Abnormal ER stress can be involved in many diseases. NOD-like receptor family pyrin domain-containing 3 (NLRP3), an intracellular receptor, can perceive internal and external stimuli. It binds to apoptosis-associated speck-like protein containing a CARD (ASC) and caspase-1 to assemble into a protein complex called the NLRP3 inflammasome. Evidence indicates that ER stress and the NLRP3 inflammasome participate in many pathological processes; however, the exact mechanism remains to be understood. In this review, we summarized the role of ER stress and the NLRP3 inflammasome in liver disorders and analyzed the mechanisms, to provide references for future related research.The endoplasmic reticulum (ER) is a key organelle responsible for the synthesis, modification, folding and assembly of proteins; calcium storage; and lipid synthesis. When ER homeostatic balance is disrupted by a variety of physiological and pathological factors—such as glucose deficiency, environmental toxins, Ca The definition of inflammasomes was first proposed by Tschopp et al. in 2002 . Inflamm2+ level changes, viral infection, oxidative stress, inflammation and hypoxia–disrupt ER homeostatic balance, ER stress can be induced, forming a large number of unfolded and misfolded proteins, calcium depletion and lipid synthesis disorders [The endoplasmic reticulum (ER) is an organelle responsible for the synthesis, folding and modification of the secretion/transmembrane protein. It also plays a key role in lipid calcium storage, detoxification and biosynthesis ,24,25. Wisorders ,27. Cellisorders . ER streisorders . Moderatisorders . Previouisorders ,33,34,35isorders . In thisNonalcoholic fatty liver disease (NAFLD) is a clinicopathological syndrome characterized by the accumulation of liver fat, excluding viral infection and excessive drinking, and includes fatty liver, nonalcoholic steatohepatitis, and liver cirrhosis. It is a common chronic liver disease in the world. Due to its high incidence rate (about 20–30%) and long-term clinical treatment, NAFLD has become a serious public health problem ,38,39. MChop using siRNA inhibited the activity of caspase-11, caspase-1 and IL-1β, but not of active caspase-3, in tunicamycin or tunicamycin + LPS-induced primary hepatocytes; this indicates that ER stress induced NLRP3 inflammasome pyroptosis to cause injury of the LPS-induced NASH model via CHOP. In conclusion, ER stress could activate the NLRP3 inflammasome and subsequent pyroptosis and apoptosis, which promoted NASH progression. Therefore, the inhibition of ER stress-dependent NLRP3 inflammasome activation and subsequent cell death might be a potential treatment for nonalcoholic hepatitis. The ER stress effectors PERK and IRE1α could both activate CHOP, thereby activating NLRP3 inflammasome, which further proved that ER stress promoted the NLRP3 inflammasome via the CHOP pathway [Nonalcoholic steatohepatitis (NASH) is the combination of lipid accumulation, hepatocyte death, inflammation, and fibrosis, which can develop into advanced fibrosis and hepatocellular carcinoma ,43. It i pathway . Reactiv pathway . In the −/− mice made the liver vulnerable to NAFLD, which led to hepatic steatosis and metabolic collapse; this was evidenced by an increase in fatty acid uptake, inhibition of β-oxidation, and a reduction in fatty acid release. Moreover, the enhanced ER stress promoted NLRP3 inflammasome activation, hepatocyte death, fibrosis, and the dysregulation of lipid homeostasis, leading to liver injury in the livers of tunicamycin-treated BI-1−/− mice. In liver biopsies obtained from NAFLD patients, the activation of the IRE1α signaling pathway was accompanied by BI-1 downregulation, suggesting that the IRE1α signaling pathway contributed to NAFLD. Moreover, the enhanced ER stress evidenced by the increasing expression of liver X-box binding protein 1 (XBP1), IRE1a, and the C/EBP homologous protein (CHOP) in HFD-fed BI-1−/− mice with NASH could also activate the NLRP3 inflammasome. Similarily, in primary mouse hepatocytes lacking BI-1, the IRE1α signaling pathway was shown to mediate NLRP3 inflammasome activation and cell death. Additionly, the inhibition of IRE1α signaling with STF-083010 counteracted the BI-1 deficiency promotion of NAFLD, indicating that BI-1 could improve NAFLD by suppressing ER stress-induced IRE1α-dependent NLRP3 inflammasome activation [Bax Inhibitor-1 (BI-1) is a negative regulator of ER stress and can improve NAFLD ,49. Cynttivation . It can Ginsenoside Rg1 (Rg1) is an active ingredient of natural medicine and has a variety of physiological functions, including anti-inflammatory, anti-apoptotic, anti-fibrosis, antioxidant and neuroprotective effects. It has been reported that Rg1 is involved in NAFLD ,52,53. YAcetylantroquinonol B (4-AAQB) is a ubiquinone from Antrodia cinnamomea and has antioxidant, anti-inflammatory and anti-hepatoma properties . The resIn summary, at present, there are few studies on the role of ER stress/NLRP3 inflammasome in NAFLD; moreover, some studies are very superficial and can only draw preliminary conclusions, which need further research to be verified. In particular, the mechanism of ER stress/the NLRP3 inflammasome involved in liver lipid metabolism remains to be further explored. In addition, in the above studies, ER stress/the NLRP3 inflammasome were inhibited to improve NAFLD. However, whether enhanced endoplasmic reticulum stress can improve NAFLD by inhibiting NLRP3 inflammatory bodies remains to be clarified. ER stress/the NLRP3 inflammasome may become a new strategy for the treatment of NAFLD.2-Induced AML12 cells; therefore, it can be deduced that ER stress inhibits NLRP3 inflammasome by clearing ROS. More and more evidence has proven that ER stress and NLRP3 inflammasome-mediated injury have a great impact on ischemia–reperfusion injury [Hepatic ischemia–reperfusion (HIR) is a physiological and pathological phenomenon, which is difficult to avoid in some types of surgery and is related to liver transplantation, liver injury and hepatectomy ,69. Whenn injury ,78, whicAcrylamide (AA) is produced by a Maillard reaction during thermal processing and a well-known potential carcinogenic compound . AA has Baicalin (BA) is one of the main bioactive components of the Chinese herbal medicine Scutellaria baicalensis. It has many pharmacological activities, such as antitumor, antibacterial, and antioxidant ,91,92. BThe farnesoid X receptor (FXR) is a member of the nuclear receptor family and exists in the intestine and liver. It helps to maintain systemic metabolic homeostasis by regulating glucose, bile acid, lipid metabolism, and energy homeostasis. Furthermore, the FXR also plays an important role in many organs, including the liver, cardiovascular system, kidney, intestine, breast, pancreas and brain ,102,103.Accumulating evidence indicates that ER stress and the NLRP3 inflammasome play an important role in liver disorders. In this review, we summarized the following: (1) ER stress could activate NLRP3 inflammasome and subsequent pyroptosis and apoptosis to lead to nonalcoholic hepatitis; (2) BI-1 could improve NAFLD by suppressing ER stress-induced IRE1a-dependent NLRP3 inflammasome activation; (3) Rg1 improved NAFLD through the inhibition of ER stress and NLRP3 inflammasome activation, which needs to be further confirmed; (4) 4-AAQB ameliorated NAFLD through the inhibition of ERS/NLRP3 inflammasome by activating the SIRT1-Nrf2 pathway, which needs to be further confirmed; (5) ORY ameliorated HIRI through the inhibition of the NLRP3 inflammasome and ER stress; (6) ER stress inhibits the NLRP3 inflammasome via the MAPK and NF-κB signaling pathways in the improvement of AA-induced hepatotoxicity using allicin, which needs to be further confirmed; (7) BA improved the PA-induced cytotoxicity of AML-12 cells through the inhibition of ER stress via the TXNIP/NLRP3 pathway, through the AMPK pathway; (8) FXR suppressed the ER stress-induced NLRP3 inflammasome via the PERK–CHOP signaling pathway in hepatocytes to improve liver injury .2S) [2S can regulate ER stress and the NLRP3 inflammasome in liver disorders is worth studying. ER stress and the NLRP3 inflammasome will become an important target for the treatment of liver disorders.It can be seen from the above summarized research that the mechanism of ER stress-regulating NLRP3 is as follows: (1) ER stress promotes the NLRP3 inflammasome via the CHOP pathway; (2) ER stress induces the NLRP3 inflammasome via IRE1a; (3) ER stress inhibits the NLRP3 inflammasome by clearing ROS; (4) ER stress suppresses the NLRP3 inflammasome via the MAPK and NF-κB signaling pathways; (5) ER stress induces the NLRP3 inflammasome through ROS/TXNIP; and (6) ER stress promotes the NLRP3 inflammasome via the PERK–CHOP signaling pathway . In the 2S) ,112. Whe


Gap junctions (GJ) and connexins play integral roles in cellular physiology and have been found to be involved in multiple pathophysiological states from cancer to cardiovascular disease. Studies over the last 60 years have demonstrated the utility of altering GJ signaling pathways in experimental models, which has led to them being attractive targets for therapeutic intervention. A number of different mechanisms have been proposed to regulate GJ signaling, including channel blocking, enhancing channel open state, and disrupting protein-protein interactions. The primary mechanism for this has been through the design of numerous peptides as therapeutics, that are either currently in early development or are in various stages of clinical trials. Despite over 25 years of research into connexin targeting peptides, the overall mechanisms of action are still poorly understood. In this overview, we discuss published connexin targeting peptides, their reported mechanisms of action, and the potential for these molecules in the treatment of disease. Gap junction channels (GJ) were first identified in the 1960s ,3 and ha2+ flux in adipocytes, astrocytes, and glioblastoma cells ,13492,13SHVR, corresponds to amino acids 64–76 of Cx43-EL1. However, this peptide also altered the channel functions of Cx32, Cx37, and Cx40 in a wide range of tissues . S. S101]. 