Liquid-liquid phase separation (LLPS) is a rapidly growing research focus due to numerous demonstrations that many cellular proteins phase-separate to form biomolecular condensates (BMCs) that nucleate membraneless organelles (MLOs). A growing repertoire of mechanisms supporting BMC formation, composition, dynamics, and functions are becoming elucidated. BMCs are now appreciated as required for several steps of gene regulation, while their deregulation promotes pathological aggregates, such as stress granules (SGs) and insoluble irreversible plaques that are hallmarks of neurodegenerative diseases. Treatment of BMC-related diseases will greatly benefit from identification of therapeutics preventing pathological aggregates while sparing BMCs required for cellular functions. Numerous viruses that block SG assembly also utilize or engineer BMCs for their replication. While BMC formation first depends on prion-like disordered protein domains (PrLDs), metal ion-controlled RNA-binding domains (RBDs) also orchestrate their formation. Virus replication and viral genomic RNA (vRNA) packaging dynamics involving nucleocapsid (NC) proteins and their orthologs rely on Zinc (Zn) availability, while virus morphology and infectivity are negatively influenced by excess Copper (Cu). While virus infections modify physiological metal homeostasis towards an increased copper to zinc ratio (Cu/Zn), how and why they do this remains elusive. Following our recent finding that pan-retroviruses employ Zn for NC-mediated LLPS for virus assembly, we present a pan-virus bioinformatics and literature meta-analysis study identifying metal-based mechanisms linking virus-induced BMCs to neurodegenerative disease processes. We discover that conserved degree and placement of PrLDs juxtaposing metal-regulated RBDs are associated with disease-causing prion-like proteins and are common features of viral proteins responsible for virus capsid assembly and structure. Virus infections both modulate gene expression of metalloproteins and interfere with metal homeostasis, representing an additional virus strategy impeding physiological and cellular antiviral responses. Our analyses reveal that metal-coordinated virus NC protein PrLDs initiate LLPS that nucleate pan-virus assembly and contribute to their persistence as cell-free infectious aerosol droplets. Virus aerosol droplets and insoluble neurological disease aggregates should be eliminated by physiological or environmental metals that outcompete PrLD-bound metals. While environmental metals can control virus spreading via aerosol droplets, therapeutic interference with metals or metalloproteins represent additional attractive avenues against pan-virus infection and virus-exacerbated neurological diseases. The recent re-classification of the eukaryotic cellular phenomena of phase separation of protein condensates as the underlying mechanism creating membraneless organelles (MLOs) for cellular compartmentalization is initiated by a liquid demixing program. Liquid-liquid phase separation (LLPS) is an evolved cellular survival strategy mediating stress-triggered environmental sensing and also nucleates the cellular self-assembly processes of biomolecular condensates (BMCs) required for many cellular processes, including signaling, cytoskeletal organization, and transcriptional regulation ,2,3,4,5.Many different viruses employ LLPS to engineer BMCs used for their replication. Various virus family-centric terms and functions are used to describe these ‘viral replication compartments’ , ‘virosomes’, ‘virus factories’ (VFs), ‘viroplasm’, ‘mini-organelles’, ‘inclusion bodies’, and ‘negri bodies’ (NBs) ,28,29,30Gag promotes Gag and Capsid (CA) domain-resistant SGs causing translational arrest [2+-dependent HIV-1 NC domain is responsible for the positioning, trafficking, and packaging of the viral genomic RNA (vRNA) during virus assembly [2+- and ZnF-dependent LLPS condensates in vitro and in living cells [2+-dependent LLPS support a pan-retrovirus-wide model of virus assembly dynamics primed by LLPS [2+-dependent manner [We have recently demonstrated that the Nucleocapsid (NC) domain of HIV-1 pr55l arrest ,51, suppl arrest ,53. Morel arrest . The Zn2ng cells . Measlesng cells . In sili by LLPS . The SARt manner ,59,60,61PrLD-containing proteins are central to dynamic protein interaction network coordinating hubs . Crystal2+ and ZnF motifs within PrLDs [2+-dependent NC condensate model may be extended to represent a fundamental underlying mechanism nucleating virus replication, and an untapped avenue for pan-virus pharmacological targeting. To elucidate a common mechanism controlling and linking LLPS, SGs, neurological aggregates, and virus assembly dynamics, we performed literature surveys demonstrating that these assemblies are promoted and inhibited by cellular, physiological, and environmental exposure to Zn2+ and Cu2+. Our findings are supported by reports that modulation in Cu/Zn ratios and altered expression of proteins maintaining cellular and physiological ion homeostasis are hallmarks of both virus infections and neurological diseases. We and others have contributed to the greater body of literature promoting the theory that towards their own replicative benefit, underlying virus infections induce neurocognitive disorders and may exacerbate neurological diseases [Indeed, a core requirement of replication and budding of bona fide infectious virus particles is the specific binding of structural viral proteins to their cognate vRNA. Since vRNA binding for retrovirus Gag proteins is dependent on Znin PrLDs , we havediseases ,70,71,72http://plaac.wi.mit.edu/), Predictor of Natural Disordered Regions  [http://mobidb.bio.unipd.it/) were used to identify and validate the positions of PrLDs in viral proteins. FASTA input sequences were obtained from the NCBI protein sequence database and analyzed using software default parameters, with VLXT and VSL2 output styles selected for PONDR, and regions having a >0.8 PONDR Score were mapped using Adobe Illustrator software. Mapped ZnF and RRM placements were determined using the NCBI protein sequence database, from supporting references listed in https://www.genome.jp/tools/motif/) and Prosite (https://prosite.expasy.org/) [http://phylot.biobyte.de) [http://itol.embl.de) [Prion-Like Amino Acid Composition  , and thesy.org/) , which wsy.org/) . Phylogrbyte.de)  and Inteembl.de) .This study did not generate new datasets or code. Protein sequences were obtained from the NCBI Reference Sequence Database, with Genbank IDs listed in Disordered metal-binding regions of proteins can be stabilized upon ion-binding for gain of structure and function . We hypoWe were interested to find which other viral proteins with functional equivalence to HIV-1 Gag in vRNA-binding and encapsidation and viral capsid architecture also possessed juxtaposing PrLDs, RRMs, and ZnFs, inducing metal-dependent condensates. In some cases where identification of functional orthologs of Gag was complicated by diverging viruses having vastly different replication intermediate steps, we broadened our inclusion criteria to accept viral proteins that bind viral RNAs, or viral proteins possessing characterized ZnFs.2+-dependence, or from their propensity to multimerize, phase-separate, and bind vRNAs [To map proximities of viral protein PrLDs, RRMs, and ZnFs, NCBI protein database amino acid sequences were analyzed using PONDR and PLAAC algorithms ,78,79,80nd vRNAs ,88,89,90nd vRNAs ,103,104.nd vRNAs ,107,108.2+ binding and pathogenesis. This virus’ ZnFs juxtapose C-terminal PrLDs as we have observed for other viral vRNA-binding proteins, while ZnF mutants have attenuated virus infectivity [The combinatorial features of PrLDs, RRMs, and ZnFs towards a chaperoned RNA-LLPS model nucleating virus assembly that we have recently described for HIV-1 is one that is dependent on the Gag NC domain  and may ectivity .High degrees of disorder thus represent an evolutionary asset providing ancient viruses, encoded by very little genetic material, a replicative advantage and protection against host-factor interference or immune recognition. Indeed, viral nucleocapsid (nucleoproteins) have also frequently been characterized as viral RNA or protein chaperones. Since RBD-containing PrLD proteins are those that nucleate SGs , to repl2+ binds to RRMs of other cellular proteins that phase-separate and assemble into SGs [In our previous work, we observed that although Spumaviruses do not have ZnFs, these, nevertheless, have an abundance of RRMs mapping to the same approximate positions juxtaposing conserved C-terminal PrLDs found for all retroviruses ,124. Theinto SGs . RRM reg2+ binding, mediate conformational changes of NC proteins, permitting binding to coiled nucleic acid grooves.Viral proteins that undergo phase separation bind vRNAs via RRMs or ZnFs within their PrLDs. Many cellular RNA-binding proteins  have arginine-rich, positively charged sequences  that drive LLPS in an ionic strength-dependent manner ,125. Pos2+, Cu2+, and magnesium (Mg2+) are the most common metal ions binding to viral proteins and participate in strand transfer during reverse transcription of the vRNA, nucleic acid annealing and integration, transcription, and vRNA maturation [Among the many roles of metal ions in biological processes, these bridge interactions between distant residues of protein domains mediate protein–ligand interactions and serve as nucleophilic catalysts of enzymatic active sites. Although many biological processes are historically accepted as metal-ion dependent, a growing body of knowledge also demonstrates that metals can be interchangeable with certain cellular mechanisms. Metals are an integral part of viral proteins and play important roles in their survival and pathogenesis. Znturation . The impturation .2+, positively influence phase separation [2+, Ca2+) negatively influence PrLD–RNA interactions and reduce LLPS, as does the metal chelator EDTA [2+ is the presumed metal binding ZnFs, ZnFs also coordinate numerous other metal ions . As thed in [2+ . Copper-d in [2+ . Cu2+ haunctions .2+-binding prevents Aβ protein β sheet conformation and aggregation [2+ binding to Aβ is more kinetically favored, and Cu2+ can bind to previously self-aggregated Aβ [2+-binding, on the other hand, induces spherical Aβ structures different from β sheet structures [2+ concentrations inhibiting Zn2+-induced aggregates [2+ and Zn2+ bind to the same Aβ histidine residues, where Zn2+ precipitates at least two peptides to induce spherical structures, whereas Cu2+ outcompetes Zn2+ to inhibit intra-protein contacts [2+), Zn2+ alone could induce Tau protein condensates, another prime causative agent of neurodegenerative disease deposits [2+ and Cu2+, with both metals inducing structural changes and decreased solubility [2+-binding induces protease resistant variants [Aβ protein has been extensively studied as a prime causative agent of neurodegenerative disease deposits . Importantly, a number of different metal ions associate with monomeric Aβ to induce its aggregation but individually cause measurable variations in the types of Aβ deposits they induce ,133. Indregation ,134 and regation ,136. Mosgated Aβ . Zn2+-biructures . Early egregates . It was contacts . Indeed,deposits . The cellubility  and difflubility ,142, whevariants ,144. Relvariants . Indeed,variants , modulat2+ versus circulating Zn2+ levels in virus-infected patients. Indeed, Zn2+ is critical for innate and adaptive immunity [2+ deficiency correlates with diminished CD4+ T cells, high viral loads, AIDS, and mortality [2+ deficiency, virus replication requires Zn2+ [An interesting conundrum is exposed in the examination of the requirement of viruses for Znimmunity  