Carbon Catabolite Repressor UvCreA is Required for Development and Pathogenicity in Ustilaginoidea virens

The rice false smut disease, caused by Ustilaginoidea virens, has emerged as a significantglobal threat to rice production. The mechanism of carbon catabolite repression plays a crucial role in theefficient utilization of carbon nutrients and enzyme regulation in the presence of complex nutritionalconditions. Although significant progress has been made in understanding carbon catabolite repression infungi such as Aspergillus nidulans and Magnaporthe oryzae, its role in U. virens remains unclear. Toaddress this knowledge gap, we identified UvCreA, a pivotal component of carbon catabolite repression,in U. virens. Our investigation revealed that UvCreA localized to the nucleus. Deletion of UvCreA resultedin decreased growth and pathogenicity in U. virens. Through RNA-seq analysis, it was found that theknockout of UvCreA led to the up-regulation of 514 genes and down-regulation of 640 genes. Moreover,UvCreA was found to be involved in the transcriptional regulation of pathogenic genes and genesassociated with carbon metabolism in U. virens. In summary, our findings indicated that UvCreA isimportant in fungal development, virulence, and the utilization of carbon sources through transcriptionalregulation, thus making it a critical element of carbon catabolite repression.

Detection of Sugar-Regulated Gene Expression and Signaling in Suspension-Cultured Rice Cells

To better understand the mechanism of sugar signaling in rice cell, the suspension-cultured rice cells were transferred from sucrose-containing (+S) to sucrose-free (-S) of MS culture medium, we found that ribosomal RNAs (rRNAs) were degraded progressively. This suggests that carbon, nitrogen, and phosphate were recycled in this process and the reduction in cellular rRNAs might lead to decreased translation to save energy in response to sugar starvation. Differential screening revealed that two groups of genes, sugar-starvation-repressed (SSR) and sugar-starvation-activated (SSA) genes, were regulated by sugar in an opposing manner. Northern-blot analysis showed that two major hybridization signals of 0.8 and 1.9 kb were induced strongly under sugar starvation. The two populations of genes corresponded with homologs of α-amylases (1.9 kb) and the glycine-rich proteins (GRPs) gene family (0.8 kb), and all were SSA genes. Expression of GRP genes was strongly induced in sugar-starved cells, which suggests that GRPs may help to protect cells against nutritional stress. Treatment of +S and -S cells with the protein kinase (PK) inhibitor staurosporine (St) and the serine/theronine phosphoprotein phosphatases 1 (PP1) and 2A (PP2A) inhibitor okadaic acid (OA) revealed that PP1 and PP2A (PPs) might be involved in increasing SSR gene expression in +S cells, and that activation of the majority of the SSA genes in -S cells might be due to PKs activity. These results suggested that PKs and PPs might be involved in the sugar regulation of SSR and SSA gene expression. An in-gel PK activity assay demonstrated that the activity of two classes of PKs (50 and 66 kDa) may be induced rapidly after transfer of +S cells to -S medium. Following transfer of -S cells to +S medium, a novel class of 38 kDa PK was induced rapidly and showed high activity. The 38 kDa PK might play a role in sugar sensing, and the 50 and 66 kDa PKs might play roles in signal sensing under sugar starvation in rice cells. These results provide valuable information on three classes of protein kinases that might play key roles in sugar sensing and signaling in rice.

tRNA cleavage: a new insight

Over the past decades,tRNA was found to be a rich hub of RNA modifications such as 1-methyladenosine and 5-methycytosine modifications and others,holding more than half of all modifications occurring in RNA molecules.Moreover,tRNA was discovered to be a source of various small noncoding RNA species,such as the stress induced angiogenin cleaved tRNA halves(tiRNA)or the miRNA like tRNA derived fragments.tRNA cleavage under stress was fist discovered in bacteria and later was found to be conserved across different species,including mammals.Under cellular stress conditions,tRNA undergoes conformational changes and angiogenin cleaves it into 3′and 5′halves.5′tiRNA halves were shown to repress protein translations.tRNA cleavage is thought of to be a cytoprotective mechanism by which cells evade apoptosis,however some data hints to the opposite;that tiRNA are cytotoxic or at least related to apoptosis initiation.tRNA cleavage also was shown to be affected by tRNA modifications via different enzymes in the cytosol and mitochondria.In this review,we will highlight the biology of tRNA cleavage,show the evidence of it being cytoprotective or a marker of cell death and shed a light on its role in disease models and human diseases as well as possible future directions in this field of RNA research.

