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Abstract Background Our previous study finds that male sterility in Salvia miltiorrhiza could result in stunted growth and reduced biomass, but their molecular mechanisms have not yet been revealed. In this article, we investigate the underlying mechanism of male sterility and its impact on plant growth and metabolic yield by using physiological analysis and mRNA sequencing (RNA-Seq). Results In this study, transcriptomic and physiological analysis were performed to identify the mechanism of male sterility in mutants and its impact on plant growth and metabolic yield. Through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, it is found that the pathways are mainly enriched in processes including organ development, primary metabolic process and secondary metabolic process. Physiological analysis show that the chloroplast structure of male sterile mutants of S. miltiorrhiza is abnormally developed, which could result in decrease in leaf gas exchange (A, E and g s), chlorophyll fluorescence (Fv, Fm and Fv/Fm), and the chlorophyll content. Expression level of 7 differentially expressed genes involved in photosynthesis-related pathways is downregulated in male sterile lines of S. miltiorrhiza, which could explain the corresponding phenotypic changes in chlorophyll fluorescence, chlorophyll content and leaf gas exchange. Transcriptomic analysis establishes the role of disproportionating enzyme 1 (DPE1) as catalyzing the degradation of starch, and the role of sucrose synthase 3 (SUS3) and cytosolic invertase 2 (CINV2) as catalyzing the degradation of sucrose in the S. miltiorrhiza mutants. The results also confirm that phenylalanine ammonialyase (PAL) is involved in the biosynthesis of rosmarinic acid and salvianolic acid B, and flavone synthase (FLS) is an important enzyme catalyzing steps of flavonoid biosynthesis. Conclusions Our results from the physiological and transcriptome analysis reveal underlying mechanism of plant growth and metabolic yield in male sterile mutants, and provide insight into the crop yield of S. miltiorrhiza.

Objective: The long-acting glucagon-like peptide-1 receptor (GLP-1R) agonist, liraglutide, stimulates insulin secretion and efficiently suppresses food intake to reduce body weight. As such, liraglutide is growing in popularity in the treatment of diabetes and chronic weight management. Within the brain, liraglutide has been shown to alter the activity of hypothalamic proopiomelanocortin (POMC) and Neuropeptide Y/Agouti-related peptide (NPY/AgRP) neurons. Moreover, the acute activities of POMC and NPY neurons have been directly linked to feeding behavior, body weight, and glucose metabolism. Despite the increased usage of liraglutide and other GLP-1 analogues as diabetic and obesity interventions, the cellular mechanisms by which liraglutide alters the activity of metabolically relevant neuronal populations are poorly understood. Methods: In order to resolve this issue, we utilized neuron-specific transgenic mouse models to identify POMC and NPY neurons for patch-clamp electrophysiology experiments. Results: We found that liraglutide directly activated arcuate POMC neurons via TrpC5 channels, sharing a similar mechanistic pathway to the adipose-derived peptide leptin. Liraglutide also indirectly increases excitatory tone to POMC neurons. In contrast, liraglutide inhibited NPY/AgRP neurons through post-synaptic GABAA receptors and enhanced activity of pre-synaptic GABAergic neurons, which required both TrpC5 subunits and K-ATP channels. In support of an additive role of leptin and liraglutide in suppressing food intake, leptin potentiated the acute effects of liraglutide to activate POMC neurons. TrpC5 subunits in POMC neurons were also required for the intact pharmacological effects of liraglutide on food intake and body weight. Thus, the current study adds to recent work from our group and others, which highlight potential mechanisms to amplify the effects of GLP-1 agonists in vivo. Moreover, these data highlight multiple sites of action (both pre- and post-synaptic) for GLP-1 agonists on this circuit. Conclusions: Taken together, our results identify critical molecular mechanisms linking GLP-1 analogues in arcuate POMC and NPY/AgRP neurons with metabolism. Keywords: Melanocortin, POMC, NPY/AgRP, Liraglutide, Patch-clamp, Electrophysiology

As mining depth gradually increases, the complex and changeable behavior state of mine production systems leads to the increasingly prominent problem of “planning is difficult to control”. The safety production situation and the difficulty of emergency management are gradually upgraded. However, the theoretical study on the behavioral trend of complex mine production systems is still an immature field. This paper proposes the scientific problem of “theory and method of time-varying computational experiments for fully mechanized mining processes in artificial system environments”. Taking the typical fully mechanized mining process in the Yushen mining area in northern Shaanxi, China, as the research object, through computer modeling, simulation and use of multiagent system theory, APSM (Agent Publish-Subscribe Model) coordination technology, multiagent cross-emergence and multilayer learning networks, the artificial fully mechanized mining system modeling, sequential mining process deduction and state transfer theory are systematically studied. First, an artificial system model equivalent to the function of the actual fully mechanized mining system is constructed. Then, under the artificial system environment, the time-varying computational experiments of the fully mechanized mining process are realized through the autonomous deduction of the fully mechanized mining agent based on a multilayer neural network and the emergence of multiagent interactions based on subscription perception; this approach aims to solve the problem of determining the overall behavior trend of the mine under the condition of “long time and large space” and to provide intellectual support and scientific basis for the “first experiment and then produce” technological model of intelligent mining.

This paper presents in detail the instrumentation for coordinated optical and current measurements of lightning discharges to a 356-m-high meteorological tower and the observation results. The tower, which is located at Shenzhen, a lower latitude coastal city in south China, was equipped with a non-inductive current shunt at its top and a high-speed camera and electromagnetic field sensors at 440 m away from the tower base. A total of 24 discharges to the tower were well documented in the period of April to June of 2019. Three of them were analyzed in detail with the focus being given to the feature of upward leaders initiated from the tower: Case 1 - a connecting upward positive leader (connecting UPL) induced by a downward negative in a downward negative discharge, Case 2 - an upward positive leader (UPL) in the initial stage of an upward negative discharge, and Case 3 - an upward negative leader (UNL) in the initial stage of an upward positive discharge. All the three leaders had a stepping feature during their initial stages, each step producing an isolated but oscillated current pulse with a time scale of 1 μs. The connecting UPL had 5 steps during its initial 2.6 ms, with a step interval ranging in 0.1~1.5 ms and a current pulse peak in 1~5 kA. The UPL had also 5 steps but during its initial 200 μs, with a step interval ranging in 20~50 μs, a step length in 0.8~2.2 m, a current pulse peak in 0.5~2.2 kA, and a leader average speed in 0.4~1.1 × 105 m/s. The UNL had 8 steps during its initial 104 μs, with a step interval ranging in 13.6~22 μs, a step length in 3.9~7.1 m, a current pulse peak in 2.3~12.5 kA, and a leader average speed in 1.9~6.6 × 105 m/s.

