Systematic Study of Heteroarene Stacking Using a Congeneric Set of Molecular Glues for Procaspase-6.

Heteroaromatic stacking interactions are important in drug binding, supramolecular chemistry, and materials science, making protein-ligand model systems of these interactions of considerable interest. Here we studied 30 congeneric ligands that each present a distinct heteroarene for stacking between tyrosine residues at the dimer interface of procaspase-6. Complex X-ray crystal structures of 10 analogs showed that stacking geometries were well conserved, while high-accuracy computations showed that heteroarene stacking energy was well correlated with predicted overall ligand binding energies. Empirically determined KD values in this system thus provide a useful measure of heteroarene stacking with tyrosine. Stacking energies are discussed in the context of torsional strain, the number and positioning of heteroatoms, tautomeric state, and coaxial orientation of heteroarene in the stack. Overall, this study provides an extensive data set of empirical and high-level computed binding energies in a versatile new protein-ligand system amenable to studies of other intermolecular interactions.

Reliability modelling and evaluating of wind turbine considering imperfect repair.

To model and evaluate the reliability of wind turbine (WT) under imperfect repair, an improved Log-linear Proportional Intensity Model (LPIM)-based method was proposed. Initially, using the three-parameter bounded intensity process (3-BIP) as the benchmark failure intensity function of LPIM, an imperfect repair effect-aware WT reliability description model was developed. Among them, the 3-BIP was used to describe the evolution process of the failure intensity in the stable operation stage with running time, while the LPIM reflected the repair effect. Second, the estimation problem for model parameters was transformed into a minimum solution problem for a nonlinear objective function, which was then solved using the Particle Swarm Optimization algorithm. The confidence interval of model parameters was finally estimated using the inverse Fisher information matrix method. Key reliability indices interval estimation based on the Delta method and point estimation was derived. The proposed method was applied to a wind farm's WT failure truncation time. The proposed method has a higher goodness of fit based on verification and comparison. As a result, it can bring the evaluated reliability closer to engineering practice.

Dissolved Oxygen and Water Temperature Drive Vertical Spatiotemporal Variation of Phytoplankton Community: Evidence from the Largest Diversion Water Source Area.

In order to study the vertical distribution characteristics of phytoplankton in the Danjiangkou Reservoir, the water source of the Middle Route Project of the South-North Water Diversion, seven sampling sites were set up in the Reservoir for quarterly sampling from 2017 to 2019, and water environment surveys were conducted simultaneously. The results showed that 157 species (including varieties) were identified, belonging to 9 phyla and 88 genera. In terms of species richness, Chlorophyta had the largest number of species, accounting for 39.49% of the total species. The Bacillariophyta and Cyanobacteria accounted for 28.03% and 13.38% of the total species, respectively. From the whole Danjiangkou Reservoir, the total phytoplankton abundance varied from 0.09 × 102 to 20.01 × 106 cells/L. In the vertical distribution, phytoplankton were mainly observed in the surface-thermospheric layer (I-II layer) and the bottom layer, while the Shannon-Wiener index showed a trend of gradually decreasing from the I-V layer. The Surfer model analysis showed that there was no significant stratification in the Q site's water temperature (WT) and dissolved oxygen (DO) levels in the water diversion area during the dynamic water diversion process. A canonical correspondence analysis (CCA) showed that DO, WT, pH, electrical conductivity (Cond), chemical oxygen demand (CODMn), total phosphorus (TP), ammonia nitrogen (NH4+-N), and total nitrogen (TN) had significant effects on the vertical distribution of phytoplankton (p < 0.05). A partial Mantel analysis showed that the vertical distribution of the phytoplankton community was related to WT, and the phytoplankton community structure at the other sites, except for Heijizui (H) and Langhekou (L), was affected by DO. This study has positive significance for exploring the vertical distribution characteristics of a phytoplankton community in a deep-water dynamic water diversion reservoir.

Plastic response of leaf traits to N deficiency in field-grown maize.

