Intensive Ambulance-Delivered Blood-Pressure Reduction in Hyperacute Stroke.

BACKGROUND: Treatment of acute stroke, before a distinction can be made between ischemic and hemorrhagic types, is challenging. Whether very early blood-pressure control in the ambulance improves outcomes among patients with undifferentiated acute stroke is uncertain. METHODS: We randomly assigned patients with suspected acute stroke that caused a motor deficit and with elevated systolic blood pressure (≥150 mm Hg), who were assessed in the ambulance within 2 hours after the onset of symptoms, to receive immediate treatment to lower the systolic blood pressure (target range, 130 to 140 mm Hg) (intervention group) or usual blood-pressure management (usual-care group). The primary efficacy outcome was functional status as assessed by the score on the modified Rankin scale (range, 0 [no symptoms] to 6 [death]) at 90 days after randomization. The primary safety outcome was any serious adverse event. RESULTS: A total of 2404 patients (mean age, 70 years) in China underwent randomization and provided consent for the trial: 1205 in the intervention group and 1199 in the usual-care group. The median time between symptom onset and randomization was 61 minutes (interquartile range, 41 to 93), and the mean blood pressure at randomization was 178/98 mm Hg. Stroke was subsequently confirmed by imaging in 2240 patients, of whom 1041 (46.5%) had a hemorrhagic stroke. At the time of patients' arrival at the hospital, the mean systolic blood pressure in the intervention group was 159 mm Hg, as compared with 170 mm Hg in the usual-care group. Overall, there was no difference in functional outcome between the two groups (common odds ratio, 1.00; 95% confidence interval [CI], 0.87 to 1.15), and the incidence of serious adverse events was similar in the two groups. Prehospital reduction of blood pressure was associated with a decrease in the odds of a poor functional outcome among patients with hemorrhagic stroke (common odds ratio, 0.75; 95% CI, 0.60 to 0.92) but an increase among patients with cerebral ischemia (common odds ratio, 1.30; 95% CI, 1.06 to 1.60). CONCLUSIONS: In this trial, prehospital blood-pressure reduction did not improve functional outcomes in a cohort of patients with undifferentiated acute stroke, of whom 46.5% subsequently received a diagnosis of hemorrhagic stroke. (Funded by the National Health and Medical Research Council of Australia and others; INTERACT4 ClinicalTrials.gov number, NCT03790800; Chinese Trial Registry number, ChiCTR1900020534.).

IMperm: a fast and comprehensive IMmune Paired-End Reads Merger for sequencing data.

The adaptive immune receptor repertoire (AIRR), consisting of T- and B-cell receptors, is the core component of the immune system. The AIRR sequencing is commonly used in cancer immunotherapy and minimal residual disease (MRD) detection of leukemia and lymphoma. The AIRR is captured by primers and sequenced to yield paired-end (PE) reads. The PE reads could be merged into one sequence by the overlapped region between them. However, the wide range of AIRR data raises the difficulty, so a special tool is required. We developed a software package for IMmune PE reads merger of sequencing data, named IMperm. We used the k-mer-and-vote strategy to pin down the overlapped region rapidly. IMperm could handle all types of PE reads, eliminate adapter contamination and successfully merge low-quality and minor/non-overlapping reads. Compared with existing tools, IMperm performed better in both simulated and sequencing data. Notably, IMperm was well suited to processing the data of MRD detection in leukemia and lymphoma and detected 19 novel MRD clones in 14 patients with leukemia from previously published data. Additionally, IMperm can handle PE reads from other sources, and we demonstrated its effectiveness on two genomic and one cell-free deoxyribonucleic acid datasets. IMperm is implemented in the C programming language and consumes little runtime and memory. It is freely available at https://github.com/zhangwei2015/IMperm.

Somatic variant analysis suite: copy number variation clonal visualization online platform for large-scale single-cell genomics.