and its immunity ,149,150,immunity ,152,153.ortality ,158,159,ortality ,163,164.res Zn2+ ,165.2+ is the most common viral protein cofactor assisting numerous replication processes of both RNA and DNA viruses [2+ also confers proper folding and varied functions to HIV-1 Integrase and Tat and Vif proteins [2+ ejectors [Zn viruses . In addi viruses , Zn2+ alproteins ,175,176.ejectors ,193,194.2+ deficiency is also common to infection by numerous other viruses, and Zn2+ supplementation as an antiviral therapy has been clinically tested against many divergent viruses, including herpes simplex virus (HSV), rhinovirus (RV), influenza, human papillomavirus (HPV), HIV-1, and hepatitus C virus (HCV) [2+-deficient plants are also more susceptible to diseases. Zn2+ deficiencies result from infection by turnip yellow and tobacco mosaic viruses, and plants supplemented with Zn2+ have ameliorated responses to pathogenic diseases [2+ was first reported to inhibit RV in 1974 [2+ against coronavirus, encephalomyocarditis virus, foot and mouth disease virus, HCV, HSV, HIV-1, HPV, Rous sarcoma virus (RSV), Semliki Forest virus (SFV), Sindbis virus, vaccinia virus, varicella-zoster virus, HEV, and arteriviruses [2+ concentrations tested exceed safe physiological ranges associated with clinical testing [Znus (HCV) . Zn2+-dediseases ,196,197. in 1974 , and zin in 1974 . Numerouiviruses ,201,202. testing ,204, pro testing ,206.2+ both promotes and inhibits virus replication, while its deficiency is associated with increased viral load, suggesting that there are major gaps in our understanding of how this metal influences viruses. While Zn2+ is required for virus replication, its chelation inhibits viral proteins  and virus replication , and Japanese encephalitis virus (JEV)) [2+ chelators, such as TPEN -1,2-ethanediamine) and 1,6-hexanediol, other drug classes with better safety profiles, including bananins, may be effective against HIV-1 and other viruses, including coronaviruses [2+ ionophores, such as pyrithione and chloroquine derivatives, among others, have also recently been shown to inhibit diverse groups of viruses, such as HSV, DENV, and SARS-CoV-2 [It is puzzling that Zns (JEV)) ,209,210.aviruses ,212. Zn2RS-CoV-2 ,214.2+- and Cu2+-regulating and -regulated cysteine-rich metallothioneins (MTs) [2+ imbalances are also present in human cancers and are also linked to regulation of HPV infection [2+ by the HPV16 E7 protein produces a compact environment leading to its self-assembly into spherical oligomers similar to those found in amyloids, whereas Zn2+ depletion results in loss of its aggregation [2+-dependent soluble agglomerates, whereas the metal chelating agent EDTA stabilizes its monomeric form and destabilizes existing agglomerates [2+ complexes also cause E6 aggregation, inhibiting its function [2+ and be regulated by Zn2+ [Another piece of the puzzle may involve Znns (MTs) . Despitens (MTs) . Metallonfection . As withnfection ,218. Notregation ,220,221.omerates , and whefunction . Recent function , a prote by Zn2+ .2+ and Zn2+-chelation both negatively affect viruses may be explained by differing research aims or experimental models from past reports. If virus biogenesis and assembly is Zn2+-dependent, then pre-formed virus capsid structure may be challenged by higher external concentrations of Zn2+ outcompeting virus-internal Zn2+, just as Zn2+ chelation would. In addition, if metal ions ultimately regulate virus core assembly (and stability) nucleated by metal-induced condensates during late stages of virus replication, then virus cores at early stages of reinfection should also be susceptible to metal ions or their chelation. Indeed, another effective antiviral agent, the zinc-finger-reactive disulfide NSC20625, ejects Zn2+ from JUNV Matrix protein RING finger motif, causing incomplete virion uncoating and release of NC into the cytoplasm [2+ bound to highly conserved ZnFs of influenza virus M1 matrix proteins is described as the determining factor of conformational transition of capsids in acidic environments, leading to their uncoating [2+ cause virus uncoating by destabilizing Zn2+-bound M1 protein [2+, leading to increased virus permeability [Confounding observations that both Znytoplasm . Zn2+ boncoating . Similarncoating . Upon in protein . Poliovieability .2+, ultimately regulating virus core assembly and stability, susceptibility of cell free viruses to environmental metal ions or their chelation should be evaluated. Indeed, virucidal agents ejecting Zn2+ from NC to inactivate HIV-1 have been developed for use as topical microbicides [2+ and Zn2+ also show virucidal activity against HIV-1 and other enveloped viruses [2+ ions and Zn2+ chelators inactivate viruses. Landmark studies relevant to the current COVID-19 pandemic tested a range of surfaces to find that copper alloys and Cu/Zn brass surfaces inactivated replication and propagation abilities of SARS-CoV [2+ is found to synergize with Cu2+ in surfaces and to increase efficacy of brass surfaces with lower percentages of Cu2+, suggesting that these ions destabilize capsids for viral genome destruction [2+ ion that affects shell stability [2+ and is also influenced by Cu2+ [2+ metallopeptidase angiotensin-converting enzyme 2 (ACE2) entry receptor, and high numbers of Zn2+-binding cysteine residues within these regions may also contribute to increasing binding affinity of the S protein to ACE2 [To determine how viruses are affected by exogenous Znobicides . Non-tox viruses . Thus, bSARS-CoV ,230 and SARS-CoV . Norovirtruction ,231. Imptability . Indeed,tability ,233. Imp by Cu2+ . Intrins to ACE2 ,235,236.2+ supports capsid assembly and structure across viruses through its interaction with ZnFs within PrLDs, difficulties in identifying effective therapeutics interfering with ZnFs or PrLDs for the elimination of virus condensates lead us to consider additional ways to control Zn2+-dependent viral condensates by examining upstream physiological or cellular regulating processes. From reports above describing Zn2+ binding site competition by other ions, such as Cu2+, that interfere with the establishment of condensates, below we describe alternative routes to control pathologic forms of LLPS, as supported by ancient evolutionary protein chaperone pathways.Despite the many instances demonstrating that Zn2+) has been used to disinfect fluids, solids, and tissues for centuries. Cu2+ was discovered by the ancient Greeks in the time of Hippocrates (400 BC), who used it to purify water and treat pulmonary disease. For their anti-fouling properties, Cu2+ vats were used to store holy water from the Ganges River, and Cu2+ strips were also used to construct ship hulls by the early Phoenicians. By the 18th century, Cu2+ was widely accepted for clinical treatment of mental disorders and pulmonary diseases. Both early American pioneers and Japanese soldiers of WWII placed Cu2+ coins to sterilize their drinking water, and NASA Apollo flights used Cu2+-based water sterilizing systems. Cu2+ has also historically been used by Africans and Asians to treat skin diseases [2+ is highly abundant and safe, with 1 mg consumed daily and excess copper released by excretion. More modern demonstrations of Cu2+ safety come from widespread and long-term use of Cu2+-based intrauterine devices (IUDs) to which human tissues have low sensitivity [2+-based IUDs correlated with 50% lower incidence of cervical cancers [Copper  [2+ concentrations lower than that required for inactivation by ethanol and where Cu2+ preserved cell viability while completely inhibiting formation of syncytia and virus production [2+ caused its cysteine-dependent oxidation [2+ in a cysteine-dependent manner, which was most notably found to cause its aggregation [Copper is a well-established biocide used against viruses and many other pathogens. In 1964, Cuiophages , followe in 1971 . In 1974nst Cu2+ . In 1992and HSV) ,245. Duroduction . More prxidation . HIV-1 pregation .2+-containing filters also effectively neutralize HIV-1 in medium and breast milk and reduce cell-associated HIV-1 in a dose-dependent manner [2+-containing filters also reduce infectious viral titers of several DNA and RNA viruses, including YFV, influenza A virus, MeV, RSV, adenovirus type 1, and cytomegalovirus [2+ in water pipes synergize with low levels of free chlorine has proven effective in inactivation of poliovirus, bacteriophage MS-2, hepatitis A virus, human rotavirus, and human adenovirus [2+ reduced survival and infectivity of waterborne viruses poliovirus, coxsackie virus types B2 and B4, echovirus 4, and simian rotavirus SA11 by 95% [The use of copper in free flow filters deactivates HIV-1 and West Nile virus, reducing infectious titers of these viruses by 5 to 6 logs . Cu2+-cot manner . Cu2+-coalovirus . Likewisenovirus ,251. The1 by 95% . Finally1 by 95% .2+ alloys can rapidly and effectively kill a wide range of microbial pathogens at a range of temperatures and under various conditions of humidity [2+ alloys is useful in preventing secondary transfer from surfaces in clinical facilities and other closed environments, including long term care facilities, public washrooms, cruise ships, and casinos [2+ in hours, while it can remain infectious on other surfaces for days [2+ surfaces [2+ also possess broad-spectrum anti-microbial and antiviral properties [Non-toxic, hybrid cured coatings containing copper also have virucidal activity against HIV-1 and other enveloped viruses . The rishumidity . Clinicahumidity ,256,257. casinos . As withfor days . This hafor days . Manufacfor days , also insurfaces . Textileoperties .2+ for their functions, while others bind to and control the uptake and delivery of Cu2+ for cellular and physiological processes. Cytosolic Cu-Zn-superoxide dismutase (SOD-1) is a ubiquitous cytosolic homodimeric isoenzyme that scavenges and catalyzes the dismutation of superoxide radicals [2+ and Zn2+ and leading to distorted SOD-1–SOD-1 interactions, leading to formation of insoluble aggregates described above [2+ chaperone [A number proteins are bound by Curadicals . Severaled above —a phenomed above . Intrigued above , entomoped above , and baced above , encode ed above . All chaed above  but may haperone . Modulathaperone ,270. Cu/haperone .2+-binding glycoprotein [2+, accounting for more than half of plasma Cu2+ [2+ from the liver and delivers it directly to cells for the synthesis of 40–70% of Cu2+-binding proteins and enzymes of various organs and tissues [2+ attributed to CP [2+ to the sites of infection to attack invading pathogens with Cu2+ toxicity [2+ homeostasis, may exacerbate the LLPS underpinnings of viral and neurological diseases.Ceruloplasmin (CP) is another Cuoprotein , mostly sma Cu2+ ,274,275. tissues ,277. A ced to CP ,279,280,toxicity . CP exprtoxicity . CP is atoxicity , and remtoxicity . CP is atoxicity ,285. Inctoxicity  and Alzhtoxicity , where itoxicity  and whertoxicity . These a2+ deficiencies associated with several diseases [Imbalances in homeostasis of physiological metals are important disease biomarkers. Plasma Cu/Zn ratios represent a common clinical assessment for Zndiseases . Elevatediseases , contribdiseases . It alsodiseases ,293 but diseases ,297,298.diseases . Signifidiseases ,301 and diseases . Increasdiseases .2+, inducing its beta-sheet conformation and aggregation [2+ and nucleic acids [sc) is induced by HIV-1 infection [2+- or Zn2+ -binding also induces PrP fold variants [2+ and Cu2+ to PrP is accompanied by structural changes and decreased solubility, where Cu2+-binding causes protease resistant PrP conformations [Fatal neurodegenerative prion diseases are caused by proteinaceous infectious particles, or “prions”. Prion disease is caused by the accumulation