2+ signaling in endothelial cells [Several other studies have described the effects of two identical peptides covering the last 10 a.a. of the Cx43-CT, referred to as TAT-Cx43CT  and TAT-al cells . In theial cells . Howeveral cells . Both TAal cells ,115. Thial cells . StudiesSRC tyrosine kinases are involved in numerous cell functions, including cell metabolism, and are found in both proliferating and postmitotic cells. The protein exists in both a cellular form (cSRC) and as an oncogenic viral homolog (vSRC) . InteracIn neural progenitor cells, cSRC is involved in survival and proliferation via the epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF-2) signaling pathways ,178. TheThe interaction between Cx43 and cSRC has also been studied in the context of neuroinflammation. In response to neuroinflammation stimuli, TAT-Cx43 266–283 reduced cSRC levels and exhibited a neuroprotective effect . In thesThe Cx43 juxtamembrane 2 (JM2) connexin-mimetic peptide, published by Calder et al. in 2015, was designed to target amino acids 231–245 of the Cx43 CT, a known microtubule-binding domain . InitialIn vivo studies in mice and rats demonstrate that JM2 treatment reduces inflammatory cell infiltration following submuscular device implant , reducesOne observation that bears mentioning in this review is the consistency with which Cx43 peptides induce PKC-associated Cx43-S368 phosphorylation . Many ofThere is conflicting information on whether Cx43 PKC-phosphorylation leads to enhanced or reduced GJ coupling ,158. PhoThe overall significance of the Cx43-PKC (S368) phosphorylation following peptide treatments remains to be fully elucidated. As described, many studies have found that peptide-induced Cx43-PKC phosphorylation was associated with improved outcomes, such as mitigated ischemic injury, reduced cellular proliferation in cancer, and improved wound healing. Thus, while highly speculative, it is possible that activating the PKC pathways may be considered cytoprotective.43Gap27 and 32Gap24 have been found to block pannexin 1 (Panx1) channels at concentrations ranging from 200 μM to 2 mM [The “Gap” peptides have shown promise of increased specificity to connexins by comparison to conventional channel blockers such as glycyrrhetinic acid . However to 2 mM . This ha to 2 mM ,187Pannexins form purine release channels that are structurally homologous to the connexins, containing four transmembrane domains, two EL, and cytoplasmic NT, IL, and CT but share little sequence homology. While connexins channels are known to form as hexamers, recent data has shown that Panx1 channels oligomerize as heptamers ,189. The2+ waves, associated with both connexin and pannexin channels. Both 37,43Gap27 and 32Gap24 can reversibly inhibit pannexin membrane currents [37,43Gap27 contains a sequence from an extracellular loop of Cx43, while 32Gap24 contains a sequence from the Cx32-IL. Interestingly, scrambled 32Gap24 showed similar levels of inhibition to its non-scrambled counterpart. Polyethyleneglycol 1500 (PEG 1500), an inert, biocompatible polymer inhibited pannexin channels similarly to the connexin-mimetic peptides. This along with the inhibitory activity of scrambled 32Gap24 peptides suggests inhibition of pannexins by mimetic peptides may function via steric block of the channel rather than peptide-protein interactions [Connexin-mimetic peptides have demonstrated effects on pannexin channels. These peptides are known to attenuate ATP release and Cacurrents . 37,43Garactions .Multiple Panx1 mimetic peptides have been developed targeting sequences on both the intracellular and extracellular loops. Peptides E1a, E1b, E1c, E2a, and E2b target Panx1-EL sequences. Both E1b and E1c can inhibit Panx1 channels by more than 10%, with E1b having the strongest inhibitory effect. The peptide 10Panx1 contains the same sequence as E1b but is three amino acids shorter. Both 10Panx1 and E1b inhibit Panx1 channels to a similar extent ,192, andThe peptide 10Panx1 inhibits Panx1 currents compared with its scrambled sequence, suggesting specificity ,196,197.In the last 25 years, several encouraging lines of evidence have demonstrated that connexin targeting peptides can improve disease outcomes in pre-clinical models of disease. Connexin proteins are attractive therapeutic targets given their prevalence in disease, although approaches to target connexins are complicated due to the ubiquitous distribution of the proteins in most tissues. Inappropriately reducing the expression of connexins or inhibiting GJ functions can be lethal. Alterations or mutations in vasculature-associated connexins, such as Cx37 and Cx40, lead to disturbances in blood vessel relaxation and have knock-on effects in other systems leading to alteration in renin secretion and increases in blood pressure. Genetic connexin mutations in humans are rare, but several have been identified. Mutations in Cx43 have been linked with cardiovascular disease ,201,202,At present, the instability of peptides severely limits their therapeutic potential. Most peptides have a half-life of seconds to minutes and are quickly degraded by changes in pH, shifts in temperature, and peptidases, making systemic delivery challenging at best . Chen etLikely, future therapeutic approaches may also require systemic delivery by encapsulation of peptides (such as in exosomes ), in a p


Inflammasomes are key intracellular multimeric proteins able to initiate the cellular inflammatory signaling pathway. NLRP3 inflammasome represents one of the main protein complexes involved in the development of inflammatory events, and its activity has been largely demonstrated to be connected with inflammatory or autoinflammatory disorders, including diabetes, gouty arthritis, liver fibrosis, Alzheimer’s disease, respiratory syndromes, atherosclerosis, and cancer initiation. In recent years, it has been demonstrated how dietary intake and nutritional status represent important environmental elements that can modulate metabolic inflammation, since food matrices are an important source of several bioactive compounds. In this review, an updated status of knowledge regarding food bioactive compounds as NLRP3 inflammasome modulators is discussed. Several chemical classes, namely polyphenols, organosulfurs, terpenes, fatty acids, proteins, amino acids, saponins, sterols, polysaccharides, carotenoids, vitamins, and probiotics, have been shown to possess NLRP3 inflammasome-modulating activity through in vitro and in vivo assays, mainly demonstrating an anti-NLRP3 inflammasome activity. Plant foods are particularly rich in important bioactive compounds, each of them can have different effects on the pathway of inflammatory response, confirming the importance of the nutritional pattern (food model) as a whole rather than any single nutrient or functional compound. Inflammasomes are key intracellular multimeric protein complexes, able to initiate inflammatory signaling through sensor receptors, defined as pattern-recognition receptors (PPR). PPRs recognize pathogen-associated molecular patterns (PAMPs), or damage-associated molecular patterns (DAMPs) generated by endogenous stress stimuli. The signal transduction continues intracellularly through an adaptor protein and an effector enzyme that cause the maturation and the secretion of pro-inflammatory cytokines. As a result, the inflammasome activation ends up with the production of caspase-1, a major mediator in the inflammatory adaptive immune cell response . FinallyAmong other PPRs, leucine-rich repeat (LRR)-containing proteins (NLR) family members (NLRP1 and 3) are widely known as components of inflammasomes. In particular, it has been reported that the NLRP3 inflammasome is crucial for adaptive cell responses towards viral, bacterial, and fungal infections. Nevertheless, when dysregulated, it has been associated with a variety of inflammatory or autoinflammatory disorders, such as diabetes, gouty arthritis, liver fibrosis, Alzheimer’s disease (AD), respiratory syndromes, atherosclerosis, and cancer initiation . The NLR2+-related signaling, Na+ influx, and K+ and chloride efflux have been reported as crucial events in the NLRP3 inflammasome [+ efflux for inflammasome assembly. It has been proposed that the activation of NLRP3 requires two steps. Firstly, the mitotic kinase NEK7 binds to NLRP3; although, this complex could not be sufficient for NLRP3 activation. Indeed, the inflammasome oligomerization requires ATP binding and thus the conversion of NACHT from an inactive to an active conformation [Many molecular mechanisms/compounds have been widely proposed for the activation of NLRP3, involving, among others, biological, physical, chemical, and metabolic agents, such as uric acid crystals, cholesterol, free fatty acids and lipids, β-amyloid (BA) protein, aluminum hydroxide, high intracellular glucose, and high levels of reactive oxygen species (ROS). Others factors, such as heme, ionic flux, lipopolysaccharide (LPS) toxin, pathogen-associated RNA, and ceramides, are also included . Likewisammasome . As a maormation . Reactivormation ,11,12.The emerging role of chronic inflammation as a determinant in the progress of the major degenerative diseases typical of modern society has promoted research to comprehend the influence of nutrition and dietary patterns on inflammatory markers. Dietary intake and nutritional status represent important environmental factors, which can modulate metabolic inflammation. In recent years, research has advanced significantly to achieve understanding of the impact of dietary components on metabolic inflammation, in the context of chronic infirmities, such as obesity, type-2 diabetes (T2D), cardiovascular diseases (CVD), and cancer . InflammIn the mid-1900s, an important concept was emerging about the relationship between the immune and metabolic response systems, indicating that insulin resistance (IR), glucose intolerance, dyslipidemia, and other metabolic abnormalities occur in the course of an infection. Later, in the 1980s, a reduced binding capacity of insulin to its receptor in isolated blood cells was found in human patients affected by acute infection . The netDrosophila melanogaster or the nematode Caenorhabditis elegans, gave similar evidence of a conventional cellular crosstalk between immune and metabolic organs. Basically, cytokines act as metabolic hormones in the adaptation to nutrient fluctuations [Drosophila, for example, the body fat acts as the liver, adipose, and immune system, serving both the functions of nutrient storage and body defense [Drosophila orthologue of TNF, and its receptor Wengen (in Drosophila) or TNFR (human); insulin and its receptors ; the TLR signaling pathways (Toll receptor in Drosophila). TNF and TLR signaling block insulin pathway or production through JNK activation and MyD88, from flies to humans, whereas activation results in abnormal metabolic homeostasis adaptation, which leads to a chronic metabolic inflammation. As a result, immunometabolic diseases often appear as clusters and promote ageing, disability, and premature death [The strong link between nutrient sensing and immune signaling is justified by an evolutionarily conserved crosstalk pathway between immune and metabolic mediators. The functional units that control key metabolic and immune functions in higher organisms have evolved from common ancestral structures. Some invertebrates, such as the fruit fly tuations . In Dros defense . During re death .It has been shown that an excess of fat mass, especially the visceral type, associated with a diet rich in saturated fats and simple sugars and low in fiber and micronutrients , which is the current Western diet, is able to promote a chronic low-grade inflammatory state, which, albeit involving molecules and signals common to the classic inflammatory response, recognizes the primary trigger in the metabolic overload . ConversMoreover, particular interest has been directed to IL-1β-mediated inflammation because of the strong inter-relationship between dietary fats, metabolic stressors, and NLRP3-mediated inflammation . MetabolDuring the feeding/fasting cycle, in physiological conditions, an intermittent, non-specific, low-grade inflammatory response occurs at the level of “metabolic tissues” , which results in a transient increase in some inflammatory proteins/cytokines in the serum. This increment reaches its maximum peak during the absorption phase  and then gradually decreases in about 2 h, when the nutrients have been distributed, metabolized, and/or accumulated in the respective cellular sites . The infExperimental data showed that the quantity and quality of fats influences the acute inflammatory response to a single meal. The study revealed that SFAs content and the ratio among polyunsaturated fatty acids (PUFAs) from ω-3 to ω-6 have proved to be the major determinants of the extent of postprandial inflammatory response ,37. In pFatty acids can affect the inflammatory response either through the synthesis of eicosanoids that regulate the transduction of