Redox-dependent gating of VDAC by mitoNEET.

MitoNEET is an outer mitochondrial membrane protein essential for sensing and regulation of iron and reactive oxygen species (ROS) homeostasis. It is a key player in multiple human maladies including diabetes, cancer, neurodegeneration, and Parkinson's diseases. In healthy cells, mitoNEET receives its clusters from the mitochondrion and transfers them to acceptor proteins in a process that could be altered by drugs or during illness. Here, we report that mitoNEET regulates the outer-mitochondrial membrane (OMM) protein voltage-dependent anion channel 1 (VDAC1). VDAC1 is a crucial player in the cross talk between the mitochondria and the cytosol. VDAC proteins function to regulate metabolites, ions, ROS, and fatty acid transport, as well as function as a "governator" sentry for the transport of metabolites and ions between the cytosol and the mitochondria. We find that the redox-sensitive [2Fe-2S] cluster protein mitoNEET gates VDAC1 when mitoNEET is oxidized. Addition of the VDAC inhibitor 4,4'-diisothiocyanatostilbene-2,2'-disulfonate (DIDS) prevents both mitoNEET binding in vitro and mitoNEET-dependent mitochondrial iron accumulation in situ. We find that the DIDS inhibitor does not alter the redox state of MitoNEET. Taken together, our data indicate that mitoNEET regulates VDAC in a redox-dependent manner in cells, closing the pore and likely disrupting VDAC's flow of metabolites

Emergency destruction system for recovered chemical munitions

At the request of the US Army Project Manager for Non-Stockpile Chemical Materiel, Sandia National Laboratories is developing a transportable system for destroying recovered, explosively configured, chemical warfare munitions. The system uses shaped charges to access the agent and burster followed by chemical neutralization to destroy them. The entire process takes place inside a sealed pressure vessel. In this paper, they review the design, operation, and testing of a prototype system capable of containing up to one pound of explosive

Technical Advisory Team (TAT) report on the rocket sled test accident of October 9, 2008.

This report summarizes probable causes and contributing factors that led to a rocket motor initiating prematurely while employees were preparing instrumentation for an AIII rocket sled test at SNL/NM, resulting in a Type-B Accident. Originally prepared by the Technical Advisory Team that provided technical assistance to the NNSA's Accident Investigation Board, the report includes analyses of several proposed causes and concludes that the most probable source of power for premature initiation of the rocket motor was the independent battery contained in the HiCap recorder package. The report includes data, evidence, and proposed scenarios to substantiate the analyses

Low flow controls on benthic and hyporheic macroinvertebrate assemblages during supra-seasonal drought

Despite the widely accepted importance of the hyporheic zone as a habitat for stream macroinvertebrates during floods, few data exist regarding community composition and distribution during periods of low flow or drought in perennial streamsi Integrating research on hyporheic invertebrates with results from a long-term study of a U K river provided the opportunity to examine how surface and hyporheic macroinvertebrate communities respond to inter-annual variability in river flow and periods of groundwater drought. Changes in the riverine macroinvertebrate community associated with low flow included a reduction in species richness and the number of individuals per sample, particularly aquatic insects. The hyporheic community was characterized by a relatively homogeneous composition during a period of severe low flow, punctuated by short-term changes associated with variation in water temperature rather than changes in discharge. We present a conceptual model of the processes influencing benthic and hyporheic invertebrates under low-flow conditions. Previous studies have seldom integrated these two assemblages and their interactions. The model presented highlights the potential importance of surface water and hyporheic zone linkages for riverine invertebrate communities under a range of flow conditions

Understanding Free Radicals: Isolating Active Thylakoid Membranes and Purifying the Cytochrome b6f Complex for Superoxide Generation Studies

All life persists in an environment that is rich in molecular oxygen. The production of oxygen free radicals, or superoxide, is a necessary consequence of the biogenesis of energy in cells. Both mitochondrial and photosynthetic electron transport chains have been found to produce superoxide associated with cell differentiation, proliferation, and cell death, thereby contributing to the effects of aging. Aerobic respiration in mitochondria consumes oxygen, whereas photosynthesis in chloroplasts or cyanobacteria produces oxygen. The increased concentration of molecular oxygen may serve to allow greater availability for the production of superoxide by cytochrome bc complexes in photosynthetic membranes compared to those of mitochondrial membranes. The isolation of well-coupled chloroplasts, containing the cytochrome b6f complex of oxygenic photosynthesis, is a vital initial step in the process of comparing the rate of production of superoxide to those of the homologous cytochrome bc1 complex of aerobic respiration. It is necessary to determine if the isolated chloroplasts have retained their oxygengenerating capability after isolation by an oxygen evolution assay with a Clark-type electrode. A necessary second step, which is the isolation of cytochrome b6f from spinach, has yet to be successfully performed. Oxygen measurements taken from chloroplasts in the presence of the uncoupler, NH4Cl, exhibited a rate of oxygen evolution over three times greater at 344 +/- 18 μmol O2/mg Chlorophyll a/hr than the rate of oxygen evolution without uncoupler at 109 +/- 29 μmol O2/mg Chlorophyll a/hr. These data demonstrate that the technique used to isolate spinach chloroplasts preserves their light-driven electron-transport activity, making them reliable for future superoxide assays