Nitrogen (N) utilization for crop production under N deficiency conditions is subject to a trade-off between maintaining specific leaf N content (SLN) important for radiation-use efficiency versus maintaining leaf area (LA) development, important for light capture. This paper aims to explore how maize deals with this trade-off through responses in SLN, LA and their underlying traits during the vegetative and reproductive growth stages. In a 10-year N fertilization trial in Jilin province, Northeast China, three N fertilizer levels have been maintained: N deficiency (N0), low N supply (N1) and high N supply (N2). We analysed data from years 8 and 10 of this experiment for two common hybrids. Under N deficiency, maize plants maintained LA and decreased SLN during vegetative stages, while both LA and SLN decreased comparably during reproductive stages. Canopy SLA (specific leaf area, cm2 g-1) decreased sharply during vegetative stages and slightly during reproductive stages, mainly because senesced leaves in the lower canopy had a higher SLA. In the vegetative stage, maize maintained LA at low N by maintaining leaf biomass (albeit hence having N content/mass) and slightly increasing SLA. These responses to N deficiency were stronger in maize hybrid XY335 than in ZD958. We conclude that the main strategy of maize to cope with low N is to maintain LA, mainly by increasing SLA throughout the plant but only during the vegetative growth phase.

Human Parvovirus B19 Nonstructural Protein 1 Regulates GATA1 Expression via the Notch Signaling Pathway in K562 Cell Line.

BACKGROUND: Human parvovirus B19 (B19) infection can affect the hematopoietic arrest in fetus by hindering the differentiation and maturation of erythroid progenitor cells. B19 nonstructural protein 1 (NS1) has been shown to inhibit the differentiation of erythroid progenitor cells. The goal of this study is to explore the role of B19 NS1 in the regulation of GATA1 and Notch signaling pathway in hematopoietic cells. METHODS: The B19 NS1 expression plasmid was reconstituted, and the possibility of NS1 regulating GATA1 and GATA2 expression modulated by Notch-Hes pathway was tested by qRT-PCR and western blot. Immunofluorescence assays were conducted to visualize pNS1 in K562 cells. RESULTS: We demonstrate that B19 NS1 inhibited GATA1 and induced Hes1/Hes5, which is involved in the activation of Notch signaling pathway. Meanwhile, NS1 exhibited promoting effects on GATA2 expression. Activation of the Notch signaling pathway up-regulated its downstream transcriptional repressor family Hes, thereby inhibiting the expression of GATA gene in K562 cells. CONCLUSIONS: The results show that B19 NS1 protein negatively regulates GATA1 related nuclear transcription and may interfere with hematopoietic cell differentiation.

Mixotrophic Cultivation Optimization of Microalga Euglena pisciformis AEW501 for Paramylon Production.

Euglena, a flagellated unicellular protist, has recently received widespread attention for various high-value metabolites, especially paramylon, which was only found in Euglenophyta. The limited species and low biomass of Euglena has impeded paramylon exploitation and utilization. This study established an optimal cultivation method of Euglena pisciformis AEW501 for paramylon production under mixotrophic cultivation. The results showed that the optimum mixotrophic conditions were 20 °C, pH 7.0, and 63 µmol photons m-2∙s-1, and the concentrations of sodium acetate and diammonium hydrogen phosphate were 0.98 g L-1 and 0.79 g L-1, respectively. The maximal biomass and paramylon content were 0.72 g L-1 and 71.39% of dry weight. The algal powder contained more than 16 amino acids, 6 vitamins, and 10 unsaturated fatty acids under the optimal cultivation. E. pisciformis paramylon was pure ß-1,3-glucan-type polysaccharide (the purity was up to 99.13 ± 0.61%) composed of linear glucose chains linked together by ß-1,3-glycosidic bonds. These findings present a valuable basis for the industrial exploitation of paramylon with E. pisciformis AEW501.

Physical and Chemical Properties of Coal Gasification Fine Slag and Its Carbon Products by Hydrophobic-Hydrophilic Separation.

