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The cyclic adenosine monophosphate (cAMP) signalling pathway plays an important role in the regulation of the development and pathogenicity of filamentous fungi. cAMP-dependent protein kinase A (PKA) is the conserved element downstream of cAMP, and its diverse mechanisms in multiple filamentous fungi are not well known yet. In the present study, gene knockout mutants of two catalytic subunits of PKA (PKA-C) in Setosphaeria turcica were created to illustrate the regulatory mechanisms of PKA-Cs on the development and pathogenicity of S. turcica. As a result, StPkaC2 was proved to be the main contributor of PKA activity in S. turcica. In addition, it was found that both StPkaC1 and StPkaC2 were necessary for conidiation and invasive growth, while only StPkaC2 played a negative role in the regulation of filamentous growth. We reveal that only StPkaC2 could interact with the transcription factor StEfg1, and it inhibited the transcription of StRAB1, a Rab GTPase homologue coding gene in S. turcica, whereas StPkaC1 could specifically interact with a transcriptional regulator StFlo8, which could rescue the transcriptional inhibition of StEfg1 on StRAB1. We also demonstrated that StRAB1 could positively influence the biosynthesis of chitin in hyphae, thus changing the filamentous growth. Our findings clarify that StPkaC2 participates in chitin biosynthesis to modulate mycelium development by targeting the Efg1-mediated transcription of StRAB1, while StFlo8, interacting with StPkaC1, acts as a negative regulator during this process.

Shengjiang Xiexin decoction, a traditional Chinese medical formula, has been utilized to alleviate the delayed-onset diarrhea induced by irino tecan. However, the chemical constituents of this formula and the activities of its constituents remain unclear. In this study, ultrahigh-performance liquid chromatography-quadrupole/orbitrap high-resolution mass spectrometry was employed to comprehensively analyze the chemical constituents of Shengjiang Xiexin decoction. A total of 270 components, including flavonoids, coumarins, triterpenoids, alkaloids, diarylheptanoids and others, were identified or characterized. Multidrug resistance-associated protein 2 is an efflux transporter responsible for regulating drug absorption. A total of 20 characteristic components from the formula were selected to evaluate their effects on the function of multidrug resistance-associated protein 2 using the vesicular transport assay. Glycyrrhizic acid and glycyrrhetinic acid were identified as potential multidrug resistance-associated protein 2 inhibitors, while 9 flavonoid aglycones increased the uptake of the substrate [

Supplementary Material 5: Supplementary Table 4. The primers used in ChIP analysis.

Supplementary Material 2: Supplementary Table 1. The sequences used for yeast one hybrid.

Supplementary Material 6

Supplementary Material 4: Supplementary Table 3. The sequences of the probes used in EMSA assay.

Supplementary Material 1: Supplementary Fig. 1 Comparison of pHIS2, the newly constructed pHIS2 prey library, and the previously constructed pHIS2 prey library. The green line indicates the two flanking sequences of the Sma I sites. (1) The map of pHIS2. (2) The prey library built in this study. (3) The prey library built in the previous study, which contains two types of pHIS2. The insertion is underlined.

Supplementary Material 3: Supplementary Table 2. The primers for the construction of the recombination Vector MBP-ERF2.

The fungal pathogen Setosphaeria turcica causes leaf blight on maize, which leads to considerable crop losses. However, how S. turcica establishes sustained systemic infection is largely unknown. Here, we report several novel factors contributing to S. turcica pathogenicity, identified using a genomic and transcriptional screen at different stages of S. turcica appressorium development. We identified two cytoskeleton regulators, SLM1 and SLM2, that are crucial for hypha and appressorium development. The SLM1 and SLM2 transcripts accumulated during germling stage but their levels were notably reduced at the appressorium stage. Deletion of SLM2 dramatically affected cell morphology, penetration ability, and pathogenicity. We also identified three different types of S. turcica glycosyl hydrolases that are critical for plant cell wall degradation. Their transcripts accumulated during the appressorium infection stage induced by cellophane and maize leaf. Most importantly, we characterized a novel and specific S. turcica effector, appressorium‐coupled effector 1 (StACE1), whose expression is coupled to appressorium formation in S. turcica. This protein is required for maize infection and induces cell death on expression in Nicotiana benthamiana. These observations suggest that the phytopathogen S. turcica is primed in advance with multiple strategies for maize infection, which are coupled to appressorium formation at the early infection stages., The phytopathogen Setosphaeria turcica is primed in advance with multiple strategies for maize infection, which are coupled to appressorium formation at the early infection stages.