Plant Long ncRNAs: A New Frontier for Gene Regulatory Control

Long non-coding RNA (lncRNA) refers to an over 200 nt functional RNA molecule that will not be translated into protein. Previously thought to be dark matters of the genome, lncRNAs have been gradually recognized as crucial gene regulators. Although tremendous progress has been made in animals and human, the study of lncRNAs in plant is still in its infancy. Here, we reviewed the biogenesis and regulation mechanisms of lncRNAs and summarized the achievements that have been made in plant lncRNA identification and functional characterization. Genome-wide identification has uncovered large amount of lncRNAs in Arabidopsis, Rice, Maize and Wheat, and more information from other plant species will be expected with the aid of deep sequencing technologies. Similar to other species, LncRNA-mediated gene regulation also widely exists in plants, even though only a few functionally characterized examples are available. Up to now, at least four divergent lncRNA-mediated regulation mechanisms have been unraveled, including target mimicry, transcription interference, PRC2 associated histone methylation and DNA methylation. lncRNAs may be involved in the regulation of flowering, male sterility, nutrition metabolism, biotic and abiotic stress response in plants.

Polycomb Repressive Complex 2-Mediated H3K27 Trimethylation Is Required for Pathogenicity in Magnaporthe oryzae

Polycomb repressive complex 2(PRC2)contributes to catalyze the methylation of histone H3 at lysine 27 and plays vital roles in transcriptional silencing and growth development in various organisms.In Magnaporthe oryzae,histone H3K27 is found to associate with altered transcription of in planta induced genes.However,it is still unknown whether and how H3K27me3 modification is involved in pathogenicity to rice and stress response.In this study,we found that core subunits of PRC2,Kmt6-Suz12-Eed,were required for fungal pathogenicity to rice in M.oryzae.Kmt6-Suz12-Eed localized in the nuclei and was necessary for the establishment of H3K27me3 modification.With ChIP-seq analysis,9.0%of genome regions enriched with H3K27me3 occupancy,which corresponded to 1033 genes in M.oryzae.Furthermore,deletion of Kmt6,Suz12 or Eed altered genome-wide transcriptional expression,while the de-repression genes in theΔkmt6 strain were highly associated with H3K27me3 occupancy.Notably,plenty of genes which encode effectors and secreted enzymes,secondary metabolite synthesis genes,and cell wall stress-responsive genes were directly occupied with H3K27me3 modification and de-repression in theΔkmt6 strain.These results elaborately explained how PRC2 was required for pathogenicity,which is closely related to effector modulated host immunity and host environment adaption.

Quantifying Growth Responses of Black Spruce and Jack Pine to Thinning within the Context of Density Management Decision-Support Systems

Models for quantifying the growth responses of black spruce (Picea mariana (Mill) BSP.) and jack pine (Pinus banksiana Lamb.) to precommercial thinning (PCT) treatments were developed. They accounted for the increased rate of stand development arising from PCT treatments through temporal adjustments to the species and site specific mean dominant height-age functions. Analytically, they utilized a relative height growth modifier consistent with observed density-dependent height repression effects. A phenotypic juvenile age-mature age correlation function was used to account for the intrinsic temporal decline in the magnitude of the PCT effect throughout the rotation. The resultant stand development patterns were in accord with theoretical and empirical expectations when the response models were integrated into algorithmic variants of structural stand density management models.