The recent advance of single-cell copy number variation (CNV) analysis plays an essential role in addressing intratumor heterogeneity, identifying tumor subgroups and restoring tumor-evolving trajectories at single-cell scale. Informative visualization of copy number analysis results boosts productive scientific exploration, validation and sharing. Several single-cell analysis figures have the effectiveness of visualizations for understanding single-cell genomics in published articles and software packages. However, they almost lack real-time interaction, and it is hard to reproduce them. Moreover, existing tools are time-consuming and memory-intensive when they reach large-scale single-cell throughputs. We present an online visualization platform, single-cell Somatic Variant Analysis Suite (scSVAS), for real-time interactive single-cell genomics data visualization. scSVAS is specifically designed for large-scale single-cell genomic analysis that provides an arsenal of unique functionalities. After uploading the specified input files, scSVAS deploys the online interactive visualization automatically. Users may conduct scientific discoveries, share interactive visualizations and download high-quality publication-ready figures. scSVAS provides versatile utilities for managing, investigating, sharing and publishing single-cell CNV profiles. We envision this online platform will expedite the biological understanding of cancer clonal evolution in single-cell resolution. All visualizations are publicly hosted at https://sc.deepomics.org.

Toxic effect of copper ions on anammox in IFFAS process filled with ZVI-10 modified carriers.

The inhibitory effects of heavy metals on anammox bacteria (AnAOB) have attracted attention worldwide. However, most are conducted in activated sludge rather than biofilm systems. The toxic effect and resistance response of anammox biofilm are not predictable from those of free-living AnAOB. Zero valent iron (ZVI) has been demonstrated to enhance anammox performance, but whether ZVI can promote AnAOB resistance to heavy metal stress remains unclear. Herein, the toxic effect of copper ions (Cu(II)) on anammox in integrated floating-film activated sludge (IFFAS) process filled with 10 wt% ZVI modified carriers (R1) was investigated. Results indicated half inhibiting concentration (IC50) of Cu(II) in R1 was 9.13 mg/L, which was much higher than that in R0 filled with conventional carriers made of high density polyethylene (HDPE) (3.94 mg/L). Long-term effect of Cu(II) demonstrated that Cu(II) concentrations less than 1.0 mg/L could not inhibit anammox biofilm significantly, whereas R1 performed better anammox process than R0 under the stress of 0.1-1.0 mg/L Cu(II). The ZVI modified carriers induced more extracellular polymeric substances (EPS) to trap Cu(II) to attenuate the toxicity to AnAOB. Besides, the activities of functional enzymes related to anammox (NIR and HDH), as well as heme-c contents, were always higher in R1 than R0 regardless of the Cu(II) dosage. Candidatus Kuenenia was identified as the predominant AnAOB, which had stronger resistance to Cu(II) stress compared to other genera in the IFFAS process. Metal resistance genes (MRGs) analysis identified AnAOB induced multi-responses to resist Cu(II) stress, such as the up-regulation of copC, cutA, cutC, cutF, cueR and cueO, to synthesize more proteins with functions of copper exocytosis, conjugation and oxidation.

Association between indicators of visceral lipid accumulation and infertility: a cross-sectional study based on U.S. women.

BACKGROUND: Evidence on the association between visceral lipid accumulation and infertility remains limited and controversial. Therefore, the current investigation is the first investigation to unveil this correlation by utilizing novel indicators of visceral lipid accumulation. METHODS: The present study utilized the NHANES 2013-2020 dataset. Researchers utilized multiple logistic regression, smoothed curve fitting, and subgroup analysis to investigate the associations of waist circumference (WC), metabolic score for visceral fat (METS-VF), lipid accumulation product (LAP), visceral adiposity index (VAI) with infertility. Additionally, the eXtreme Gradient Boosting (XGBoost) algorithm model was utilized to evaluate the relative importance of the factors. RESULTS: After adjusting for potential factors that could influence the results, researchers discovered that all these four indicators of visceral lipid accumulation exhibited strong positive correlations with the probability of infertility. The subgroup analysis demonstrated that the correlations remained consistent in the majority of subgroups (P for interaction > 0.05). The results of XGBoost algorithm model indicate that METS-VF is the most meaningful factor in infertility. The ROC curve research revealed that while METS-VF had the greatest AUC values, there was no variation in the AUC value of different markers of visceral fat accumulation (P > 0.05). CONCLUSIONS: The present investigation discovered that increased WC, METS-VF, LAP, and VAI were associated with a heightened prevalence of infertility.