There Is Always a Wild Side to an Innocent Face

There Is Always a Wild Side to an Innocent Face, taken by Katie Stofleth on the Linfield College January Term Program in Southeast Asia. 3rd place, People division, in the 2015 Linfield College Study Abroad Photo Contest.https://digitalcommons.linfield.edu/intl_photos2015/1005/thumbnail.jp

Complications à long terme chez les patients traités pour un neuroblastome dans l’enfance, étude d’une cohorte régionale française

Background. Neuroblastoma is a childhood cancer treated with surgery or multimodal therapy including chemotherapy and radiations. Long term complications can occur many years after treatment. The aim of this study was to describe physical long-term sequelae for adult neuroblastoma survivors and to evaluate their correlation with treatment. Methods. A prospective multicentric regional study was conducted to evaluate long-term complications for all childhood cancer survivors in Rhone-Alpes Region. 40 neuroblastoma survivors treated between 1987 and 1999 were seen in long term follow-up clinical consultation. Results. The median follow-up was 20.5 years (range 16.9-25.7). Received treatments were surgery alone for 13 patients, surgery and standard chemotherapy for 21 patients and multimodal intensive therapy for 6 patients. A total of 94 physical long-term complications were observed in 32 patients (80%) with a median of 2 per patient (range 0-9). The main long-term complications were metabolism disorder (42.5%), neurological impairment (32.5%), musculoskeletal impairment (27.5%), hearing loss (20%) and endocrine complications (17.5%). The number of sequelae increased after multimodal intensive therapy (p < 0.0001), as well as their severity (p < 0.0001). Psychiatric disorders were found in 52.9% of patients. Conclusion. A large proportion of patients treated for neuroblastoma in childhood experience long term complications, especially after an intensive treatment. This underscores the need for long term follow-up for childhood cancer survivors.Contexte. Le neuroblastome est un cancer pédiatrique traité par chirurgie seule qui peut être associée à une chimiothérapie standard ou haute dose, ou une radiothérapie en fonction du niveau de risque de la maladie. Des complications à long terme peuvent survenir, y compris plusieurs années après le traitement. Notre objectif était de décrire les séquelles à long terme chez des adultes survivants d’un neuroblastome et d’évaluer leur corrélation avec le traitement reçu. Méthodes. Une étude prospective multicentrique a été menée afin d’évaluer les complications survenant chez les patients traités pour un neuroblastome en région Rhône-Alpes. 40 patients traités entre 1987 et 1999 ont été reçus en consultation de suivi. Résultats. La durée médiane de suivi était de 20.5 ans (16.9-25.7). 13 patients avaient reçu une chirurgie seule, 21 une chimiothérapie standard et 6 un traitement combiné intensif. 94 complications à long terme ont été rapportées chez 32 patients (80%) avec une médiane de 2 (0-9). Les principales séquelles étaient métaboliques (42.5%), neurologiques (32.5%), musculosquelettiques (27.5%), auditives (20%) et endocriniennes (17.5%). Leur nombre était significativement augmenté en cas de traitement combiné intensif (p < 0.0001), de même que celui des complications sévères (p < 0.0001). Des troubles psychiatriques étaient retrouvés chez 52.9% des patients. Conclusion. Une grande proportion de patients traités pour un neuroblastome souffraient de complications à long terme et particulièrement ceux ayant reçu un traitement intensif. Cette étude souligne la nécessité d’un suivi à long terme pour les adultes survivants d’un cancer dans l’enfance

Talking About Mindfulness: Portrayals in Diverse Communication Communities

Thesis (Ph.D.)--University of Washington, 2017-08Mindfulness as a concept and a practice has gained significant recognition, referenced and employed in academic studies and the popular press, among other communities. Within these literatures, broad conclusions are often drawn about mindfulness, its relationships to other variables, and its impacts or benefits. Given the surge of writings about mindfulness in these contexts, and the complexity of the mindfulness construct itself, it is not surprising that there is variation in its conceptualization across sources even as the same name “mindfulness” is used. It is therefore important to discern “what are different groups talking about, when talking about mindfulness?” to assess, among other things, the varying meanings and values imbued within the term by these different communities. Taking an Ethnography of Communication approach, this study performs a communication codes analysis on two communication communities (popular press articles and academic journal articles) to determine if there are particular communication codes associated with either community and, if so, what distinguishes them both within and across the communities. This analysis has implications for academic understanding on as well as practical application of the topic/practice