Three species, Calocybe lilacea, C. longisterigma, and C. subochraceus, were newly discovered in Inner Mongolia, Hunan, and Liaoning provinces of China. Calocybe lilacea is mainly characterized by its small-sized basidiocarp, brownish-orange pileus, lilac gray to dull violet stipe, and noncellular epicutis. The main characteristics of C. longisterigma are its light brown to brownish-orange pileus, long sterigmata, and noncellular epicutis. The main morphological features of C. subochraceus are its small-sized basidiocarp, pale-orange pileus, adnexed lamellae, tortuous stipe, and noncellular epicutis. These morphological features confirmed that the three species of Calocybe all belonged to Sect. Carneoviolaceae. Phylogenetic analysis based on a combined dataset (ITS–nrLSU–RPB2) determined that the three species belong to the genus Calocybe and form a distinct lineage. The morphological differences between the three new species and other related species of Calocybe are also discussed.

Objective: Hypothalamic Pro-opiomelanocortin (POMC) and Neuropeptide Y/Agouti-Related Peptide (NPY/AgRP) neurons are critical nodes of a circuit within the brain that sense key metabolic cues as well as regulate metabolism. Importantly, these neurons retain an innate ability to rapidly reorganize synaptic inputs and electrophysiological properties in response to metabolic state. While the cellular properties of these neurons have been investigated in the context of obesity, much less is known about the effects of exercise training. Methods: In order to further investigate this issue, we utilized neuron-specific transgenic mouse models to identify POMC and NPY/AgRP neurons for patch-clamp electrophysiology experiments. Results: Using whole-cell patch-clamp electrophysiology, we found exercise depolarized and increased firing rate of arcuate POMC neurons. The increased excitability of POMC neurons was concomitant with increased excitatory inputs to these neurons. In agreement with recent work suggesting leptin plays an important role in the synaptic (re)organization of POMC neurons, POMC neurons which express leptin receptors were more sensitive to exercise-induced changes in biophysical properties. Opposite to effects observed in POMC neurons, NPY neurons were shunted toward inhibition following exercise. Conclusions: Together, these data support a rapid reorganization of synaptic inputs and biophysical properties in response to exercise, which may facilitate adaptations to altered energy balance and glucose metabolism. Keywords: Melanocortin, Energy balance, Leptin receptor, Exercise, Patch-clamp, Electrophysiology

For the liquid epoxy nanosuspensions with both fibril and spherical polypyrrole (PPy) nanostructures, a stronger PPy nanofibers/epoxy interaction and more temperature stable behavior with a lower flow activation energy of nanosuspensions with nanofibers (54.34 kJ mol-1) than that with nanospheres (71.15 kJ mol-1) were revealed by rheological studies. As well as the common enhancing mechanism of limiting crack propagation in the polymer matrix, the nanofibers further initiated the shear bands in the epoxy resin to give a higher tensile strength (90.36 MPa) than that of pure epoxy (70.03 MPa) and even that of the epoxy nanocomposites with nanospheres (84.53 MPa). With a larger specific surface area, the nanofibers rather than nanospheres were observed to reduce the flammability of epoxy more efficiently by assisting more char formation of the epoxy resin. The hydroxyl groups formed between the protons of the doped acid in the PPy nanofillers and the epoxy broke the conjugate structure of PPy, leading to a higher bandgap in the nanocomposites (Eg1 ¼ 3.08 eV for 1.0 wt% PPy nanofibers) than that of pure nanofillers (1.8 eV for PPy nanofibers and 1.2 eV for PPy nanospheres). Due to the high aspect ratio, the PPy nanofibers could form the conductive path more easily than the PPy nanospheres to provide a lower percolation threshold value. The real permittivity was observed to increase with increasing the PPy nanofiller loading, and the enhanced permittivity was interpreted by the interfacial polarization. [ABSTRACT FROM AUTHOR]

A 56-year-old male was diagnosed with right lung upper lobe squamous cancer with right hilar and mediastinum lymph node metastasis. After four cycles of neoadjuvant immunochemotherapy, reexamination by computed tomography showed progressive disease of the primary lesion. Then, the patient underwent a right lung upper lobectomy, and hilar and mediastinum lymph node dissection. Surgical pathology showed a partial response to immunochemotherapy. Single-cell RNA sequencing was used to characterize the infiltrating immune cell atlas after neoadjuvant immunochemotherapy; the most common infiltrating immune cell types were cytotoxic CD8+ T cells, monocyte-derived dendritic cells, and macrophages. Imaging mass cytometry revealed a transformation from cold to hot tumor after neoadjuvant immunochemotherapy. In this case study, we are the first to report a case of neoadjuvant immunochemotherapy pseudoprogression, proved by surgical pathology, single-cell RNA sequencing, and imaging mass cytometry. Both single-cell RNA sequencing and imaging mass cytometry revealed an activated immune microenvironment after neoadjuvant immunochemotherapy.

Background Dynamic visual attention is important in basketball because it may affect the performance of players and thus the match outcome. The goals of this study were to investigate the difference in dynamic visual attention characteristics between highly skilled basketball players and nonathletic college students and to explore the relationship between visual attention and game-related performance among the basketball players. Methods In total, 24 highly skilled basketball players and 24 nonathletic college students participated in a multiple object tracking task. The task was conducted so that either the number of targets that were visually tracked or the speed at which a given number of tracked targets moved was altered to examine the difference in dynamic visual attention characteristics between the basketball players and nonathletic college students. The relationship between visual tracking speed (VTS) and game-related statistics, including assists, steals, mistakes, fouls and points scored recorded for every match during the season, was assessed among the basketball players by using Pearson correlations. Results A significant main effect of target tracking load was observed (P < 0.001), with visual tracking performance significantly decreased as target number increased. In addition, the speed at which the targets moved had a significant effect on visual tracking performance (P < 0.001), with tracking performance significantly decreased as target speed increased. However, no significant difference was observed in the abilities of basketball players and nonathletic college students to simultaneously track up to six targets. By contrast, a significant interaction between group and target speed was found (P < 0.001), with the visual tracking accuracy of basketball players significantly greater than that of college students at the higher target speeds examined (P < 0.001). Among basketball players, there were positive, large, and statistically significant correlations in the accuracy in VTS trials and the number of assists (P < 0.001) and between the accuracy in VTS trials and the number of steals (P < 0.001). Conclusion The advantage of skilled basketball players to handle dynamic visual information in a multiple object tracking task was not attributable to the target number but to the target speed. Those athletes with greater dynamic visual attention were more likely to successfully assist or to steal the ball, enhancing performance of the athlete as well as contributing to a more successful team match. These findings may inform basketball training programs to improve player and team performances during matches.

