146 research outputs found

    Leucine Biosynthesis Is Involved in Regulating High Lipid Accumulation in Yarrowia lipolytica

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    The yeast Yarrowia lipolytica is a potent accumulator of lipids, and lipogenesis in this organism can be influenced by a variety of factors, such as genetics and environmental conditions. Using a multifactorial study, we elucidated the effects of both genetic and environmental factors on regulation of lipogenesis in Y. lipolytica and identified how two opposite regulatory states both result in lipid accumulation. This study involved comparison of a strain overexpressing diacylglycerol acyltransferase (DGA1) with a control strain grown under either nitrogen or carbon limitation conditions. A strong correlation was observed between the responses on the transcript and protein levels. Combination of DGA1 overexpression with nitrogen limitation resulted in a high level of lipid accumulation accompanied by downregulation of several amino acid biosynthetic pathways, including that of leucine in particular, and these changes were further correlated with a decrease in metabolic fluxes. This downregulation was supported by the measured decrease in the level of 2-isopropylmalate, an intermediate of leucine biosynthesis. Combining the multi-omics data with putative transcription factor binding motifs uncovered a contradictory role for TORC1 in controlling lipid accumulation, likely mediated through 2-isopropylmalate and a Leu3-like transcription factor

    Accumulation of Succinyl Coenzyme A Perturbs the Methicillin-Resistant Staphylococcus aureus (MRSA) Succinylome and Is Associated with Increased Susceptibility to Beta-Lactam Antibiotics

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    Penicillin binding protein 2a (PBP2a)-dependent resistance to ÎČ-lactam antibiotics in methicillin-resistant Staphylococcus aureus (MRSA) is regulated by the activity of the tricarboxylic acid (TCA) cycle via a poorly understood mechanism. We report that mutations in sucC and sucD, but not other TCA cycle enzymes, negatively impact ÎČ-lactam resistance without changing PBP2a expression. Increased intracellular levels of succinyl coenzyme A (succinyl-CoA) in the sucC mutant significantly perturbed lysine succinylation in the MRSA proteome. Suppressor mutations in sucA or sucB, responsible for succinyl-CoA biosynthesis, reversed sucC mutant phenotypes. The major autolysin (Atl) was the most succinylated protein in the proteome, and increased Atl succinylation in the sucC mutant was associated with loss of autolytic activity. Although PBP2a and PBP2 were also among the most succinylated proteins in the MRSA proteome, peptidoglycan architecture and cross-linking were unchanged in the sucC mutant. These data reveal that perturbation of the MRSA succinylome impacts two interconnected cell wall phenotypes, leading to repression of autolytic activity and increased susceptibility to ÎČ-lactam antibiotics. IMPORTANCE mecA-dependent methicillin resistance in MRSA is subject to regulation by numerous accessory factors involved in cell wall biosynthesis, nucleotide signaling, and central metabolism. Here, we report that mutations in the TCA cycle gene, sucC, increased susceptibility to ÎČ-lactam antibiotics and was accompanied by significant accumulation of succinyl-CoA, which in turn perturbed lysine succinylation in the proteome. Although cell wall structure and cross-linking were unchanged, significantly increased succinylation of the major autolysin Atl, which was the most succinylated protein in the proteome, was accompanied by near complete repression of autolytic activity. These findings link central metabolism and levels of succinyl-CoA to the regulation of ÎČ-lactam antibiotic resistance in MRSA through succinylome-mediated control of two interlinked cell wall phenotypes. Drug-mediated interference of the SucCD-controlled succinylome may help overcome ÎČ-lactam resistance

    Structural properties of disk galaxies. II. Intrinsic shape of bulges

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    (Abridged) The structural parameters of a magnitude-limited sample of 148 unbarred S0-Sb galaxies were analyzed to derive the intrinsic shape of their bulges. We developed a new method to derive the intrinsic shape of bulges based on the geometrical relationships between the apparent and intrinsic shapes of bulges and disks. The equatorial ellipticity and intrinsic flattening of bulges were obtained from the length of the apparent major and minor semi-axes of the bulge, twist angle between the apparent major axis of the bulge and the galaxy line of nodes, and galaxy inclination. We found that the intrinsic shape is well constrained for a subsample of 115 bulges with favorable viewing angles. A large fraction of them is characterized by an elliptical section (B/A<0.9). This fraction is 33%, 55%, and 43% if using their maximum, mean, or median equatorial ellipticity, respectively. Most are flattened along their polar axis (C<(A+B)/2). The distribution of triaxiality is strongly bimodal. This bimodality is driven by bulges with Sersic index n>2, or equivalently, by the bulges of galaxies with a bulge-to-total ratio B/T>0.3. In particular, bulges with n\leq2 and with B/T\leq0.3 show a larger fraction of oblate axisymmetric (or nearly axisymmetric) bulges, a smaller fraction of triaxial bulges, and fewer prolate axisymmetric (or nearly axisymmetric) bulges with respect to bulges with n>2 and with B/T>0.3, respectively. According to predictions of the numerical simulations of bulge formation, bulges with n\leq2, which show a high fraction of oblate axisymmetric (or nearly axisymmetric) shapes and have B/T\leq0.3, could be the result of dissipational minor mergers. Both major dissipational and dissipationless mergers seem to be required to explain the variety of shapes found for bulges with n>2 and B/T>0.3.Comment: 16 pages, 12 figures; accepted for publication in A&