Efficacy of Modified Yiwei Shengyang Decoction in Combination with FOLFOX4 Chemotherapy for Advanced Gastric Cancer.

Objective: To explore the clinical efficacy of modified Yiwei Shengyang Decoction combined with FOLFOX4 chemotherapy regimen in patients with advanced gastric cancer. Methods: Ninety patients with advanced gastric cancer, admitted to Cangzhou Central Hospital from January 2021 to December 2022, were randomized 1:1 into control and study groups. The control group received FOLFOX4 chemotherapy alone, while the study group received additional modified Yiwei Shengyang Decoction. Chinese medicine (TCM) symptom scores (TCM symptoms refer to the signs and manifestations of imbalances or disharmony within the body according to the principles of Traditional Chinese Medicine. These symptoms are assessed and diagnosed based on a holistic understanding of the individual's physical, mental, and emotional state. TCM symptoms may include various indicators such as pulse characteristics, tongue appearance, body temperature, complexion, energy levels, sleep patterns, appetite, digestion, pain, and specific subjective experiences reported by the patient, such as fatigue, anxiety, or insomnia), gastric cancer biomarkers such as serum CEA and CA199 levels, immune function, clinical efficacy, and side effects were compared. Results: Before treatment, both groups had similar TCM symptom scores. Post-treatment, the study group showed significantly greater reductions in appetite, epigastric pain, nausea, vomiting, and diarrhea scores compared to the control group (P < .001). After treatment, CEA and CA199 levels decreased significantly in both groups, with the study group exhibiting significantly lower levels than the control group (P = .001, .001). Post-treatment, CD3+ and CD4+ levels were higher in the study group, while CD8+ levels were lower than in the control group (P < .001). Treatment efficiency was significantly higher in the study group (62.33%) than in the control group (37.78%) (P = .02). Conclusion: Modified Yiwei Shengyang Decoction combined with FOLFOX4 chemotherapy regimen is a promising option for patients with gastric cancer. It significantly improves immune indicators and appetite, reduces adverse symptoms including epigastric pain, nausea, vomiting, and diarrhea, and substantially enhances quality of life. Moreover, traditional Chinese medicine treatment is safe and merits promotion in clinics.

Highly Efficient Capture of Volatile Iodine by Conjugated Microporous Polymers Constructed Using Planar 3- and 4-Connected Organic Monomers.

The effective capture and recovery of radioiodine species associated with nuclear fuel reprocessing is of significant importance in nuclear power plants. Porous materials have been proven to be one of the most effective adsorbents for the capture of radioiodine. In this work, we design and synthesize a series of conjugated microporous polymers (CMPs), namely, TPDA-TFPB CMP, TPDA-TATBA CMP, and TPDA-TECHO CMP, which are constructed based on a planar rectangular 4-connected organic monomer and three triangular 3-connected organic monomers, respectively. The resultant CMPs are characterized using various characterization techniques and used as effective adsorbents for iodine capture. Our experiments indicated that the CMPs exhibit excellent iodine adsorption capacities as high as 6.48, 6.25, and 6.37 g g-1 at 348 K and ambient pressure. The adsorption mechanism was further investigated and the strong chemical adsorption between the iodine and the imine/tertiary ammonia of the CMPs, 3D network structure with accessible hierarchical pores, uniform micromorphology, wide π-conjugated structure, and high-density Lewis-base sites synergistically contribute to their excellent iodine adsorption performance. Moreover, the CMPs demonstrated good recyclability. This work provides guidance for the construction of novel iodine adsorbent materials with high efficiency in the nuclear power field.

A demonstration concentrating solar power plant in China: Carbon neutrality, energy renewability and policy perspectives.