signals at cell membrane or cytoplasmic level, by modulating the activity of transcription factors involved in the inflammatory process.An examination of data from the National Health and Nutrition Examination Survey (NHANES 99-00) found that levels of SFAs in serum phospholipids of male civil servant workers (40–69 years) positively correlated with some inflammatory markers, such as HS-CRP (high sensitivity C-reactive protein) and fibrinogen; by contrast, phospholipid PUFA levels were inversely associated with HS-CRP ,36. It h-Phosphorylation of insulin receptor substrate-1 (IRS-1) in serine and its degradation and elimination through the ubiquitin pathway, thus blocking insulin signal transmission;-Activation of the transcription factor NF-kB, by its dissociation from the cytoplasmic inhibitor (IkB) and transfer to the nucleus, with over-regulation of the pro-inflammatory gene expression.In vitro studies have shown that SFAs, such as lauric, myristic, and palmitic acids, are able to activate the Toll-like receptors (TLR) of adipocytes and macrophages . The SFAIn conclusion, SFAs impair insulin signaling through TLR4, activating the NF-κB pathway and NLRP3 inflammasome that promotes the conversion of pro-IL-1β into mature IL-1β ,31,32. IFurthermore, the ω-3 PUFAs, by binding to the G-protein-coupled receptor 120 , block the NFκB activation pathway. Finally, EPA would be able to inhibit the activity of the Δ-6 desaturase enzyme, thus reducing AA formation ,44. SomeDiets with relatively high glycemic index (GI) and glycemic load (GL) have been linked to elevated risk of T2D, coronary heart disease, and stroke, particularly among overweight individuals . GI and Recent clinical trials indicate that targeting the prototypic pro-inflammatory cytokine IL-1β ameliorates the outcomes of cardiovascular disease, which is the first cause of death in T2D patients. Several T2D-related metabolic factors, including reactive oxygen species, glyco/lipoxidation end products, and cholesterol crystals, have been involved in the pathogenesis of diabetes complications (diabetic kidney disease and diabetic retinopathy) and in the progression of atherosclerosis and NAFLD .2•−), hydrogen peroxide (H2O2)) by normal or malfunctioning mitochondria, or indirectly by other activators of NLRP3. In particular, it was suggested that increased ROS are sensed by a complex of redox-dependent inhibition of thioredoxin (TRX) and TRX-interacting protein (TXNIP) and cause the dissociation of the complex described above. Following an increase in cellular ROS concentration, this complex dissociates and TXNIP binds to the LRR region of NLRP3, with the consequent NLRP3 activation [High levels of glucose and non-esterified fatty acids (NEFAs) can cause oxidative stress due to both the decoupling of oxidative phosphorylation and to the increased mitochondrial β-oxidation. Lee et al. found that hyperglycemia-induced elevated mitochondrial reactive oxygen species in myeloid cells of T2D patients are associated with increased production of inflammasome-dependent cytokines IL-1β and IL-18 . Some dativation .Chronic hyperglycemia (associated or not with increased triglycerides and NEFAs) promotes the formation of advanced glycation end-products (AGEs). The interaction between AGEs and receptors for advanced glycation end-products (RAGEs), the activation of the polyols pathway and the auto-oxidation of glucose lead to increased production of ROS which, in turn, induce the activation of transcription factors, such as NF-kB . The resConsumption of whole grains, through increased fiber intake, is associated with a reduction in serum concentrations of inflammation biomarkers ; although, the mechanisms still need to be elucidated, and a synergistic effect of fiber, minerals, vitamins, or phytochemicals, such as lignans and phenolic acids, could take place ,36.Several studies in recent years have demonstrated the role of specific nutrients in the primary or secondary prevention of inflammation-related diseases, such as metabolic syndrome and diabetes. In this regard, some nutraceuticals and bioactive compounds  have been widely studied and have been recognized for their ability to inhibit the synthesis of TNF-α in monocytes and macrophages and downregulate the expression of TLR2 and TLR4 in human monocytes .However, due to the reciprocal synergistic or antagonistic interactions between nutrients and/or other food compounds , it is clear that, in order to understand the role of diet in the modulation of the inflammatory state and oxidative stress, it is necessary to evaluate the nutritional pattern in its entirety. It was found that diet characterized by an abundant consumption of vegetables, fruit, nuts, olive oil, legumes, and fish, with moderate amounts of alcohol and low intake of red meat, processed meats, refined grains, and dairy products, has a protective effect against immunometabolic diseases . These aThe literature search was carried out in August 2021 in official scientific databases, using combinations of keywords “NLRP3”, “NLRP3 inflammasome”, “food”, “nutrients”, and “bioactive compound” in the literature published during the last five years. Study selection and data were collected according to the 2009 Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines , Figure Polyphenols are secondary metabolites produced by the metabolic processes of plants. Although more than 8000 polyphenols have been identified up to now, all these compounds arise from a common intermediate—phenylalanine or shikimic acid. Generally, they occur in conjugated forms, with one or more sugar residues, organic acids, amines, lipids, and other phenols . PolyphePhenolic acids are one of the main compounds of polyphenol class. Phenolic acids are characterized by a phenolic ring with a carboxylic acid function and can be further divided into two main groups, benzoic acid or cinnamic acid derivatives, based on the C1–C6 or C3–C6 backbones . SeveralSinapic acid is a hydroxycinnamic acid widely found in many foodstuffs, such as vegetables, spices, citrus, berry fruits, cereals, oilseed crops, wine, and vinegar. Sinapic acid has shown antioxidant and anti-inflammatory properties , as wellFerulic acid is an another hydroxycinnamic acid commonly found in fruits, vegetables, grains, beans, leaves, seeds, nuts, grasses, flowers, and in some plants, such as corn and wheat, and in the spice turmeric . A totalChlorogenic acid is another phenolic acid, widely studied for its anti-inflammatory properties . ChlorogLastly, among cinnamaldehyde-related compounds, only cinnamaldehyde and 2-methoxycinnamaldehyde limited the expression of NLRP3 and pro-IL-1β at 25–100 µM, whereas cinnamic acid, cinnamyl alcohol, cinnamyl acetate, and α-methylcinnamaldehyde were ineffective, thus demonstrating the importance of the propenal group in the side chain ,138.N-oxide). Isoorientin, a flavone identified in Gentiana roots, has shown to possess anti-NLRP3 inflammasome activation in in vivo and in vitro models of hyperuricemia by using 5 and 10 mg/kg (0.4 and 0.8 mg/kg HED) dosages in male ICR mice for in vivo studies and 25, 50, 100, 200, and 400 µM isoorientin for in vitro assays (MXC207 cells). In both cases, a dose-dependent reduction in inflammasome expression was observed, through the inhibition of xanthine oxidase activity and interleukin release [Among flavones, apigenin, largely present in common fruits and vegetables, such as parsley, onions, oranges, tea, etc., has shown to be effective in reducing NLRP3 inflammasome activation in two different studies. In the first one, in vivo and in vitro models of high-fat diet-induced non-alcoholic fatty liver disease (HFD-induced NAFLD) have been treated with apigenin . In part release . In a ve release . Luteoli release .Regarding flavanones, hesperidin methylchalcone and naringin have been demonstrated to be effective in reducing NLRP3 inflammasome activation. In particular, hesperidin methylchalcone, particularly present in citrus fruits, has shown to reduce the NLRP3, ASC, pro-caspase-1, and pro-IL-1β mRNA expression in induced gout arthritis Swiss mice models when orally administrated at 30 mg/kg (2.4 mg/kg HED) . In a mo−/− mice to observe the potential reduction in atherosclerotic inflammation, where NLRP3 inflammasome is largely involved. In the study, an important decrease in pro-IL-1β and IL-1β was registered [E. coli-infected cells has also been carried out to demonstrate the anti-inflammasome activity of quercetin during infection. In particular, the 12 h treatment with 200 µM quercetin before infection has shown to strongly reduce NLRP3, caspase-1, and IL-1β expression, together with an enhanced ROS scavenger activity [Among polyphenols, quercetin, largely present in several vegetables and fruits, has been the most considered compound for the study of its anti-NLRP3 inflammasome activity. In particular, in an alcohol-induced acute liver injury in male SPF-Wistar mice, the oral administration of 100 mg/kg quercetin (8 mg/kg HED) for 14 days caused a reduction in the injury evolution, thanks to a lower expression of inflammasome factors including ROS, NF-κB, NLRP3 inflammasome, IL-1β, and IL-18, measured by ELISA assays in the liver tissue . The samgistered . The effgistered , since aactivity . Similaractivity . Other tactivity . It is nactivity .O-β-glucoside (C3G), a phenolic molecule mainly present in red–violet fruits. In particular, 4-hydroxyhexenal-induced inflammation in human retinal pigment epithelial cells has shown to be less severe when cells have been pre-treated for 2 h with 50 and 100 µM C3G, showing a dose-dependent decrease in NLRP3, IL-18, IL-β, and caspase-1 [Curcumin is the main active compound of turmeric and its anti-inflammatory activities have been largely demonstrated in many inflammatory diseases, in which the inhibition of NLRP3 inflammasome represents an important mechanism of action . This acaspase-1 . C3G hasaspase-1 . Green taspase-1 . In partaspase-1 . Resveraaspase-1 . In partFood extracts characterized by the presence of polyphenols mixtures have also shown to be effective in the inhibition of NLRP3 inflammasome. In this case, it is not possible to have an idea about the contribute of single molecules that are present in the mixture, also in consideration of the synergistic, additive, and antagonistic effects that can occur . Anyway,Zea mays L.) and common bean (Phaseolus vulgaris L.) snacks are food products, rich in polyphenols, such as gallic acid and other compounds, such as butyric acid and verbascose, that present an in vitro anti-inflammatory activity. Regarding the NLRP3 inflammasome complex, the FNDF pure components inhibited the NLRP3 assemblage, decreasing caspase-1 activity, IL-1β, and apoptosis in THP-1 cells and differentiated Caco-2 cells after NLRP3 inflammasome activation [Fermented non-digestible fraction (FNDF) of baked corn  being the most abundant one. In a recent work, the green tea polyphenols (GTPs) have reported a modulating activity against the NLRP3 inflammasome activation . In partIn another study, the GTPs were administrated to the same mice, in a dose (100–200 mg/kg) (8 and 16 mg/kg HED) comparable with the normal drinking tea levels consumed by humans, in order to determine their protective effects against the inflammasome activation. The results confirmed that GTPs inhibited the NLRP3, ASC, and caspase-1 p20 expression in a dose-dependent manner .Soy isoflavones represent an interesting flavonoid choice for the treatment of many inflammation disorders. A recent study conducted on dextran sodium sulphate (DSS)-treated mice reported