As one of the promising MAX phase materials for high-temperature applications, Cr2AlC is considered as a potential substitution bond coat material in thermal barrier coating systems. In this paper, the microstructure evolution and elemental diffusion behavior near the interface of the diffusion couple composed of Cr2AlC and single crystal superalloy DD5 were investigated at 1100 °C, 1150 °C, and 1200 °C. Elemental interdiffusion between Cr2AlC and DD5 occurs significantly, resulting in the formation of a thick layer of Kirkendall holes after 20 h heat treatment at 1100 °C and higher temperatures. The outward diffusion of Ni into Cr2AlC and the inward diffusion of Al into DD5 alloy causes the formation of β-NiAl matrix embedded with dispersed Cr7C3 phase. Simultaneously, the precipitation of σ-TCP phase and degradation of the γ/γ′ matrix occurs in the alloy. Additionally, TaC, M2C (where M = Ta, W, Cr), and M23C6 (M = Cr, Re, W) compounds are formed near the interface along with the dissolution of σ-TCP phases. It is further found that Al in Cr2AlC exhibits the highest average effective diffusion coefficient among the four dominant diffusing elements. It also displays the lowest diffusion activation energy which is due to its relatively weak CrAl and AlAl bonds.

Summary: The identification of cell surface proteins on stem cells or stem cell derivatives is a key strategy for the functional characterization, isolation, and understanding of stem cell population dynamics. Here, using an integrated mass spectrometry- and microarray-based approach, we analyzed the surface proteome and transcriptome of cardiac progenitor cells (CPCs) generated from the stage-specific differentiation of mouse and human pluripotent stem cells. Through bioinformatics analysis, we have identified and characterized FZD4 as a marker for lateral plate mesoderm. Additionally, we utilized FZD4, in conjunction with FLK1 and PDGFRA, to further purify CPCs and increase cardiomyocyte (CM) enrichment in both mouse and human systems. Moreover, we have shown that NORRIN presented to FZD4 further increases CM output via proliferation through the canonical WNT pathway. Taken together, these findings demonstrate a role for FZD4 in mammalian cardiac development.

The effects of the surface functional groups on the electrochemical properties and electrochemical energy storage were systematically studied by employing the as-received multi-walled carbon nanotubes (MWNTs) and two other MWNTs modified with ether and carboxylic acid functionalities (-C-O-C and -COOH), and ester functional groups (-COOR). The structure and morphology of the films prepared by electropolymerizing aniline onto MWNTs- coated carbon paper substrate were characterized by scanning electron microscopy and Fourier transform infrared spectroscopy. The band gaps obtained from ultraviolet-visible spectroscopy suggested slight difference in the structure of the nanocomposite films induced by the functional groups in the MWNTs. Electrochemical measurements revealed that all the nanocomposite films with MWNTs exhibited an enhanced capacitance compared to the film grown onto pure carbon paper. An energy density as high as 30 µWh/cm² corresponding to a power density of around 8 mW/cm ² was obtained even at a high current density of 20 mA/cm ² for the nanocornposite films. An equivalent circuit consisting of a combined resistance and a capacitor is well fitted with the electrochemical impedence spectroscopy data. Stability study showed that the nanocomposite films retained around 80% of the discharge capacity even after 1000 charge-discharge cycles, rendering these films promising for supercapacitor applications. [ABSTRACT FROM AUTHOR]

Collision problem of two centered rarefaction waves for one-dimensional isentropic magnetogasdynamics is studied. When there is no vacuum in the initial data, it is proven that the vacuum never appears in the interaction domain, while it appears only when the collision time is large enough. Further, the existence and uniqueness of smooth solution for the Goursat problem developed on the penetration domain is obtained, and the collision results are shown. Finally, some numerical results exhibiting the collision process are presented. [ABSTRACT FROM AUTHOR]

The early developmental toxicity in zebrafish exposed by Phenanthrene (Phe) and Fluorene (H) was investigated by determining mortality and morphological malformation endpoints, such as no inflation of swimming bladder (no ISB), axial malformations (AM, crooked/clubbed), pen- cardial edema (PE) and yolk sac edema (YSE). The lethal concentration to cause 50% mortality (LC50) in embryos was 78.632 μmol L-1 for Phe and 236.930 μmol L-1 for Fl, respectively. Embryos exposed to 40 μmol L-1 Phe exhibited a reduction of blood stream speed and sharply increasing of axial malformation rate. When exposed to 80 μmol L-1 Phe, the significant (at p<0.05) increase of PE or YSE index was observed. Embryos exposed to H had significant axial malformations, but didn't cause significant PE, YSE and speed reduction of blood stream as those exposed to Phe. The joint effect of Phe and Fl was antagonistic. The 42-h and 114-h Phe-exposure results showed similar mortality and malformation rates. All of the toxic endpoints showed the significant differences (p<0.05) between 18-h and 114-h exposure, suggesting that 42-h exposure produced very serious and irreversible toxicological effects, but that 18-h Phe-exposure only generated slight, reversible toxic effects. At low concentrations (3.12-12.5 μmol L-1) of Phe producing no obviously morphological abnormality or mortality, the locomotion analysis on the larval zebrafish showed that Phe-exposed groups have significantly (p<0.05) higher speed movements (>15 mmls) than control groups. [ABSTRACT FROM AUTHOR]

The existence of either eccentricity or slenderness has a significant effect on the mechanical properties of a structure or member. These properties can change the working mechanism, failure mode, and bearing capacity of the structure or member. A concrete-filled, glass fibre-reinforced, polymer tube composite column has the same problem. We carried out experiments on the influences of eccentricity and slenderness on the mechanical properties of concrete-filled, glass fibre-reinforced, polymer tube composite columns. The experimentally recorded stress–strain relationships are presented graphically, and the ultimate axial stresses and strains and the FRP tube hoop strains at rupture were tabulated. The results indicate that the influences of slenderness and eccentricity on the composite columns were significant with regard to the axial strain, hoop strain, ultimate bearing capacity, lateral displacement, and failure mode. Based on the existing research literature and the results reported in this paper, the bearing capacity formula of a composite slender column under an eccentric load was established. The theoretical results were in good agreement with the experimental results.

Emergence and intercontinental spread of highly pathogenic avian influenza A(H5Nx) virus clade 2.3.4.4 is unprecedented. H5N8 and H5N2 viruses have caused major economic losses in the poultry industry in Europe and North America, and lethal human infections with H5N6 virus have occurred in Asia. Knowledge of the evolution of receptor-binding specificity of these viruses, which might affect host range, is urgently needed. We report that emergence of these viruses is accompanied by a change in receptor-binding specificity. In contrast to ancestral clade 2.3.4 H5 proteins, novel clade 2.3.4.4 H5 proteins bind to fucosylated sialosides because of substitutions K222Q and S227R, which are unique for highly pathogenic influenza virus H5 proteins. North American clade 2.3.4.4 virus isolates have retained only the K222Q substitution but still bind fucosylated sialosides. Altered receptor-binding specificity of virus clade 2.3.4.4 H5 proteins might have contributed to emergence and spread of H5Nx viruses.