Concentrating solar power (CSP) is considered as a promising renewable electricity source due to its superiority in providing dispatchable and base-load electricity. This study performs a systems process analysis to quantify the carbon emissions and nonrenewable energy costs induced by a state-of-art demonstration CSP plant located in the Tibetan plateau. Estimated to induce 111.2 g CO2 eq/kWh carbon emissions and 1.42 MJ/kWh non-renewable energy consumption, the CSP plant is considered to have extremely high carbon neutrality (88.8%) and energy renewability (86.4%). The prominent performance of carbon emissions reduction and energy conservation induced by the CSP plant shed light on its superiority of reliable power supply and environmental benefits. The plant is expected to cumulatively fulfill 3.4 million tons of carbon emissions reduction over its life cycle. In contrast to coal-based power and other renewable energy technologies, CSP technology is shown to be a promising solution to the low-carbon energy transition. Besides, a scenario analysis indicates that the incremental employment of CSP technologies will play a critical role in coping with climate change and energy security in China. Moreover, multiple policies to facilitate the development of the CSP system in China are elaborated, such as the promotion of integrated solar combined-cycle systems. The empirical finding draws a holistic picture of the carbon neutrality and energy sustainability performance of CSP technologies, and the systematic analysis in this study provides comprehensive policy perspectives for energy policy in the Tibetan region as well as in China in the context of global climate change.

Global pastureland use as reflected in inter-regional supply chain.

Pastureland used for livestock grazing is globally much bigger than arable land. This study investigates the pastureland use embedded in global supply chains by using multi-regional systems input-output anlysis, tracing embodied pastureland use from source of exploitation to sink of final consumption in the global economy. The world's pastureland resources is shown reallocated through the supply chain mainly to the four major economies: EU, the United States, China, and Japan. These four economies are responsible for driving more than half of the global pastureland exploitation. Major supply chains responsible for the global reallocation of pastureland use include the cattle supply chain from Other Asia & Pacific to the United States, China, and Japan, and the cattle supply chain from Africa to EU and the Middle East. This paper demonstrates the nature and scale of the global reallocation of pastureland resources through the supply chain, highlighting the fact that the global shift of pastureland use from nature-based to economic-based may exacerbate ecological inequity between world regions. It is proposed that future policies and regulations should encourage sustainability goals not only on a regional level but on a global scale, finding pathways to sustainable and equitable livestock production by inter-regional collaboration.

The CHY-Type Zinc Finger Protein FgChy1 Regulates Polarized Growth, Pathogenicity, and Microtubule Assembly in Fusarium graminearum.

Microtubules (MTs), as transport tracks, play important roles in hyphal-tip growth in filamentous fungi, but MT-associated proteins involved in polarized growth remain unknown. Here, we found that one novel zinc finger protein, FgChy1, is required for MT morphology and polarized growth in Fusarium graminearum. The Fgchy1 mutant presented curved and directionless growth of hyphae. Importantly, the conidia and germ tubes of the Fgchy1 mutant exhibited badly damaged and less-organized beta-tubulin cytoskeletons. Compared with the wild type, the Fgchy1 mutant lost the ability to maintain polarity and was also more sensitive to the anti-MT drugs carbendazim and nocodazole, likely due to the impaired MT cytoskeleton. Indeed, the hyphae of the wild type treated with nocodazole exhibited a morphology consistent with that of the Fgchy1 mutant. Interestingly, the disruption of FgChy1 resulted in the off-center localization of actin patches and the polarity-related polarisome protein FgSpa2 from the hyphal-tip axis. A similar defect in FgSpa2 localization was also observed in the nocodazole-treated wild-type strain. In addition, FgChy1 is also required for conidiogenesis, septation, sexual reproduction, pathogenicity, and deoxynivalenol production. Overall, this study provides the first demonstrations of the functions of the novel zinc finger protein FgChy1 in polarized growth, development, and virulence in filamentous fungi.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

SCYN: single cell CNV profiling method using dynamic programming.