the inhibiting capacity of soy isoflavones on NLRP3 inflammasome complex expression, reducing the NLRP3, caspase-1 p20, and ASC protein levels and suppressing the release of IL-1β and IL-18 .Red raspberry polyphenol extracts are rich in anthocyanins, ellagic acid, myricetin, (−)-epicatechin, and (+)-catechin. The extract was administrated to C57BL/6 mice revealing the attenuation of NLRP3 inflammasome activation in adipose tissue macrophages and epididymal white adipose tissue. The downregulation of NLRP3 inflammasome activation, leading to reduced expression levels of IL-1β, IL-18, and NLRP3 proteins, was also observed in vitro on C3H10T1/2 cells .Allium  and Brassica  genera [Allium genus vegetables, methylsulfonylmethane, has shown to have anti-NLRP3 inflammasome activity in in vitro assays on human and mouse macrophages by using different methylsulfonylmethane solution concentrations  [Organosulfur compounds (OSCs) are a class of molecules usually present in several food sources, such as cereals, legumes, vegetables, and fruits. However, the main food sources of OSCs belong to the ) genera  and are ) genera ,144. Amo) genera . In both) genera . When be) genera . Finallynd 8.0%) . In partRosmarinus and Salvia plants, has been demonstrated to reduce the evolution of DSS-induced colitis in male Balb/c mice daily gavaged with 50 or 100 mg/kg (4 and 8 mg/kg HED) of carnosic acid for 10 days by inhibiting the activation of caspase-2 and the release of pro-inflammatory cytokines and ROS in the colon tissue [X. aethiopica, has shown to possess a dose-dependent  triggering effect of NLRP3 inflammasome in BALB/c mice macrophages infected with L. amazonensis promastigotes by increasing the production of the inflammatory mediators NO and IL-1β [Terpenes are a class of volatile hydrocarbon compounds derived from two or more isoprene units and represent the main components of vegetable essential oils. In recent years, a paramount interest for these molecules has emerged, since several biological activities, such as antitumoral, anti-inflammatory, antibacterial, antiviral, antimalarial, and cardiovascular modulation have been demonstrated . In partn tissue . Moreoven tissue . The addnd IL-1β .Ainsliaea yunnanensis [Although not present in natural sources used as foods, other diterpenoids, sesquiterpenes, iridoids, phytocannabinoids, and derivatives typical of plants used in Eastern and Western medicine, namely α-bisabolol, aucubin, abscisic acid (phytohormone), cannabidiol, phytanyl amine, triptolide, tanshinone IIA, sodium tanshinone IIA sulfonate, paclitaxel, phorbol myristate acetate, andrographolide, oridonin, glaucocalyxin A, and teuvincenone F have shown to be effective in the prevention of NLRP3 inflammasome activation by blocking the release of inflammasome enhancers, mainly IL-1β, IL-4, IL-6, IL-12, IL-18, caspase-1, and ROS ,152,153.nanensis , have shIn general, most of the evidence highlighted that SFAs acted as priming signals of inflammasome activation, whereas MUFAs and PUFAs have been shown to impair this activation, but these assumptions have frequently been challenged . To bett+, K+-ATPase transmembrane protein, resulting in an increase in K+ efflux, which is considered a NLRP3 activator [Different research groups attempted to investigate the mechanisms underlying SFA-mediated NLRP3 inflammasome activation. Wen et al. demonstrated palmitate acid inhibits AMP-activated protein kinase in LPS-primed bone marrow-derived macrophages followed by the accumulation of mitochondrial ROS, thus activating the NLRP3-ASC inflammasome and causing caspase-1, IL-1β, and IL-18 production . Accordictivator .However, few studies reported the anti-inflammatory effects displayed by different SFAs. As an example, virgin coconut oil (VOC), mainly composed of medium chain saturated fatty acids (6–12 carbons), reduced IL-1β protein, caspase-1, and NLRP3 genes expression at different doses in AD (receiving Amyloid-β) and in high-fat diet (HFD) models both in vitro and in vivo . The antIn the literature, most of the studies on NLRP3 inflammasome and fatty acids are focused on PUFAs. In particular, fish oil, a matrix particularly enriched in ω-3 DHA and EPA, has been extensively studied. All data reported the anti-inflammatory properties of EPA and DHA resulting in a downregulation of NLRP3 gene expression and in a decreased secretion of pro-inflammatory cytokines (IL-1β and IL-18) in obesity models ,103, in Interestingly, different properties have been reported between ω-3 and ω-6 PUFAs. Schuster et al. investigated the anti-inflammatory activity of different fatty acids  in macrophages, blood monocytes, and hepatocytes, confirming the data described above. Nevertheless, when compared with ω-6 PUFAs, ω-3 were more potent in inhibiting ATP-mediated NLRP3 inflammasome . ConversLycium barbarum) has been confirmed, demonstrated the ability to counteract ethanol-induced hepatic damage in a murine model of AFLD targeting the converged AMPK-FoxO3a mitophagy and NPLR3 pathway combined to P2X7 and adiponectin receptor 1 on the hepatocyte membrane [w/w) to the normal diet [Recently, only two carotenoids were studied in vivo for the modulation of NLPR3 inflammasome. Zeaxanthin dipalmitate, a lipophilic antioxidant whose primary presence in some functional fruits , and RDP3 , were identified and isolated from the aqueous extract of the shelled fruits of  effects ,108. The effects  study waRegarding the hyperuricemia and its associated kidney inflammation, the effects and biological mechanisms of tuna meat oligopeptides (TMOP) were investigated in mice . The conConcerning the intestinal diseases, the ability of an isoflavone-free soy protein concentrate (SPC) to prevent inflammation and loss of intestinal barrier function have been examined . The cytOtherwise, a gluten-derived peptide with high content of glutamine and proline residues, α-gliadin 31–43, has been identified, which appears to induce an innate immune response in the gut . Mice inBy considering that the redox state of the cell can mediate the production of placental cytokines, which are responsible for the function and development of the placenta and that the NLRP3 inflammasome represents the first line of innate immunity , the NLRNLRP3 inflammasome has been reported to play an essential role in the inflammation responses during acute lung injury . RegardiSince cholinergic degeneration plays a major role in the pathophysiology of AD, it has been investigated whether choline supplementation during adulthood delays the progression of this disease . The stuGinseng is considered as a highly valued herb and widely used in dietary supplements and herbal medicines, as it exerts a regulation on endocrine system, nerves, metabolism, and other physiological functions ,165. The3-PPD , a ginsenoside isolated from Panax ginseng, relieves liver damage, by inducing apoptosis of activated hepatic stellate cells [3-PPD  against hepatic fibrosis and inflammation in thioacetamide (TAA)-stimulated mice by activating the LXRs (liver X receptors) pathway [3-PPD has a hepatoprotective effect against liver fibrosis and reduces inflammation by regulating P2X7R-mediated NLRP3 inflammasome.It has been confirmed that 25-OCHte cells . Conside pathway . Study rPanax notoginseng in ameliorating NLFD through inhibition of inflammasome activation. The saponin extract, compared with conventional red ginseng, contained significantly increased amounts of ginsenosides Rh1 (10.34-fold) and Rg2 (7.1-fold), which are the main components highlighting the pharmacological activity of ginseng. In this study mice were fed a fast-food diet for 16 weeks to induce NLFD and then treated with saponin extract (50 or 150 mg/kg) for 9 weeks to determine the effects of the saponin. Particularly, it has been observed that Rh1 and Rg2 ginsenosides exerted anti-inflammatory effects and inhibited NLRP3 inflammasome by promoting mitophagy and alleviating mtROS production.Wang et al.  analyzedThe characteristics of the ginsenoside compound K (CK) are peculiar, a final metabolite of panaxadiol ginsenosides. CK seems to be involved in the stimulation of insulin secretion and its protective effect against diabetic nephropathy has been elucidated in inhibiting the oxidative stress, NLRP3 inflammasome, and NF-κB/p38 ,168.Numerous studies have demonstrated the neuroprotective action and antidiabetic properties of CK, indicating its potential in the treatment of memory disorders and cognitive dysfunctions related to diabetes mellitus; in particular, a study tried to assess the effects of CK against memory impairment and cognitive dysfunction in diabetic db/db mice treated with 10 mg/kg of CK for 12 weeks . The resGlycyrrhiza glabra, acts as a hepatoprotective agent in the immune and anti-inflammatory modulation of liver disease [Magnesium isoglycyrrhizinate, a magnesium salt of 18α-glycyrrhizic acid stereoisomer extracted from the roots of the plant  disease . The eff disease . Rats we disease .Physalis alkekengi L. var. franchetii, displayed anti-inflammatory activity in intestinal ischemia [Physalin B, one of the main active withanolides existing in ischemia . The Phyischemia ; mice weMoringa oleifera, β-sitosterol, was studied to evaluate its anti-inflammatory activity in two cell lines [A plant phytosterol isolated from ll lines . Tween 8Trametes orientalis, used for the treatment of pulmonary disease, is a polysaccharide composed of galactose, glucose, mannose, and arabinose with molar ratios of 5.79:5.77:3.45:1.20 (average MW 63000), which proved to inhibit the activation of NLRP3 inflammasome and the release of pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6, by the detection of expression levels of proteins in lung tissue involved in the NLRP3 inflammatory pathway [Dendrobium officinale (DOPS) was likely to via suppressing mRNA expression of NLRP3, ASC, caspase-1, IL-1β, and IL-18 both in vivo, in DSS-induced acute ulcerative colitis mice, and in vitro, in LPS-stimulated NCM460 cells model, through a possible downregulation of β-arrestin1 expression (which could positively regulate NLRP3) [Ganoderma lucidum polysaccharides (GLPS), isolated from a Chinese medicinal mushroom, proved to markedly inhibit liver inflammatory factors via suppression of NLRP3 in liver tissue. Indeed, GPLS-treated acute liver injury mice exhibited a significantly decreased protein expression levels of NLRP3, ASC, and caspase-1 [Armillariella tabescens (AT), that belongs to the family of Tricholomataceae, is a well-known traditional medicinal mushroom, characterized by 86.52% of polysaccharides . AT possesses potent antioxidant and anti-inflammatory properties by possibly involving the repression of the TXNIP/NLRP3 inflammasome pathway in the liver of T2D mice [Polysaccharides from different sources  have proved to exert an anti-inflammatory role via affecting the NLRP3 inflammasome pathway. However, a detailed mechanistic study on how polysaccharides modulate NLRP3 activation remained to be explored. In particular, several polysaccharides involved in the modulation of NLRP3 inflammasome belong to traditional Chinese medicine, but they are also present in dietary plants. As an example, the major bioactive component of  pathway . Liang ee NLRP3) . A recene NLRP3) . Ganoderaspase-1 . ArmillaT2D mice . Among tT2D mice . The NLRT2D mice .Recently, Castro-Alves et al. investigated the activity of non-digestible carbohydrates (NDCs) from chayote fruit, which consist mainly of pectic homogalacturonan and highly branched rhamnogalacturonan-II, as well as hemicellulosic material including glucomannan, xyloglucan, and