The nucleocapsid protein facilitates p53 ubiquitination-dependent proteasomal degradation via recruiting host ubiquitin ligase COP1 in PEDV infection.

Porcine epidemic diarrhea virus (PEDV) is a highly contagious enteric pathogen of the coronavirus family and caused severe economic losses to the global swine industry. Previous studies have established that p53 is a host restriction factor for PEDV infection, and p53 degradation occurs in PEDV-infected cells. However, the underlying molecular mechanisms through which PEDV viral proteins regulate p53 degradation remain unclear. In this study, we found that PEDV infection or expression of the nucleocapsid protein downregulates p53 through a post-translational mechanism: increasing the ubiquitination of p53 and preventing its nuclear translocation. We also show that the PEDV N protein functions by recruiting the E3 ubiquitin ligase COP1 and suppressing COP1 self-ubiquitination and protein degradation, thereby augmenting COP1-mediated degradation of p53. Additionally, COP1 knockdown compromises N-mediated p53 degradation. Functional mapping using truncation analysis showed that the N-terminal domains of N protein were responsible for interacting with COP1 and critical for COP1 stability and p53 degradation. The results presented here suggest the COP1-dependent mechanism for PEDV N protein to abolish p53 activity. This study significantly increases our understanding of PEDV in antagonizing the host antiviral factor p53 and will help initiate novel antiviral strategies against PEDV.

An activatable small-molecule fluorogenic probe for detection and quantification of beta-amyloid aggregates.

Extracellular accumulation of ß amyloid (Aß) peptides in the brain is thought to be a pathological hallmark and initial event before the symptom starts of Alzheimer's patients. Herein, we developed two series of benzo[d]thiazole-based small-molecule compounds (BM1-BM4, BPM1-BPM4) with a donor-acceptor (D-A) or donor-π-acceptor (D-π-A) architecture, respectively, based on structure-activity relationship. Among them, the optimized BPM1 not only displayed the highest binding affinity to Aß aggregates over other proteins or Aß monomers, but was readily activated its fluorescence with 10-fold fluorescence enhancement, allowing for specifically and sensitively detecting Aß aggregates. BPM1 also exhibits several other advantages including low molecular weight, low cytotoxicity and excellent biological stability. Besides, cell staining results confirmed that SK-N-BE(2) cells can be fluorescently lighted up as well as cell permeability and damage when treated with BPM1-bound Aß1-42 aggregates.

[Applicability of Seven Glomerular Filtration Rate Evaluation Formulas in Dose Adjustment of High Concentration of Methotrexate Chemotherapy in Children with ALL].

OBJECTIVE: To investigate the diagnostic efficacy of seven glomerular filtration rate (GFR) evaluation formulas Schwartz2009, Schwartz1976, Counahan-Barratt, Filler, CKD-EPIscysc, Cockrofi-Gault, CKD-EPIScysC-Scr in high concentration of methotrexate (HDMTX) chemotherapy dose adjusted cut-off point (GFR ≤85 ml/min) in children with acute lymphoblastic leukemia (ALL). METHODS: One hundred and twenty-four children with ALL were included in the study. GFR determined by renal dynamic imaging (sGFR) was used as the standard to evaluate the accuracy, consistency of eGFR calculated by seven formulas and sGFR, and the diagnostic efficacy of each formula when the sGFR ≤85 ml/min boundary. RESULTS: All of the accuracy of eGFR estimated by Schwartz2009 were greater than 70% in the 0-3, >4 and ≤6, >6 and ≤9, >9 and ≤16 years old group and male group, and the consistency exceeded the professional threshold. When the sensitivity of the ROC curve sGFR ≤85 ml/min was 100% of CKD-EPIscysc in the 0-3, >3 and ≤4 years old group, Filler in the >3 and ≤4 years old group, and Cockrofi-Gault in the >6 and ≤9 years old group, the specificity was 73.02%, 78.95%, 78.95%, 69.32%, respectively, and the AUC under the ROC curve was the largest (P<0.05). CONCLUSION: Schwartz2009 formula predicts the highest accuracy of eGFR in the 7 glomerular filtration rate. CKD-EPIscysc, Filler, and Cockrofi-Gault formulas have more guiding signi-ficance for the adjustment of HDMTX chemotherapy in pre-adolescence in children with ALL when sGFR ≤85 ml/min.