    Subtyping patients with heroin addiction at treatment entry: factor derived from the Self-Report Symptom Inventory (SCL-90)

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    <p>Abstract</p> <p>Background</p> <p>Addiction is a relapsing chronic condition in which psychiatric phenomena play a crucial role. Psychopathological symptoms in patients with heroin addiction are generally considered to be part of the drug addict's personality, or else to be related to the presence of psychiatric comorbidity, raising doubts about whether patients with long-term abuse of opioids actually possess specific psychopathological dimensions.</p> <p>Methods</p> <p>Using the Self-Report Symptom Inventory (SCL-90), we studied the psychopathological dimensions of 1,055 patients with heroin addiction (884 males and 171 females) aged between 16 and 59 years at the beginning of treatment, and their relationship to age, sex and duration of dependence.</p> <p>Results</p> <p>A total of 150 (14.2%) patients with heroin addiction showed depressive symptomatology characterised by feelings of worthlessness and being trapped or caught; 257 (24.4%) had somatisation symptoms, 205 (19.4%) interpersonal sensitivity and psychotic symptoms, 235 (22.3%) panic symptomatology, 208 (19.7%) violence and self-aggression. These dimensions were not correlated with sex or duration of dependence. Younger patients with heroin addiction were characterised by higher scores for violence-suicide, sensitivity and panic anxiety symptomatology. Older patients with heroin addiction showed higher scores for somatisation and worthlessness-being trapped symptomatology.</p> <p>Conclusions</p> <p>This study supports the hypothesis that mood, anxiety and impulse-control dysregulation are the core of the clinical phenomenology of addiction and should be incorporated into its nosology.</p

    Minimal residual disease in breast cancer: an overview of circulating and disseminated tumour cells

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    Managing Spent Nuclear Fuel at the Idaho National Laboratory NATO Advanced Research Workshop, Safety-Related Issues of Spent Nuclear Fuel Storage MANAGING SPENT NUCLEAR FUEL AT THE IDAHO NATIONAL LABORATORY SHORT TITLE: MANAGING SPENT NUCLEAR FUEL AT INL

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    Abstract: The Idaho National Laboratory (INL) has a large inventory of diverse types of spent nuclear fuel (SNF). This legacy derives from the history of the INL as the National Reactor Testing Station, and from its mission to recover HEU from SNF and to test and examine SNF after irradiation. The INL also has a large diversity of SNF storage facilities, some 50 years old. SNF at INL has many forms-from intact assemblies down to metallurgical mounts, and some fuel has been wet stored for over 40 years. SNF is stored bare or in metal cans under water, or dry in vaults, caissons or casks. Inspection shows varying corrosion and degradation of the SNF and its storage cans. SNF has been stored in 10 different facilities: 5 pools, one cask storage pad, one vault, two generations of caisson facilities, and one modular Independent Spent Fuel Storage Installation (ISFSI). The pools range in age from 40 years old to the most modern in the US Department of Energy (DOE) complex. The near-term objective is to move SNF from older pools to interim dry storage, allowing shutdown and decommissioning of the older facilities. This move involves drying methods that are dependent on fuel type. The long-term objective is to have INL SNF in safe dry storage and ready to be shipped to the National Repository. The unique features of the INL SNF requires special treatments and packaging to meet the proposed repository acceptance criteria and SNF will be repackaged in standardized canisters for shipment and disposal in the National Repository. Disposal will use the standardized canisters that can be co-disposed with High Level Waste glass logs to limit the total fissile material in a repository waste package. The DOE standardized canister also simplifies the repository handling of the multitude of DOE SNF sizes and shapes

    Capillary Electrophoresis–Nanoelectrospray Ionization–Selected Reaction Monitoring Mass Spectrometry via a True Sheathless Metal-Coated Emitter Interface for Robust and High-Sensitivity Sample Quantification

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    A new sheathless transient capillary isotachophoresis (CITP)/capillary zone electrophoresis (CZE)–MS interface, based on a commercially available capillary with an integrated metal-coated ESI emitter, was developed in this study aiming at overcoming the reproducibility and ruggedness problems suffered to a certain degree by almost all the available CE–MS interfaces, and pushing the CE–MS technology suitable for routine sample analysis with high sensitivity. The new CITP/CZE–MS interface allows the electric contact between ESI voltage power supply and the CE separation liquid by using a conductive liquid that comes in contact with the metal-coated surface of the ESI emitter, making it a true sheathless CE–MS interface. Stable electrospray was established by avoiding the formation of gas bubbles from electrochemical reaction inside the CE capillary. Crucial operating parameters, such as sample loading volume, flow rate, and separation voltage, were systematically evaluated for their effects on both CITP/CZE separation efficiency and MS detection sensitivity. Around one hundred CITP/CZE–MS analyses can be easily achieved by using the new sheathless CITP/CZE interface without a noticeable loss of metal coating on the ESI emitter surface, or degrading of the ESI emitter performance. The reproducibility in analyte migration time and quantitative performance of the new interface was experimentally evaluated to demonstrate a LOQ below 5 attomole
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