The molecular mechanisms underlying acute leptin and serotonin 2C receptor-induced hypophagia remain unclear. Here, we show that neuronal and pro-opiomelanocortin (Pomc)-specific loss of transient receptor potential cation 5 (TrpC5) subunits is sufficient to decrease energy expenditure and increase food intake resulting in elevated body weight. Deficiency of Trpc5 subunits in Pomc neurons is also sufficient to block the anorexigenic effects of leptin and serotonin 2C receptor (Ht2Cr) agonists. The loss of acute anorexigenic effects of these receptors is concomitant with a blunted electrophysiological response to both leptin and Ht2Cr agonists in arcuate Pomc neurons. We also demonstrate that the Ht2Cr agonist lorcaserin-induced improvements in glucose and insulin tolerance are blocked by TrpC5 deficiency in Pomc neurons. Together, our results link TrpC5 subunits in the brain with leptin- and serotonin 2C receptor-dependent changes in neuronal activity, as well as energy balance, feeding behavior, and glucose metabolism.

Medical titanium alloys remain the most successful commercial bone implant material, but efforts are demanded to improve the biocompatibility. Silk fibroin (SF) is a natural structural protein with tunable mechanical properties and biocompatibility. Herein we established a facile method of physical depositing SF coatings via HFIP solvent onto Ti6Al4V (TC4) biometals. To stabilize SF coatings, ethanol induced physical crosslinking and EGDE chemical crosslinking was applied. The coating-substrate system revealed tightly integrated interface between SF and TC4. Importantly, the introduction of SF coatings significantly increased the water wettability of the TC4 surface by decreasing the water contact angle from 93° to 67°. Furthermore, the interfacial bonding strength, measured by advanced centrifugal measurement, was affected by the surface roughness of the substrate in a parabolic fashion and the bonding strength increased with reduced thickness of the coating. An appropriate roughness was optimized at Ra~1 μm and 7 MPa adhesion strength was achieved. Cytotoxicity using L929 cell line and cell viability and cytoskeleton organization using MG63 cell line of the SF-TC4 hybrid were investigated. The study provides a facile way of physical deposition of biopolymer coatings on inert medical metals as well as a basis for further improving the adherence of biometals and polymers. Keywords: Metal-polymer interface, Surface roughness, Structural protein, Biocompatibility, Adhesion strength, Bone tissue engineering

Background Distinguishing among species in the genus Lepista is difficult because of their similar morphologies. Methods To identify a suitable DNA barcode for identification of Lepista species, we assessed the following five regions: internal transcribed spacer (ITS), the intergenic spacer (IGS), nuclear ribosomal RNA subunit, mitochondrial small subunit rDNA, and tef1. A total of 134 sequences from 34 samples belong to eight Lepista species were analyzed. The utility of each region as a DNA barcode was assessed based on the success rates of its PCR amplification and sequencing, and on its intra- and inter-specific variations. Results The results indicated that the ITS region could distinguish all species tested. We therefore propose that the ITS region can be used as a DNA barcode for the genus Lepista. In addition, a phylogenetic tree based on the ITS region showed that the tested eight Lepista species, including two unrecognized species, formed eight separate and well-supported clades.

Abstract Radical resection is the most effective method for malignant tumor treatments. However, conventional imaging cannot fully satisfy the clinical needs of surgical navigation. This study presents a novel triple‐modality positron emission tomography (PET)–Cerenkov radiation energy transfer (CRET)–confocal laser endomicroscopy (CLE) imaging strategy for intraoperative tumor imaging and surgical navigation. Using clinical radiopharmaceuticals and fluorescein sodium (FS), this strategy can accurately detect the tumor and guide the tumor surgery. The FS emission property under Cerenkov radiation excitation is investigated using 2‐deoxy‐2‐18F‐fluoroglucose and 11C‐choline. Performances of the PET–CRET–CLE imaging and the CRET–CLE image‐guided surgery are evaluated on mouse models. The CRET signal at 8 mm depth is stronger than the Cerenkov luminescence at 1 mm depth in phantoms. In vivo experiments indicate that 0.5 mL kg−1 of 10% FS generates the strongest CRET signal, which can be observed immediately after FS injection. A surgical navigation study shows that the tumors are precisely detected and resected using intraoperative CRET–CLE. In summary, a PET–CRET–CLE triple‐modality imaging strategy is developed. This strategy can detect the tumors and precisely guide the tumor resection using clinical pharmaceuticals. This triple‐modality imaging shows high potential in surgical navigation research and clinical translation.

Influenza A virus (IAV) neuraminidase (NA) receptor-destroying activity and hemagglutinin (HA) receptor-binding affinity need to be balanced with the host receptor repertoire for optimal viral fitness. NAs of avian, but not human viruses, contain a functional 2nd sialic acid (SIA)-binding site (2SBS) adjacent to the catalytic site, which contributes to sialidase activity against multivalent substrates. The receptor-binding specificity and potentially crucial contribution of the 2SBS to the HA-NA balance of virus particles is, however, poorly characterized. Here, we elucidated the receptor-binding specificity of the 2SBS of N2 NA and established an important role for this site in the virion HA-NA-receptor balance. NAs of H2N2/1957 pandemic virus with or without a functional 2SBS and viruses containing this NA were analysed. Avian-like N2, with a restored 2SBS due to an amino acid substitution at position 367, was more active than human N2 on multivalent substrates containing α2,3-linked SIAs, corresponding with the pronounced binding-specificity of avian-like N2 for these receptors. When introduced into human viruses, avian-like N2 gave rise to altered plaque morphology and decreased replication compared to human N2. An opposite replication phenotype was observed when N2 was combined with avian-like HA. Specific bio-layer interferometry assays revealed a clear effect of the 2SBS on the dynamic interaction of virus particles with receptors. The absence or presence of a functional 2SBS affected virion-receptor binding and receptor cleavage required for particle movement on a receptor-coated surface and subsequent NA-dependent self-elution. The contribution of the 2SBS to virus-receptor interactions depended on the receptor-binding properties of HA and the identity of the receptors used. We conclude that the 2SBS is an important and underappreciated determinant of the HA-NA-receptor balance. The rapid loss of a functional 2SBS in pandemic viruses may have served to balance the novel host receptor-repertoire and altered receptor-binding properties of the corresponding HA protein.

Cerenkov Luminescence Tomography (CLT) is a non-invasive three-dimensional in vivo detection technology. However, due to the ill-posedness of CLT, the reconstructed result has many artifacts, which will mislead the researchers to make a wrong diagnostic decision. Enlightened by the development of artificial neural networks, we proposed a Fuzzy Autoencoder Clustering method to eliminate these artifacts and improve the reconstruction quality. To assess the performance of our method, several numerical simulation experiments and real physical phantom experiments are conducted. Compared with the raw reconstruction results and the commonly used manual threshold processed ones, it is demonstrated that our method is capable of filtering the artifact areas effectively, making reconstruction results clearer. It is anticipated that the method presented in this paper will help advance the CLT technology and promote the clinic translation of CLT technology.