Coal gasification fine slag is a kind of solid waste with low resource utilization rate. The complex embedding of residual carbon and inorganic minerals (ash materials) is the main reason restricting the efficient resource separation and utilization of residual carbon or ash materials. Hydrophobic-hydrophilic separation (HHS) is a separation technology in which mineral particles with different surface hydrophobicity values are enriched in the water phase and oil phase under the action of mechanical stirring. The water on the surface of hydrophobic particles is replaced by the oil phase to form flocs, which are enriched in the hydrophobic liquid phase, while hydrophilic particles are dispersed into the aqueous phase. In this study, the HHS process was used to separate the carbon/ash from the fine gasification slag produced by a Shenning gasifier, Texaco gasifier, and GSP gasifier of Ningxia Coal Industry Co., Ltd. The physicochemical properties of the original sample and the residual carbon products obtained by hydrophobic-hydrophilic separation were analyzed. The results show that HHS can separate the carbon/ash in the three kinds of fine slag to varying degrees. The carbon element is enriched into the hydrophobic phase to form the concentrates, while the silicon element, oxygen element, and metal element enter the tailings. The spherical ash with different particle sizes distributed on the surface of residual carbon and the gap of the matrix is basically removed, while the ash in the carbon-ash melt is difficult to remove. The ash contents of the concentrate and tailings of fine slag of the Shenning gasifier are 22.58 and 96.28%, respectively, which reach the best ash index compared with that of the other two gasifiers. From the change of mineral surface properties after HHS, the distribution of oxygen-containing groups, benzene rings, Si-O, and clay minerals or carbonate minerals in the three kinds of fine slag residual carbon products is basically similar. Compared with the other two gasifier products, the GSP gasifier concentrate has a larger specific surface area and less ash material, more amorphous carbon structures (less graphitic), and more active sites, resulting in a stronger combustion activity.

Respiratory Syncytial Virus Nonstructural Protein 1 Promotes 5-Lipoxygenase via miR-19a-3p.

Background: Respiratory syncytial virus (RSV) infection can regulate the expression of a wide range of noncoding microRNAs (miRNAs), in which mir-19a-3p can participate in airway inflammatory response by regulating 5-lipoxygenase (5-LO) pathway. RSV nonstructural protein (NS) 1 is involved in the airway hyperresponsiveness during RSV infection. Methods: The expression levels of miR-19a-3p and inflammatory signaling-related indicators were detected using quantitative real-time PCR and western blot analyses on the A549 cells transfected with NS1 expression plasmids (pNS1). The 5-LO-mediated inflammatory signaling pathway was assessed when the miR-19a-3p or 5-LO was inhibited. Results: The immunofluorescence analysis showed that the plasmid-mediated NS1 protein was observed in both the cytoplasm and nucleus. The expression level of miR-19a-3p was significantly upregulated in the pNS1 or RSV-treated cells, which was reversed by the NS1 small interfering RNA. In addition, pNS1 also upregulated the expression of 5-LO, interleukin-5 (IL-5), and leukotriene B4 (LTB4), which was also significantly inhibited by the miR-19a-3p antagonists. The 5-LO inhibitor MK886 prevented the increase in the expression level of IL-5 induced by pNS1. Conclusions: These results suggested that the RSV NS1 might play an important role in the pathogenesis of RSV by activating the 5-LO and subsequent inflammatory cytokines through miR-19a-3p.

Instrumentation-Compact Digital Microfluidic Reaction Interface-Extended Loop-Mediated Isothermal Amplification for Sample-to-Answer Testing of Vibrio parahaemolyticus.