On Satan in Paradise Lost from the View of Milton's Loss of Freedom

The motive forces of literary creation are closely related to an author’s personal experiences.Milton experienced great losses during his lifetime,which produced deep infl uences on his poetic writing.This thesis,based on Freud’s"repression"and"compensation"theory,analyzes Milton’s psychological state in the creation of Paradise Lost,aiming at exploring infl uences of the poet’s experience of loss on the creation of Satan in his masterpiece.

Isolation and Characterization of Nickel Uptake by Nickel Resistant Bacterial Isolate (NiRBI)

Objective Bioremediation technology has gained importance because microbes could be the convenient source of bio-absorption/bioaccumulation of metals from effluent streams. Methods The nickel-resistant bacterial isolates （NiRBI） were selected from various bacterial isolates from industrial effluent and grown in nutrient broth containing different concentrations of nickel sulfate （0.3-3.0 mmol/L） and their capability of accumulating metal from the medium. Results Well-defined growth of NiRBI was observed in the medium containing up to 2.5 mmol/L of nickel. The isolate was identified using 16S rRNA and closely related to Pseudomonas fragi. Maximum accumulation of nickel （0.59 mg/g dry weight of bacterial cells） was observed when NiRBI was grown in media containing 2 mmol/L of nickel. The protein profile of the NiRBI cellular extract by SDS-PAGE showed two metal stress-induced proteins of molecular weight 48 KD and 18 KD with a simultaneous down regulation of four proteins of 46.7 KD, 42.2 KD, 19.7 KD, and 4.0 KD. Conclusion 48 KD and 18 KD proteins play a role in metal resistance mechanism by NiRBI.

EFFECT OF HIGH TEMPERATURE REPRESSING ON PHOSPHORUS SEGREGATION IN 93W-4.5Ni-2.5Fe ALLOY

The influence of high temperature repressing treatment on the segregation of phosphorus in the 93W alloy was carefully investigated by means of Auger electron spectroscope,EPMA, TEM and SEM.The segregation of phosphorus has been observed at the tungsten-tungsten grain boundaries,particularly at the tungsten-matrix interphases when the specimens were kept at the temperature in the range of 1200—1500℃ ,followed by furnace cooling.However, no segregation of phosphorus was observed at the interfaces after the specimens were re- pressed at the range of temperature,followed by furnace cooling.After investigation,the dis- locations in the matrix phase of as-repressed specimens directly influenced the phosphorus segregation to the interfaces.After annealing,the mechanical properties were reduced because of the phosphorus segregation at the interface boundaries.

Characterization of Zm26Sub5 and Its Potential Roles in Regulating Pollen Fertility of S-CMS in Maize

Mitochondrial orf355-orf77 and the nuclear fertility restorer locus Rf3/rf3 mutually control the fertility of male gametes in CMS-S in maize （Zea mays L.）. A fragment of gene Zm26Sub5 was identified through cDNA-AFLP from a set of Rf3/rf3 near-isogenic lines in a previous study. In the present study, real-time PCR analysis revealed that Zm26Sub5 expression levels were much higher in pollen of S-Mo17Rf3Rf3 than in pollen of S-Mo17rf3rf3, and also higher than in fresh leaves, mature leaves, and roots of both S-Mo17Rf3Rf3 and S-Mo17rf3rf3. In silico cloning of full-length cDNA was successfully implemented. Gene Zm26Sub5 was 1 451 bp in size, of which 1 329 bp encoded a protein with 443 amino acids. The structure of this gene was analyzed by comparing its full length cDNA to homologous genomic DNA sequence （GenBank accession： Ac209463.3）. Subsequent sequence analysis led to sub cellular localization of protein ZM26SUB5, and construction of a phylogenetic tree. In silico mapping indicated that Zm26Sub5 was located on chromosome 5 and closed to a reported starch-filled pollen ratio QTL. ZM26SUB5, therefore might have potential roles in repressing mitochondrial PCD which is associated with sterile activity in pollen in S-type cytoplasm.