Removal of oxytetracycline and ofloxacin in wastewater by microalgae-bacteria symbiosis for bioenergy production.

The development of microalgae-bacteria symbiosis for treating wastewater is flourishing owing to its high biomass productivity and exceptional ability to purify contaminants. A nature-selected microalgae-bacteria symbiosis, mainly consisting of Dictyosphaerium and Pseudomonas, was used to treat oxytetracycline (OTC), ofloxacin (OFLX), and antibiotic-containing swine wastewater. Increased antibiotic concentration gradually reduced biomass productivity and intricately changed symbiosis composition, while 1 mg/L OTC accelerated the growth of symbiosis. The symbiosis biomass productivity reached 3.4-3.5 g/L (5.7-15.3 % protein, 18.4-39.3 % carbohydrate, and 2.1-3.9 % chlorophyll) when cultured in antibiotic-containing swine wastewater. The symbiosis displayed an excellent capacity to remove 76.3-83.4 % chemical oxygen demand, 53.5-62.4 % total ammonia nitrogen, 97.5-100.0 % total phosphorus, 96.3-100.0 % OTC, and 32.8-60.1 % OFLX in swine wastewater. The microbial community analysis revealed that the existence of OTC/OFLX increased the richness and evenness of microalgae but reduced bacteria species in microalgae-bacteria, and the toxicity of OFLX to bacteria was stronger than that of OTC.

Is Distributional Justice Equivalent to Prosocial Sharing in Children's Cognition?

Distribution and sharing are social preference behaviors supported and shaped by selection pressures, which express individuals' concern for the welfare of others. Distributive behavior results in distributive justice, which is at the core of moral justice. Sharing is a feature of the prosocial realm. The connotations of distribution and sharing are different, so the principles, research paradigms, and social functions of the two are also different. Three potential causes of confusion between the two in the current research on distribution and sharing are discussed. First, they share common factors in terms of individual cognition, situation, and social factors. Second, although they are conceptually different, prosocial sharing and distribution fairness sensitivity are mutually predictive in individual infants. Similarly, neural differences in preschoolers' perception of distribution fairness predict their subsequent sharing generosity. Finally, similar activation regions are relevant to distribution and sharing situations that need behavioral control on a neural basis.

Microalgae cultivation for antibiotic oxytetracycline wastewater treatment.

Microalgae-based technology provides a potential approach to biologically treating oxytetracycline (OTC) wastewater due to its environmental friendliness, low cost, and high efficiency. However, the OTC degradation and transformation characteristics by microalgae are still unclear and need further exploration. This study used microalgae Chlorella sorokiniana MB-1 for OTC wastewater treatment. The OTC with an initial concentration less than 50 mg L-1 promoted microalgae growth, while OTC with a concentration higher than 100 mg L-1 inhibited microalgae growth significantly. More than 99% OTC was removed with the biomass productivity up to 1.8 g L-1 when treated OTC with 10 mg L-1 initial concentration for 7 days. Chlorophyll and total sugar contents decreased, while protein and lipid contents increased compared to the control without OTC. The malondialdehyde content firstly reduced but subsequently enhanced when increased OTC concentration, while superoxide dismutase content gradually enhanced, manifesting that traces of OTC stimulate microalgae antioxidant capacity, while the increasing OTC caused further oxidative damage to microalgae cells. The removal pathways of OTC mainly include photolysis (75.8%), biodegradation (17.8%), biosorption (3.6%), and hydrolysis (2.7%). Overall, removing OTC by microalgae was confirmed to be an excellent technology for treating antibiotics wastewater whilst accumulating microalgae biomass.

Biobased carbon dots production via hydrothermal conversion of microalgae Chlorella pyrenoidosa.