The polymer separator is part of the gel polymer electrolyte (GPE) and plays a crucial role in battery. In this study, a novel PES-based separator for lithium-oxygen batteries was prepared using non-solvent-induced phase separation (NIPS) technique firstly and modified by doping Al2O3 nanoparticles. It is proved that the properties of the PES/Al2O3 separator are affected by the presence of Al2O3 nanoparticles. The results show that the PES/Al2O3 separator with Al2O3 content of 2 wt% and 4 wt% has lower crystallinity, higher electrolyte uptake, and better mechanical properties and thermal stability than PES-based separator without Al2O3. The resulting PES/Al2O3 separator containing 4 wt% Al2O3 has higher ionic conductivity (0.49 mS cm−1) and lithium ions transference number (0.28). Consequently, the assembled batteries with the PES/Al2O3 separator containing 4 wt% Al2O3 exhibit excellent cycling performance (75 cycles, 1000 mAh g−1 at 0.05 mA cm−2). Therefore, PES/Al2O3 separator has a significance meaning in the research of high-safety and long-cycle lithium-oxygen battery.

RNA polymerase II (RNAPII) is an essential machinery for catalyzing mRNA synthesis and controlling cell fate in eukaryotes. Although the structure and function of RNAPII have been relatively defined, the molecular mechanism of its assembly process is not clear. The identification and functional analysis of assembly factors will provide new understanding to transcription regulation. In this study, we identify that RTR1, a known transcription regulator, is a new multicopy genetic suppressor of mutants of assembly factors Gpn3, Gpn2, and Rba50. We demonstrate that Rtr1 is directly required to assemble the two largest subunits of RNAPII by coordinating with Gpn3 and Npa3. Deletion of RTR1 leads to cytoplasmic clumping of RNAPII subunit and multiple copies of RTR1 can inhibit the formation of cytoplasmic clump of RNAPII subunit in gpn3-9 mutant, indicating a new layer function of Rtr1 in checking proper assembly of RNAPII. In addition, we find that disrupted activity of Rtr1 phosphatase does not trigger the formation of cytoplasmic clump of RNAPII subunit in a catalytically inactive mutant of RTR1. Based on these results, we conclude that Rtr1 cooperates with Gpn3 and Npa3 to assemble RNAPII core.

In this paper, a gel polymer electrolyte based on sulfonated polyetherimide (SPEI)/polyetherimide (PEI) blending separator has been fabricated through non-solvent-induced phase separation (NIPS) method. The performance of the SPEI/PEI blending separator is investigated through the morphology and electrochemical analysis. The addition of SPEI polymer has great effect on the morphology of separator’s top and bottom surface. When the SPEI mass ratio increases, the SPEI/PEI blending separator shows high porosity (up to 72.80%), ion conductivity (up to 2.63 mS cm−1), and low tensile strength (down to 7.98 MPa). The discharge specific capacity, capacity retention rate, and average capacity attenuation rate of lithium-sulfur battery using SPEI-2 (separator with the weight ratio of SPEI/PEI = 2:8) are 736.1 mAh g−1, 57.2%, and 0.21% after 200 cycles, respectively. The result shows that the SPEI/PEI blending separator has lower capacity decays compared with SPEI-0 (separator with the weight ratio of SPEI/PEI = 0:10) and the commercial Celgard 2320 separator, which indicates that the existence of sulfonic acid group is conducive to the stability of the lithium-sulfur battery cycle. Therefore, it is concluded that the SPEI/PEI blending separator prepared by NIPS method is a potential separator for high-performance rechargeable lithium-sulfur battery.