Isolation of enriched-yielders and fed-batch production of alkaline protease from the newly isolated <i>Bacillus</i>sp. BHA

An alkalophilc and thermophilic Bacillus sp. BHA that produced a thermostable alkaline protease was isolated from decaying protein substrates. The isolate was found to grow in pH range 7 - 11 with an optimum pH 9.0 and temperature up to 55℃. The activity of alkaline protease of Bacillus sp. BHA (68.98 APU/ml) was found higher than the standard strains of Bacillus amyloliquefaciens MTCC 610 (8.98 APU/ml) and Bacillus subtilis MTCC 8349 (12.14 APU/ml, used in this study, and was comparable (68.98 APU/ml, equivalent to 30.38 APU/mg) to the activity of the commercially produced standard protease procured from Novo Nordisk, Denmark (30.35 APU/mg). Hence, the proteolytic activity produced by this isolate was further investigated in batch and fed-batch process. Sucrose was the best carbon source for the production of protease activity by that isolate. Different organic nitrogen sources (casein, peptone and beef extract) at 1% (w/v) with varying levels of sucrose (1% - 4% w/v) initially repress enzyme synthesis. The duration and extent of repression decreased with increased concentration of sucrose. Maximum protease activity was found in basal medium with 4% (w/v) sucrose and 1% (w/v) yeast extract. Yeast-extract was thought to be an inducer of enzyme synthesis. Further, the basal medium was unique with respect to the enzyme production, as protease production was growth associated with the peak enzyme production being detected at the time of maximum growth. Interestingly, a rise in 34.2% (104.86 APU/ml) of protease activity was detected at incubation temperature of 50℃ and when culture filtrate was assayed at 60℃, signifying a high temperature stability of the produced protease by this isolate. Additional studies on the enzyme characterization were resulted in recognition of highly significant properties of the activity towards casein at pH 9.0 and stability at high temperature with retention of 96% the enzyme activity at 60℃. The parametric study under feed intervals had enabled improvement in the maximum protease activities attainable from batch cultures in excess of 21.78% and 26.32% via two feeding strategies. A small continual increase in enzyme activity (132.46 APU/ml during 24 h - 120 h) and enhancement in protease production in excess of 36.84% was observed by fed-batch process than the batch experiment.

Infected cell protein 0 functional domains and their coordination in herpes simplex virus replication

Herpes simplex virus 1(HSV-1) is a ubiquitous human pathogen that establishes latent infection in ganglia neurons. Its unique life cycle requires a balanced "conquer and compromise" strategy to deal with the host anti-viral defenses. One of HSV-1 α(immediate early) gene products, infected cell protein 0(ICP0), is a multifunctional protein that interacts with and modulates a wide range of cellular defensive pathways. These pathways may locate in different cell compartments, which then migrate or exchange factors upon stimulation, for the purpose of a concerted and effective defense. ICP0 is able to simultaneously attack multiple host pathways by either degrading key restrictive factors or modifying repressive complexes. This is a viral protein that contains an E3 ubiquitin ligase, translocates among different cell compartments and interacts with major defensive complexes. The multiple functional domains of ICP0 can work independently and at the same time coordinate with each other. Dissecting the functional domains of ICP0 and delineating the coordination of these domains will help us understand HSV-1 pathogenicity as well as host defense mechanisms. This article focuses on describing individual ICP0 domains, their biochemical properties and their implication in HSV-1 infection. By putting individual domain functions back into the picture of host anti-viral defense network, this review seeks to elaborate the complex interactions between HSV-1 and its host.

Interpretation on Heathcliff in Wuthering Heights In Light of Psychoanalysis

This thesis aims to analyze the main character of the novel Wuthering Heights from a psychoanalytical perspective.By using techniques adapted from psychoanalysis,this thesis finds that the complex character Heathcliff at first reading is easier to understand,and indeed fits into some of the basic victim-persecutor patterns identified in traditional psychoanalytical work.From Heathcliff we can see the importance of one's psychology and the power of psychoanalysis.