Objective: Adiponectin receptors (AdipoRs) are located on neurons of the hypothalamus involved in metabolic regulation – including arcuate proopiomelanocortin (Pomc) and Neuropeptide Y/Agouti-related peptide (NPY/AgRP) neurons. AdipoRs play a critical role in regulating glucose and fatty acid metabolism by initiating several signaling cascades overlapping with Leptin receptors (LepRs). However, the mechanism by which adiponectin regulates cellular activity in the brain remains undefined. Methods: In order to resolve this issue, we utilized neuron-specific transgenic mouse models to identify Pomc and NPY/AgRP neurons which express LepRs for patch-clamp electrophysiology experiments. Results: We found that leptin and adiponectin synergistically activated melanocortin neurons in the arcuate nucleus. Conversely, NPY/AgRP neurons were inhibited in response to adiponectin. The adiponectin-induced depolarization of arcuate Pomc neurons occurred via activation of Phosphoinositide-3-kinase (PI3K) signaling, independent of 5′ AMP-activated protein kinase (AMPK) activity. Adiponectin also activated melanocortin neurons at various physiological glucose levels. Conclusions: Our results demonstrate a requirement for PI3K signaling in the acute adiponectin-induced effects on the cellular activity of arcuate melanocortin neurons. Moreover, these data provide evidence for PI3K as a substrate for both leptin and adiponectin to regulate energy balance and glucose metabolism via melanocortin activity. Author Video: Author Video Watch what authors say about their articles Keywords: Melanocortin, Obesity, Diabetes, Energy balance, Patch-clamp, Electrophysiology

Proliferating progenitor cells undergo changes in competence to give rise to post-mitotic progeny of specialized function. These cell-fate transitions typically involve dynamic regulation of gene expression by histone methyltransferase (HMT) complexes. However, the composition, roles, and regulation of these assemblies in regulating cell-fate decisions in vivo are poorly understood. Using unbiased affinity purification and mass spectrometry, we identified the uncharacterized C2H2-like zinc finger protein ZNF644 as a G9a/GLP-interacting protein and co-regulator of histone methylation. In zebrafish, functional characterization of ZNF644 orthologs, znf644a and znf644b, revealed complementary roles in regulating G9a/H3K9me2-mediated gene silencing during neurogenesis. The non-overlapping requirements for znf644a and znf644b during retinal differentiation demarcate critical aspects of retinal differentiation programs regulated by differential G9a-ZNF644 associations, such as transitioning proliferating progenitor cells toward differentiation. Collectively, our data point to ZNF644 as a critical co-regulator of G9a/H3K9me2-mediated gene silencing during neuronal differentiation.

Background: Blind and excessive application of fertilizers was found during the cultivation of Panax notoginseng in fields, as well as increase in root rot disease incidence. Methods: Both “3414” application and orthogonal test designs were performed at Shilin county, Yunnan province, China, for NPK (nitrogen, phosphorus, and potassium) and mineral fertilizers, respectively. The data were used to construct the one-, two-, and three-factor quadratic regression models. The effect of fertilizer deficiency on root yield loss was also analyzed to confirm the result predicted by these models. A pot culture experiment was performed to observe the incidence rate of root rot disease and to obtain the best range in which the highest yield of root and saponins could be realized. Results: The best application strategy for NPK fertilizer was 0 kg/667 m2, 17.01 kg/667 m2, and 56.87 kg/667 m2, respectively, which can produce the highest root yield of 1,861.90 g (dried root of 100 plants). For mineral fertilizers, calcium and magnesium fertilizers had a significant and positive effect on root yield and the content of four active saponins, respectively. The severity of root rot disease increased with the increase in soil moisture. The best range of soil moisture varied from 0.56 FC (field capacity of water) to 0.59 FC, when the highest yield of root and saponins could be realized as well as the lower incidence rate of root disease. Conclusion: These results indicate that the amount of nitrogen fertilizer used in these fields is excessive and that of potassium fertilizer is deficient. Higher soil moisture is an important factor that increases the severity of the root rot disease.

Ytterbium silicate coatings were deposited on SiCf/SiC ceramics matrix composite (CMC) substrates by plasma spray-physical vapor deposition (PS-PVD), and the microstructures and phase constituents of the coatings were studied. The results show that the Yb2SiO5 coating prepared with high power and low pressure (65 kW/2 mbar) had quasi-columnar structure, mainly deposited from the vapor phase, whereas the coating prepared with low power and high pressure (40 kW/10 mbar) had a typical layered structure, mainly deposited from the liquid phase. The deposition efficiency of parameter A (~2 μm/min) was also significantly lower than that of parameter B (~20 μm/min). After annealing at 1300 °C for 20 h, the coating prepared by 65 kW/2 mbar was mainly composed of ytterbium disilicate phase (77.2 wt %). The coating also contained some silicon-rich phase. The coating prepared by 40 kW/10 mbar basically consisted of ytterbium monosilicate (63.6 wt %). In addition, a small amount of silicon-rich phase and ytterbium-rich phase were also present in the coating. Accompanied with calculation results by the FactSage software, the cause of deviations in phase compositions was analyzed.

Various Ytterbium-Silicate-Mullite composites were successfully fabricated by adding Yb2SiO5 into mullite ceramics and then using pressureless sintering at 1550 °C. The influence of Yb2SiO5 addition on the microstructure, mechanical properties, and thermal conductivity of ytterbium-silicate-mullite composites was investigated. Results show that the composites mainly consisted of a mullite matrix and some in situ formed Yb2Si2O7 and Al2O3 phases. By the addition of Yb2SiO5, the Vickers hardness of composites reached ~9.0 at an additive concentration of 5 mol %. Fracture toughness increased to ~2.7 MPa·m1/2 at the additive concentration of 15 mol %, owing to the trans-granular fracture and crack deflection of the pinning effect of the Al2O3 phase in the composites. With the increase of the Al2O3 phase in the composite, the thermal conductivity for the 15YbAM reached around 4.0 W/(m·K) at 1200 °C. Although the thermal conductivity increased, it is still acceptable for such composites to be used as environmental barrier coatings.