BACKGROUND: Copy number variation is crucial in deciphering the mechanism and cure of complex disorders and cancers. The recent advancement of scDNA sequencing technology sheds light upon addressing intratumor heterogeneity, detecting rare subclones, and reconstructing tumor evolution lineages at single-cell resolution. Nevertheless, the current circular binary segmentation based approach proves to fail to efficiently and effectively identify copy number shifts on some exceptional trails. RESULTS: Here, we propose SCYN, a CNV segmentation method powered with dynamic programming. SCYN resolves the precise segmentation on in silico dataset. Then we verified SCYN manifested accurate copy number inferring on triple negative breast cancer scDNA data, with array comparative genomic hybridization results of purified bulk samples as ground truth validation. We tested SCYN on two datasets of the newly emerged 10x Genomics CNV solution. SCYN successfully recognizes gastric cancer cells from 1% and 10% spike-ins 10x datasets. Moreover, SCYN is about 150 times faster than state of the art tool when dealing with the datasets of approximately 2000 cells. CONCLUSIONS: SCYN robustly and efficiently detects segmentations and infers copy number profiles on single cell DNA sequencing data. It serves to reveal the tumor intra-heterogeneity. The source code of SCYN can be accessed in https://github.com/xikanfeng2/SCYN .

Fluorescent hollow ZrO2@CdTe nanoparticles-based lateral flow assay for simultaneous detection of C-reactive protein and troponin T.

Highly fluorescent hollow ZrO2@CdTe nanoparticles (NPs) were synthesized efficiently via the hydrothermal method. By changing the hydrothermal time of ZrO2@CdTe NP, the peaks of fluorescence spectra measured at fluorescent excitation of 330 nm were at 540 nm, 590 nm, and 640 nm, respectively. Hollow ZrO2 NPs have a uniform core-shell structure with the size of 178 ± 10 nm and shell of 19 ± 4 nm. The as-prepared yellow-ZrO2@CdTe NPs were used to develop lateral flow assay (LFA) for the sensitive and qualitative detection of C-reactive protein (CRP). The visual limit of detection of the LFA for the CRP antigen was 1 µg/L within 20 min, which is 1000-fold lower than that of colloidal gold-based LFA. In addition, a multiplex lateral flow assay (mLFA) was developed using the as-prepared green and red-ZrO2@CdTe NPs for the simultaneous, specific, sensitive, and qualitative detection of CRP and troponin T (cTnT). The visual limits of detection of CRP and cTnT in mLFA were 10 µg/L and 0.1 mg/L, respectively. The excellent performance of ZrO2@CdTe NPs should facilitate their application in point-of-care technology for the detection of other biomarkers.

Vacant Parking Slot Detection in the Around View Image Based on Deep Learning.

Due to the complex visual environment, such as lighting variations, shadows, and limitations of vision, the accuracy of vacant parking slot detection for the park assist system (PAS) with a standalone around view monitor (AVM) needs to be improved. To address this problem, we propose a vacant parking slot detection method based on deep learning, namely VPS-Net. VPS-Net converts the vacant parking slot detection into a two-step problem, including parking slot detection and occupancy classification. In the parking slot detection stage, we propose a parking slot detection method based on YOLOv3, which combines the classification of the parking slot with the localization of marking points so that various parking slots can be directly inferred using geometric cues. In the occupancy classification stage, we design a customized network whose size of convolution kernel and number of layers are adjusted according to the characteristics of the parking slot. Experiments show that VPS-Net can detect various vacant parking slots with a precision rate of 99.63% and a recall rate of 99.31% in the ps2.0 dataset, and has a satisfying generalizability in the PSV dataset. By introducing a multi-object detection network and a classification network, VPS-Net can detect various vacant parking slots robustly.

FgPrp4 Kinase Is Important for Spliceosome B-Complex Activation and Splicing Efficiency in Fusarium graminearum.