Background. With the prevalence of mental issues worldwide, more and more people are suffering from psychological torture. Mental Health Gap Action Program (mhGAP) has been introduced to improve the life quality of humans. Objectives. To explore and synthesize evidence of participants’ experience of mental health first aid (MHFA) training course. Method. Peer-reviewed qualitative evidence was systematically reviewed and thematically synthesized. Cumulative Index to Nursing and Allied Health Literature (CINAHL), Medical Literature Analysis and Retrieval System Online (MEDLINE), Psychological Information (PsycINFO), PubMed, Psych ARTICLES, Web of Science, Joanna Briggs Institute (JBI), and National Institute for Health and Care Excellence (NICE) databases were searched for the inception of the present study. The study’s quality was appraised using the Critical Appraisal Checklist for Qualitative Research of Joanna Briggs Institute (JBI) appraisal tool. All the participants who have attended the MHFA training course (excluding instructors) setting were included. Results. Six papers published between 2005 and 2019 were included for thematic synthesis. The review indicated that MHFA had been a positive experience for participants. Conclusions. MHFA courses can provide participants with professional knowledge of mental health counseling and improve their knowledge, practice, and attitudes towards their patients. Professional MHFA training courses should therefore be popularized and promoted among other populations.

Yeast is one of the best-understood biological systems for genetic research. Over the last 40 years, geneticists have striven to search for homologues of tumor suppressors in yeast to simplify cancer research. The star tumor suppressor p21, downstream target of p53, is one of the primary factors on the START point through negatively regulating CycD/E-CDK, the yeast counterpart Cln3-Cdk1. Not like yeast Whi5 that was identified as the analog of the retinoblastoma tumor suppressor protein (Rb) and hence promoted to uncover the mechanism of its cancer suppression, homologue of p21 had not been found in yeast. Our lab identified Cip1 in budding yeast as a novel negative regulator of G1-Cdk1 and proposed that Cip1 is an analog of human p21. Recently, we demonstrated a dual repressive function of Cip1 on START timing via the redundant Cln3 and Ccr4 pathways. This work in yeast may help clarify the complex regulation in human p53-p21 signaling cascade. In this review, we will discuss the yeast paralogs of star tumor suppressors in the control of G1/S transition and present the new findings in this field.

RNA polymerase II is one of the most vital macromolecular complexes in eukaryotes and the assembly of such complete enzyme requires many factors. Three members of GPN-loop GTPase family Npa3/Gpn1, Gpn2, and Gpn3 participate in the biogenesis of RNA polymerase II with nonredundant roles. We show here that rapid degradation of each GPN protein in yeast leads to cytoplasmic accumulation of Rpb1 and defects in the assembly of RNA polymerase II, suggesting conserved functions of GPN paralogs for RNA polymerase II biogenesis as in humans. Taking advantage of a multicopy genetic screening, we identified GPN3 and assembly factor RBA50 among others as strong suppressors of npa3ts mutants. We further demonstrated that Npa3 interacts with Gpn3 and Rba50, similarly human Gpn1 physically interacts with Gpn3 and RPAP1 (human analog of Rba50). Moreover, a mutual dependency of protein levels of Npa3 and Gpn3 was also clearly presented in yeast using an auxin-inducible degron (AID) system. Interestingly, Rpb2, the second largest subunit of RNA polymerase II was determined to be the subunit that interacts with both Gpn1 and Rba50, indicating a close association of Npa3 and Rba50 in Rpb2 subcomplex assembly. Based on these results, we conclude that Npa3 interacts with Gpn3 and Rba50, for RNA polymerase II biogenesis. We therefore propose that multiple factors may coordinate through conserved regulatory mechanisms in the assembly of RNA polymerase complex.

The high performance and safety of lithium-sulfur (Li-S) batteries are the focus of research in recent years. The separator is an important part of the battery, whose performance stability directly affects the performance of the battery. What’s more, there are still many defects in the commercial polyolefin separator. In this work, a polyethylene/polyetherimide (PE/PEI) composite separator is designed by surface coating and non-solvent-induced phase separation (NIPS) method. The film breakage temperature and anti-puncture strength of the PE/PEI composite separator (181.96 °C, 7.90 N) are higher than those of the commercial PE separator (153.00 °C, 6.20 N), which will greatly ensure the safety problems caused by internal self-heating and external short circuit of Li-S batteries. The PE/PEI composite separator–based Li-S battery delivers a high initial specific capacity of 1265 mAh g−1 at 0.2 C and exhibits excellent rate capability compared with the commercial PE separator. Therefore, the PE/PEI composite separator is a candidate separator for the high-performance and high-safety Li-S battery.