glucurono(arabino)xylan, in human THP-1 macrophage-like cells. Results showed that NDCs indirectly inhibit NLRP3 activation through the interaction between the NDCs and other pattern-recognition receptors that are essential to induce priming signals required for NLRP3 activation .A. rollestoni), could improve the release of cytokines and the expression of NLRP3 in primary macrophages, confirmed by the elevated expression of NLRP3, cleaved caspase-1, and ASC proteins [Several papers highlighted the anti-inflammatory activity of different polysaccharides probably ascribed to NLRP3 inhibition by decreased protein levels of NLRP3, ASC, caspase-1, and IL-1β ,173,174;proteins .The importance of lipophilic vitamin E and its derivatives, which mainly occur as abundant nutrients in oily nuts and seeds, in inflammation and oxidation processes has been elegantly reviewed by Wallert and collaborators . DiffereVitamin D and its derivatives were also studied, alone or in association with pro-biotics and microelements as food supplements, for their effects on the modulation of inflammasome halting the priming step required for NLRP3 (and NLRP1) activation in immune cells, in placental explants from preeclamptic and normotensive pregnant women, in clinical trials involving COVID-19 patients , in the Lactobacillus paracasei KW3110 (1 × 106 cells/mL), Bifidobacterium infantis (2 × 108 CFU/mL) in combination with the prebiotic xylooligosaccharide , Enterococcus faecium NCIMB 10415 (1 × 107 CFU/mL), and heat-killed cells of Enterococcus faecalis  were recently proposed in different in vitro and in vivo disease models of DSS-induced ulcerative colitis [5 CFU of Lactobacillus rhamnosus GR-1 and 3 × 105–2 × 107 CFU of Lactobacillus johnsonii L531) with the aim of limiting the inflammatory damage of bacterial infections induced by Escherichia coli in porcine mammary epithelial cells, MAC-T cells and in a mouse mastitis model, or Salmonella typhimurium in IPEC-J2 cells [Lactic acid bacteria (LAB) are currently used as food supplements for human health as probiotics due to their modulatory effects on intestinal conditions, immunomodulatory function, and host metabolism. The effect of each probiotic is dependent on the cell line and species used. To test the anti-inflammatory action of LAB or their main components  via the inhibition of inflammasome activation,  colitis , HFD-ind colitis , cerulei colitis , and col colitis , as well colitis . The benJ2 cells ,185,186.Apart from table sugar, foods are “complex matrices” made up of different components, divided into macronutrients , micronutrients , and water. The presence of other minority components also increases the complexity of food matrices. Some of them are defined as “bioactive compounds” because, based on scientific evidence, it is assumed that they can have a positive effect on our organism and can play a fundamental and important role in modulating inflammasomes ,30,35.The “bioactive compounds” belong to many different categories based on their chemical nature and on the type of positive effect they could exert on human health. Commonly, they are not considered nutrients and are referred as “essential and non-essential compounds” that occur in nature, are part of the food chain, and can help to protect ourselves from illnesses and improve our quality life ,187. AltCertainly, the most numerous classes of “bioactive compounds” are represented by phytochemicals: molecules of various kinds such as carotenoids, tocopherols, phytosterols, phenolic compounds, etc., but, in any case, these are of vegetable origin. Among all these different metabolites that are important for their health benefits, the phenolic compounds have been studied most extensively ,191. TheThe intake of “bioactive compounds” in the human diet varies enormously in relation to the type, quantity, and quality of foods consumed. Generally, the most important food sources of bioactive nutrients are vegetable, fruits, chocolate, tea, wine, olives, and spices; however, their concentration in the same food varies, often significantly, in relation to the cultivation techniques and food processing, the degree of ripeness, and the time elapsed between harvesting and consumption ,203. ThuUsually, to improve the sensory attributes and make a product more attractive for taste, consistency, and appearance, many foods are submitted to culinary treatments. Different domestic practice and cooking techniques  have been compared to define the optimal process to reduce the degradation of the “bioactive compounds”. It was observed that food nature, cooking time, temperature, addition of acidulant, spices, or other condiments might influence both the amount of water-soluble “bioactive compounds” and of those fixed to the lipidic matrix . Each foAlthough there is evidence showing the beneficial effects of “bioactive compounds” present in foods, more studies suggest that their health benefits instead depend on the synergy and interactions among different molecules . TherefoDespite the growing amounts of studies, definitive conclusions supporting the bioavailability of most “bioactive compounds” from a wide variety of food for optimal nutrition and health well-being are difficult to obtain. Consumers’ demand for well-being pushes towards an increasing request for food supplements and innovative new market segments have been developed to meet their needs. Alternatively, dietary supplements, also differentiated into nutraceutical or functional foods, can be supplied to consumers to deliver a specific bioactive compound or a group of them ,189. GenNowadays, a great variety of nutraceuticals and functional foods are provided to the consumers; however, their use is largely unregulated, or they are treated differently according to local jurisdictions. Generally, they are more the subject of marketing hype, and for many of these “supplements”, it is not even yet known whether they provide more benefits than risks to consumers. Currently the term “bioactive compound” is not defined in the European regulations; however, the European Food Safety Authority (EFSA) is carrying out scientific evaluations to assess the safety and the toxicity of “bioactive compounds” that can be found in foods . Due to Finally, food consumed as such, or after cooking processes, as well as water, may also contain internal or accidental components with a stimulating effect on NLRP3 inflammasome. Among these, mycotoxins , trace metals , environmental and food contaminants , and acrylamide are few examples of NLRP3 activators, endowed with harmful inflammatory effects ,219,220.Eating is something we have always done and which, from birth to old age, strongly affects life quality and physical and psychological well-being. Nowadays, consumers are increasingly interested in lifestyles and nutritional habits that can prevent diseases since different studies have evidenced that a relationship occurs between these factors and pathological consequences based on inflammatory processes. In this regard, it should be emphasized that these effects are related, not only to the quantity, but, above all, to the variety of food choices and, in particular, to the inclusion of fruit and vegetables. Plant foods are particularly rich in important bioactive compounds, each of them can produce different effects on the pathway of inflammatory response, confirming the importance of the nutritional pattern (food model) as a whole, rather than the single nutrient or functional compound.


Pd NPs/N00Cl-100, was fullycharacterized both in the liquid phase and in the solid state, andits deactivation upon recovery was analyzed.Twenty-onegreen solvents, including glycerol-derived ethers, andtheir eutectic mixtures with two renewable ammonium salts, were usedfor the straightforward synthesis, stabilization, and immobilizationof palladium nanoparticles (Pd NPs). The nature of the solvent allowstuning of the characteristics and properties of resulting catalyticsystems in terms of particle size and morphology, stability, reactivity,and recoverability. Pd NPs immobilized in glycerol-based solventswere applied in the catalytic hydrogenation of alkenes, alkynes, andcarbonyl compounds, as well as in the selective semihydrogenationof alkynes to alkenes. The optimal experimental parameters and theinfluence on the reactivity of the physicochemical properties of solvent,mainly the viscosity, were studied. Moreover, the most active andrecoverable catalytic system,  Glyceryl monoether-derived deep eutectic solvents exhibitadequate properties for the preparation, immobilization, and stabilizationof highly active and recoverable Pd-NP catalytic systems for hydrogenationreactions of a wide scope of substrates. For this reason, the enhancement of their stabilityis a key aspect that can be kinetically achieved by introducing steric,electronic, or electrosteric repulsion forces.7 Organic ligands, surfactants, and polymers have been widely usedfor NPs stabilization.9 Once stabilized, NPs can be useddirectly or supported, blending the main features of both homogeneousand heterogeneous catalysts.10 The immobilization,either on solids or liquids, facilitates the recovery and reuse ofmetal NPs, and above all, increases their stability.11 Apart from solid supports such as clays, zeolites, oxides,polymers, and carbonaceous materials,13 structurally organized liquid phases are being successfullyapplied for the immobilization of NPs.15Metallicnanocatalysts have attracted huge interest in fields suchas nanomaterials, optics, catalysis, and biomedicine, due to theirelectronic properties and high specific surface areas.16 Among them, biobasedsolvents such as polyols,17 water,4 ionic liquids (ILs),18 deep eutectic solvents (DESs),19 andsupercritical fluids4 have already beenused for the synthesis of NPs. In particular, glycerol has shown valuableproperties for the preparation and stabilization of metallic species,20 due to its hydrogen-bond supramolecular structure.21 On account of the current bioglycerine surplus,22 a plethora of DESs have been described using glycerol asa hydrogen-bond donor (HBD), in combination with different ammoniumsalts such as choline chloride (ChCl).24 Some glycerolderivatives, including carbonates, ketals, esters, and ethers, havealso been studied as useful and versatile biosolvents.25 In fact, green methods for the synthesis of monoethers and diethersof glycerol with multiple substitution patterns have been recentlydescribed.30 Thanks to their interesting physicochemical properties and theirvery low eco-toxicity,31 these glycerol-derivedsolvents are being used in novel applications,32 as chemical precursors,33 andalso for replacing glycerol as the HBD component in eutectic solvents.34 Nevertheless, none of them has been used asa solvent for the synthesis of NPs, and only for the immobilizationof preformed Pd NPs in a very recent work.35In thepast decade, the development of green solvents has enabledtheir intensive application owing to advantageous physicochemicalproperties such as both moderated volatility and flammability, biodegradabilityand low eco-toxicity.36 palladium, platinum, and nickel catalysts have been profuselyapplied in industrial hydrogenations for the large-scale productionof different petrochemicals, foods, drugs, and fertilizers.39 In this context, the use of metal nanoparticles immobilized ontosolid supports has demonstrated their interest, in particular forrecycling purposes.1 However, liquid phaseshave been less considered as supports in the immobilization of NPsfor catalytic applications.40Metal-catalyzed hydrogenation