Interactions of influenza A virus (IAV) with sialic acid (SIA) receptors determine viral fitness and host tropism. Binding to mucus decoy receptors and receptors on epithelial host cells is determined by a receptor-binding hemagglutinin (HA), a receptor-destroying neuraminidase (NA) and a complex in vivo receptor-repertoire. The crucial but poorly understood dynamics of these multivalent virus-receptor interactions cannot be properly analyzed using equilibrium binding models and endpoint binding assays. In this study, the use of biolayer interferometric analysis revealed the virtually irreversible nature of IAV binding to surfaces coated with synthetic sialosides or engineered sialoglycoproteins in the absence of NA activity. In addition to HA, NA was shown to be able to contribute to the initial binding rate while catalytically active. Virus-receptor binding in turn contributed to receptor cleavage by NA. Multiple low-affinity HA-SIA interactions resulted in overall extremely high avidity but also permitted a dynamic binding mode, in which NA activity was driving rolling of virus particles over the receptor-surface. Virus dissociation only took place after receptor density of the complete receptor-surface was sufficiently decreased due to NA activity of rolling IAV particles. The results indicate that in vivo IAV particles, after landing on the mucus layer, reside continuously in a receptor-bound state while rolling through the mucus layer and over epithelial cell surfaces driven by the HA-NA-receptor balance. Quantitative BLI analysis enabled functional examination of this balance which governs this dynamic and motile interaction that is expected to be crucial for penetration of the mucus layer and subsequent infection of cells by IAV but likely also by other enveloped viruses carrying a receptor-destroying enzyme in addition to a receptor-binding protein.

In 2014, novel highly pathogenic avian influenza A H5N2, H5N5, H5N6, and H5N8 viruses caused outbreaks in Asia, Europe, and North America. The H5 genes of these viruses form a monophyletic group that evolved from a clade 2.3.4 H5N1 variant. This rapid emergence of new H5Nx combinations is unprecedented in the H5N1 evolutionary history.

DCAMKL1 (doublecortin and CaM kinase-like 1) has been found to be overexpressed and function as an oncogene in several types of cancer, but there are limited reports on the role of DCAMKL1 in bladder cancer. The messenger RNA and protein expression of DCAMKL1 in bladder cancer tissues and cell lines was measured by quantitative reverse transcription polymerase chain reaction, western blotting, or immunohistochemistry. The correlation between DCAMKL1 protein expression and clinicopathological characteristics was analyzed. Univariate and multivariate Cox regression models were adopted to evaluate prognostic significance of DCAMKL1 in bladder cancer patients. In our results, DCAMKL1 messenger RNA and protein were overexpressed in bladder cancer tissues compared with adjacent normal tissues. DCAMKL1 protein overexpression was positively associated with clinical stage, muscularis invasion, lymph node metastasis, and distant metastasis. The univariate and multivariate analyses suggested DCAMKL1 protein overexpression was an unfavorable prognostic factor in bladder cancer patients. In conclusion, DCAMKL1 is an independent poor prognostic factor for bladder cancer patients.

Chromatin regulation is driven by multicomponent protein complexes, which form functional modules. Deciphering the components of these modules and their interactions is central to understanding the molecular pathways these proteins are regulating, their functions, and their relation to both normal development and disease. We describe the use of affinity purifications of tagged human proteins coupled with mass spectrometry to generate a protein-protein interaction map encompassing known and predicted chromatin-related proteins. On the basis of 1,394 successful purifications of 293 proteins, we report a high-confidence (85% precision) network involving 11,464 protein-protein interactions among 1,738 different human proteins, grouped into 164 often overlapping protein complexes with a particular focus on the family of JmjC-containing lysine demethylases, their partners, and their roles in chromatin remodeling. We show that RCCD1 is a partner of histone H3K36 demethylase KDM8 and demonstrate that both are important for cell-cycle-regulated transcriptional repression in centromeric regions and accurate mitotic division.

Hydrogen peroxide (H2O2) and nitric oxide (NO) are key signaling molecules in cells whose levels are increased in response to various stimuli and are involved in plant secondary metabolite synthesis. In this paper, the roles of H2O2 and NO on salvianolic acid B (Sal B) production in salicylic acid (SA)-induced Salvia miltiorrhiza cell cultures were investigated. The results showed that H2O2 could be significantly elicited by SA, even though IMD (an inhibitor of NADPH oxidase) or DMTU (a quencher of H2O2) were employed to inhibit or quench intracellular H2O2. These elicited H2O2 levels significantly increased NO production by 1.6- and 1.46 fold in IMD+SA and DMTU+SA treatments, respectively, and induced 4.58- and 4.85-fold Sal B accumulation, respectively. NO was also markedly elicited by SA, in which L-NNA (an inhibitor of NO synthase) and cPTIO (a quencher of NO) were used to inhibit or quench NO within cells, and the induced NO could significantly enhance H2O2 production by 1.92- and 1.37-fold in L-NNA+SA and cPTIO+SA treatments, respectively, and 3.27- and 1.50-fold for Sal B accumulation, respectively. These results indicate that elicitation of SA for either H2O2 or NO was independent, and the elicited H2O2 or NO could act independently or synergistically to induce Sal B accumulation in SA-elicited cells.

In this study, three-dimensional numerical studies have been performed to investigate the performance of a rotating detonation combustor with a diverging nozzle downstream. The effects of a diverging nozzle on the formation and propagation process of a detonation wave and typical flow field parameters in a rotating detonation combustor are mainly discussed. The results indicate that the diverging nozzle downstream is an important factor affecting the performance and design of a rotating detonation combustor. The diverging nozzle does not affect the formation and propagation process of the rotating detonation wave, while the time of two key wave collisions are delayed during the formation process of the detonation wave. With increases of the diverging angle, the rotating detonation combustor with the diverging nozzle can still maintain a certain pressure gain performance. Both the diverging nozzle and diverging angle have great influence on the flow field parameters of the rotating detonation combustor, including reducing the high pressure and temperature load, making the distribution of the outlet parameters uniform, and changing the local supersonic flow at the outlet. Among them, the outlet static pressure is reduced by up to 88.32%, and the outlet static temperature is reduced by up to 32.12%. This evidently improves the working environment of the combustor while reducing the thermodynamic and aerodynamic loads at the outlet. In particular, the diverging nozzle does not affect the supersonic characteristics of the outlet airflow, and on this basis, the Mach number becomes coincident and enhanced.

Fluorescence molecular tomography (FMT) allows the detection and quantification of various biological processes in small animals in vivo, which expands the horizons of pre-clinical research and drug development. Efficient three-dimensional (3D) reconstruction algorithm is the key to accurate localization and quantification of fluorescent target in FMT. In this paper, 3D reconstruction of FMT is regarded as a sparse signal recovery problem and the compressive sampling matching pursuit (CoSaMP) algorithm is adopted to obtain greedy recovery of fluorescent signals. Moreover, to reduce the modeling error, the simplified spherical harmonics approximation to the radiative transfer equation (RTE), more specifically SP3, is utilized to describe light propagation in biological tissues. The performance of the proposed reconstruction method is thoroughly evaluated by simulations on a 3D digital mouse model by comparing it with three representative greedy methods including orthogonal matching pursuit (OMP), stagewise OMP(StOMP), and regularized OMP (ROMP). The CoSaMP combined with SP3 shows an improvement in reconstruction accuracy and exhibits distinct advantages over the comparative algorithms in multiple targets resolving. Stability analysis suggests that CoSaMP is robust to noise and performs stably with reduction of measurements. The feasibility and reconstruction accuracy of the proposed method are further validated by phantom experimental data.