A promising green hydrothermal process was used to produce biobased nanomaterials carbon dots (CDs) by using microalgae Chlorella pyrenoidosa (CP) and its main model compounds (i.e., glucose, glycine, and octadecanoic acid). The possible reaction pathway including hydrolysis, Amadori rearrangement, cyclization/aromatization, and polymerization was first proposed for the hydrothermal process to produce microalgae-based CDs. Interactions among carbohydrates and proteins in microalgae were vital intermediate reactions in the generation of CDs. The mass yield of CDs reached 7.2% when the CP was hydrothermally treated with 20:1 of liquid-to-solid ratio at 230 °C for 6 h. It was confirmed that nitrogen, sulfur, phosphorous, and potassium were doped onto CP-based CDs (CD-CP) successfully without additional reagents or treatments. The CD-CP yield was 4.0-24.3 times higher than that of model compound-based CDs. Regarding morphology, CD-CP was constituted by many spherical nanoparticles smaller than 20 nm. These CDs emitted blue fluorescence under ultraviolet light, and the fluorescence quantum yield of CD-CP was 4.7-9.4 times higher than that of CP model compound-based CDs. Last, CD-CP displayed broad application prospects as a sensor for Fe3+ detection in wastewater with high sensitivity.

m5C Regulator-Mediated Methylation Modification Patterns and Tumor Microenvironment Infiltration Characterization in Papillary Thyroid Carcinoma.

Recently, immune response modulation at the epigenetic level is illustrated in studies, but the possible function of RNA 5-methylcytosine (m5C) modification in cell infiltration within the tumor microenvironment (TME) is still unclear. Three different m5C modification patterns were identified, and high differentiation degree was observed in the cell infiltration features within TME under the above three identified patterns. A low m5C-score, which was reflected in the activated immunity, predicted the relatively favorable prognostic outcome. A small amount of effective immune infiltration was seen in the high m5C-score subtype, indicating the dismal patient survival. Our study constructed a diagnostic model using the 10 signature genes highly related to the m5C-score, discovered that the model exhibited high diagnostic accuracy for PTC, and screened out five potential drugs for PTC based on this m5C-score model. m5C modification exerts an important part in forming the TME complexity and diversity. It is valuable to evaluate the m5C modification patterns in single tumors, so as to enhance our understanding towards the infiltration characterization in TME.

Crystal structure of 4-(anthracen-9-yl)pyridine.

The title compound, C19H13N, which crystallizes in the monoclinic C2/c space group with one half-mol-ecule in the asymmetric unit, was synthesized by Suzuki-Miyaura cross-coupling reaction of 9-bromo-anthracene with pyridin-4-ylboronic acid and purified by column chromatography on silica gel. Light-yellow crystals of 4-(anthracen-9-yl)-pyridine suitable for X-ray diffraction were collected by the solvent evaporation method. In the crystal, pairs of mol-ecules are connected by inter-molecular C-H⋯π (pyridine) inter-actions [d(H7⋯Cg) = 2.7391 (2) Å], forming cyclic centrosymmetric dimers, further resulting in an infinite one-dimensional linear chain along the c-axis direction. Weak face-to-face π-π stacking inter-actions [d(Cg⋯Cg) = 3.6061 (2) Å] link neighboring lamellar networks into the supra-molecular structure.

The Effect of Selenium on CYP450 Isoform Activity and Expression in Pigs.

Selenium is an essential nutrient in diets; however, the effects of selenium on enzyme metabolic activation are not currently clear. Cytochromes P450 (CYP450) are major phase I metabolic enzymes involved in the biotransformation of xenobiotics and endogenous compounds to form electrophilic reactive metabolites. To investigate the effect of selenium on CYP450 isoform activity, the Landrace pigs were divided into three groups: the control group (containing Se 0.15 mg/kg), the Se-deficient group (Se 0.03 mg/kg), and the Se-supply group (Se 0.35 mg/kg). After 1 week of administration, a mixed solution (20 mg/kg of dextromethorphan, phenacetin, chlorzoxazone, and 10 mg/kg of testosterone in a CMC-Na solution) was intravenously injected into all pigs. The mixed solution content and pharmacokinetic parameters were assayed by HPLC and DAS, respectively. To investigate the effect of selenium on CYP450 isoform expression, RNA-Seq analysis, Western boltting, and qPCR were used. Results showed that Se-supply group significantly increased the activity and expression of CYP1A2 and CYP2D25, and decreased CYP3A29. Se-deficient group decreased the activity of CYP1A2, CYP2D25, and CYP2E1. These results demonstrated that selenium content affecting the activity or expression of the CYP450 isoform may lead to a food-drug interaction.

