36 research outputs found

    Characterization of diverse natural variants of CYP102A1 found within a species of Bacillus megaterium

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    An extreme diversity of substrates and catalytic reactions of cytochrome P450 (P450) enzymes is considered to be the consequence of evolutionary adaptation driven by different metabolic or environmental demands. Here we report the presence of numerous natural variants of P450 BM3 (CYP102A1) within a species of Bacillus megaterium. Extensive amino acid substitutions (up to 5% of the total 1049 amino acid residues) were identified from the variants. Phylogenetic analyses suggest that this P450 gene evolve more rapidly than the rRNA gene locus. It was found that key catalytic residues in the substrate channel and active site are retained. Although there were no apparent variations in hydroxylation activity towards myristic acid (C14) and palmitic acid (C16), the hydroxylation rates of lauric acid (C12) by the variants varied in the range of >25-fold. Interestingly, catalytic activities of the variants are promiscuous towards non-natural substrates including human P450 substrates. It can be suggested that CYP102A1 variants can acquire new catalytic activities through site-specific mutations distal to the active site

    Pyruvate Dehydrogenase Kinase Is a Metabolic Checkpoint for Polarization of Macrophages to the M1 Phenotype

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    Metabolic reprogramming during macrophage polarization supports the effector functions of these cells in health and disease. Here, we demonstrate that pyruvate dehydrogenase kinase (PDK), which inhibits the pyruvate dehydrogenase-mediated conversion of cytosolic pyruvate to mitochondrial acetyl-CoA, functions as a metabolic checkpoint in M1 macrophages. Polarization was not prevented by PDK2 or PDK4 deletion but was fully prevented by the combined deletion of PDK2 and PDK4; this lack of polarization was correlated with improved mitochondrial respiration and rewiring of metabolic breaks that are characterized by increased glycolytic intermediates and reduced metabolites in the TCA cycle. Genetic deletion or pharmacological inhibition of PDK2/4 prevents polarization of macrophages to the M1 phenotype in response to inflammatory stimuli (lipopolysaccharide plus IFN-γ). Transplantation of PDK2/4-deficient bone marrow into irradiated wild-type mice to produce mice with PDK2/4-deficient myeloid cells prevented M1 polarization, reduced obesity-associated insulin resistance, and ameliorated adipose tissue inflammation. A novel, pharmacological PDK inhibitor, KPLH1130, improved high-fat diet-induced insulin resistance; this was correlated with a reduction in the levels of pro-inflammatory markers and improved mitochondrial function. These studies identify PDK2/4 as a metabolic checkpoint for M1 phenotype polarization of macrophages, which could potentially be exploited as a novel therapeutic target for obesity-associated metabolic disorders and other inflammatory conditions

    Global, regional, and national burden of disorders affecting the nervous system, 1990–2021: a systematic analysis for the Global Burden of Disease Study 2021