As a valuable and powerful platform technology, yeast surface display (YSD) has been widely used in various biotechnological fields, but little in agricultural applications. Xylanase can be used as an elicitor in the plant–pathogen interaction system and induce corn defence response. In this study, the YSD technology was used to express the xylanase protein family from Setosphaeria turcica, which is a novel strategy for plant protection. First, five genes encoding xylanase were selected from the S. turcica genome database. The conserved domains of these genes were analyzed, which revealed that these genes were highly conserved. Here, we selected two of the five genes and expressed them by a prokaryotic expression system, but the two xylanase genes were expressed in inclusion bodies and showed no activity. To make them available in the application, we successfully constructed them in a YSD system. The products were detected for enzyme activities and the activities of two xylanases were 16.72 and 19.12 U/mL, respectively. We finally proved that the yeast-displaying xylanase was an efficient strategy to induce corn defence response, as four defence genes in corn, PR-1, PR-4, SOD and CAT were induced after treatment with the yeast displaying xylanases.

This paper outlines analysis the process gg → H → WW → llνν Monte Carlo, collision events with MH=125GeV. The discovery of the Higgs boson perfected the standard model and proposed more processes of the production, decay of the Higgs and the Higgs Mechanism. In this analysis, ggH process is reconstructed. Analyzed and calculated the probability of decay WW pairs as productions of decay process. Simulated Monte Carlo signal samples and background are compared to data from Atlas.

The novel male sterility line 12-14A of Wucai (Brassica campestris) was generated by backcrossing. In this study, morphological, molecular, and cytological analyses were performed to characterize this line. The phenotypes of flower organs of 12-14A were different from those of its maintainer line 12-14B, as the flowers of 12-14A were completely infertile. Molecular analysis confirmed that the male sterility line 12-14A was classified as the ogu CMS type. An analysis of anther development in the CMS line demonstrated that two types of abortion occur simultaneously: pollen mother cell abortion and tetrad-to-mononuclear microspore abortion. In addition, the mature anthers of 12-14A showed typical abortive features, such as appearing dry, shrunken, and scaly. A heterosis test of hybrids from CMS 12-14A and four inbred lines was carried out, and the F1 hybrids exhibited clear heterosis in plant weight, as well as in the protein, soluble sugar, and vitamin C content, suggesting high yields and high quality potential. These results indicate that this novel CMS line may have powerful potential in Wucai hybrid breeding.

We developed a novel Agrobacterium-mediated anther transformation for Wucai in planta, and in this procedure, the male germ line was the predominant target. Wucai (Brassica campestris L.), a variant of non-heading Chinese cabbage, is widely cultured in China and only improved by classic breeding methods. Here, a novel and efficient in planta Agrobacterium-mediated anther transformation method is developed based on the optimization of several factors that affect anther transformation. After optimization, transformation with the manual pollination application led to increased transient GUS expression in anthers (reaching 91.59%) and the transformation efficacies in planta (0.59–1.56% for four commercial cultivars). The stable integration and inheritance of the transgenes were further examined by molecular and genetic analyses. Three T2 transgenic lines presented a segregation ratio of 3:1, which was consistent with the Mendelian feature of a single dominant gene. In addition, the GUS histochemical assay and genetic crossing analysis revealed that the male germ line was the predominant target in this transformation. This optimized transformation system could provide a useful tool for both the improvement of cultivar qualities and investigation of functional genes in Wucai.

Traumatic cataract is a common complication of ocular contusion. Caused by various forms of trauma, the course and prognosis of traumatic cataract are also different. Intraocular lens (IOL) implantation is the most effective treatment at the present time and can provide good visual quality and near, medium, and far vision. In this study, an adaptive medical ultrasound image denoising algorithm was constructed based on the biological vision mechanism and was applied to observe IOL implantation and postoperative care of 30 patients with traumatic cataract. Results showed that eyesight could be restored to the original healthy state after IOL implantation, and eyesight was close to normal 3 months after surgery. The corresponding noise pixel block of the noise reduction algorithm was obviously the lowest. A total of 30 patients (9 patients with contusion, 13 patients with intraocular foreign body, and 8 patients with perforation) were treated with aspheric and multifocal IOL to achieve the expected outcome. The noise reduction algorithm constructed in this study effectively reduced the speckle noise of ultrasound images, improved the clarity of ultrasound images, and achieved the effective observation of medical nursing outcome after traumatic cataract IOL implantation. After IOL implantation, the retinal nerves of patients with traumatic cataract were affected to different degrees.