A novel allosteric site in casein kinase 2α discovered using combining bioinformatics and biochemistry methods.

Casein kinase 2 (CK2) is a highly pleiotropic serine-threonine kinase, which catalyzed phosphorylation of more than 300 proteins that are implicated in regulation of many cellular functions, such as signal transduction, transcriptional control, apoptosis and the cell cycle. On the other hand, CK2 is abnormally elevated in a variety of tumors, and is considered as a promising therapeutic target. The currently available ATP-competitive CK2 inhibitors, however, lack selectivity, which has impeded their development in cancer therapy. Because allosteric inhibitors can avoid the shortcomings of conventional kinase inhibitors, this study was aimed to discover a new allosteric site in CK2α and to investigate the effects of mutations in this site on the activity of CK2α. Using Allosite based on protein dynamics and structural alignment, we predicted a new allosteric site that was partly located in the αC helix of CK2α. Five residues exposed on the surface of this site were mutated to validate the prediction. Kinetic analyses were performed using a luminescent ADP detection assay by varying the concentrations of a peptide substrate, and the results showed that the mutations I78C and I78W decreased CK2α activity, whereas V31R, K75E, I82C and P109C increased CK2α activity. Potential allosteric pathways were identified using the Monte Carlo path generation approach, and the results of these predicted allosteric pathways were consistent with the mutation analysis. Multiple sequence alignments of CK2α with the other kinases in the family were conducted using the ClustalX method, which revealed the diversity of the residues in the site. In conclusion, we identified a new allosteric site in CK2α that can be altered to modulate the activity of the kinase. Because of the high diversity of the residues in the site, the site can be targeted using rational drug design of specific CK2α inhibitors for biological relevance.

Atorvastatin treatment is associated with increased BDNF level and improved functional recovery after atherothrombotic stroke.

BACKGROUND: Statins have a positive impact on ischemic stroke outcome. It has been reported that statin have neuroprotective function after ischemic stroke in addition to lipid-lowering effect in animal model. However, the neuroprotective function of statin after stroke has not been confirmed in clinical studies. The aim of this study was to evaluate in a clinical model if statins induce neuroprotection after stroke. We, therefore, assessed serum brain-derived neurotrophic factor (BDNF) levels and functional recovery in atherothrombotic stroke patients and investigated their relationship with atorvastatin treatment. METHODS: Seventy-eight patients with atherothrombotic stroke were enrolled and randomly assigned to atorvastatin treatment group or placebo control group. Neurological function after stroke was assessed with the National Institutes of Health Stroke Scale, modified Rankin Scale (mRS) and Barthel Index (BI). The serum BDNF levels were both measured at 1 day and 6 weeks after stroke. Linear regression was used to assess the association between BDNF levels and neurological function scores. RESULTS: The mRS and BI were markedly improved in the atorvastatin group when compared to placebo at 6 weeks after stroke. The serum BDNF levels in atorvastatin group were significantly elevated by 6 weeks after stroke and higher than the BDNF levels in controls. In addition, the serum BDNF levels significantly correlated with mRS and BI after stroke. Our results demonstrated that atorvastatin treatment was associated with the increased BDNF level and improved functional recovery after atherothrombotic stroke. CONCLUSION: This study indicates that atorvastatin-related elevation in the BDNF level may promote functional recovery in stroke patients.

Crystal Structures of PI3Kα Complexed with PI103 and Its Derivatives: New Directions for Inhibitors Design.