of abnormally folded isoforms of the cellular prion protein , where the unstructured amyloidogenic region of PrP preferentially binds to Curegation . Recent regation . Antimicic acids ,306. PRNic acids ,310,311,ic acids . PrP is ic acids ,314. Notnfection ,315. As variants ,142, sugrmations ,143,144.2+ and Cu2+ metal ions differentially influence formation of BMCs and protein aggregates of neurological diseases, and then extends to how viral infections induced metal imbalances may exacerbate neurological diseases (We were interested in gaining a better understanding of mechanistic intersections and fundamental features of proteins commonly associated with pan-viral engineered BMCs and neuropathologies. To propose a simplified model illustrating how viral infections may exacerbate neurological diseases, a bioinformatics-coupled meta-analysis system biology framework investigated how cellular, physiological, and environmental metals may govern both essential BMCs and pathological protein aggregates. The model illustrates how physiological Zndiseases A. Such i2+ bioavailability to inactivate viruses, possibly by outcompeting Zn2+-dependent processes during both early and late stages of virus replication. In early stage virus binding, entry, and uncoating, high Cu2+ concentrations may prematurely disrupt pre-formed virus capsid integrity to cause premature vRNA exposure and its subsequent degradation. In late stages of virus replication, high Cu2+ concentrations may interfere with the biogenesis of Zn2+-mediated viral BMCs required for nuclear transcription, export, translation, and trafficking and vRNA packaging by NC proteins. Indeed, many more viral proteins than those analyzed in 2+ [2+ concentrations. High Cu2+ that outcompetes Zn2+ for correct folding, activity, or multimerization of essential cellular proteins as BMCs may rather nucleate their aggregation as insoluble pathological plaques of neurodegenerative diseases (The model first illustrates how virus infections cause increases in both Cu/Zn ratios and expression of ancient antimicrobial prion proteins, such as Aβ protein. For one, this suggests that an evolutionary conserved anti-viral mechanism promotes heightened Cued in 2+ ,175,176.diseases A.2+ supplementation has been successfully used as treatment against many viruses [2+ supplementation is also described as a paradigm-shifting practice for neurodegenerative disease associated with Zn2+ and/or Cu2+ homeostasis abnormalities [2+ for BMC formation regulating many cellular processes, suggests that prolonged altered Cu/Zn ratios may represent an underappreciated and underlying feature of diseases (2+ metalloproteins and transporters are also modified by viral infections [2+ supplementation may assist in resolving its associated morbidities.Indeed, virus infections have been shown to exacerbate neurodegenerative disease phenotypes ,70,71,72 viruses , Zn2+ sumalities . Elevatediseases . In addifections ,326,327.2+. Cu2+ has historically been an easily procured effective antimicrobial and its anti-viral usage is supported by many demonstrations that this metal effectively inactivates all viruses tested. The current COVID-19 pandemic highlights an urgent need for methods reducing virus transmission. Mandatory face masks protect against virus aerosol droplets, but with evidence of SARS-CoV-2’s survival on certain surfaces for hours to days, and with copper surfaces showing the fastest inactivation [2+ coating of reused face masks and shields may be more protective (2+ mediates virus inactivation is by its outcompeting virus-internalized structure-lending Zn2+, leading to premature virus uncoating and degradation of the consequently exposed vRNA (2+ coatings to touch surfaces may reduce transmission of current and future dangerous viruses while targeting drugs or vaccines become approved and available. Considering the global economic arrest caused by COVID-19, simpler strategies such as Zn2+ supplementation or Cu2+ surface coatings may keep future pandemics at bay.The model extends itself to propose countering viral transmission using environmental Cutivation , Cu2+ cootective B. Finallsed vRNA C. Theref


Sinapic acid (SA) is a naturally occurring phenolic compound with antioxidant properties. It also has a wide range of pharmacological properties, such as anti-inflammatory, anticancer, and hepatoprotective properties. The present study aimed to evaluate the potential pharmacological effects of SA against hypertrophic responses in neonatal rat cardiomyocytes. In order to evaluate the preventive effect of SA on cardiac hypertrophy, phenylephrine (PE)-induced hypertrophic cardiomyocytes were treated with subcytotoxic concentrations of SA. SA effectively suppressed hypertrophic responses, such as cell size enlargement, sarcomeric rearrangement, and fetal gene re-expression. In addition, SA significantly inhibited the expression of mitogen-activated protein kinase (MAPK) proteins as pro-hypertrophic factors and protected the mitochondrial functions from hypertrophic stimuli. Notably, SA activated Sirt3, a mitochondrial deacetylase, and SOD2, a mitochondrial antioxidant, in hypertrophic cardiomyocytes. SA also inhibited oxidative stress in hypertrophic cardiomyocytes. However, the protective effect of SA was significantly reduced in Sirt3-silenced hypertrophic cardiomyocytes, indicating that SA exerts its beneficial effect through Sirt3/SOD signaling. In summary, this is the first study to reveal the potential pharmacological action and inhibitory mechanism of SA as an antioxidant against cardiac hypertrophy, suggesting that SA could be utilized for the treatment of cardiac hypertrophy. Cardiac hypertrophy is a type of cardiac remodeling that results in the thickening of the heart walls in response to many pathological stimuli, including myocardial infarction and hypertension . It occuSilent information regulator 2 (SIR2) is a family of histone deacetylases (HDACs) that catalyzes the deacetylation of both histone and non-histone lysine residues . SIR2 prMany studies have reported that Sirt3 plays a crucial role in the pathogenesis of various cardiac diseases, such as cardiac hypertrophy, heart failure, and doxorubicin-induced cardiomyopathy . Sirt3 aMany functional food compounds have been reported to possess cardiac protective effects as regulators of the epigenome, which is closely associated with cardiovascular diseases (CVD) . Indeed,2,2-diphenyl-1-picrylhydrazyl (DPPH) -2,5-diphenyltetrazolium bromide  assay. In brief, NRVMs were seeded onto 96-well plates (2000 cells/well) and treated with 100 μM PE only, or were pretreated with various concentrations of SA, such as 50, 100, 200, 400, and 800 μM, for 24 h, and then treated with 100 μM PE for another 24 h. After 24, 48, and 72 h, the cardiomyocytes were further incubated with 0.5 mg/mL MTT reagent. After incubation at 37 °C for 2 h, the supernatants were removed, and formazan crystals were dissolved in 100 μL DMSO. The absorbance was measured at 570 nm, using a spectrophotometer . The cell viability was evaluated by comparison with control cells treated with 0.1% DMSO.After treatment with SA and/or PE, NRVMs were cultured on collagen-coated coverslips and fixed with 4% paraformaldehyde for 10 min, permeabilized with 0.5% Triton X-100 in phosphate-buffered saline for 10 min, and further blocked with 5% bovine serum albumin (BSA), for 1 h, at room temperature. The cells were then incubated with a primary antibody against α-actinin , at 4 °C, overnight, followed by incubation with an Alexa 488-conjugated secondary antibody , for 1 h, at room temperature. Immunofluorescence staining was observed under a fluorescence microscope . Cell-surface areas were measured by using NIH ImageJ software.Total RNA was isolated from NRVMs, using the TRI reagent (Sigma) to measure the mRNA expression levels of hypertrophic markers, such as atrial natriuretic factor (ANF), brain natriuretic peptide (BNP), and β-myosin heavy chain (β-MHC), and of mitochondrial biogenesis genes, such as nuclear respiratory factor-1 (NRF-1), peroxisome proliferator-activated receptor α (PPARα), estrogen-related receptor α (ERRα), and peroxisome proliferator-activated receptor γ coactivator-1β (PGC-1β). Total RNA was reverse transcribed, using ImProm II Reverse Transcriptase  with oligo-dT priming. Then qRT-PCR was performed, using SYBR qPCR Master Mix , in the TaKaRa Thermal Cycler Dice Real Time System Single TP 815 . GAPDH was used as a loading control. The primer sequences are shown in NRVMs treated with SA and/or PE were harvested and lysed in RIPA buffer  containing a protease inhibitor cocktail  and a phosphatase inhibitor cocktail (Sigma). Protein homogenates from the cardiomyocytes were separated on SDS–PAGE and transferred onto PVDF membranes . After blocking for 1 h with 5% BSA in TBST buffer , the membranes were incubated overnight, at 4 °C, with the appropriate primary antibodies against MAPK/ERK kinase , phosphorylated MEK , extracellular signal-regulated kinase 1/2 , phosphorylated ERK1/2 , c-Jun N-terminal kinase , p-JNK , mitochondrial subunit A of complex II , Sirt3 , SOD2 , acetylated K122-SOD2 , acetylated K68-SOD2 , and β-actin . The membranes were then incubated with the appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies . Protein bands were developed by using an enhanced chemiluminescence kit  and a UVITEC Mini HD9 system . Equal protein loading was confirmed by probing for β-actin on the same membrane. The intensity of each protein was quantified by using NIH ImageJ software.ATP levels were measured, using the ATP Colorimetric/Fluorescence Assay Kit , as described previously, with minor modification . In brieIntracellular ROS production was detected by the fluorescence intensity of DCFH-DA . Briefly, the cells were resuspended in DCF-DA solution and incubated for 30 min, at 37 °C. After the cells were washed with PBS, the absorbances were measured at 488 nm excitation and 525 nm emission wavelength, on a microplate reader . The malondialdehyde (MDA) and catalase levels were measured by using colorimetric/fluorometric assay kits according to the manufactures’ instructions . The absorbances for MDA and catalase were 532 and 5570 nm, respectively, using a spectrophotometer .Sirt3 siRNAs and the Sirt3-mimic negative control were purchased from Dharmacon CO. . Four individual Sirt3 siRNAs were mixed. NRVMs were cultured in serum-free medium for 24 h and transfected with 50 nM Sirt3 siRNA mixture and Sirt3-mimic siRNAs (negative control), using DharmaFECT-1 reagent for 48 h, according to the manufacturer’s instructions (Dharmacon). The cells were then pretreated with SA for 24 h, followed by exposure to 100 μM PE for another 24 h, to stimulate hypertrophic responses. The Sirt3 siRNA sequences were as follows: Sirt3 siRNA #1: GCUCAUGGGUCCUUUGUAU, Sirt3 siRNA #2: GGAU GGACAGGACG GUAA, Sirt3 siRNA #3: CAGCAAGGUUCUUACUACA, and Sirt3 siRNA #4: CAGGUUGUCUGAAUCGGUA.P < 0.05 was considered statistically significant.All data are presented as the mean ± standard error of the mean (SEM). Statistical significance was assessed by one-way analysis of variance (ANOVA) with a Bonferroni post hoc test for multiple comparisons, using Prism 5.03 software . To elucidate the cytotoxic effects of SA in cardiomyocytes, the viability of the cardiomyocytes was determined by using the MTT assay after treatment with 50, 100, 200, 400, and 800 μM SA for 24, 48, and 72 h. The results showed that treatment with 50, 100, 200, and 400 μM SA for any duration did not significantly reduce the cell viability . On the To determine whether SA could attenuate cardiomyocyte hypertrophy in NRVMs, the cardiomyocytes were treated with 100, 200, and 400 μM SA for 24 h and further treated with 