High-copy rescue genetic screening is a powerful strategy for the identification of suppression genetic interactions in the model eukaryotic organism Saccharomyces cerevisiae (budding yeast). The strain carrying the mutant allele of interest is transformed with a genomic library cloned in a high-copy plasmid. Each clone carries a genomic fragment insertion of around 10 kb, typically containing one to three complete genes under their own promoters. The high-copy vector favors the accumulation of high levels of the corresponding protein, aimed at suppressing the mutant phenotype. Typically, high-copy genetic screens select for viable clones under conditions restrictive or lethal for the query mutant strain. Here, we describe in detail the procedure to generate a high-copy genomic library and a protocol for rescue genetic screening and identification of the suppressor clones.

High-copy rescue genetic screening is a powerful strategy for the identification of suppression genetic interactions in the model eukaryotic organism Saccharomyces cerevisiae (budding yeast). The strain carrying the mutant allele of interest is transformed with a genomic library cloned in a high-copy plasmid. Each clone carries a genomic fragment insertion of around 10 kb, typically containing one to three complete genes under their own promoters. The high-copy vector favors the accumulation of high levels of the corresponding protein, aimed at suppressing the mutant phenotype. Typically, high-copy genetic screens select for viable clones under conditions restrictive or lethal for the query mutant strain. Here, we describe in detail the procedure to generate a high-copy genomic library and a protocol for rescue genetic screening and identification of the suppressor clones.

The objective of this study was isolation of humanized single-chain variable fragment (scFv) antibodies against IL1RAP, a promising cancer marker in leukemia and other tumors. Here, we screened for antibody fragments that recognized IL1RAP using a phage-display library of humanized scFv through several rounds of bio-panning and further FACS selection of enriched yeast-display sub-library. Two specific scFv clones 02 and 61 were isolated with high affinity against IL1RAP. Selected IL1RAP scFv antibodies containing Fc segment were expressed and purified from 293F cells. scFv-02 expressed and purified from mammalian cells exhibited higher IL1RAP binding affinity on IL1RAP-positive 293F cell surface compared to scFv-61. In addition, scFv-02 was significantly specific to IL1RAP, as no binding signal of IL1RAP was detected on the surface of IL1RAP CRISPR-Cas9 edited cell line generated in this study. Two scFv antibodies targeting IL1RAP were isolated by phage and yeast display screening. scFv-02 could serve as a promising material for further development of IL1RAP targeted immunotherapy of leukemia and other diseases. Our work also modified the process for the selection of scFv against targeted antigen by stepwise using phage display, yeast display and mammalian cell display techniques.

Fibrauretine is the main active ingredient in rattan stems of Fibraurea recisa Pierre. The aim of this study was to evaluate the cognitive-enhancing effects and underlying molecular mechanisms of fibrauretine compatibilized with ginsenosides on Alzheimer's disease (AD) induced in mice with amyloid β-protein (Aβ

The Cu-based catalysts supported on different materials (CeO2, ZrO2, CeO2-ZrO2) were tested towards syngas production from CO2 reforming of ethanol in order to determine the role of support. The physicochemical properties of the as-prepared catalysts were intensively explored by various techniques such as BET、TPR、Raman、SEM and TEM etc. Noticeably, the key features such as metal-support interaction, the reducibility and the oxygen vacancies decreased in the following order: CuCeZr > CuCe > Cu/Zr. Among the investigated catalysts, analysis of ethanol conversion as well as evaluation of the product distribution suggested CuCeZr sample as the most appropriate system for ethanol dry reforming. More precisely, CuCeZr catalyst exhibited full ethanol conversion at 700 °C and the H2/CO ratio was close to the theoretical one (1.0). It also showed the promising stability as long as 90 h time-on-stream. In addition, the reliance of ethanol turnover frequency (TOF) on the supports was noted and CuCeZr sample still presented the highest value versus that of the others. Characterization results of the used catalysts indicated that both the smaller extent of Cu sintering and the less deactivating coke formation was observed for CuCeZr catalyst after the stability test, because of its abundant oxygen defects and/or sufficient metal-support interfacial area. It should be noted that the nature of carbonaceous deposits on the used CuCeZr catalysts were mainly amorphous structure. In contrast, the encapsulating coke which was detrimental for catalyst behavior was obviously detected. Herein, the present work revealed that support played an important role in ethanol dry reforming and might provide some valuable insight for the development of efficient catalysts.