The Comparison Between Two Spinsters in Winesburg, Ohio

Sherwood Anderson is highly skilled in portraying grotesques distorted by the industrialized society. This paper is to unfold the repression and inner struggle of the two typical spinster images in the novel Winesburg Ohio, the masculine small town, which entraps women with the conventional codes. They achieve economic independence in their earlier years which is their first attempt to overthrow the yoke of man-dominated world. They employ different means to release the deep buried passion and eagerness to love and to be loved so that they can shake off loneliness and repression. However, in the climatic moments, they fail in their seek for self-fulfillment and are doomed to be tossed between traditional women roles and modern awakening.

Strategies that regulate LSD1 for novel therapeutics

Histone methylation plays crucial roles in regulating chromatin structure and gene transcrip-tion in epigenetic modifications.Lysine-specific demethylase 1(LSD1),the first identified histone de-methylase,is universally overexpressed in various diseases.LSD1 dysregulation is closely associated with cancer,viral infections,and neurodegenerative diseases,etc.,making it a promising therapeutic target.Several LSD1 inhibitors and two small-molecule degraders(UM171 and BEA-17)have entered the clinical stage.LSD1 can remove methyl groups from histone 3 at lysine 4 or lysine 9(H3K4 or H3K9),resulting in either transcription repression or activation.While the roles of LSD1 in transcrip-tional regulation are well-established,studies have revealed that LSDl can also be dynamically regulated by other factors.For example,the expression or activity of LSD1 can be regulated by many proteins that form transcriptional corepressor complexes with LSD1.Moreover,some post-transcriptional modifica-tions and cellular metabolites can also regulate LSD1 expression or its demethylase activity.Therefore,in this review,we will systematically summarize how proteins involved in the transcriptional corepressor complex,various post-translational modifications,and metabolites act as regulatory factors for LSD1 ac-tivity.

Achieving robust nitritation in a modified continuous-flow reactor: From micro-granule cultivation to nitrite-oxidizing bacteria elimination

In this study,a modified continuous-flow nitrifying reactor was successfully operated for rapid cultivation of micro-granules and achieving robust nitritation.Results showed that sludge granulation with mean size of ca.100μm was achieved within three weeks by gradually increasing settling velocity-based selection pressure from 0.48 to 0.9 m/hr.Though Nitrospira like nitrite-oxidizing bacteria(NOB)were enriched in the micro-granules with a ratio between ammonia-oxidizing bacteria(AOB)and NOB of 5.7%/6.5% on day 21,fast nitritation was achieved within one-week by gradually increasing of influent ammonium concentration(from 50 to 200 mg/L).Maintaining ammonium in-excess was the key for repressing NOB in the micro-granules.Interestingly,when the influent ammonium concentration switched back to 50 mg/L still with the residual ammonium of 15–25 mg/L,the nitrite accumulation efficiency increased from 90%to 98%.Experimental results suggested that the NOB repression was intensified by both oxygen and nitrite unavailability in the inner layers of micro-granules.Unexpectedly,continuous operation with ammonium in excess resulted in overproduction of extracellular polysaccharides and overgrowth of some bacteria(e.g.,Nitrosomonas,Arenimonas,and Flavobacterium),which deteriorated the micro-granule stability and drove the micro-granules aggregation into larger ones with irregular morphology.However,efficient nitritation was stably maintained with extremely high ammonium oxidation potential(>50 mg/g VSS/hr)and nearly complete washout of NOB was obtained.This suggested that smooth and spherical granule was not a prerequisite for achieving NOB wash-out and maintaining effective nitritation in the granular reactor.Overall,the microgranules exhibited a great practical potential for high-rate nitritation.