100 μM PE for another 24 h. Immunofluorescence staining against α-actinin revealed that, compared with the vehicle-treated cardiomyocytes, sarcomeric rearrangement was evoked in the PE-induced hypertrophic cardiomyocytes. However, compared with PE treatment only, SA pretreatment of hypertrophic cardiomyocytes prior to PE treatment blocked the sarcomeric rearrangement A. Cell-sPrevious studies have reported that the activation of the mitogen-activated protein kinase (MAPK) signaling pathway, including MEK, ERK1/2, and JNK proteins, plays a critical role in the induction of cardiac hypertrophy ,41. TherPrevious studies have shown that the impairment of the mitochondrial structure and functions are closely associated with cardiac hypertrophy ,43. TherSirt3, a mitochondrial deacetylase enzyme, has a preventive effect against cardiac hypertrophy through regulating the mitochondrial function by the deacetylation of mitochondrial proteins, particularly SOD2 ,44. TherTo determine whether SA can inhibit oxidative stress in hypertrophic cardiomyocytes, we determined ROS production and levels of oxidative-stress-related proteins, such as MDA and catalase. The DCFH-DA as a ROS sensor dye significantly increased in the PE-only treated cardiomyocytes (65% increase in the PE-only treated cells vs. the vehicle-treated control cells). However, the increased level of DCFH-DA in the PE-only treated cells was inhibited on pretreatment with SA in a dose-dependent manner A. The leTo further assess whether the Sirt3/SOD2 signaling pathway is required for the anti-hypertrophic effect of SA in hypertrophic cardiomyocytes, we silenced the expression of Sirt3 by Sirt3 siRNA mixture transfection in SA-pretreated or vehicle-pretreated hypertrophic cardiomyocytes treated with PE. We first determined whether the Sirt3 siRNA mixture can effectively block Sirt3 expression. We found that the expression level of Sirt3 significantly decreased in the cardiomyocytes transfected with the Sirt3 siRNA mixture for 48 h (0.2-fold decrease in the Sirt3 siRNA-transfected cells vs. the negative control siRNA-transfected cells) A. To assAs cardiac hypertrophy is considered to be a major risk factor for cardiovascular morbidity and mortality ,46, manyTo elucidate the protective effect of SA against cardiac hypertrophy, in vitro approaches using NRVMs were used. NRVMs are relatively easy to isolate and survive for a long period in culture system compare with adult cardiomyocytes . Especia2O2- or antimycin A–treated adult cardiomyocyte and also blocked ROS production in I/R injured heart [In a previous study, the pharmacological effect of SA was evaluated on myocardial ischemia–reperfusion injury (I/R injury) and oxidative stress, a major response to I/R injury, using both ex vivo and in vitro approaches in the Langendorff-isolated heart system and H9c2 cardiomyoblast cells. That study showed that SA, a phenolic compound, protects cardiac cells from I/R injury by inhibiting oxidative stress due to its antioxidant ability . Additioed heart . To dateDue to the high consumption of energy in the heart, the mitochondria play an important role in maintaining the cardiac function through energy production . TherefoWe also determined the activities of MAPK proteins, such as MEK, ERK1/2, and JNK, in SA-pretreated hypertrophic cardiomyocytes. MAPK proteins are involved in various cellular functions, including proliferation, differentiation, and apoptosis . They alThe mitochondria are the main sources of ROS, which cause oxidative stress. At a normal state, the production of ROS and intracellular antioxidants is balanced, resulting in the clearance of ROS. However, an imbalance between ROS and antioxidants causes oxidative stress and cellular damage. In particular, the heart is susceptible to oxidative stress because a larger number of mitochondria and fewer antioxidant proteins exist in cardiac cells than in other cells. Many studies have shown that the pathogenesis of many cardiac diseases, including cardiac hypertrophy, is closely associated with the generation of oxidative stress from the mitochondria within cardiomyocytes. SOD2, also known as manganese-dependent superoxide dismutase (MnSOD), is an antioxidant present in the mitochondria. SOD2 activity is mediated by post-translational modification through reversible lysine acetylation at K68 and K122 ,60. In pMany natural compounds have been shown to have beneficial effects against oxidative stress. Especially, barley beta-glucan (BBG) as a natural polysaccharide exhibited the cardiac protective effect in oxidative-stress-induced human umbilical vein endothelial cells (HUVECs) and zebrafish model through promoting SOD2 expression and angiogenesis . In addiIn conclusion, the present study revealed that SA attenuates cardiac hypertrophic responses through the preservation of mitochondrial functions. Furthermore, these inhibitory effects of SA are mediated by Sirt3 and SOD2, a mitochondrial antioxidant. We propose that SA has the potential to prevent and treat cardiac hypertrophy. The beneficial effect of SA against cardiac hypertrophy should be further determined by the physiological aspects, using adult cardiomyocytes, and verified by in vivo study, using pressure-overload-induced mouse animal model to overcome limitation of presented results, using NRVMs.


Chlorocebus pygerythrus), sampled near Lake Nabugabo, Masaka District, Uganda. Sequencing of the complete viral genomes and subsequent phylogenetic analysis identified LNMV as a distinct member of species Kunsagivirus C, in the undercharacterized picornavirid genus Kunsagivirus.We report the discovery and sequence-based molecular characterization of a novel virus, lanama virus (LNMV), in blood samples obtained from two wild vervet monkeys ( Picornavirales  is the se virus . The divse virus ,5,6.Coracias garrulus Linnaeus, 1758), a carnivorous migratory bird [Kunsagivirus  [Kunsagivirus A [Eidolon helvum Kerr, 1792), sampled in Southwest Cameroon [Papio cynocephalus Linnaeus, 1766), sampled in Tanzania in 1986 [Kunsagivirus B and Kunsagivirus C, respectively [In 2013, a novel picornavirus was discovered in feces of a wild European roller , sampled near Lake Nabugabo, Uganda.Here, we report the discovery and sequence-based molecular characterization of a novel kunsagivirus, lanama virus (LNMV), in blood samples obtained from two wild vervet monkeys  or tantalus monkeys  [g) and stored in liquid nitrogen until processing. RNA was extracted and converted to cDNA, which was sequenced on an Illumina MiSeq instrument according to previously published methods [One male and two female apparently healthy wild adult vervet monkeys were sampled for virus discovery at Lake Nabugabo, a satellite lake of Lake Victoria, in Masaka District, Central Region, Uganda. The sampled monkeys were members of a habituated group of vervet monkeys, studied since May 2011 .  ,11. In ky, 1841) ,13. Bloo methods . Briefly methods .Translated amino-acid sequences were aligned using Clustal W software  as imple2). The initial inocula were recovered and sequentially added in the same manner to human cervical carcinoma HeLa cells (ATCC CCL-2) and baby hamster kidney BHK-21 cells (ATCC CCL-10). Next, 200 µL of prewarmed media was added to each well, and cells were incubated and observed daily for cytopathic effect for 5 days. Media were removed from cells, replaced with 200 µL per well of fresh media, and cells were lysed by three freeze/thaw rounds at −80 °C. The resulting lysates were collected and pooled with the respective cell supernatants. For Passage 2, 300 µL of pooled supernatants and lysates from each sample condition was added to fresh cells in six-well plates under gentle agitation. Then, 2 mL of prewarmed media was added to each well, and cells were incubated and observed daily for cytopathic effect. On Day 0, Day 3, and Day 5 after inoculation, 200 µL of media was collected from each well and mixed with 600 µL of TRIzol LS (Thermo Fisher Scientific) for virus inactivation and downstream sequencing analysis. Removed volumes were replaced with prewarmed media.Virus isolation was attempted in Vero E6 cells (American Type Culture Collection (ATCC), Manassas, VA, USA; CRL-1586), seeded into wells of a 96-well plate according to ATCC culture recommendations. An amount of 50 µL of primate plasma or mock plasma  was added to each well and incubated for 1 h at 37 °C and 5% COKunsagivirus C), they appear to lack one of the 2A proteins (possibly 2A2) . All virus isolation attempts were unsuccessful.Deep sequencing yielded, after quality trimming, 4,203,737, 3,185,890, and 5,375,995 sequences of 155, 154, and 164 nucleotide average length for vervet monkeys 1, 2, and 3, respectively. Analysis of these sequences revealed the presence of a novel picornavirid in the blood of the two female vervet monkeys (vervet monkeys 2 and 3), whereas the male vervet monkey (vervet monkey 1) appeared to be uninfected. There were also putative hits to varied retrovirids, small circular DNA viruses, and hepadnavirids, but these were not investigated further due to ambiguous taxonomic assignments and low depth of coverage. In silico analyses enabled the assembly of near-complete picornavirid genomes  from both individuals (GenBank #MW218667 and #MW218668), with 337 and 134 reads from vervet monkeys 2 and 3, respectively, mapping to the picornavirid. Comparison of the obtained genome sequences to published picornavirid genomes enabled the identification of picornavirid-typical cleavage sites in the encoded polyprotein and, thus, the description of the genomic organization . Sequencbly 2A2) . The estKunsagiviruses A1 and B1 were discovered in feces samples of birds and bats, respectively ,7. Thus,


Psoriasis patients are associated with dysbiosis of gut microbiota and the deficiency of vitamin D or selenium. We herein present the update information regarding the stimulatory or regulatory effects of nutrients or food on psoriasis and the possible alleviation of psoriasis by nutritional strategies.Psoriasis is a chronic inflammatory skin disease characterized by accelerated tumor necrosis factor-α/interleukin-23/interleukin-17 axis, hyperproliferation and abnormal differentiation of epidermal keratinocytes. Psoriasis patients are frequently associated with obesity, diabetes, dyslipidemia, cardiovascular diseases, or inflammatory bowel diseases. Psoriasis patients often show unbalanced dietary habits such as higher intake of fat and lower intake of fish or dietary fibers, compared to controls. Such dietary habits might be related to the incidence and severity of psoriasis. Nutrition influences the development and progress of psoriasis and its comorbidities. Saturated fatty acids, simple sugars, red meat, or alcohol exacerbate psoriasis via the activation of nucleotide-binding domain, leucine-rich repeats containing family, pyrin domain-containing-3 inflammasome, tumor necrosis factor-α/interleukin-23/interleukin-17 pathway, reactive oxygen species, prostanoids/leukotrienes, gut dysbiosis or suppression of regulatory T cells, while  Obesitycontrols . Psoriascontrols . In vivocontrols . It is sobiotics .In this article, we review the recent studies regarding the stimulatory or regulatory effects of individual nutrients or food on psoriasis, and the possible alleviation of psoriatic symptoms by nutritional strategies.SFAs, such as palmitic acid or stearic acid, are rich in butter or red meat. The increased intake of SFAs is proposed to be a risk factor of obesity, dyslipidemia, or cardiovascular diseases . The hign-6 PUFA linoleic acid is rich in vegetable oils or margarines. Relations of linoleic acid consumption and psoriasis are still controversial. Linoleic acid undergoes metabolic conversion to arachidonic