reactions of unsaturated compoundsrepresent efficient and sustainable procedures, due to its total atomeconomy and low waste generation. Since the first Ni-catalyzed hydrogenationreactions described by Sabatier and Senderens,42 In this case, the ILacts as the reaction medium, as well as the NP stabilizer, ligand,and support.18 Moreover, the selectivitycan be modulated by changing the structure of the solvent.18 Despite their chemical similarity to ILs, DESshave been barely applied in hydrogenation reactions. Just a few workshave appeared, such as the hydrogenation of methyl cinnamate in urea-carbohydratemixtures using the Wilkinson’s catalyst,43 or the use of Pd NPs immobilized in choline-based DESsin turn microencapsulated in polysilanes for the hydrogenation ofunsaturated compounds.44 Finally, Gómezand co-workers have used polyols, mainly glycerol, as media for theimmobilization of Ni NPs, Pd NPs, and bimetallic Pd/Cu NPs for hydrogenationprocesses.46To date, metal NPs in different ILs have been efficientlyusedin selective hydrogenations.In this work, we study the utilityof glycerol-derived solventssuch as mono-, di-, and triethers of glycerol, as well as eutecticmixtures thereof, both as media for the sustainable synthesis, stabilization,and immobilization of Pd NPs and also for the selective hydrogenationof different unsaturated substrates. Morphology, stability, reactivity,and recoverability of the catalytic systems have been analyzed, discussingthe relevance of the nature of the solvent.Supporting Information (SI). All themanipulations were performed using Schlenk techniques under an argonatmosphere, unless otherwise stated. High-pressure reactions werecarried out in a Top Industrie Autoclave. Conversions and yields weredetermined by gas chromatography coupled to mass spectrometry (GC-MS),using a PerkinElmer Clarus 500 chromatograph equipped with a flameionization detector (FID), and a PerkinElmer Clarus MS 560 spectrometeras the mass detector. GC experimental conditions are detailed in the SI.The chemicals used in this work as reagent gradeare listed inthe SI.Glycerol-based solvents, catalyticsystems, and reaction productswere fully characterized. Experimental and characterization detailsare gathered in the R00 monoethers were selectivelyprepared from glycidol andthe corresponding alcohol by means of basic catalysis, following ourpreviously described methodology.27 Glycerylsymmetric R0R diethers and nonsymmetric R0R′ ones were synthesized starting from epichlorohydrin and the correspondingalcohol or alcohols, according to our previous works.30 Glycerol RRR triethers were obtained by methylationof the glycerol diether or monoether (1 mol) with iodomethane (1.4mol), prior to ether deprotonation with NaH (1.2 mol) in dry THF (150mL). After heating at 60 °C for 30–90 min, the reactionmixture was poured into cold water (133 mL) and quenched with HCl. Glycerol triethers were extracted with diethyl ether(3 × 66 mL), and the combined organic extracts were washed withan aqueous solution of sodium thiosulfate (5 wt %) to eliminate iodinetraces. Then, the organic phase was dried over MgSO4 andfiltered off, and the diethyl ether was removed under reduced pressureto furnish the desired solvents. All the glycerol-derived solventswere completely purified by vacuum distillation and dried at 80 °Covernight under vacuum prior to use.Glycerol 2 and 111 mg of poly-N-vinylpyrrolidone  were completely dissolved,under an argon atmosphere, in 5 mL of the glycerol ether dried asdescribed above. That system was placed under vacuum, pressurizedwith hydrogen (3 bar), and stirred at 80 °C for 10 h. Then, theresulting black colloidal solution (10 mM of Pd NPs) was cannulatedand kept under argon prior to use in catalysis.In a Fisher–Porterbottle, 11.2 mg (0.05 mmol) of Pd(OAc)2 weredissolved in 5 mL of the dried glycerol-based DES  and stirred under argon at 80 °C for10 h. The black colloidal suspension obtained (10 mM or 1 mol % Pd)was kept under argon.In a Schlenkflask, 11.2 mg (0.05 mmol) of Pd(OAc)N00Cl-100 solvent).After theabove-described synthesis, NPs in glycerol-derived solvents were transferredto a centrifugation tube and ethanol (5 mL) was added. Centrifugationwas carried out at 3000 rpm for 5 min, and then the solution was separatedby decantation. This process was repeated twice with ethanol (5 mL),followed by two additional washings with acetone (2 mL). The remainingblack powder was dried under vacuum at 80 °C overnight. Elementalanalysis (Pd determined by ICP-AES): Pd 88.9%, C 5.29%, N 0.95%, H0.46% (in n-pentane(5 × 5 mL) at 40 °C using vigorous stirring and periodsof 15 min between each extraction. Purification of products was carriedout by column chromatography or Kugelrohr distillation. Upon extractionof the organic products, the catalytic system was dried at 80 °Cunder vacuum for 5 h prior to a new run.The corresponding substrate (1 mmol)and the suspension of Pd NPs/glycerol-derivedsolvent  were placed into a Fisher–Porterbottle or in an autoclave vessel (for hydrogenations working from 10 to 55bar pressure) and stirred under an argon atmosphere. The reactionmixture was put under vacuum and pressurized with hydrogen at thedesired pressure. The system was stirred and heated at 80 °Cin a silicone oil bath for the specified reaction times. Then, thereaction system was cooled down to room temperature and then depressurized.The organic compounds were extracted with 34 Some features of these greenmedia such as their low eco-toxicity, low volatility, viscosity, andhigh solubilizing ability for organic and inorganic compounds34 encouraged us to use them for the synthesisof original catalytic systems based on palladium nanoparticles immobilizedin liquid phases (Pd NPs/solvent). With this aim, theproperties of the glycerol-derived medium not only allow the reductionof the palladium salt used as a zerovalent Pd NPs precursor owingto the solubilization of H2 but also enable the formationand stabilization of the metal NPs. Given the importance of Pd nanocatalyststoward surface-assisted reactions,17 wedecided to apply the Pd NPs/solvent systems in differenthydrogenation catalytic processes. In addition, the solubility oforganic substrates and the recovery of reaction products from theglycerol-based reaction medium was also assessed in terms of simplemanipulation and high reproducibility.As we havepreviously described, a great variety of differently substitutedglycerol ethers can be synthesized in an eco-friendly manner in orderto achieve a portfolio of renewable solvents with interesting physicochemicalproperties.000), glyceryl monoethers(R00), diethers (symmetric R0R and nonsymmetric R0R′), and triethers (RRR), as well astheir eutectic mixtures with two biobased ammonium salts, namely,choline chloride (ChCl) and N,N,N-triethyl-2,3-dihydroxy-propan-1-aminiumchloride  and the glyceryl monoethers 100, 3i00, and 3F00 were used as the HBD componentin combination with the two above-mentioned salts, ChCl and N00Cl, as the HBA component, according to a 1:2HBA/HBD molar ratio. Also, the ternary mixture N00Cl-100-3F03F, including the symmetric diether 3F03F, was preparedin a 1:1:1 molar ratio , glycerol monoethers , glyceryldiethers ,and glycerol triethers  as solvents, as well as PVP as the stabilizer of thenanoparticles, were used.In this work, Pd NPs were synthesized bythe reduction of a palladium precursor, Pd(OAc)20Under mild conditions, at a hydrogenpressure of 3 bar and a temperature of 80 °C, the reduction ofPd(II) could be evidenced in the glycerol-derived media after 30 min,thanks to a color change of the resulting suspension, from orangeto Pd(0) black 2. In ord000), the dispersionand homogeneity of the NPs were limited by its high viscosity (1200cP versus 0.7–68 cP for the studied glycerol ethers at roomtemperature). All TEM micrographs and size distributions are detailedin the Supporting Information, except forsolvents 101 and 111, which could not berecorded in liquid media due to the higher volatility of both solvents.Interestingly, the amount and mean diameter of NPs showed a directdependence on the nature of the glycerol-derived solvent used in thesynthesis. In general, NPs prepared in more polar and viscous mediaincreased their particle size. As it can be observed in 3F03F < 100 < 3F00, the meanparticle size was respectively 1.3, 1.4, and 1.8 nm, albeit with relativelylarge size distributions. When comparing the influence of the polarityand hydrogen bond ability of the solvent on the morphology of thesynthesized NPs, using two differently substituted glycerol etherssuch as 3F00 and 3F03F, it can be observedthat the higher polarity and HBD ability of the monoether seems tobe related to the formation of NPs presenting higher mean sizes. Otheraspects with minor influence on polarity, such as the ramificationor the length of the ether alkyl chain, did not seem to have a significantinfluence on the NPs’ shape or size. For instance, in the casesof using 200 and 3i00, with a similar polarity,both solvents provided similar sized Pd NPs (1.5 ± 0.7 nm).Transmission electron microscopy (TEM) analyses of these colloidalsuspensions in glycerol ethers revealed spherical-shaped and well-dispersedPd NPs 2.HoweveThe synthesis of Pd NPs in the glycerol-derivedDES, using the same palladium precursor, was achieved by heating themixture at 80 °C under an argon atmosphere in the absence ofany additional stabilizers 3. The io48 The resulting Pd NP black colloidal suspensions obtained using thisstraightforward procedure were stable. In particular, NPs synthesizedin these glycerol-derived eutectic media showed different morphologies,populations, and size distributions than in the case of using glycerolethers  into zerovalent palladium species.lethers 3.It canN00Cl-3F00 and ChCl-3F00, presented bigger meandiameters (ca. 2.2 nm) than in the case of the lesspolar solvents, i.e., 1.8 ±0.9 nm for N00Cl-100 and 1.9 ± 0.9 nm for N00Cl-3i00. In addition,the high viscosity of the solvent also influenced the Pd NPs dispersion.An inhomogeneous distribution of nanoparticles in solvents N00Cl-000 and N00Cl-3F00, with viscosity values of 2693 and 553cP, respectively, was observed. Although similar particle diametersand size distributions were found for Pd NPs synthesized in DES composedof the same glyceryl ether and different HBA salt, an influence ofthe nature of the ammonium salt on the NPs’ stability was identified.Thus, the presence of ChCl instead of N00Cl triggered some palladium agglomeration in the case of solvents ChCl-000, ChCl-3i00, and ChCl-3F00. Nevertheless, when using the fluorinated ether 3F00, similar trends were observed with both ammonium salts. Moreover,Pd NPs in N00Cl-100-3F03F showed a good dispersion ofhomogeneous particles with an average size of 2.0 ± 0.8 nm. Theinterest in these ternary mixture systems is based on the possibilityof fine-tuning the characteristics of metal NPs for specific catalyticapplications. As expected, the size was intermediate between thoseof N00Cl-100 and N00Cl-3F00, as a consequenceof the intermediate property values of this medium.Pd NPs synthesized in fluorinated media, such as Pd NPs/N00Cl-100 was chosen as a homogeneously dispersed nanoparticlesuspension in a DES for full characterization both in the liquid phaseand in the solid state . FT-IR analysis of Pd NPs in the solid state suggestedthe presence of remaining solvent N00Cl-100 in the isolatedNPs , as a result of thestrong interaction between the ionic solvent and palladium duringthe Pd NPs synthesis, thus demonstrating the major role of DES asan electrosteric stabilizer. Both elemental and