The phosphatidylinositol 3-kinase (PI3K) signaling pathway plays important roles in cell proliferation, growth, and survival. Hyperactivated PI3K is frequently found in a wide variety of human cancers, validating it as a promising target for cancer therapy. We determined the crystal structure of the human PI3Kα-PI103 complex to unravel molecular interactions. Based on the structure, substitution at the R1 position of the phenol portion of PI103 was demonstrated to improve binding affinity via forming a new H-bond with Lys802 at the bottom of the ATP catalytic site. Interestingly, the crystal structure of the PI3Kα-9d complex revealed that the flexibility of Lys802 can also induce additional space at the catalytic site for further modification. Thus, these crystal structures provide a molecular basis for the strong and specific interactions and demonstrate the important role of Lys802 in the design of novel PI3Kα inhibitors.

The mechanism of allosteric inhibition of protein tyrosine phosphatase 1B.

As the prototypical member of the PTP family, protein tyrosine phosphatase 1B (PTP1B) is an attractive target for therapeutic interventions in type 2 diabetes. The extremely conserved catalytic site of PTP1B renders the design of selective PTP1B inhibitors intractable. Although discovered allosteric inhibitors containing a benzofuran sulfonamide scaffold offer fascinating opportunities to overcome selectivity issues, the allosteric inhibitory mechanism of PTP1B has remained elusive. Here, molecular dynamics (MD) simulations, coupled with a dynamic weighted community analysis, were performed to unveil the potential allosteric signal propagation pathway from the allosteric site to the catalytic site in PTP1B. This result revealed that the allosteric inhibitor compound-3 induces a conformational rearrangement in helix α7, disrupting the triangular interaction among helix α7, helix α3, and loop11. Helix α7 then produces a force, pulling helix α3 outward, and promotes Ser190 to interact with Tyr176. As a result, the deviation of Tyr176 abrogates the hydrophobic interactions with Trp179 and leads to the downward movement of the WPD loop, which forms an H-bond between Asp181 and Glu115. The formation of this H-bond constrains the WPD loop to its open conformation and thus inactivates PTP1B. The discovery of this allosteric mechanism provides an overall view of the regulation of PTP1B, which is an important insight for the design of potent allosteric PTP1B inhibitors.

Liquefaction of bamboo shoot shell for the production of polyols.

Bamboo (Dendrocalamus latiflorus Munro) shoot shell (BSS) was liquefied in polyethylene glycol 400 (PEG400) and ethylene glycol (EG) catalyzed by sulfuric acid under atmospheric pressure. The effects of liquefaction conditions such as liquid-solid ratio, temperature, time, catalyst, solvents ratio, and material size on the liquefaction yield of BSS have been investigated. Methods including Elemental analysis, Thermogravimetric analysis, Fourier transform infrared spectroscopy, nuclear magnetic resonance and gas chromatography-mass spectrometry were selected to analyze the characteristics of products in three fractions: an aqueous fraction (AQ), an acetone-soluble fraction (AS) and a residue (RS), respectively. Results showed that the highest liquefaction percentage was 99.79% under the optimal conditions (liquid-solid ratio 6:1; temperature 150°C; reaction time 80min; raw size more than 40 mesh; catalyst mass percentage of solvent 4%; solvent volume ratio 3:1). Polyols could be obtained effectively by the liquefaction of BSS, an agricultural by-product.

[Genetic diversity and phylogeny of bradyrhizobia associated with Desmodium spp].

OBJECTIVE: Genetic diversity and phylogeny of bradyrhizobial strains associated with Desmodium spp. in subtropic and temperate regions of China were analyzed. METHODS: We studied 29 desmodia isolates from different regions with BOX-PCR fingerprinting and multilocus sequence analysis (nifH, nodC and recA gene) to describe the genotypic characteristics and phylogenetic relationships. RESULTS: We achieved 25 genotypes with BOX-PCR genomic fingerprinting analysis, indicating that the tested strains had a great diversity at genomic level. The representative bradyrhizobial strains were located in three phylogenic branches with multilocus sequence analysis (niffH, nodC and recA gene), that was closely related to Bradyrhizobium elkanii, Bradyrhizobium japonicum and Bradyrhizobium yuanmingense, respectively. CONCLUSION: The desmodia bradyrhizobia had abundantly diversity. Diverse symbiotic genes including nifH and nodC genes were also found in these strains that indicated that the symbiotic genes were mainly maintained by vertical transfer in these bradyrhizobial populations and coevolved with housekeeping genes.