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    BackgroundDisorders affecting the nervous system are diverse and include neurodevelopmental disorders, late-life neurodegeneration, and newly emergent conditions, such as cognitive impairment following COVID-19. Previous publications from the Global Burden of Disease, Injuries, and Risk Factor Study estimated the burden of 15 neurological conditions in 2015 and 2016, but these analyses did not include neurodevelopmental disorders, as defined by the International Classification of Diseases (ICD)-11, or a subset of cases of congenital, neonatal, and infectious conditions that cause neurological damage. Here, we estimate nervous system health loss caused by 37 unique conditions and their associated risk factors globally, regionally, and nationally from 1990 to 2021.MethodsWe estimated mortality, prevalence, years lived with disability (YLDs), years of life lost (YLLs), and disability-adjusted life-years (DALYs), with corresponding 95% uncertainty intervals (UIs), by age and sex in 204 countries and territories, from 1990 to 2021. We included morbidity and deaths due to neurological conditions, for which health loss is directly due to damage to the CNS or peripheral nervous system. We also isolated neurological health loss from conditions for which nervous system morbidity is a consequence, but not the primary feature, including a subset of congenital conditions (ie, chromosomal anomalies and congenital birth defects), neonatal conditions (ie, jaundice, preterm birth, and sepsis), infectious diseases (ie, COVID-19, cystic echinococcosis, malaria, syphilis, and Zika virus disease), and diabetic neuropathy. By conducting a sequela-level analysis of the health outcomes for these conditions, only cases where nervous system damage occurred were included, and YLDs were recalculated to isolate the non-fatal burden directly attributable to nervous system health loss. A comorbidity correction was used to calculate total prevalence of all conditions that affect the nervous system combined.FindingsGlobally, the 37 conditions affecting the nervous system were collectively ranked as the leading group cause of DALYs in 2021 (443 million, 95% UI 378–521), affecting 3·40 billion (3·20–3·62) individuals (43·1%, 40·5–45·9 of the global population); global DALY counts attributed to these conditions increased by 18·2% (8·7–26·7) between 1990 and 2021. Age-standardised rates of deaths per 100 000 people attributed to these conditions decreased from 1990 to 2021 by 33·6% (27·6–38·8), and age-standardised rates of DALYs attributed to these conditions decreased by 27·0% (21·5–32·4). Age-standardised prevalence was almost stable, with a change of 1·5% (0·7–2·4). The ten conditions with the highest age-standardised DALYs in 2021 were stroke, neonatal encephalopathy, migraine, Alzheimer's disease and other dementias, diabetic neuropathy, meningitis, epilepsy, neurological complications due to preterm birth, autism spectrum disorder, and nervous system cancer.InterpretationAs the leading cause of overall disease burden in the world, with increasing global DALY counts, effective prevention, treatment, and rehabilitation strategies for disorders affecting the nervous system are needed

    Influence of Load Test Scale on Prediction of Ultimate Bearing Capacity of Aggregate Pier Reinforced Clay

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    Predicting the bearing capacity of spread footings on aggregate pier-reinforced soil is particularly important for ground improvement. Various methods for predicting the bearing capacity of spread footings supported on aggregate pier-reinforced clay have been proposed. In addition, many fields or laboratory load tests have been conducted to identify the improvement effects of aggregate pier. However, no study has quantitatively compared the effect of the experimental scale on the prediction of bearing capacity. In this study, multiple linear regression analyses were performed for field and laboratory load tests, and the effects of the load test scale on bearing capacity prediction were identified. The sensitivity analysis showed that the prediction model for the laboratory load test (MLRL) exhibited a higher sensitivity for the undrained shear strength than that of the field load test model (MLRF). However, the sensitivity of the area replacement ratio of MLRL was less than half that of the MLRF. As the undrained shear strength increased, the predicted bearing capacity of the MLRL was larger than that of the MLRF owing to the influence of boundary conditions on the experimental equipment

    Evaluation of the Soil Thrust on Continuous Tracks Considering Independent Soil Shearing by Grousers

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    Soil thrust is a traction force of an off-road tracked vehicle. Existing soil thrust assessment methods assume that a continuous shear failure plane exists along the soil-track interface. However, recent experimental works revealed that the soil beneath the track is divided into soil blocks; each soil block is sheared independently. This study proposed a new total soil thrust assessment method based on the block-shaped failure mechanism, and the total soil thrust predicted by the proposed and the existing methods was compared. The results showed that the existing method showed a greater maximum soil thrust at a lower slip rate than the proposed method. When the soil exhibited a hardening behavior, the total soil thrust in the existing method was always greater than that in the proposed method. However, when the soil exhibited softening behavior or hump behavior, the total soil thrust in the existing method was greater than that in the proposed method until a specific slip ratio, after which the latter became greater. Unlike the existing method, the proposed method can consider the difference in soil thrust with the grouser shape ratio, showing that the total soil thrust increased as the grouser height increased and the shape ratio decreased