PRP4 encodes the only kinase among the spliceosome components. Although it is an essential gene in the fission yeast and other eukaryotic organisms, the Fgprp4 mutant was viable in the wheat scab fungus Fusarium graminearum. Deletion of FgPRP4 did not block intron splicing but affected intron splicing efficiency in over 60% of the F. graminearum genes. The Fgprp4 mutant had severe growth defects and produced spontaneous suppressors that were recovered in growth rate. Suppressor mutations were identified in the PRP6, PRP31, BRR2, and PRP8 orthologs in nine suppressor strains by sequencing analysis with candidate tri-snRNP component genes. The Q86K mutation in FgMSL1 was identified by whole genome sequencing in suppressor mutant S3. Whereas two of the suppressor mutations in FgBrr2 and FgPrp8 were similar to those characterized in their orthologs in yeasts, suppressor mutations in Prp6 and Prp31 orthologs or FgMSL1 have not been reported. Interestingly, four and two suppressor mutations identified in FgPrp6 and FgPrp31, respectively, all are near the conserved Prp4-phosphorylation sites, suggesting that these mutations may have similar effects with phosphorylation by Prp4 kinase. In FgPrp31, the non-sense mutation at R464 resulted in the truncation of the C-terminal 130 aa region that contains all the conserved Prp4-phosphorylation sites. Deletion analysis showed that the N-terminal 310-aa rich in SR residues plays a critical role in the localization and functions of FgPrp4. We also conducted phosphoproteomics analysis with FgPrp4 and identified S289 as the phosphorylation site that is essential for its functions. These results indicated that FgPrp4 is critical for splicing efficiency but not essential for intron splicing, and FgPrp4 may regulate pre-mRNA splicing by phosphorylation of other components of the tri-snRNP although itself may be activated by phosphorylation at S289.

Two Cdc2 Kinase Genes with Distinct Functions in Vegetative and Infectious Hyphae in Fusarium graminearum.

Eukaryotic cell cycle involves a number of protein kinases important for the onset and progression through mitosis, most of which are well characterized in the budding and fission yeasts and conserved in other fungi. However, unlike the model yeast and filamentous fungi that have a single Cdc2 essential for cell cycle progression, the wheat scab fungus Fusarium graminearum contains two CDC2 orthologs. The cdc2A and cdc2B mutants had no obvious defects in growth rate and conidiation but deletion of both of them is lethal, indicating that these two CDC2 orthologs have redundant functions during vegetative growth and asexual reproduction. However, whereas the cdc2B mutant was normal, the cdc2A mutant was significantly reduced in virulence and rarely produced ascospores. Although deletion of CDC2A had no obvious effect on the formation of penetration branches or hyphopodia, the cdc2A mutant was limited in the differentiation and growth of infectious growth in wheat tissues. Therefore, CDC2A plays stage-specific roles in cell cycle regulation during infectious growth and sexual reproduction. Both CDC2A and CDC2B are constitutively expressed but only CDC2A was up-regulated during plant infection and ascosporogenesis. Localization of Cdc2A- GFP to the nucleus but not Cdc2B-GFP was observed in vegetative hyphae, ascospores, and infectious hyphae. Complementation assays with chimeric fusion constructs showed that both the N- and C-terminal regions of Cdc2A are important for its functions in pathogenesis and ascosporogenesis but only the N-terminal region is important for its subcellular localization. Among the Sordariomycetes, only three Fusarium species closely related to F. graminearum have two CDC2 genes. Furthermore, F. graminearum uniquely has two Aurora kinase genes and one additional putative cyclin gene, and its orthologs of CAK1 and other four essential mitotic kinases in the budding yeast are dispensable for viability. Overall, our data indicate that cell cycle regulation is different between vegetative and infectious hyphae in F. graminearum and Cdc2A, possibly by interacting with a stage-specific cyclin, plays a more important role than Cdc2B during ascosporogenesis and plant infection.

FgCDC14 regulates cytokinesis, morphogenesis, and pathogenesis in Fusarium graminearum.

Members of Cdc14 phosphatases are common in animals and fungi, but absent in plants. Although its orthologs are conserved in plant pathogenic fungi, their functions during infection are not clear. In this study, we showed that the CDC14 ortholog is important for pathogenesis and morphogenesis in Fusarium graminearum. FgCDC14 is required for normal cell division and septum formation and FgCdc14 possesses phosphatase activity with specificity for a subset of Cdk-type phosphorylation sites. The Fgcdc14 mutant was reduced in growth, conidiation, and ascospore formation. It was defective in ascosporogenesis and pathogenesis. Septation in Fgcdc14 was reduced and hyphal compartments contained multiple nuclei, indicating defects in the coordination between nuclear division and cytokinesis. Interestingly, foot cells of mutant conidia often differentiated into conidiogenous cells, resulting in the production of inter-connected conidia. In the interphase, FgCdc14-GFP localized to the nucleus and spindle-pole-body. Taken together, our results indicate that Cdc14 phosphatase functions in cell division and septum formation in F. graminearum, likely by counteracting Cdk phosphorylation, and is required for plant infection.

