cell comparison of relative fweLose and fweubi levels ultimately determines which cell undergoes apoptosis. This “extracellular code” may constitute an ancient mechanism to terminate competitive conflicts among cells