acid, which is incorporated in the cell membrane. When tissues are exposed to stimuli like injuries or cytokines, arachidonic acid is released from membranes and converted into lipid mediators by various enzymes. Prostanoids and leukotrienes (LTs) are the main lipid mediators derived from arachidonic acid, and might promote psoriasis [2 (TXA2) binds TXA2 receptor (TP), and the level of TXA2 is increased in murine IMQ-induced psoriasiform dermatitis [2 synthase ameliorated the IMQ-induced psoriasiform dermatitis and suppressed the IL-17 production by γδT17 cells [The essential soriasis . Among prmatitis . Stimula17 cells . TP-defi17 cells .2 (PGE2) also promotes the IMQ-induced psoriasiform dermatitis; PGE2 produced by fibroblasts promotes the production of IL-23 in dendritic cells (DCs) [2, which acts back on the PGE2 receptors, EP2 and EP4 in these cells and enhances IL-23-induced expression of IL-23R by activating signal transducer and activator of transcription 3 (STAT3), cyclic AMP-responsive element binding protein 1, and NF-κB through cyclic AMP-protein kinase A signaling [2 signaling also induces the expression of various inflammatory Th17 signature genes like IL17a, IL17f, IL18r1. In IL-23-induced psoriasis model mice, the deletion of EP2 and EP4 in T cells inhibited the accumulation of Th17 cells and abolished the dermatitis [Another prostanoid, prostaglandin Els (DCs) . IL-23 iignaling . This PGrmatitis ,21.4 binds its receptor BLT1 on neutrophils, inducing their infiltration in the IMQ-induced psoriasiform dermatitis. The deletion of BLT1 or inhibition of LTB4 synthesis impaired the neutrophil infiltration and ameliorated the IMQ-induced dermatitis [Among LTs, LTBrmatitis .n-3 PUFAs, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), rich in fish are metabolized into resolvin E1 and D1, respectively. Barrea et al. reported that psoriasis patients showed lower intake of n-3 PUFAs compared to healthy controls [n-3 PUFA level in psoriasis patients is inversely correlated with psoriasis area and severity index (PASI) [n-3 PUFAs have been suspected to have anti-psoriatic effects. It is reported that n-3 PUFAs inhibit Th17 differentiation: DHA-treated DCs showed the decreased expression of costimulatory molecules, CD40, CD80, CD86, reduced IL-12, IL-23, IL-6 secretion, and exerted a reduced ability to induce Th1/Th17 differentiation [4-induced IL-23 production and migration of DCs, Th17 cells and γδT17 cells. These antagonistic effects of resolving E1 were mediated by binding to LTB4 receptor, BLT1. Resolvin D1 alleviated the IMQ-induced psoriasiform dermatitis and reduced the expression of IL-23, IL-17A, IL-17F, IL-22, and TNF-α in the lesions [controls . Serum nx (PASI) . n-3 PUFntiation . Resolvi lesions . The eff lesions .Cutibacterium acnes also produce SCFAs which might also exert a similar regulatory function in the skin [Aire) in medullary thymic epithelial cells via GPR41 [+ T cells into peripheral Tregs through histone H3 acetylation of the Foxp3 gene intronic enhancer by inhibiting histone deacetylase [IL-10 and Foxp3 transcripts [SCFAs are the fermentation products of dietary fiber in the colon, and enter circulation and reach the organs outside of intestines, including the skin. SCFAs like butyrate, propionate and acetate, regulate inflammation in the intestines. Skin commensals like the skin . SCFAs athe skin . SCFAs ithe skin , which cthe skin ,30. SCFAia GPR41 . SCFAs acetylase . Topicalnscripts . Tregs inscripts . Lesionanscripts . The rednscripts . Sodium nscripts : sodium nscripts .The excessive intake of simple sugars like sucrose may exacerbate psoriasis . High-frCxcl2, Ly6g mRNAs, Gr-1 protein and developed more Munro’s micro abscesses, and higher expression of NLRP3 and IL-1β in response to IMQ. The Western diet-fed mice showed the increased baseline expression of IL-17a without IMQ treatment. The mice fed with a high-fat, low-sugar diet showed more body weight gain, but less inflammation in response to IMQ than Western diet-fed mice, indicating that obesity is not sufficient and sugar content of the Western diet is a critical factor for the enhancement of dermatitis.Feeding with Western diet containing high levels of fat and of simple sugars in mice for 12 to 16 weeks aggravated the IMQ-induced psoriasiform dermatitis while high-fat, low-sugar diet did not [E. coli, decreased protective bacteria like Firmicutes, decreased SCFA production and its receptor GPR43 expression in the colon, and decreased forkhead box P3 (Foxp3)+ Tregs in mesenteric lymph nodes, leading to the enhanced systemic inflammation as well as colon [The short-time (4 weeks) feeding with Western diet promoted the accumulation of IL-17A-producing γδT cells with enriched IL-23 receptor expression in IMQ-induced psoriasiform dermatitis . The shoas colon . Long-teas colon . BA sequas colon , while tas colon . BAs binas colon . Such crThe 2003–2006 National Health and Nutrition Examination Survey data in USA showed lower intake of simple carbohydrates in psoriasis patients compared to controls . In contTraf3ip2 mutation, ameliorated symptoms of psoriasis-like dermatitis, scratching behaviors, and increased the secretion of mucin in ileum and of IgA in cecum, with alteration of the composition of gut microbiota [Dietary fiber represents carbohydrates that are resistant to digestion in the small intestine and that undergo varying degrees of fermentation in the colon. Glucose-based resistant starch is abundant in grains or legumes, and highly fermentable. The dietary supplementation of fibers is reported to show systemic and intestinal anti-inflammatory effects . The intcrobiota .FLG gene mutations. R501X and 2282del4 null mutations in FLG gene are associated with AD susceptibility among northern European populations with low sun exposure, but not among Greek and Egyptian cohorts with high sun exposure, indicating that FLG mutations are negatively selected in populations with high UV exposure and resultant high vitamin D synthesis in the skin [There are two ways to meet vitamin D requirements in mammals: via dietary intake and via synthesis in skin by sun exposure. The food sources of vitamin D are cod liver oil, swordfish, salmon, tuna, sardines, beef liver, egg, or cheese. A whole-body exposure to UVB radiation inducing the minimal erythema dose for 15–20 min leads to the production of up to 10,000 IU of vitamin D while the recommended dietary allowance of vitamin D for adults ≤70 years is 600 IU/day . The disthe skin .It is reported that serum vitamin D levels are reduced in patients with psoriasis or PSA compared to controls ,53. In aVitamin D is a key modulator of inflammation. Vitamin D acts on monocytes/macrophages and down-regulates their production of TNF-α, IL-1β, IL-6, or IL-8 . VitaminVitamin B12 is enriched in fish/shellfish  or liver . Vitamin B12 scavenges nitric oxides and ROS and thus protects various cells from inflammatory oxidative stress ,58. VitaFoxp3 promoter and expression of Foxp3 protein in CD4+ T cells driven by mucosal DCs through costimulation via CD28 and dependently on TGF-β1 [+ Tregs in peripheral CD4+ T cells by enhancing TGF-β signaling via increasing the expression and phosphorylation of Smad3, and simultaneously inhibits the development of Th17 cells by inhibiting the expression of IL-6Rα and IL-23R [The dietary sources of vitamin A (retinol) are livers, fish, eggs, or butter while provitamin A like β-carotene, which is converted to vitamin A after intestinal absorption, is enriched in green/yellow vegetables like carrots or spinach. Dietary vitamin A is absorbed in the intestine, delivered mainly to the liver, and to a lesser extent, to kidney, adipose tissues, or bone marrow . Most ofn TGF-β1 . RAs enhd IL-23R .Treatment with retinoids might be associated with potential adverse effects, hyperostosis or tissue calcification . It is rIL-1β, IL-6, IL-8, TNF-α, VEGFA , CCL2, and IL-23 mRNA in TNF-α-stimulated HaCaT cells [Soybean is suggested to be a potential anti-psoriasis agent . IsoflavaT cells . GenisteaT cells .CXCL10, IL-6, STAT3, NFKB1, CCL4 while stimulated the transcription of gene repressed in psoriasis, IL-1RN in peripheral blood cells or lesional skin [Genistein also suppressed the ROS generation induced by TNF-α or lipopolysaccharide (LPS) in HaCaT cells by scavenging free radicals, enhancing the activity of antioxidant enzymes, and reducing the production of hydrogen peroxide . It is tnal skin .E. coli-induced mastitis, a selenium-sufficient diet decreased pro-inflammatory TXB2, PGE2, and LTB4 secretion in milk compared to cows with a selenium-deficient diet [4 and LTC4 [2 production, which was mediated via reducing ROS by selenoproteins GPxs or TrxRs [2 synthase and subsequent production of 15-deoxy-Delta12,14-prostaglandin J2, an anti-inflammatory eicosanoid which inhibits NF-κB, while suppressed the expression of NF-κB-dependent genes, microsomal PGE2 synthase and TXA2 synthase and subsequent production of PGE2 and TXA2 [Selenium is included broadly in fish/shellfish, eggs, poultry, or grains. Selenium is an essential trace element having anti-oxidative and immunoregulatory properties ,77. Seleent diet . The oveand LTC4 . In macror TrxRs ,80,81. Tor TrxRs . Seleniuand TXA2 .It is reported that serum selenium levels in psoriasis patients are lower than those in controls , indicatBacteroides and increasing Proteobacteria, Fusobacteria and potentially pathogenic bacteria from Prevotellaceae, Enterobacteriaceae, Vellionellaceae, or Streptococcaceae, which may contribute to intestinal hyperpermeability and endotoxemia, and lead to systemic inflammation including the skin [Psoriasis may be triggered or worsened by alcohol intake ,36,85. Athe skin . Chronicthe skin . The chrThere is a link between increased alcohol intake and increased anxiety and depression . PsoriasEnterobacteriaceae and E. coli while reduced Firmicutes and Lactobacillus in gut microbiota, and reduced the synthesis of butyrate which induces Tregs in colon, and exacerbated dextran sodium sulfate-induced colitis [Red meat  abundantly contains SFAs, inducing NLRP3 inflammasome and IL-23/IL-17 pathway, and heme . Dietary colitis , which mLactobacillus and Bifidobacterium which can induce Tregs [Bacteroides fragilis, Faecalibacterium prausnitzii, Clostridium cluster VI and XIVa) and their metabolites  promote the activity and number of Tregs [Lactobacillus brevis SBC8803 in human subjects decreased transepidermal water loss and increased corneal hydration [Lactobacillus johnsonii protects hairless mice against UV-induced contact hypersensitivity with reduction of epidermal Langerhans cells and increase of systemic IL-10 levels [Coprococcus species. Gut microbiota in patients with psoriasis showed decrease in beneficial species Parabacteroides and Coprobacillus, which is common to IBD, while patients with PSA showed decrease in Akkermansia and Ruminococcus, protective taxa producing SCFAs. Another study reported that the gut of psoriasis patients showed less abundant Faecalibacterium prausnitzii producing butyrate and exerting anti-inflammatory action, compared to controls [Probiotics are living microorganisms that confer health benefits to the host when administered in adequate amounts . Most mice Tregs . The admce Tregs . Certainof Tregs ,95,96,97ydration . Oral ad0 levels . It is r0 levels : the gutcontrols . DecreasLactobacillus pentosus GMNL-77 administration reduced skin scores in IMQ-induced psoriasiform dermatitis and decreased the expression of TNF-α, IL-6, IL-23, IL-17A, IL-17F, and IL-22 in the skin lesions and decreased the numbers of Th17 and Th22 cells in the spleen [Bifidobacterium infantis 35624 administration in psoriasis patients reduced plasma levels of TNF-α and CRP [Bifidobacterium longum CECT 7347, B. lactis CECT 8145 and Lactobacillus rhamnosus CECT 8361) for 90 psoriasis patients together with topical corticosteroid and calcipotriol [p < 0.05). In the probiotic group but not in placebo group, there was a disappearance of Rhodococcus, a bacterium related to septicemia and biofilm production, and an increase in Collinsella and Lactobacillus, genera associated with better gut health, comparing initial and final time-points. The results supported that the probiotic blend alleviated psoriasis symptoms, and confirmed its efficacy in modulating the microbiota composition. Future studies using synbiotics, probiotics combined with dietary fibers, are promising.Several studies show that supplementation with probiotics ameliorates skin inflammation in psoriasis model mice: e spleen . The eff and CRP . A randoipotriol . At 12-wThe analysis of dietary habits in adult Japanese psoriasis patients revealed that they showed higher body mass indices (BMIs), higher intakes of fish/shellfish, pulses, vitamin B12, and vitamin D, and lower intake of meat, compared to those of healthy controls . The resn-3 PUFAs like DHA or EPA might be recommended. Patients with obesity might be considered to take low-calorie diet to reduce the weight. Psoriatic patients with low serum levels of vitamin D or selenium might be considered to take vitamin D or selenium supplementation, respectively. These dietary changes should be considered to complement the therapeutic effects of standard first line therapy for psoriasis or PSA, and also in parallel with the standard therapy for comorbidities like cardiometabolic diseases or IBDs.Based on the stimulatory or regulatory effects of nutrients or food on psoriasis, it is proposed that dermatologists might evaluate the psoriatic patient′s current diet and nutritional status by consulting nutritionists, at the time of diagnosis. It is considered to guide the patients to the appropriate dietary plan if needed. The diets with appropriate calorie and balanced composition of nutrients might be proposed for the individual patients. Generally, psoriatic patients are proposed to take diet with appropriate composition of fat and sugar with sufficient intake of fish/shellfish, soybean, and dietary fibers, avoiding excess intake of red meat, simple sugars, and alcohol. The supplementation of Several trials of dietary changes for the alleviation of psoriasis have been done. Based on the review of nutritional strategies ,106, diep < 0.001), and PASI 75 was achieved in 66.7% in intervention group vs 29% in control group (p < 0.001). However, after discontinuation of cyclosporine, 80% of patients in intervention group returned to baseline PASI levels at week 52, despite the continued low-calorie diet, indicating that improvement was dependent on medication or medication adherence rather than diet. Several other controlled and uncontrolled clinical trials of low-calorie diet provided conflicting results [Obesity is a chronic low-grade inflammation status, and is associated with the incidence and severity of psoriasis. Thus, low-calorie diet reducing weight may be helpful to improve psoriasis for the patients with obesity. A 24-week randomized controlled investigator-blinded clinical trial was conducted in 61 patients . The eff results ,111,112,n-3 PUFA dietary supplementation was theorized to aid in psoriasis patients. n-3 PUFA-supplemented diet improved PASI from 7.7 (baseline) to 5.3 at 3 months and 2.6 at 6 months compared to control  [Fish oil or  < 0.05) . However < 0.05) ,116,117  < 0.05) ,119,120.p < 0.001). Other trials, however, report small sample sizes and inconsistent results [To explore the efficacy of oral vitamin D in psoriasis, an open-label study instructed 85 patients to ingest 0.5 µg of calcitriol daily followed by a 0.5 µg dose increase every two weeks for a total 6 months to three years . Mean PA results ,123,124.p < 0.05) [Owing to low plasma levels of selenium in psoriasis patients, the efficacy of selenium supplementation was investigated by randomized, controlled trials and a clinical trial of selenium supplementation together with UVB therapy. These studies, however, showed no PASI improvements by selenium supplementation ,126. On  < 0.05) . Furtherp < 0.001) [Several studies documented the association of psoriasis and gluten sensitivity, celiac disease . To inve< 0.001) . After d< 0.001) . Due to n-3 PUFAs, vitamin D, vitamin B12, dietary fibers, SCFAs, genistein, selenium or probiotics ameliorate psoriasis or its comorbidities via the suppression of above inflammatory signaling pathways or the induction of Tregs. The manipulation of the disease-regulatory effects of nutrients or food may be useful for the management of psoriasis. Personalized diets could be proposed for individual patients based on their nutritional status and conditions of psoriasis and its comorbidities. Nutrition can be a key factor for the development and progress of psoriasis.We have reviewed the studies regarding the stimulatory or regulatory effects of nutrients or food on psoriasis and possible alleviation of psoriasis by nutritional strategies. SFAs, red meat, simple sugars, or alcohol exacerbate psoriasis and its comorbidities via the activation of NLRP3 inflammasome cascade or TNF-α/IL-23/IL-17 axis, generation of ROS, prostanoids/LTs, gut dysbiosis or suppression of Tregs. In contrast,


The 26S proteasome, a master player in proteolysis, is the most complex and meticulously contextured protease in eukaryotic cells. While capable of hosting thousands of discrete substrates due to the selective recognition of ubiquitin tags, this protease complex is also dynamically checked through diverse regulatory mechanisms. The proteasome’s versatility ensures precise control over active proteolysis, yet prevents runaway or futile degradation of many essential cellular proteins. Among the multi-layered processes regulating the proteasome’s proteolysis, deubiquitination reactions are prominent because they not only recycle ubiquitins, but also impose a critical checkpoint for substrate degradation on the proteasome. Of note, three distinct classes of deubiquitinating enzymes—USP14, RPN11, and UCH37—are associated with the 19S subunits of the human proteasome. Recent biochemical and structural studies suggest that these enzymes exert dynamic influence over proteasome output with limited redundancy, and at times act in opposition. Such distinct activities occur spatially on the proteasome, temporally through substrate processing, and differentially for ubiquitin topology. Therefore, deubiquitinating enzymes on the proteasome may fine-tune the degradation depending on various cellular contexts and for dynamic proteolysis outcomes. Given that the proteasome is among the most important drug targets, the biology of proteasome-associated deubiquitination should be further elucidated for its potential targeting in human diseases. This all β-ring . Its outl β-ring . The 19Sl β-ring ,5.Considering the strict requirement of ubiquitin modification for proteasomal recognition of substrates, it is striking that the human proteasome contains three distinct classes of deubiquitinating enzymes (DUBs)—two cysteine proteases, USP14 Ubp6 in yeast) and UCH37/UCH-L5, and the metalloprotease RPN11/PSMD14/POH1  in yeast. In contRecent cryo-electron microscopy (cryo-EM) studies have revealed a multitude of conformational states of the proteasome both in substrate-free and substrate-bound forms ,15. ThesRoughly 11–40% of isolated proteasomes are transiently associated with Ubp6/USP14, suggesting the dynamic influence of this DUB over proteolysis ,21,22. AApart from activation by proteasome, Ubp6/USP14 can, in turn, allosterically regulate the proteasome activity; this enzyme induces CP gate opening, stimulates the ATPase activity of RP, and inhibits substrate degradation through a non-catalytic mechanism ,19,32,33Emerging evidence has clearly demonstrated that specific inhibition of USP14 by small-molecules accelerates the degradation of a subset of substrates by enhancing proteasomal activity ,13,34,35DEUBAD domain represses UCH37’s DUB activity, primarily because the FRF hairpin in NFRKBDEUBAD blocks the ubiquitin docking site directly [Proteasome-bound UCH37 is known to exhibit poor DUB activity for isopeptide-linked ubiquitin–protein conjugates, despite showing predominant activity over USP14 against ubiquitin adducts with small leaving groups, such as ubiquitin amidomethylcoumarin (UbAMC) ,37. Simidirectly ,39. In adirectly ,39.The substrate specificity of UCH37 remains elusive see also , but rec2+-dependent deubiquitinase and also an essential subunit of the proteasome [Unlike USP14 and UCH37, RPN11 is an ATP- and Znoteasome ,10. Thisoteasome ,41. Remaoteasome ,26,30. TAnother striking feature of RPN11 is the conformational switch in its Ins-1 loop near the catalytic site ,43. In aUbp6/USP14 is transiently associated with the proteasome and can be easily lost during the process of proteasome isolation ,22,23,46It was once widely assumed that proteasome-bound USP14 progressively shortens the chains, one ubiquitin moiety at a time, from the substrate distal tip—the so called “distal chain trimming model” ,47. AlthAlong with its catalytic impact on the substrate, Ubp6/USP14 also exerts multiple allosteric effects on the proteasome, mainly through a non-catalytic mechanism ,33,50,51J) mutant phenotype in mice [J phenotypes in the nervous system [USP14 is often associated with oncogenic function, since this enzyme is overexpressed in several cancers and is positively correlated with tumor recurrence and poor prognosis ,55,56,57 in mice ,60,61,62s system ,64,65,66SC [Finley and colleagues have identified highly selective USP14 inhibitors, IU1 and its derivatives, by high-throughput screening and subsequent structure–activity relationship studies ,34,37, aSC ,34,67,68SC , Parkin SC , innate SC ,71, andrSC , metabolSC ,74, and DEUBAD and NFRKBDEUBAD  is a proteasome RP-associated thiol protease . As discKBDEUBAD B. In addA recent study proposed that proteasome-associated UCH37 can debranch ubiquitin chains to promote substrate degradation . EmerginAs with USP14, UCH37 is associated with oncogenesis, although this DUB has been also implicated in many different cellular processes . ElevateRPN11, which is also known as PSMD14 or POH1, is a JAMM metalloprotease class of DUB and also an integral subunit of the 26S proteasome that resides on the lid subcomplex of 19S RP . This enAs expected, RPN11 plays an essential role in proteasome integrity and cell viability across species ,102,103.The critical contribution of the proteasome to healthy cellular proteostasis has been increasingly recognized in recent years ,112. To