Thioredoxins are involved in the activation of the PMK1 MAP kinase pathway during appressorium penetration and invasive growth in Magnaporthe oryzae.

In Magnaporthe oryzae, the Mst11-Mst7-Pmk1 MAP kinase pathway is essential for appressorium formation and invasive growth. To determine their roles in Pmk1 activation and plant infection, we characterized the two thioredoxin genes, TRX1 and TRX2, in M. oryzae. Whereas the Δtrx1 mutants had no detectable phenotypes, deletion of TRX2 caused pleiotropic defects in growth, conidiation, light sensing, responses to stresses and plant infection progresses. The Δtrx1 Δtrx2 double mutant had more severe defects than the Δtrx2 mutant and was non-pathogenic in infection assays. The Δtrx2 and Δtrx1 Δtrx2 mutant rarely formed appressoria on hyphal tips and were defective in invasive growth after penetration. Pmk1 phosphorylation was barely detectable in the Δtrx2 and Δtrx1 Δtrx2 mutants. Deletion of TRX2 affected proper folding or intra-/inter-molecular interaction of Mst7 and expression of the dominant active MST7 allele partially rescued the defects of the Δtrx1 Δtrx2 mutant. Furthermore, Cys305 is important for Mst7 function and Trx2 directly interacts with Mst7 in co-IP assays. Our data indicated that thioredoxins play important roles in intra-cellular ROS signalling and pathogenesis in M. oryzae. As the predominant thioredoxin gene, TRX2 may regulate the activation of Pmk1 MAPK via its effects on Mst7.

Germ stem cells are active in postnatal mouse ovary under physiological conditions.

STUDY HYPOTHESIS: Are active ovarian germ stem cells present in postnatal mouse ovaries under physiological conditions? STUDY FINDING: Active ovarian germ stem cells exist and function in adult mouse ovaries under physiological conditions. WHAT IS KNOWN ALREADY: In vitro studies suggested the existence of germ stem cells in postnatal ovaries of mouse, pig and human. However, in vivo studies provided evidence against the existence of active germ stem cells in postnatal mouse ovaries. Thus, it remains controversial whether such germ stem cells really exist and function in vivo in postnatal mammalian ovaries. STUDY DESIGN, SAMPLES/MATERIALS, METHODS: Octamer-binding transcription factor 4 (Oct4)-MerCreMer transgenic mice were crossed with R26R-enhanced yellow fluorescent protein (EYFP) mice to establish a tamoxifen-inducible tracing system so that Oct4-expressing potential ovarian germ stem cells in young adult mice (5-6 weeks old) can be labeled with EYFP. The germ cell activities of DNA replication, mitotic division, entry into meiosis and progression to primordial follicle stage were investigated by means of immunofluorescent staining of ovarian tissues collected at different time points post-tamoxifen injection (1 day, 3 days, 2 months and 4 months). Meiosis entry and primordial follicle formation were also measured by EYFP-labeled single-cell RT-PCR. Germ cell proliferation and mitotic division were examined through 5-bromodeoxyuridine triphosphate incorporation assay. At each time point, ovaries from two to three animals were used for each set of experiment. MAIN RESULTS AND THE ROLE OF CHANCE: By labeling the Oct4-expressing small germ cells and tracing their fates for up to 4 months, we observed persistent meiosis entry and primordial follicle replenishment. Furthermore, we captured the transient processes of mitotic DNA replication as well as mitotic division of the marked germ cells at various time periods after tracing. These lines of evidence unambiguously support the presence of active germ stem cells in postnatal ovaries and their function in replenishing primordial follicle pool under physiological conditions. Moreover, we pointed out that Oct4(+) deleted in azoospermia-like (Dazl)(-) but not Oct4(+)Dazl(+) or Oct4(+) DEAD (Asp-Glu-Ala-Asp) Box Polypeptide 4 (Ddx4)(+) cells contain a population of germ stem cells in mouse ovary. LIMITATIONS, REASONS FOR CAUTION: This study was conducted in mice. Whether or not the results are applicable to human remain unclear. The future work should aim at identifying the specific ovarian germ stem cell marker and evaluating the significance of these stem cells to normal ovarian function. WIDER IMPLICATIONS OF THE FINDINGS: Clarifying the existence of active germ stem cells and their functional significance in postnatal mammalian ovaries could provide new insights in understanding the mechanism of ovarian aging and failure. LARGE SCALE DATA: Not applicable. STUDY FUNDING/COMPETING INTERESTS: This work was supported by the National Key Basic Research Program of China (grant number 2012CBA01300) and the National Natural Science Foundation of China to P.Z. (31571484). No competing interests are reported.