Protein O-GlcNAcylation homeostasis regulates facultative heterochromatin to fine-tune sog-Dpp signaling during Drosophila early embryogenesis

Protein O-GlcNAcylation is a monosaccharide post-translational modification maintained by two evolutionarily conserved enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Mutations in human OGT have recently been associated with neurodevelopmental disorders, although the mechanisms linking O-GlcNAc homeostasis to neurodevelopment are not understood. Here, we investigate the effects of perturbing protein O-GlcNAcylation using transgenic Drosophila lines that overexpress a highly active OGA. We reveal that temporal reduction of protein O-GlcNAcylation in early embryos leads to reduced brain size and olfactory learning in adult Drosophila. Downregulation of O-GlcNAcylation induced by the exogenous OGA activity promotes nuclear foci formation of Polycomb-group protein Polyhomeotic and the accumulation of excess K27 trimethylation of histone H3 (H3K27me3) at the mid-blastula transition. These changes interfere with the zygotic expression of several neurodevelopmental genes, particularly short gastrulation (sog), a component of an evolutionarily conserved sog-Decapentaplegic (Dpp) signaling system required for neuroectoderm specification. Our findings highlight the importance of early embryonic O-GlcNAcylation homeostasis for the fidelity of facultative heterochromatin redeployment and initial cell fate commitment of neuronal lineages, suggesting a possible mechanism underpinning OGT-associated intellectual disability.

Tax Incentives and Maturity Mismatch between Investment and Financing:Evidence from China

This paper examines the effects of China's accelerated depreciation policy(ADP)on the maturity mismatch between investment and financing.Using panel data for China's A-share nonfinancial listed companies from 2010 to 2019 and a staggered difference-in-differences approach,we found the following.First,ADP significantly aggravated the degree of corporate maturity mismatch,and this result was robust across multiple checks.Second,due to an insufficient long-term loan supply,firms had to finance the fixed investments induced by ADP with short-term debts,leading to maturity mismatches.Third,the positive policy effects were mainly significant for firms with high policy exposure,high-risk preferences,a high degree of information asymmetry,and firms with weak long-term financing capacity.Finally,maturity mismatch exacerbated corporate financial risks.Our research findings indicate that passive maturity mismatch is prevalent among Chinese companies and emphasize the need to address financial repression in order to mitigate the potential financial risks that may arise from tax incentives.

Profiling proteomic responses to hexokinase-II depletion in terpene-producing Saccharomyces cerevisiae

Hexokinase II(Hxk2)is a master protein in glucose-mediated transcriptional repression signaling pathway.De-grading Hxk2 through an auxin-inducible protein degradation previously doubled sesquiterpene(nerolidol)pro-duction at gram-per-liter levels in Saccharomyces cerevisiae.Global transcriptomics/proteomics profiles in Hxk2-deficient background are important to understanding genetic and molecular mechanisms for improved nerolidol production and guiding further strain optimization.Here,proteomic responses to Hxk2 depletion are investi-gated in the yeast strains harboring a GAL promoters-controlled nerolidol synthetic pathway,at the exponential and ethanol growth phases and in GAL80-wildtype and gal80Δbackgrounds.Carbon metabolic pathways and amino acid metabolic pathways show diversified responses to Hxk2 depletion and growth on ethanol,including upregulation of alternative carbon catabolism and respiration as well as downregulation of amino acid synthesis.De-repression of GAL genes may contribute to improved nerolidol production in Hxk2-depleted strains.Seven-teen transcription factors associated with upregulated genes are enriched.Validating Ash1-mediated repression on the RIM4 promoter shows the variation on the regulatory effects of different Ash1-binding sites and the syner-gistic effect of Ash1 and Hxk2-mediated repression.Further validation of individual promoters shows that HXT1 promoter activities are glucose-dependent in hxk2Δbackground,but much weaker than those in HXK2-wildtype background.In summary,inactivating HXK2 may relieve glucose repression on respiration and GAL promoters for improved bioproduction under aerobic conditions in S.cerevisiae.The proteomics profiles provide a better genetics overview for a better metabolic engineering design in Hxk2-deficient backgrounds.