Vibrio parahaemolyticus is usually spread via consumption of contaminated seafood and causes vibriosis. By combination of digital microfluidic (DMF) and loop-mediated isothermal amplification (LAMP), we provided an automated instrumentation-compact DMF-LAMP device for sample-to-answer detection of V. parahaemolyticus. For the first time, how much the proper mixing might facilitate the DMF-LAMP process is explored. The results illustrated that increasing the number of flow configurations and decreasing the fluid-reversibility will extend the interfacial surface available for diffusion-based mass transfer within a droplet microreactor, thus contributing to the overall amplification reaction rate. Noticeably, the DMF-LAMP amplification plateau time is shortened by proper mixing, from 60 min in static mixing and traditional bulk LAMP to 30 min in 2-electrode mixing and 15 min in 3-electrode mixing. The device achieved much higher detection sensitivity (two copies per reaction) than previously reported devices. V. parahaemolyticus from spiked shrimps is detected by Q-tip sampling associated with 3-electrode mixing DMF-LAMPs. The detectable signal occurs within only 3 min at a higher concentration and, at most, is delayed to 18 min, with a detection limit of <0.23 × 103 CFU/g. Thus, the developed DMF-LAMP device demonstrates potential for being used as a sample-to-answer system with a quick analysis time, high sensitivity, and sample-to-answer format.

Respiratory syncytial virus nonstructural protein 1 breaks immune tolerance in mice by downregulating Tregs through TSLP-OX40/OX40L-mTOR axis.

Respiratory syncytial virus (RSV) infection in early life is associated strongly with the subsequent development and exacerbation of asthma, however, the mechanism is still ambiguous. In this study, we identified that RSV nonstructural protein (NS) 1 plays a critical role. Plasmid-mediated overexpression of NS1 induced significant airway hyperresponsiveness, eosinophilia, and mucus hyperproduction in mice. In the pNS1 group, there were markedly elevated proportions of Th2 and Th17 cells, while Th1 and Foxp3+ regulatory T cells (Tregs) significantly declined compared with the control group. Serum concentrations of interleukin (IL)-4, IL-5, IL-6, IL-17, transforming growth factor-beta, and tumor necrosis factor-alpha increased but levels of interferon-gamma and interleukin-10 declined in pNS1 group. Besides, NS1 caused a significant rise of serum thymic stromal lymphopoietin (TSLP) and OX40L levels, and a neutralizing mAb anti-OX40L was capable of promoting RSV clearance and attenuating the airway allergic inflammation caused by pNS1. Otherwise, OX40L-blocking counteracts the inhibitory effect of pNS1 on Tregs in the spleen. RSV NS1 caused elevated levels of phospho-AKT, phospho-mTOR, and phospho-S6K1, which were partially attenuated by anti-OX40L. Moreover, a specific inhibitor of mTORC1 significantly relieved the inhibition of Foxp3 expression and Tregs differentiation. Together, the data indicate that RSV NS1 protein breaks immune tolerance and induces airway inflammation and hyperresponsiveness in mice. In this process, NS1-stimulated TSLP and OX40L play a major role by inhibiting the induction of Tregs, which is at least partially mediated by modulating AKT-mTOR signaling pathways.

miRNAs and Leukotrienes in Respiratory Syncytial Virus Infection.

MicroRNAs (miRNAs) are small, non-coding RNAs that regulate posttranscription by binding to 3'-untranslated regions of target mRNAs. Recent functional studies have elucidated mechanisms that miRNAs regulate leukotriene synthesis by perturbing arachidonic acid metabolism. Both microarrays and high-throughput sequencing revealed distinct differential expression of miRNAs in children with respiratory syncytial virus (RSV) infection compared with healthy controls. Abnormal miRNA expression may contribute to higher leukotriene levels, which is associated with airway hyperreactivity. Targeting miRNAs may benefit to restore the homeostasis of inflammatory reaction and provide new strategies to alleviate airway hyperreactivity induced by RSV. In this article, we provide an overview of the current knowledge about miRNAs modulating leukotrienes through regulation of arachidonic acid metabolism with a special focus on miRNAs aberrantly expressed in children with RSV infection.

A SARS-CoV-2 familial cluster infection reveals asymptomatic transmission to children.