The goal of this study was to determine whether Levey-Jennings charts, which are widely used in clinical laboratories, can be used to create standardized internal quality controls (IQCs) for prenatal molecular diagnosis.Aneuploid amniocyte lines with trisomy 13, 21, and 18, and 47,XXY were established by transfection with SV40LTag-pcDNA3.1(−)and combined at different ratios to generate aneuploidy chimeric quality-control cell mixtures A to H. These quality-control cells were then used to calculate the Methods for constructing aneuploid amniocyte lines were developed and a set of quality-control cells (A-H) were prepared. The This method resolves the problems of a shortage of quality-control materials and a lack of quality-control charts in prenatal molecular diagnosis such as NIPT, NGS, aCGH/SNP, PCR, and FISH. Levey-Jennings chart-based IQCs for prenatal diagnosis such as FISH can be used to easily monitor whether IQC results are within acceptable limits, and then infer whether the diagnostic results for clinical samples are reliable. We expect that this standardized IQC will be useful for a wide range of molecular diagnostic laboratories. With the widespread development of molecular diagnostic technologies such as aneuploidy detection by fluorescence in situ hybridization (FISH) , noninvaIn 1931, Shewhart proposed the use of control charts for monitoring quality in manufacturing operations . He defiIn the current study, we developed a set of aneuploidy chimeric quality-control cells (A-H) by immortalizing aneuploid amniocytes via SV40LTtransfection to address the lack of quality-control materials for methods used in prenatal diagnosis such as NIPT, NGS, and FISH –8. More The SV40LTag-pcDNA3.1(−) recombinant vector was constructed and transfected into PT67 cells (ATCC CL-12284) using liposome transfection reagent to immortalize the cells . The SV45/ml. The trisomy 13 and 21 cell suspensions were mixed at ratios of 1:9, 3:7, 7:3, and 9:1 to prepare the quality-control cells A, B, C, and D containing 10, 30, 70, and 90% trisomy 13 and 90, 70, 30, and 10% trisomy 21 cells, respectively. Similarly, the trisomy 18 and 47,XXY cell suspensions were mixed to prepare the quality-control cells E, F, G, and H containing 10, 30, 70, and 90% trisomy 18 and 90, 70, 30, and 10% 47,XXY cells, respectively.The trisomy 13, 18, and 21, and 47,XXY amniotic fluid cell lines were suspended at a density of 1 × 10A FISH kit was purchased from Jinpujia Company . In this kit, GLP13 (green) is located at 13q14 and GLP21 (red) is located at 21q22. The 18/X/Y trichrome probes localize to the centromeres of chromosomes 18, X, and Y and show sky blue, green, and red fluorescence, respectively. The cells were fixed in a mixture of methanol and acetic acid (3:1) and then stained according to the manufacturer’s instructions. Technicians from the prenatal diagnosis laboratory of the Women’s Hospital, School of Medicine, Zhejiang University, and the Department of Clinical Laboratory Research, Shulan Hospital, Zhejiang University, counted 100 cells with obvious target hybridization (fluorescence) signals under a fluorescence microscope for each group of clinical samples (primary amniocytes) and quality-control cells. The quality-control cells (A-D) ratios for trisomy 13 or 21 hybridization signals were determined each day for 20 consecutive days to generate 20 datasets. Similarly, the quality-control cells (E-H) ratios for trisomy 18 or 47,XXY hybridization signals were determined. The x-axis was numbered 1–31 to represent 31 days, and the y-axis was labeled y-axis to generate the Levey-Jennings chart-based IQC for trisomy 13 and then selected for in G418-containing medium (50–400 mg/L) for 7–15 days to obtain positive cell clones  were prepared from amniocyte lines that had been immortalized, passaged, and frozen at − 196 °C in liquid nitrogen. In total, there are eight quality control samples: A, B, C, D, E, F, G, and H  were validated using a 13/21 two-color probe and an 18/X/Y trichrome probe. These probes both yielded positive fluorescence signals  for detection with the 13/21 probe and quality-control cells (E–H) for detection with the 18/X/Y probe. Quality-control cells A, B, C, and D contained 10, 30, 70, and 90% trisomy 13 cells and 90, 70, 30, and 10% trisomy 21 cells, respectively, while quality-control cells E, F, G, and H were composed of 10, 30, 70, and 90% trisomy 18 cells and 90, 70, 30, and 10% 47,XXY cells, respectively. Quality-control cells (A–H) and the test samples were analyzed 20 times under the same conditions to obtain datasets , the entclass1pt{minimaentclass1pt{minimaQuality-control cell (A) and clinical samples were analyzed for 18 days under the same conditions Table . The resentclass1pt{minimaentclass1pt{minimamentclasspt{minimaentclass1pt{minimaentclass1pt{minimamentclasspt{minimaetal [Aneuploidy is one of the most common birth defects, and is mainly detected by prenatal molecular diagnosis –3. The ietal . indicatetal  proposedetal .To overcome the shortage of quality control materials –8, we deOur main purpose was to create a novel use for Levey-Jennings charts in molecular biology laboratories by developing characterized paired quality-control cells. Levey-Jennings charts, which were initially used by Westgard to monitor the quality of manufacturing operations, were recommended for use in clinical laboratories by Levey and Jennings to judge whether the observed control measurements (or observations) represent reliable or unreliable performance of the analytical method, which in turn may suggest certain sources of error, and so aid in problem solving , 10. HenWe used FISH as a representative method to study the feasibility of establishing Levey-Jennings chart-based IQCs for monitoring molecular diagnostic quality, because FISH is widely used to detect aneuploidies after amniocentesis and to perform chromosome analysis of leukemic blood samples and other tissues –24. In gx-axis and the numerical value of the quality control test is represented on the y-axis. Moreover, it contains several reference lines, including one for the mean and three on either side of the mean representing standard deviation limits  [As shown in Fig. nd 3 SD) –12. As s limits) –12, whic limits) –14.The Levey-Jennings chart-based IQC for aneuploid detection can be used to monitor in real-time whether the quality control results are outside of acceptable limits and then determine whether the test results for the clinical samples are reliable, which should be widely applicable for IQC in prenatal molecular diagnostical laboratory.


Plant adaptations to changing environments rely on integrating external stimuli into internal responses. Brassinosteroids (BRs), a group of growth-promoting phytohormones, have been reported to act as signal molecules mediating these processes. BRs are perceived by cell surface receptor complex including receptor BRI1 and coreceptor BAK1, which subsequently triggers a signaling cascade that leads to inhibition of BIN2 and activation of BES1/BZR1 transcription factors. BES1/BZR1 can directly regulate the expression of thousands of downstream responsive genes. Recent studies in the model plant Arabidopsis demonstrated that BR biosynthesis and signal transduction, especially the regulatory components BIN2 and BES1/BZR1, are finely tuned by various environmental cues. Here, we summarize these research updates and give a comprehensive review of how BR biosynthesis and signaling are modulated by changing environments and how these changes regulate plant adaptive growth or stress tolerance. Most higher plants are rooted in their habitats and cannot move around to select favorable growth conditions. They must deal with their living situations by constantly perceiving, processing, and responding to ever-changing environmental cues, such as light, temperature, water, nutrients, and microbes ,2. Many CPD expression can be regulated by a putative transcriptional coregulator, BREVIS RADIX (BRX) [CPD [DWF4, whose encoding protein catalyzes a rate-limiting step in BR biosynthesis, is modulated by two bHLH transcription factors, TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR1 (TCP1) and PHYTOCHROME INTERACTING FACTOR 4 (PIF4) [DWF4 and positively regulate its expression [To date, more than 70 BRs have been isolated in plants . These BIX (BRX) . It was RX) [CPD . In addi4 (PIF4) ,38. Bothpression . Previoupression ,41,42,43pression ,45,46 , photoreceptors such as UV RESISTANCE LOCUS 8 (UVR8), CRYPTOCHROME1 (CRY1) and PHYTOCHROME B (PHYB), and an autophagy receptor DOMINANT SUPPRESSOR OF KAR2 (DSK2) [2O2) and reduced by a thioredoxin TRXh5 [BL and its precursor CS are perceived by a cell-surface receptor kinase complex consisting of BRASSINOSTEROID INSENSITIVE 1 (BRI1) as the receptor and BRI1-ASSOCIATED KINASE 1 (BAK1) as the coreceptor, both of which belong to the leucine-rich repeat receptor-like kinase (LRR-RLK) protein family ,48,49. B2 (DSK2) ,91,92,93in TRXh5 . It was in TRXh5 .Light acts not only as an energy source, but also as an important environmental signal to regulate plant growth and development . Light qbas1-D, suppresses the long hypocotyl phenotype caused by mutations in the photoreceptor phytochrome B (phyB) [bri1-5, an intermediate mutant allele of BRI1 [Several lines of evidence suggest a crosstalk between light and BR signaling. Firstly, mutants defective in either BR biosynthesis or signaling exhibit constitutive photomorphogenic phenotypes, including shortened hypocotyls, opened and expanded cotyledons, when grown in darkness . SecondlB (phyB) . Thirdly of BRI1 . CPD, DWF4, and BR6OX2, could be induced in the dark and repressed in the light [PIF4 is tightly controlled by the circadian clock, and the protein stability of PIF4 is regulated by light [CPD and DWF4 [pifq, a quadruple mutant of PIFs, the fluctuated expression patterns of these BR biosynthetic genes were completely blocked, suggesting that PIFs are essential to diurnal BR biosynthesis [DWF4 mRNA accumulation in root tips and therefore is important for root growth [BAS1 (CYP734A1) and SOB7 (CYP72C1) [Previous reports demonstrated that the expression of several BR biosynthetic genes, he light ,102,103.he light . The expby light . PIF4 caand DWF4 . In pifqt growth . AnotherCYP72C1) .BES1 and BZR1 are elevated at night and decreased during the day [Light also affects BR signaling A. The tr the day . Interes the day . In fact the day . A group the day . In addi the day ,89,90,92 the day ,89,90,92According to these studies, under normal light conditions, the photoreceptors are activated and associated with BES1 and BZR1, BR signaling is therefore repressed . Under aTemperature is an important environmental factor that governs the seasonal and diurnal behavior of plants grown in temperate geographical zones . Many plCPD, DWF4, and YUCCA5 [bes1-D showed reduced sensitivity to warm-induced root growth acceleration [In thermomorphogenesis, the photo receptor phytochrome B (phyB) was also identified as a major temperature sensor ,115. Besd YUCCA5 ,102,117.d YUCCA5 . In contd YUCCA5 . A recenleration . The stuleration .COLD-REGULATED (COR) genes play fundamental roles in plant tolerance to cold stress, many of which encode osmolytes and cryoprotective proteins that protect cells from freezing damage [COR genes contain a conserved cis-element termed C-REPEAT/DEHYDRATION-RESPONSIVE ELEMENT (CRT/DRE), which is directly bound by the CRT/DRE BINDING FACTOR (CBF/DREB) family transcription factors [COR genes, therefore, is transcriptionally activated by CBFs [COR genes that are cold-regulated but in a CBF-independent manner [CBFs is tightly controlled by cold, with induction within 15–30 min, peaks at 1–3 h, and declines afterwards [CBFs [CBFs by directly binding to their promoters under cold stress [ICE1 expression is slightly upregulated by cold, its protein stability is predominantly controlled by posttranslational modifications including phosphorylation, ubiquitination, and SUMOylation, in response to cold stress [Temperature below the growth-permissive range leads to cold stresses, including chilling stress (0–15 °C) and freezing stress (<0 °C) . Plants g damage . Some CO factors . The exp by CBFs . There at manner . Expressterwards . Severalds [CBFs . For exad stress . Althougd stress ,128,129.CBFs and hence induce the CBF-dependent COR gene expression [COR genes to modulate plant response to cold stress [CBF expression and was considered as a strategy for balancing plant growth and cold stress responses [BRs were shown to play a promotive role in plant resistance to the cold environment as they increase chilling tolerance when externally applied . Moreovepression ,133. Actd stress . BIN2 ind stress . BIN2-acesponses .Heat stress is caused by high temperature (above the growth-permissive range), which affects photosynthesis, alters membrane fluidity, and disrupts the overall balance of metabolic processes . Heat stbri1-5) or treatment with a BR biosynthetic inhibitor, BRZ [+-ATPase AHA2. The authors proposed that this may contribute to the strong hydrotropism observed in Ws [Water is vital for plants due to its irreplaceable roles in various physiological processes. Water in soil, which is greatly affected by climate change, is an important constraint for plant growth and distribution . Plants tor, BRZ . The stued in Ws ,141. WheRD26 is one of the target genes of BES1 and is repressed by BR signaling [Drought is one of the most common abiotic stresses against plant growth and even survival . Severalignaling . Droughtignaling .2+ signal can be sensed by diverse Ca2+ binding proteins such as calmodulins (CaM) [2+/CaM promotes BR biosynthesis through interacting with and activating DWARF1, a Ca2+-dependent calmodulin-binding protein responsible for converting 24-methylenecholesterol to CR, the initial BR biosynthetic precursor [2+, thus initiating a Ca2+ signaling cascade in the cytosol [2+ signaling [Mineral nutrients, mainly taken up from the soil, are essential for normal plant growth and development . Importans (CaM) . A previrecursor . Percept cytosol . Furtherignaling . This noignaling . Similarignaling . Alteredignaling . The BR ignaling . The repignaling . ulp1a mutants were salt-tolerant [Except for its involvement in coordinating plastic growth in response to mineral nutrients, it was demonstrated that BRs help plants to tolerate various ion stresses, such as salinity . Exogenotolerant . Taken tMicrobial pathogens constitute an important aspect of biotic stress challenging plant growth and development. To effectively combat the invasion of pathogens, multiple layers of defense mechanisms have been developed by plants during evolution, including preformed structural barriers and inducible defense mechanisms . Once pabri1 and bin2-1 [PR1, PR2, and PR5, is remarkably lower in a BR biosynthetic gene mutant cpd than that in wild type; (2) Exogenous application of BRs conferred diverse crops resistance to a broad array of pathogens [BRs are increasingly reported to be involved in plant response to pathogen attack D. The rod bin2-1 ,180. Othathogens ,178. Altathogens . In a reathogens . Interesathogens . The flgSince BRI1-mediated BR signaling and FLS2-mediated PTI response share BAK1, BIK1, and BSK1 as their common regulatory components, it is logical to speculate that these proteins may contribute to the crosstalk between BR signaling and PTI response ,170,171.MAPK cascades, consisting of MAPK kinase kinases (MAPKKKs), MAPK kinases (MKKs), and MAPKs, play important roles in diverse biological processes . For exaDespite these aforementioned studies suggesting an interaction between BR signaling and PTI response at the level of the receptor or downstream cytoplasmic kinases, it was speculated that this interaction might only make a minor contribution to the suppression of PTI . The traBRs have been reported to function in a wide spatial-temporal territory during the entire life span of plants . However


Malignant peripheral nerve sheath tumor (MPNST) is a rare sarcoma. Owing to the lack of specific histological criteria, immunohistochemical, and molecular diagnostic markers, several differential diagnoses must be considered. Advances in molecular testing can provide significant insights for management of rare tumor.The patient was a 50-year-old man with a history of lumpectomy on the right back 30 years ago. He felt a stabbing pain at the right iliac fossa and went to the local hospital.By immunohistochemistry, the tumor cells stained positively for S-100 , CD34 (strong +++) and Ki-67 (20%), and negatively for smooth muscle actin, pan-cytokeratin, neurofilament, pan-cytokeratin-L, GFAP, CD31, STAT6, ERG, myogenin, and MyoD1. Combined with the histopathology and immunohistochemistry results, our initial diagnosis was solitary fibrous tumor (SFT) or MPNST. The tissue biopsy was sent for next-generation sequencing. neurofibromatosis type 1 Q1395Hfs∗22 somatic mutation, neurofibromatosis type 1 D483Tfs∗15 germline mutation, and amplifications of BTK, MDM2, ATF1, BMPR1A, EBHA2, GNA13, PTPN11, RAD52, RPTOR, and SOX9, as well as TJP1-ROS1 fusion, CDKN2A-IL1RAPL2 fusion and CDKN2A/UBAP1 rearrangement were identified. Given that NAB2-STAT6 fusion, a specific biomarker of SFT, was not identified in our patient's tumor, the SFT was excluded by through genetic testing results. Therefore, our finally diagnosis was a MPNST by 2 or more pathologists.Subsequently, the patient received crizotinib therapy for 2 months and showed stable disease. However, after crizotinib continued treatment for 4 months, the patient's disease progressed. Soon after, the patient stopped crizotinib treatment and died in home.To our knowledge, this is the first report of the TJP1-ROS1 fusion, which expands the list of gene fusions and highlights new targets for targeted therapy. Also, our case underlines the value of multi-gene panel next-generation sequencing for diagnosis of MPNST. Because of morphological and immunohistochemical complexity, especially outside the NF1 context, the diagnosis of MPNST remains challenging.Malignant peripheral nerve sheath tumors (MPNSTs) are rare spindle cell sarcomas arising from peripheral nerve cells, including Schwann cells. Advances in NGS technology have made it possible to identify key gene fusions and identify novel therapeutic targets across different types of tumors. With the development of molecular biology, targeted therapy has achieved remarkable success, which have better therapeutic effects and less adverse reactions.,8Breakthrough of next generation sequencing (NGS) platforms throughout this decade provides affordable and reliable high-throughput sequencing for evaluation of functional DNA variations in many diseases. especially for patients with unclear diagnosis by traditional immunohistochemical detection, as well as morphology and history. At present, tumor gene mutation detection based on NGS technology can achieve satisfactory results in establishing a diagnosis and the corresponding targeted therapy based on driver gene mutations.In addition, NGS genomic profiling may be of great significance in determining the tumor tissue origin,Here, a 50-year-old man was finally diagnosed with MPNST according to histopathology, immunohistochemistry and NGS results. This patient harbored a novel TJP1-ROS1 fusion and NF1 germline mutation identified by NGS, whom achieved stable disease (SD) under crizotinib treatment.2A 50-year-old man underwent lumpectomy on the right back 30 years ago, and the histopathology results showed neurofibroma. A mass (4 × 4 cm) in the right iliac fossa was found during the patient's physical examination 20 years ago, however, the patient did not receive any treatment. An egg-sized mass at the right ilium was found in April 2018. Due to no obvious discomfort or pain, he did not see a doctor. He felt a stabbing pain at right iliac fossa two months later and went to the local hospital on September 24, 2018. A computerized tomography (CT) scan revealed multiple masses in the right psoas major, iliopsoas and para-piriformis and right hip front, most probably neurofibroma. For further diagnosis and treatment, he was transferred to our hospital on October 20, 2018.The patient did not have palpitations, chest distress, low fever, night sweat, or recent body weight changes. Chest CT showed masses in the middle upper lobe of the right lung, and multiple soft tissue nodules in the bilateral parenchyma Fig. A. CombinTo make a definite diagnosis and obtain the optimal treatment, the tissue biopsy of patient was prepared for NGS  on November 11, 2018. An NF1 Q1395Hfs∗22 somatic mutation, NF1 D483Tfs∗15 germline mutation, as well as amplifications of BTK, MDM2, ATF1, BMPR1A, EBHA2, GNA13, PTPN11, RAD52, RPTOR, and SOX9 were identified. Importantly, a TJP1-ROS1 fusion , CDKN2A (NM_058195)-IL1RAPL2 fusion , and CDKN2A/UBAP1 rearrangement were also identified by NGS. These genomic alterations were also verified by RNA-sequencing in addition to an EPHB1-MECOM fusion . Since the NAB2-STAT6 fusion is a specific biomarker for SFT, the SFT was excluded by through genetic testing results. Therefore, the patient was finally diagnosed with a MPNST by 2 or more pathologists.In November 2018, the patient started crizotinib therapy . During a reexamination in January 2019, CT scan and MRI showed shrunken local lesion, and the efficacy was evaluated as SD  in East Asian patients with ROS1 positive advanced NSCLC reported an ORR of 71.7% and median PFS of 15.9 months in ROS1 fusion patients treated with crizotinib. Another clinical trial conducted in European patients with ROS1-rearranged lung cancer who had undergone documented crizotinib therapy achieved an ORR of 80%, median PFS of 9.1 months, and disease control rate of 86.7%. Considering the patient harbored a ROS1 fusion, he started crizotinib therapy and showed SD after treatment for two months. However, after crizotinib continued treatment for 4 months, the patient's disease progressed. Soon after, the patient stopped crizotinib treatment and died in home.Crizotinib was the first targeted agent approved by the US Food and Drug Administration for the treatment of ROS1-rearranged non-small cell lung cancer (NSCLC) in 2016, which was based on a phase I expansion cohort crizotinib trial (NCT00585195). The trial demonstrated a median progression-free survival (PFS) of 19.2 months and objective response rate (ORR) of 72% in patients with advanced ROS1-rearranged NSCLC. Germline mutation in NF1 gene has been identified in several tumors, including glioma, MPNST, neurofibroma, and so on.–21 The germline mutation rate of the NF1 gene  is about ten-fold higher than in other inherited disease genes; although the underlying reasons for this new high mutation rate are unclear. This NF1 germline mutation in our case turned 483rd coding aspartic acid into threonine, and terminated the coding at the 15th coding of the new open reading frame, which may cause nonsense mutation-mediated mRNA degradation in the cell, leading to functional deletion of NF1 gene. Given that NF1 D483Tfs∗15 is a heritable germline variation, it may be involved in the relapse of the patient.Other important finding is that NF1 D483Tfs∗15 mutation detected in our case is a heritable germline variation, which may be involved in the relapse of the patient. NF1 is a tumor suppressor gene located on chromosome band 17q11.2. NF1 encodes neurofibromin, which is a member of the GTPase activating protein family. It can activate the GTPase activity of the RAS protein, and then negatively regulate the KRAS pathway. It has been shown that there are more than 500 different NF1 mutations, of which most are unique to the particular kindred.To our knowledge, this is the first case of MPNST with a TJP1-ROS1 fusion, raising interesting questions regarding treatment. Also, the NF1 germline mutation in this patient may be related to relapse. In view of the inheritance of NF1 germline mutations, it is necessary to adjust preventive measures for our patient and his family members, and to offer predictive testing to healthy relatives.Conceptualization: Juming Li, Lingxiang Liu, Yongzhong Wei.Data curation: Qi Zhang, Yumin Huang.Investigation: Juming Li, Lingxiang Liu.Methodology: Yihong Zhang, Xiaoyan Gan, Siqin Liu, Zhen Yue.Validation: Yongzhong Wei.Writing – original draft: Juming Li, Lingxiang Liu.Writing – review & editing: Qi Zhang, Yumin Huang, Yihong Zhang, Xiaoyan Gan, Siqin Liu, Zhen Yue, Yongzhong Wei.