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    Evaluation of the Soil Thrust on Continuous Tracks Considering Independent Soil Shearing by Grousers

    No full text
    Soil thrust is a traction force of an off-road tracked vehicle. Existing soil thrust assessment methods assume that a continuous shear failure plane exists along the soil-track interface. However, recent experimental works revealed that the soil beneath the track is divided into soil blocks; each soil block is sheared independently. This study proposed a new total soil thrust assessment method based on the block-shaped failure mechanism, and the total soil thrust predicted by the proposed and the existing methods was compared. The results showed that the existing method showed a greater maximum soil thrust at a lower slip rate than the proposed method. When the soil exhibited a hardening behavior, the total soil thrust in the existing method was always greater than that in the proposed method. However, when the soil exhibited softening behavior or hump behavior, the total soil thrust in the existing method was greater than that in the proposed method until a specific slip ratio, after which the latter became greater. Unlike the existing method, the proposed method can consider the difference in soil thrust with the grouser shape ratio, showing that the total soil thrust increased as the grouser height increased and the shape ratio decreased

    Deep Learning-Based Ultrasonic Testing to Evaluate the Porosity of Additively Manufactured Parts with Rough Surfaces

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    Ultrasonic testing (UT) has been actively studied to evaluate the porosity of additively manufactured parts. Currently, ultrasonic measurements of as-deposited parts with a rough surface remain problematic because the surface lowers the signal-to-noise ratio (SNR) of ultrasonic signals, which degrades the UT performance. In this study, various deep learning (DL) techniques that can effectively extract the features of defects, even from signals with a low SNR, were applied to UT, and their performance in terms of the porosity evaluation of additively manufactured parts with rough surfaces was investigated. Experimentally, the effects of the processing conditions of additive manufacturing on the resulting porosity were first analyzed using both optical and scanning acoustic microscopy. Second, convolutional neural network (CNN), deep neural network, and multi-layer perceptron models were trained using time-domain ultrasonic signals obtained from additively manufactured specimens with various levels of porosity and surface roughness. The experimental results showed that all the models could evaluate porosity accurately, even that of the as-deposited specimens. In particular, the CNN delivered the best performance at 94.5%. However, conventional UT could not be applied because of the low SNR. The generalization performance when using newly manufactured as-deposited specimens was high at 90%

    Long-Term Compressive Strength Development of Steel Fiber Shotcrete from Cores Based on Accelerator Types at Tunnel Site

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    In this study, cement minerals, aluminates, and alkali-free accelerators incorporated with steel fiber were used to scrutinize the influence of accelerating agents on the long-term performance of tunneling shotcrete. Performance tests were identified based on the core compressive strength of mix shotcrete specimens with different types of accelerating agents throughout timeframes of 1, 3, 6, and 12 months. Here, 37 kg of steel fiber was incorporated into the cement mineral and aluminate mixes, and 40 kg of steel fiber was incorporated in an alkali-free mix for the shotcrete mix design. The KSF 2784 and ASTM 214 standards were followed for specimen fabrication and core cutting. For all specimens, shotcrete test panels of 250 × 600 × 500 mm were manufactured for core compressive strength tests conducted using 100, 75 and 55 mm diameter cylindrical molds and a length-to-diameter ratio of 2. The 1-month compressive strength of all test variables satisfied the Korea Expressway Co. standard of 21 MPa. The core compressive strength of the shotcrete specimens showed a tendency to increase with age, but a strength reduction occurred in 6 months and increased again at 12 months. Moreover, the impact of the diameter changes in the shotcrete core specimens was analyzed based on the mixing. For 12 months, a large increase in the core compressive strength occurred, particularly in the alkali-free specimens. The comparison also focused on the relative strength compared with a cast concrete mold and shotcrete core specimens. It is necessary to use alkali-free accelerators considering the long-term performance of tunnels and worker safety
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