Spatially targeted optical microproteomics (STOMP) is a novel proteomics technique for interrogating micron-scale regions of interest (ROIs) in mammalian tissue, with no requirement for genetic manipulation. Methanol or formalin-fixed specimens are stained with fluorescent dyes or antibodies to visualize ROIs, then soaked in solutions containing the photo-tag: 4-benzoylbenzyl-glycyl-hexahistidine. Confocal imaging along with two photon excitation are used to covalently couple photo-tags to all proteins within each ROI, to a resolution of 0.67 µm in the xy-plane and 1.48 µm axially. After tissue solubilization, photo-tagged proteins are isolated and identified by mass spectrometry. As a test case, we examined amyloid plaques in an Alzheimer's disease (AD) mouse model and a post-mortem AD case, confirming known plaque constituents and discovering new ones. STOMP can be applied to various biological samples including cell lines, primary cell cultures, ex vivo specimens, biopsy samples, and fixed post-mortem tissue.

We conducted a retrospective analysis to assess the feasibility of involved field irradiation (IFI) in elderly patients with esophageal squamous cell cancer (ESCC).We performed a retrospective review of the records of elderly patients (≥ 70 years) with unresectable ESCC and no distant metastases who received treatment with radiotherapy between January 2009 and March 2013. According to the irradiation volume, patients were allocated into either the elective nodal irradiation (ENI) group or the IFI group. Overall survival (OS), progression-free survival (PFS) and treatment-related toxicities were compared between the two groups.A total of 137 patients were enrolled. Fifty-four patients (39.4%) were allocated to the ENI group and 83 patients (60.6%) to the IFI group, the median doses in the two groups were 60 Gy and 59.4 Gy, respectively. For the entire group, the median survival time (MST) and PFS were 16 months and 12 months, respectively. The median PFS and 3-year PFS rate in the ENI group were 13 months and 20.6%, compared to 11 months and 21.0% in the IFI groups (p = 0.61). The MST and 3-year OS rate in the ENI and IFI groups were 17 months and 26.4% and 15.5 months and 21.7%, respectively (p = 0.25). The rate of grade ≥ 3 acute irradiation esophagitis in the ENI group was significantly higher than that in the IFI group (18.5% vs. 6.0%; p = 0.027). Other grade ≥ 3 treatment-related toxicities did not significantly differ between the two groups.IFI resulted in decreased irradiation toxicities without sacrificing OS in elderly patients with ESCC.

Recently, the simplified spherical harmonics equations (SPN) model has attracted much attention in modeling the light propagation in small tissue geometries at visible and near-infrared wavelengths. In this paper, we report an efficient numerical method for fluorescence molecular tomography (FMT) that combines the advantage of SPN model and adaptive hp finite element method (hp-FEM). For purposes of comparison, hp-FEM and h-FEM are, respectively applied to the reconstruction process with diffusion approximation and SPN model. Simulation experiments on a 3D digital mouse atlas and physical experiments on a phantom are designed to evaluate the reconstruction methods in terms of the location and the reconstructed fluorescent yield. The experimental results demonstrate that hp-FEM with SPN model, yield more accurate results than h-FEM with diffusion approximation model does. The phantom experiments show the potential and feasibility of the proposed approach in FMT applications.

Salicylic acid (SA) is an elicitor to induce the biosynthesis of secondary metabolites in plant cells. Hydrogen peroxide (H2O2) plays an important role as a key signaling molecule in response to various stimuli and is involved in the accumulation of secondary metabolites. However, the relationship between them is unclear and their synergetic functions on accumulation of secondary metabolites are unknown. In this paper, the roles of SA and H2O2 in rosmarinic acid (RA) production in Salvia miltiorrhiza cell cultures were investigated. The results showed that SA significantly enhanced H2O2 production, phenylalanine ammonia-lyase (PAL) activity, and RA accumulation. Exogenous H2O2 could also promote PAL activity and enhance RA production. If H2O2 production was inhibited by NADPH oxidase inhibitor (IMD) or scavenged by quencher (DMTU), RA accumulation would be blocked. These results indicated that H2O2 is secondary messenger for signal transduction, which can be induced by SA, significantly and promotes RA accumulation.

OBJECTIVES:This study is to investigate if polymerase I and transcript release factor (PTRF) acts as a modulator in glioblastoma (GBM) chemoresistance. METHODS:Multidrug resistant (MDR) GBM cell line U251AR was established by exposing the U251 cell line to imatinib. The 2D-DIGE and MALDI-TOF/TOF-MS were performed on U251 and U251AR cell lines to screen MDR-related proteins. The expression of PTRF was determined by Western blot and quantitative RT-PCR analyses. RESULTS:When compared with the parental U251 cells, expression of 21 proteins was significantly altered in U251AR cells. Among the 21 differentially expressed proteins, the expression of PTRF was up-regulated by 2.14 folds in U251AR cells when compared with that in the parental U251 cells. Knockdown of PTRF in GBM cell lines significantly increased chemosensitivity of cells to various chemical drugs and decreased the expression levels of caveolin1, a major structural component of caveolae. Expression levels of PTRF and caveolin1 were significantly up-regulated in the relapsed GBM patients. The mRNA level of PTRF and caveolin1 showed a positive correlation in the same GBM specimens. CONCLUSIONS:Our results indicate that PTRF acts as a modulator in GBM chemoresistance.

FGF21 is a novel secreted protein with robust anti-diabetic, anti-obesity, and anti-atherogenic activities in preclinical species. In the current study, we investigated the signal transduction pathways downstream of FGF21 following acute administration of the growth factor to mice. Focusing on adipose tissues, we identified FGF21-mediated downstream signaling events and target engagement biomarkers. Specifically, RNA profiling of adipose tissues and phosphoproteomic profiling of adipocytes, following FGF21 treatment revealed several specific changes in gene expression and post-translational modifications, specifically phosphorylation, in several relevant proteins. Affymetrix microarray analysis of white adipose tissues isolated from both C57BL/6 (fed either regular chow or HFD) and db/db mice identified over 150 robust potential RNA transcripts and over 50 potential secreted proteins that were changed greater than 1.5 fold by FGF21 acutely. Phosphoprofiling analysis identified over 130 phosphoproteins that were modulated greater than 1.5 fold by FGF21 in 3T3-L1 adipocytes. Bioinformatic analysis of the combined gene and phosphoprotein profiling data identified a number of known metabolic pathways such as glucose uptake, insulin receptor signaling, Erk/Mapk signaling cascades, and lipid metabolism. Moreover, a number of novel events with hitherto unknown links to FGF21 signaling were observed at both the transcription and protein phosphorylation levels following treatment. We conclude that such a combined "omics" approach can be used not only to identify robust biomarkers for novel therapeutics but can also enhance our understanding of downstream signaling pathways; in the example presented here, novel FGF21-mediated signaling events in adipose tissue have been revealed that warrant further investigation.