Aurora-A modulates MMP-2 expression via AKT/NF-κB pathway in esophageal squamous cell carcinoma cells.

Esophageal squamous cell carcinoma (ESCC) is one of the most common malignancies. It is necessary to identify new markers for predicting tumor progression and therapeutic molecular targets. It has been reported that overexpressions of Aurora-A and matrix metalloproteinases 2 (MMP-2) may promote the malignant development of tumor. However, the relationship between Aurora-A and MMP-2 expression in tumor patients has not been investigated. In addition, the underlying mechanisms that Aurora-A regulates MMP-2 expression are still not fully elucidated. In this study, we demonstrated that Aurora-A and MMP-2 were overexpressed in ESCC tissues compared with paired normal adjacent tissues (P < 0.0001). Overexpression of Aurora-A was associated with the lymph node metastasis of ESCC (P = 0.01). Significantly, Aurora-A protein expression was positively correlated with MMP-2 protein expression in ESCC tissues (r = 0.66, P < 0.0001) as well as in ESCC cell lines. The level of Aurora-A expression was also positively correlated with the invasion capability of ESCC cells. Furthermore, Aurora-A overexpression significantly increased ESCC cell invasion by the upregulation of MMP-2 expression. In addition, Aurora-A overexpression promoted nuclear factor-kappaB (NF-κB) activation, and Aurora-A-mediated MMP-2 upregulation was abrogated by NF-κB inhibitor. Further analysis showed that activation of NF-κB was severely attenuated by AKT inhibitor in cells overexpressing Aurora-A. Taken together, these data indicate that Aurora-A overexpression upregulates MMP-2 expression through activating AKT/NF-κB signaling pathway in ESCC cells. These findings reveal that Aurora-A may be used as an important indicator for the judgment of malignant behavior of ESCC, and may be an attractive target for cancer therapy.

The natural polycyclic tetramate macrolactam HSAF inhibit Fusarium graminearum through altering cell membrane integrity by targeting FgORP1.

Fusarium graminearum is a dominant phytopathogenic fungus causing Fusarium head blight (FHB) in cereal crops. Heat-stable antifungal factor (HSAF) is a polycyclic tetramate macrolactam (PoTeM) isolated from Lysobacter enzymogenes that exhibits strong antifungal activity against F. graminearum. HSAF significantly reduces the DON production and virulence of F. graminearum. Importantly, HSAF exhibited no cross-resistance to carbendazim, phenamacril, tebuconazole and pydiflumetofen. However, the target protein of HSAF in F. graminearum is unclear. In this study, the oxysterol-binding protein FgORP1 was identified as the potential target of HSAF using surface plasmon resonance (SPR) combined with RNA-sequence (RNA-seq). The RNA-seq results showed cell membrane and ergosterol biosynthesis were significantly impacted by HSAF in F. graminearum. Molecular docking showed that HSAF binds with arginine 1205 and glutamic acid 1212, which are located in the oxysterol-binding domain of FgORP1. The two amino acids in FgORP1 are responsible for HSAF resistance in F. graminearum though site-directed mutagenesis. Furthermore, deletion of FgORP1 led to significantly decreased sensitivity to HSAF. Additionally, FgORP1 regulates the mycelial growth, conidiation, DON production, ergosterol biosynthesis and virulence in F. graminearum. Overall, our findings revealed the mode of action of HSAF against F. graminearum, indicating that HSAF is a promising fungicide for controlling FHB.