Information on SARS-CoV-2 asymptomatic infection and infectivity in children is limited. In this study, we aimed to report the epidemiological and clinical characteristics of a familial cluster infection including children with SARS-CoV-2. On February 1, 2020, two children(case 1 and case 2), an 8-year-old girl and a 9-year-old boy, were admitted to the isolation ward in Xiangyang Central Hospital, Hubei province, China, with the diagnosis of COVID-19. Before admission, they had been staying at home with their father and never contacted with any confirmed patients except their mother (case 3) who returned from Wuhan on January 22. Both case 1 and case 2 got mild symptoms. Case 3 did not develop any symptoms until February 6, 2020, with an asymptomatic period of 15 days. She was transferred to ICU and administered multiple treatment according to the disease progression and chest CT manifestations. Her nucleic acid test turned positive until Feb 21, 2020, 15 days after symptoms onset, 30 days after her return from Wuhan. Our data showed that patients with SARS-CoV-2 may have the ability to transmit during their asymptomatic period even with the negative of viral nucleic acid in pharyngeal swabs.

Enhanced in vitro antitumor efficacy of a polyunsaturated fatty acid-conjugated pH-responsive self-assembled ion-pairing liposome-encapsulated prodrug.

The development of clinical chemotherapeutics is always challenging due to the toxicity and side effects of drugs not only for tumor cells but also for normal cells. Therefore, nano-drug delivery systems and prodrug strategies have been applied to address this challenge. Herein, we report a liposome-encapsulated small-molecule prodrug nanosystem, self-assembled by doxorubicin (DOX) and mixed polyunsaturated fatty acid (MPUFA) ion-pairing (MPUFAs-DOX@Liposomes), which has a high omega-3 PUFA content. The increased lipophilicity of ion-paired MPUFAs-DOX can significantly improve the drug loading efficiency (∼97%). Electrostatic interaction, the hydrophobic effect and hydrogen bonding between the ion-pairing agents led to superior pH-responsive release of DOX from liposomes over DOX-loaded liposomes (DOX@Liposomes), with a more rapid release rate at pH 5.0 than at pH 7.4, which is beneficial for decreasing the toxicity of DOX under physiological conditions. Finally, the in vitro antitumor effects were investigated for two tumor cell types, A549 and MCF-7, and the results demonstrated that MPUFAs-DOX@Liposomes showed the highest cytotoxicity compared with free DOX and DOX@Liposomes because of the ready uptake under the effect of PUFAs. Hence, liposomes loaded with ion-paired MPUFAs-DOX is a promising formulation for combination cancer therapy.

Crystal structure of hydroxyquinol 1,2-dioxygenase PnpC from Pseudomonas putida DLL-E4 and its role of N-terminal domain for catalysis.

Hydroxyquinol 1,2-dioxygenase is a key enzyme in the hydroxyquinol pathway of p-nitrophenol (PNP) degradation, and catalyzes the ring cleavage of benzenetriol to maleylacetate. Here, we report the first structure of a hydroxyquinol 1,2-dioxygenase from the Gram-negative bacterium Pseudomonas putida DLL-E4 (PnpC) at the resolution of 2.1 Å. The tertiary structure of PnpC resembles that of the homologous intradiol dioxygenases. The catalytic Fe(III) is pentacoordinated by the conserved Tyr160, Tyr194, His218 and His220, the citrate anion and one water molecule. Among the residues expected to interact with the substrate, structural comparison with the (chloro)catechol dioxygenases suggested that Asp80, Thr81 and Val248 are responsible for the substrate specificity. Moreover, truncation of the N-terminal α-helix of PnpC suggested the N-terminal domain is required for its soluble expression and enzyme catalysis. Our results might provide insights in the substrate recognition and rational design of this enzyme class to be used in bioremediation.

Assessment of neonatal brain volume and growth at different postmenstrual ages by conventional MRI.