Objective. This study evaluated the long-term effects and clinical significance of latent abnormal pathology on elder living donor kidney graft function after renal transplantation in China. Methods. One-hundred and thirty-eight living donor renal transplantations have been carried out at our hospital in recent years. Of these, 72 Time-Zero biopsies were performed and used in this analysis. Clinical data were retrospectively measured at 3, 6, 12, and 24 months after renal transplants. Relationships and effects from biopsy results taken from implanted donor kidney grafts were analyzed. Results. Time-Zero biopsy pathology results from donor kidneys showed that 48.61% of donor kidneys had latent abnormal changes; arterial lesions of donor kidneys had significant effects on the renal function of grafts after 2 years' transplantation; correlations between donor age and arterial lesions were significant; and Time-Zero biopsy pathology results could help predict the long-term function of a renal graft. Conclusions. Existing latent pathological changes of an elder living donor kidney before transplantation could affect long-term renal function. Whether a senior donor is used should be very carefully considered.

Based on the Scaled conjugate gradient (SCALCG) method presented by Andrei (2007) and the projection method presented by Solodov and Svaiter, we propose a SCALCG method for solving monotone nonlinear equations with convex constraints. SCALCG method can be regarded as a combination of conjugate gradient method and Newton-type method for solving unconstrained optimization problems. So, it has the advantages of the both methods. It is suitable for solving large-scale problems. So, it can be applied to solving large-scale monotone nonlinear equations with convex constraints. Under reasonable conditions, we prove its global convergence. We also do some numerical experiments show that the proposed method is efficient and promising.

A natural male sterile mutant of Salvia miltiorrhiza (Labiatae, Sh-B) was found during field survey in 2002. Our objective was to analyze its genetic mechanism for producing F1 hybrid seeds and to develop a molecular marker linked to male sterile gene for selection of a hybrid parent line. The segregation ratios of male sterile plants to fertile plants in the progenies of both testcross and backcross were 1:1 in continuous experiments conducted in 2006-2009. The male sterile Sh-B was heterozygous (Msms). The male sterile plants could capture most pollen (2 granule/cm(2)·24 h) with row ratio (female:male 2:1) within 45-cm distance and harvest the largest amount of 6495 g hybrid seeds per hectare. We also developed DNA markers linked to the male sterile gene in a segregating population using bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) techniques. The segregating population was subjected to BSA-AFLP with 128 primer combinations. One out of fourteen AFLP markers (E11/M4208) was identified as tightly linked to the dominant male sterile gene with a recombination frequency of 6.85% and at a distance of 6.89 cM. This marker could be converted to PCR-based assay for large-scale selection of fertile plants in MAS (marker-assisted selection) at the seedling stage. Blastn analysis indicated that the male sterile gene sequence showed higher identity with nucleotides in Arabidopsis chromosome 1-5, and was more likely to encode S-adenosylmethionine-dependent methyltransferase, in which DNA methylation regulated the development of plant gametogenesis.

This paper studies the finite element (FE) approximation to a second-type variational inequality. The supe rclose and superconvergence results are obtained for conforming bilinear FE and nonconforming EQrot FE schemes under a reasonable regularity of the exact solution u∈H5/2(Ω), which seem to be never discovered in the previous literature. The optimal L2-norm error estimate is also derived for EQrot FE. At last, some numerical results are provided to verify the theoretical analysis.

Pharmacodynamic (PD) biomarkers are an increasingly valuable tool for decision-making and prioritization of lead compounds during preclinical and clinical studies as they link drug-target inhibition in cells with biological activity. They are of particular importance for novel, first-in-class mechanisms, where the ability of a targeted therapeutic to impact disease outcome is often unknown. By definition, proximal PD biomarkers aim to measure the interaction of a drug with its biological target. For kinase drug discovery, protein substrate phosphorylation sites represent candidate PD biomarkers. However, substrate phosphorylation is often controlled by input from multiple converging pathways complicating assessment of how potently a small molecule drug hits its target based on substrate phoshorylation measurements alone. Here, we report the use of quantitative, differential mass-spectrometry to identify and monitor novel drug-regulated phosphorylation sites on target kinases. Autophosphorylation sites constitute clinically validated biomarkers for select protein tyrosine kinase inhibitors. The present study extends this principle to phosphorylation sites in serine/threonine kinases looking beyond the T-loop autophosphorylation site. Specifically, for the 3'-phosphoinositide-dependent protein kinase 1 (PDK1), two phospho-residues p-PDK1(Ser410) and p-PDK1(Thr513) are modulated by small-molecule PDK1 inhibitors, and their degree of dephosphorylation correlates with inhibitor potency. We note that classical, ATP-competitive PDK1 inhibitors do not modulate PDK1 T-loop phosphorylation (p-PDK1(Ser241)), highlighting the value of an unbiased approach to identify drug target-regulated phosphorylation sites as these are complementary to pathway PD biomarkers. Finally, we extend our analysis to another protein Ser/Thr kinase, highlighting a broader utility of our approach for identification of kinase drug-target engagement biomarkers.

Influenza A viruses (IAVs) quickly adapt to new environments and are well known to cross species barriers. To reveal a molecular basis for these phenomena, we compared the Ser/Thr and Tyr phosphoproteomes of murine lung epithelial cells early and late after infection with mouse-adapted SC35M virus or its non-adapted SC35 counterpart. With this analysis we identified a large set of upregulated Ser/Thr phosphorylations common to both viral genotypes, while Tyr phosphorylations showed little overlap. Most of the proteins undergoing massive changes of phosphorylation in response to both viruses regulate chromatin structure, RNA metabolism and cell adhesion, including a focal adhesion kinase (FAK)-regulated network mediating the regulation of actin dynamics. IAV also affected phosphorylation of activation loops of 37 protein kinases including FAK and several phosphatases, many of which were not previously implicated in influenza virus infection. Inhibition of FAK proved its contribution to IAV infection. Novel phosphorylation sites were found on IAV-encoded proteins, and the functional analysis of selected phosphorylation sites showed that they either support (NA Ser178) or inhibit (PB1 Thr223) virus propagation. Together, this data allow novel insights into IAV-triggered regulatory phosphorylation circuits and signaling networks. [ABSTRACT FROM AUTHOR]