We Rise by Lifting Others

We Rise by Lifting Others, taken by Katie Stofleth on the Linfield College January Term Program in Southeast Asia. 3rd place, Judges\u27 Award division, in the 2015 Linfield College Study Abroad Photo Contest.https://digitalcommons.linfield.edu/intl_photos2015/1002/thumbnail.jp

Connecting Iron-Sulfur Transport at the Cytosol and Mitochondria Interface

There exist three different NEET family proteins in human cells. Named for their shared characteristic amino acid sequence (Asp-Glu-Glu-Thr), they were first identified as pioglitazone drug targets, one of the thiazolidinedione class of insulin-sensitizing therapeutics for Type 2 diabetes. Although each share similar structural organization and coordinate two [2Fe-2S] clusters with a unique 3Cys:1His coordination chemistry, their cellular localization varies and their functional roles are dissimilar. MitoNEET (mNT, CISD1), was the first characterized and is an outer mitochondrial membrane-anchored dimer with the C-terminal iron-sulfur cluster domain in the cytosol. Nutrient Deprivation Autophagy Factor 1 (NAF-1, Miner1, CISD2) is anchored to the mitochondrial-associated membranes of the endoplasmic reticulum (ER) with the iron-sulfur cluster domain in the cytosol. The third human NEET family protein is Mitochondrial Inner NEET (MiNT, Miner2, CISD3), is distinguished among the three, as it is the only soluble NEET, exists as a monomer, and is found inside the mitochondrial matrix. The NEET proteins are critical for cellular iron regulation and reactive oxygen species (ROS) protection, redox sensing, metabolic regulation, and are critical for maintenance of respiratory electron transfer machinery. A missense mutation in NAF-1 causes Wolfram syndrome 2 and other NEET pathologies include neurodegeneration, diabetes, premature aging, as well as play a role in the proliferation of several cancers. Collectively these indicated a crucial underlying cellular role for iron management, redox regulation, and metabolism. Until recently the role that mNT and NAF-1 play in the cell was unresolved. Though plenty of in situ and in vivo findings were published, elucidation of the precise cellular role of these proteins proved fruitless to scientific investigation. The major clues to their putative role were that 3Cys:1His iron-sulfur coordination is utilized by other iron-sulfur cluster transfer proteins, and a NEET family protein found in Arabidopsis thaliana (At-NEET) transfers iron into the mitochondria. Furthermore, RNAi knockdown of At-NEET caused free iron overload in the cell and led to ROS accumulation. Both mNT and NAF-1 have recently been identified as key links between the iron-sulfur cluster (ISC) biogenesis pathways in the mitochondria and the cytosol by transferring [2Fe-2S] clusters to human anamorsin, an early key piece of machinery that is responsible for providing [2Fe-2S] to the cytosolic iron-sulfur assembly (CIA) pathway. It has also been shown that mNT can transfer clusters to NAF-1, establishing that these proteins connect the flow of iron-sulfur away from the mitochondria to cytosolic targets.The method by which iron-sulfur clusters are exported out of the mitochondrion to the cytosol is currently now well characterized. To determine whether human mNT could translocate iron-sulfur clusters into the mitochondrion through the outer membrane voltage dependent anion channel (VDAC), we examined the ability of mNT to control membrane potential across a bilayer embedded with VDAC. We characterized the mNT-VDAC binding with microscale thermophoresis (MST) and hydrogen-deuterium exchange mass spectrometry (HDX-MS) in addition to computational fragment-docking direct-coupling analysis (Fd-DCA) to propose a model for mNT-VDAC binding. Through these data, our model suggests that mNT interacts measurably with the inner VDAC channel in addition to controlling VDAC gating in a redox-dependent manner, while NAF-1 does neither.To further identify and characterize cytosolic iron-sulfur protein transfer partners, we selected human glutaredoxin-3 (GRX3), a protein recently determined to carry [2Fe-2S] clusters and contribute to the CIA pathway. We identified that GRX3 selectively transfers clusters to mNT, but not NAF-1, further establishing diverged cellular roles for these two proteins. This is also the only protein reported to date that can transfer [2Fe-2S] clusters to mNT. These findings further solidify the role of the NEET proteins link the ISC and CIA, and demonstrate that there is directionality of iron-sulfur transfer at the interface of the ISC-CIA pathways. This directionality suggests a path for feedback and recycling of cytosolic iron-sulfur