Data regarding neonatal brain volumes represent a basis for monitoring early brain development, and large sample of neonatal brain volume data has not been well described. This study was focused on neonatal brain volumes at different postmenstrual ages (PMA) and postnatal age (PNA).A cohort of 415 neonates with PMA 30 to 43 weeks were recruited for the determination of brain volumes. Intracranial cavity (ICC), total brain tissue (TBT), and cerebrospinal fluid (CSF) were evaluated on the basis of T1-weighted sagittal plane magnetic resonance images. Brain magnetic resonance imaging was assessed using maturation scoring system and multiple linear regression analysis was conducted to forecast the effect factors of brain volumes.TBT volume reached a peak growth at 39 to 40 weeks, ICC volume presented peak growth later at around 43 to 44 weeks, and CSF had a cliff fallen at 37 to 38 weeks PMA at scan. The maturation score increased along with PMA, and the TBT and CSF volumes were significantly different between higher and lower gestational age (GA) groups. The ICC and TBT volumes in higher GA group were larger than lower GA group. Most infants in higher GA group had higher TMS than those in lower GA group. Gender, PMA, PNA, and birth weight were predictors of TBT and ICC volumes.Our results showed that premature volumes of ICC and TBT enlarged with the increasing PMA, while volumes of CSF decreased at 37 weeks. Premature earlier to leave the uterus can lead to brain mature retard although they had the same GA compared with those later birth neonates.

High-dose of intravenous immunoglobulin modulates immune tolerance in premature infants.

BACKGROUND: Intravenous immunoglobulin (IVIG) is commonly used to improve the immunomodulatory effects, although its regulatory effect on premature Treg cells is unclear. The purpose of this study is to study the effect of high dose of IVIG (HD-IVIG) on Treg cells expression and cytokine profile in premature birth. METHODS: Fifty-two premature infants were enrolled in this study and thirty-one premature infants who were suspected to have intrauterine infection received HD-IVIG (1-2 g/kg) at the first day of birth; the remaining 21 premature infants were assigned as the control group. The peripheral blood CD4 + T and foxp3+ Treg cells were checked by flow cytometry, and cytokine concentrations were detected by cytometric bead array. RESULTS: With the gestational age growth, peripheral blood CD4 + T and foxp3+ Treg cells of prematurity gradually declined from 50% to 35% and from 8% to 6%, respectively. Meanwhile, HD-IVIG increased the percentage of CD4 + T and foxp3+ Treg cells compared with their baseline levels (p < 0.001). HD-IVIG demonstrated different regulating effects on cytokines secretion, increased IL-17 and TGF-ß, and inhibited IL-6 secretion. CONCLUSION: Our results demonstrated that HD-IVIG not only enhanced the premature immune tolerance, but also suppressed the excessive inflammation response mediated by IL-6. TRIAL REGISTRATION: This study was under the clinical study registration (ChiCTR-ORC-16008872, date of registration, 2016-07-21).

Profile and clinical characterization of seizures in hospitalized children.

INTRODUCTION: Seizure is the commonest pediatric neurological disorder, which is frightening to caretakers. The current study aims to determine profile, clinical spectrum and analyze the commonest etiology of seizures in children admitted to a tertiary hospital in Central China. METHODS: This was a hospital based retrospective study carried out in Zhongnan Hospital of Wuhan University, China. Computerized data was collected from January 2012 to May 2015. Variables collected were demographics, clinical presentations and laboratory tests; brain imaging studies, electroencephalography, diagnosis, prognosis, outcome and duration of hospitalization. RESULTS: A total of 200 patients were admitted with seizures. There were 109 (54.5%) males and 91 (45.5%) females. Among these patients, 193 (96.5%) were aged 1 month to 5 years and 182 (91.0%) presented with seizures and fever. Generalized tonic-clonic seizure was the most common seizure type in 196 (98.0%) children. Febrile seizure was the leading etiology of seizure in 175 (87.5%) children followed by epilepsy in 11 (5.5%) children. There were only 3 (2%) children with central nervous system infections. Abnormal brain images were noted in 10 (20%) out of 50 patients. Among 193 children tested for different infections, 49 (25.4%) had positive results. Viral infections were commonest infections by 49.0%, atypical bacterial 34.7% and 16.3% coinfections. CONCLUSION: Seizure was the commonest neurological condition of children admitted in our hospital, febrile seizures being the commonest etiology. The prognosis and outcomes were good but there were prolonged days of hospitalization. Children with unprovoked seizures require brain-imaging studies for better understanding of seizure etiology.