ICP-AES analyses showeda consistent palladium percentage (ca. 89%). As couldbe expected, the rest of the mass seems to correspond to the above-mentionedsolvent N00Cl-100 stabilizing the NPs. As shown in 5/2 and 3d3/2 peaks, respectively).49 Furthermore,the XPS analysis revealed a N/Cl ratio of 3:8 (see Table S1 in the SI). This fact could be explained by the partialdegradation of the ammonium salt of DES (N00Cl), in theform of volatile amines (Hofmann elimination), with X-ray irradiation.In orderto evidence the structure of these immobilized Pd NPs, NPs see S21, as aFigure S19 and Figure S22 in the SI). The five maincrystallographic diffraction planes of crystalline Pd(0) can be appreciatedin 46Powder X-ray diffraction (PXRD) analysis allowed calculation ofa crystallite size of 3.9 ± 0.9 nm for Pd(0), which was consistentwith the mean diameter of 3.3 ± 2.1 nm calculated in the solidstate using TEM microscopy (see E)-4-phenylbut-3-en-2-one-4-phenylbut-3-en-2-one (1) catalyzed by Pd NPs immobilized both in glycerol . For this reaction medium, the best experimentalconditions were 1 bar of H2 and heating at 80 °C for2 h . In terms of recycling, only PdNPs/N00Cl-100 gave excellent conversion and selectivity inthe second run. Thus, this system showed the highest TON and TOF values. For the rest of the catalyticsystems, conversions did not exceed 43% in the second run.The influence of the reaction solvent was also assessedin orderto determine the most active Pd NPs/solvent system. Thus, we evaluatedthe catalytic activity at a shorter reaction time  to glyceryl ether-derived DES(12.5–21.4 cP at 80 °C), and even more for the DES composedof pure glycerol .34 The representation of the accumulated TON values versusthe dynamic viscosity of the solvent  in In a previous work, it was demonstratedthat the high viscosityof glycerol (with a dynamic viscosity of 31.9 cP at 80 °C) hindersthe diffusion of both the reagents and catalyst.N00Cl-100 < N00Cl-3i00 < N00Cl-3F00 < N00Cl-000, the inverse trend in the catalytic productivity(3920 > 1940 > 1880 > 900) can be observed. A similar behaviorinthe case of mixtures with ChCl and in the case of glyceryletherscan be also implemented. Other physicochemical properties such asdensity, surface tension, refractivity, or heat capacity of the solventhave also been considered.34 In the caseof surface tension, a weak relationship with reactivity was found,probably due to the same reasons above explained for viscosity.For instance, along the growing viscosity sequence 9 Therefore, viscosity differences are not enoughto explain catalytic differences, for example the highest reactivityfor Pd NPs immobilized in N00Cl-100 and N00Cl-100–3F03F, with respect to monoethers such as 100. Thus, we observedthat systems with a higher number of smaller NPs (mean diameter around1–2 nm) presented enhanced reactivity, as in the case of Pd NPs/3F03F, due to a larger active surface. Conversely,systems with less-spherical and bigger NPs, dispersed in an inhomogeneousmanner, such as ChCl-3F00 or N00Cl-3F00,would present less total active surface with respect to N00Cl-100, in agreement with a decrease in the TON. In addition, the formationof agglomerates notably reduces the catalytic active surface, whichwas probably the case for ChCl-000 and ChCl-3F00 solvents. Thus, Pd NPs’ morphology is modified by fine-tuningof the solvent, triggering an effect on catalytic reactivity. Forexample, the size of NPs in N00Cl-100-3F03F in betweenthose of N00Cl-100 and N00Cl-3F00 resultedin an intermediate TON value (2740), with respect to those of N00Cl-100 (3920) and N00Cl-3F00 (1880). Furthermore,the observed catalytic behavior also indicates that the DES supramolecularstructure impacts the size of PdNPs (effect on the PdNPs formationmechanism) and dispersion (effect on stabilization), N00Cl-100 being the best solvent to lead to small PdNPs and avoid their agglomeration,even after catalysis. On the basis of reported studies related tonanoparticles/solvent interactions, the electronic state of the PdNPs in the different solvents should be quite similar.51In addition to the properties of the reaction medium, the sizeand morphology of Pd NPs directly impact the catalytic reactivity.N00Cl-100, was used to perform additional recycling studiesfor the benchmark hydrogenation reaction  or in the monoether 100 started in the secondcycle. In the case of using the system Pd NPs/N00Cl-100, quantitative conversions were achieved for three consecutive cycles.Notably, Pd NPs suspended in the eutectic medium displayed greaterstability and recyclability than in the cases of the monoether orpure glycerol, even using a lower palladium load.Furthermore, the solvent giving the best catalytic results, reaction 4. With tPd NPs/N00Cl-100 was reused in two reaction cycles with quantitative conversionsand two additional runs with 61% of conversion. Unfortunately, cycles5–7 showed a progressive deactivation of the catalytic system.Attempts to regenerate the catalyst, by hydrogen and thermal treatment,or by the addition of solvent, were not successful. TEM analysis ofthe reused Pd NPs immobilized in N00Cl-100 . Theas-recovered catalyst was redispersed in fresh N00Cl-100 and used in the hydrogenation of (E)-4-phenylbut-3-en-2-one(1), exhibiting an important deactivation (<10% conversion).This behavior denotes a major effect of DESs on the stabilizationof Pd NPs during their synthesis.Additionally, the solvent stabilityduring the recycling was studiedby Pd NPs/N00Cl-100, was successfullyapplied in the hydrogenation of different compounds-4-phenylbut-3-en-2-one (1), 2-vinylnaphthalene (2), and 2,3,4,5-tetrahydro-1,1′-biphenyl(3), was quantitatively achieved using low Pd loadingsand mild reactionconditions (4), the hydrogenation product 4H and different isomerization products were obtained at shortreaction times (4 toward n-dodecane (4H) was only achieved at longerreaction times . All these results pointout that the catalytic activity of Pd NPs/N00Cl-100 isat least comparable to that of other Pd NPs described in glycerolfor similar substrates and reaction conditions.46 Additionally, this catalytic system required harsher reaction conditionsin the reduction of ketones to the corresponding alcohols. Therefore,the hydrogenation of substrate 1 under 20 bar of H2 pressure afforded the total hydrogenation of the double bondas well as partial reduction of the carbonyl group , Pd NPs/N00Cl-100 provided its semihydrogenationwith total conversion and selectivity toward (Z)-5-decene. However, at longer reaction times, full hydrogenation wasobserved (7) and 1-phenylpropyne (8), semihydrogenationtoward 7H and 8H was not fully accomplishedusing this catalyst (9) allowedcompletion of the conversion of the alkyne to an 87:13 9H/9S mixture .In addition, the complete hydrogenationof the differently activatedalkynes Pd NPs/N00Cl-100 wasalso studied toward hydrodehalogenation reactions of haloarenes (see Scheme S1 in the SI), exhibiting moderate tocomplete conversions, thus broadening the efficiency of this NP systemin hydrogen-mediated catalytic processes.Finally, the catalytic system ChCl and N00Cl), have been effectively used for the straightforwardsynthesis, immobilization, and stabilization of palladium nanoparticles(Pd NPs). In the case of the DES, no additional stabilizers nor reductiveagents are necessary for the reduction of the palladium precursor,Pd(OAc)2, thus improving the sustainability of Pd NPs preparation.The strong coordination of DES to Pd(0) has demonstrated the NP stabilizationrole of the solvent. The morphology of Pd NPs strongly depended onthe physicochemical properties of the used solvent, thus allowingthe tuning of size, distribution, and homogeneity of the NPs.Inthis work, 21 green solvents such as glycerol and glyceryl monoethers,diethers, and triethers, as well as the eutectic solvents resultingfrom the mixture with two bioammonium salts (N00Cl-R00 solvents, N00Cl-100 being the best media. Finally,the physicochemical properties of glycerol-derived solvents as liquidsupports for nanocatalysis play a key role in the activity outcomesand the recyclability of the catalytic systems.Palladium nanoparticles immobilized in glycerol-based solventshave shown high catalytic activity in the hydrogenation of conjugatedand nonconjugated alkenes, alkynes, and carbonyl compounds, as wellas relevant selectivity in the semihydrogenation of alkynes to alkenes.Best results are obtained in pure glyceryl ethers and in


Schwannomas are neoplasms that typically arise from the myelin sheath of peripheral nerves and rarely originate within the brain parenchyma. Some case reports present schwannomas arising from the brainstem, but regrowth of the tumor and the efficacy of postoperative irradiation have not been examined. In addition, the genetic background of schwannomas arising from the brainstem has not been investigated.NF2, SMARCB1, and LZTR1, and microsatellite analysis for loss of 22q did not show any somatic variants or 22q loss.A 21-year-old male presented with diplopia, dysphagia, and left-sided hemiparesis, dysesthesia, and ataxia. Intracranial imaging showed a heterogeneous mass with a cystic lesion in the pontomedullary junction. Since the tumor caused obstructive hydrocephalus, the patient underwent subtotal tumor resection. A histopathologic evaluation aided a diagnosis of brainstem intraparenchymal schwannoma. Gradual postoperative mass regrowth was recognized. Three-dimensional conformal radiotherapy was performed on the residual mass and surgical cavity. No tumor regrowth was observed 4 years after surgery. To investigate the genetic background of the tumor, target sequences for 36 genes, including We suggest that brainstem schwannomas might differ from conventional schwannomas in their genetic background. Schwannomas arise from the myelin sheath of peripheral nerves and account for 8–10% of all intracranial tumors –4. In ad18F-fluorodeoxyglucose positron emission tomography, uptake in the lesion was non-significant, indicating minimal or no malignancy. A thorough survey of malignancy in the trunk did not reveal any lesions. Preoperative diagnostic angiography did not show tumor staining, which suggested a differential diagnosis of pilocytic astrocytoma. The fourth ventricle partially collapsed due to the mass effect, and signs of intracranial hypertension caused by obstructive hydrocephalus gradually emerged; thus, urgent tumor resection was planned.A 21-year-old male without any notable past medical history presented to our hospital with numbness in the left upper extremity. Symptoms emerged a few months before the patient presented at our hospital and had been gradually deteriorating. Upon admission, a neurological examination revealed bilateral abducent palsy with partial impairment of adduction in the right eye when the patient looked to the left , dysphagia, and left-sided hemiparesis, dysesthesia, and ataxia. His family history, including schwannomas and meningiomas, was unremarkable. Brain magnetic resonance imaging (MRI) showed a 4-cm mass in the dorsal pontomedullary area adjacent to the fourth ventricle  10. The ventrocaudal residual mass was followed up with gadolinium-enhanced MRI  as the manufacturer's protocols. Primary antibodies used were as follows: anti-S100 , anti-epithelial membrane antigen (EMA) , anti-glial fibrillary protein (GFAP) , Ki-67 . S100 was evaluated in nuclei and cytoplasm. EMA was evaluated at the cell membrane. GFAP was evaluated in cytoplasm. Ki-67 labeling index was calculated as percent positive tumor nuclei.NF2, SMARCB1, and LZTR1, using HaloPlexHS targeted capture . Targeted genes were NF2, SMARCB1, LZTR1, ALPK2, APC, ARID1A, ARID1B, BAP1, BRAF, BRCA2, CABIN1, CAST, CDKN2A, CDKN2B, COQ6, DDR1, EPB41L3, GLI1, MEN1, MUTYH, PIK3CA, PTCH1, PTEN, SDHD, SHH, SMARCE1, SMO, SUFU, TAB3, TERT, TP53, TRAF7, TSC1, TSC2, and WRN. These genes are reported as pertinent in schwannomas and other related tumors [To reveal the genetic background of the tumor, target sequences were performed on all exons in the 36 selected genes from the tumor and germline DNA, including Sequencing data was generated using the Illumina HiSeq 2000 sequencer and the 100-base pair paired-end sequencing protocol across rapid flow cell lanes. An analysis of the number of reads of each amplicon in the HaloplexHS data was performed with a mean coverage of 119 × with barcodes for the tumor sample and 278 × with barcodes for the blood sample. Somatic variants were initially called following GATK best practice using Mutect2, and false positives were filtered out by GATK FilterMutectCalls. On the other hand, germline variants were initially called following GATK best practice using Haplotypecaller, and potential false positives were filtered out by FilterVariantTranches after CNNScoreVariants with default parameters. Funcotator including ClinVar database was applied to annotate the remained variants. Sequencing reads were aligned to the human genome (hg19). Detailed conditions will be provided upon a reasonable request.NF2 were selected from the Genome Database and included D22S268, D22S1163, D22S929, D22S280, and D22S282. Detailed conditions of this analysis will be provided upon a reasonable request.Microsatellite analysis was also performed to detect 22q loss. Five microsatellite polymorphic markers flanking The final pathology report suggested brainstem intraparenchymal schwannoma. The specimen had typical characteristics of a schwannoma, such as Antoni A and B regions Fig. a, b. TheNF2, SMARCB1, LZTR1, or the other targeted genes. The 22q loss was also not detected . To clarify the genetic background of brainstem schwannomas, genome-wide comprehensive genetic analysis is essential. In addition, epigenetic analysis could also be important. Multiple studies on vestibular schwannomas showed that hypermethylation of the enomenon , 33. Howocations . EpigeneNF2 and 22q loss. This case study suggests that brainstem schwannomas may differ from conventional schwannomas in their genetic background.We experienced the first case of brainstem schwannoma recurrence after resection, which was successfully controlled by irradiation. To the best of our knowledge, this study is the first to report the results of a genetic examination of brainstem intraparenchymal schwannoma. Our genetic analysis revealed that the tumor did not have any somatic variants pertinent to schwannoma, such as in


The proteome revealed that post-transcriptional modification may be widely involved in lignification development. Overall, our results revealed that xylem development is a rapid thickening process in P. tomentosa, and expression patterns varied temporally from cell division to cell death.Xylem development plays an important role in the wood formation of plants. In this study, we found that xylem development was a rapid thickening process characterized by initially rapid increases in the number of tracheary elements and fiber cells and the thickness of the secondary walls that later plateaued. Transcriptome analysis showed that the xylan and lignin biosynthetic pathways, which are involved in the early rapid thickening of the xylem, were mainly upregulated in the second month. The expression of a total of 124 transcription factors (TFs), including 28 NAC TFs and 31 MYB TFs, peaked in 2- and 3-month-old plants compared with 1-month-old plants. Based on previous studies and the key  Wood in perennial angiosperms is formed by the differentiation of angiogenic layers into xylem cells. In angiosperms, it is mainly composed of axially elongated vascular elements and fibers and radially elongated ray cells . The forArabidopsis is mediated by a three-layer transcriptional regulation model  in Populus trichocarpa. Previous research has shown that PtrWND2B and PtrWND6B, which are functional orthologs of Arabidopsis SND1 and VND7, function as master switches activating the secondary wall biosynthetic program in fibers and vessels  and v-myb avian myeloblastosis viral oncogene homolog (MYB) transcription factors (TFs) forming a hierarchical network that gradually regulates the synthesis of cellulose, hemicellulose, and lignin during secondary wall formation . Previouon model . Recent on model , 2018. C vessels , 2011. I PtrWNDs . In addiickening . These sPopulus have shown that gene expression is tightly controlled  and transmission electron microscopy (TEM) analysis were used to study the secondary wall thickening of fiber cells over the 3 months . We calcTo investigate xylem cell PCD during xylem development, we examined cellular degradation using TEM. No degradation of cellular components, including mitochondria, plastids, vesicles, and membrane-like vesicles, could be observed in mature fiber cells, and few cellular components were observed in the TE cells of WT-1M. Only a few cellular components, such as mitochondria, plastids, and non-degraded nuclei were observed in the cytoplasm of fiber cells in WT-2M plants. The cellular components had completely disappeared in the fiber cells of WT-3M plants. In TE cells of WT-2M or WT-3M plants, the cellular contents had completely disappeared. These results showed that WT-2M plants were in a period of rapid xylem growth.P-value < 0.01) in at least one of the three time points. Compared with WT-1M plants, 2,494 and 4,844 genes were upregulated and downregulated in the WT-2M plants, respectively  was performed, and DEGs were divided into 12 clusters based on their expression patterns. We divided the 12 clusters into four groups with similar expression patterns . In grouTo identify DEGs involved in xylem development, all DEGs in the 12 clusters were subjected to GO enrichment analysis. In cluster 3, the DEGs were significantly upregulated in WT-2M plants compared with WT-1M plants, but their expression was relatively stable in WT-3M plants . GO enriCluster 8 was mainly enriched in ribosome-related genes, cluster 5 was mainly enriched in vacuolar-related genes, and cluster 9 was mainly enriched in nucleus-related genes . These rCYCD3, NIT4, and E2F2, were expressed most strongly in WT-1M compared with WT-2M or WT-3M (PtoXTH8.1/PtoXTH8.2 (XTH8), PtoEXPA8/PtoEXPA12 (EXPA8), were most strongly expressed in WT-1M plants , PtoRNS3 (RNS3), PtoSAG102 (SAG102), and PtoSCPL45.1/PtoSCPL45.2 (SCPL45), were highly expressed in WT-2M plants compared with WT-1M and WT-3M plants. This result suggests that the second month may be the period during which vacuole rupture and nuclear disintegration and these genes may be involved in these processes during the early stages of PCD. A total of 24 DEGs, such as PtoRD21.1 (RD21B), PtoβVPE2/PtoβVPE1 (βVPE), and PtoXSP1.2 (XSP1), were highly expressed in WT-3M plants compared with WT-1M and WT-2M plants and may be involved in protein degradation in the later stages of PCD.We also identified 50 protease genes that may be involved in the regulation of the PCD process . A totalPopulus trichocarpa  wood-associated NAC domain TFs (PtoWNDs) or MYB TFs during xylem development. Our results showed that 98 cell wall biosynthetic genes  and 47 PCD-related genes, have SNBEs or SMREs (cis-regulatory element TGTG[T/G/C] bound to GmNAC30 (ortholog of ATAF1) and GmNAC81 (ortholog of AtNAC036) was present in the promoters of PCD-related genes (PtoNAC072.1/PtoNAC072.2 (ortholog of GmNAC81), or PtoNAC045 (ortholog of GmNAC30).Two regulatory elements, namely secondary wall NAC-binding elements (SNBE) and secondary wall MYB-responsive elements (SMREs), which are considered to be key cis-acting elements, exist independently or cooperatively in the promoters of genes related to secondary wall formation , 2013. Tand SMRE , 4. Thisor SMREs . A conseed genes . Our resed genes , suggestPtrWNDs directly binds to the SNBE sites present in the promoters of several of the WND-regulated TFs involved in secondary wall biosynthesis, cell wall modification, and PCD  of this TRN. Based on the key cis-acting elements (SNBEs), 13 TFs including 4 MYB TFs, which may act as the second master switches involved in xylem development, were in the second layer (PtoNAC072.1/PtoNAC072.2 (ortholog of GmNAC81), and PtoNAC045 (ortholog of GmNAC30) (PtoWNDs and MYBs and were in the fourth layer (Previous studies show that  and PCD . MYB46 aCD genes  were usend layer . Based ond layer . In addind layer . Based oGmNAC30) . PCD-relth layer .PtoWND1A, PtoWND1B, PtoNAM3, and PtoNAC3.2) and four MYB TFs , were measured in WT-1M, WT-2M, and WT-3M plants using qRT-PCR. Expression patterns were similarly based on RNA-seq and qRT-PCR, thus validating the transcriptome data  in at least one of the three time points  and nuclease RNS3 (PtoRNS3) were highly expressed in WT-2M compared with WT-1M and WT-3M and might be involved in DNA degradation and vacuole rupture  have been shown to activate various downstream TFs, including SND2, SND3, MYB20, MYB42, MYB43, MYB46, MYB52, MYB54, MYB58, MYB63, MYB69, MYB83, MYB85, MYB103, and KNAT7 , involveM plants . This sucis-acting elements SNBEs, SMREs, W-box, and TGTG[T/G/C]. We also elucidated the regulatory network of TFs with known functions in this TRN. Arabidopsis AtMYB61, Poplar MYB61 (POPTR_0015s09430) positively regulated the lignin and cellulose biosynthesis (PtoMYB61.2 (POPTR_0005s00340) have SNBEs and SMREs, PtoMYB61.3 (POPTR_0002s18700) only has SNBE elements and is an ortholog of AtMYB61, suggesting that PtoMYB61.3 may be targeted by PtoWNDs and act as the second master switches involved in the lignin and cellulose biosynthesis  medium  in a growth chamber with a 14 h light/10 h dark photoperiod at 25°C. We selected the stems from 1, 2, and 3-month-old poplar plants showing good growth for analyses.The poplars used in the study were P. tomentosa at three different ages . The specific experimental methods followed those of a previous study . The sample preparation method of TEM follows that of 1. Statistical differences were determined using Student’s t-test.Roots up to the fourth stem node of P. tomentosa tissue-cultured seedlings in different months up to the fourth stem node 1∼2 mm thick. The sample preparation method of SEM followed that of a previous study  were validated, and the primers are shown in P. tomentosa actin gene was used as the internal control. All reactions were run in triplicate for each sample. Relative expression levels of candidate genes were calculated by the 2–ΔΔCt method.Total RNA was extracted from the xylem of poplar stems in different months using the RNAprep Pure Plant Kit  following the instructions of the manufacturer. Quantitative real-time PCR (qRT–PCR) was performed using gene-specific primers and FastFire qPCR PreMix (Probe)  on a CFX Maestro software Real-Time System . Eight genes including NAC and MYB TFs  was determined using Fisher’s exact test (P ≤ 0.05). There were three biological replicates for each sample. The PTM site of proteins was predicted using Plant PTM Viewer 2.The cortex and phloem were peeled off the stems of 1, 2, and 3-month-old us study . We depoThe datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/CZ and HLu designed the experiments, analyzed the data, and wrote the manuscript. CZ, JZ, YL, XL, XG, HLi, DL, and HLu performed the experiments. CZ performed the bioinformatics analysis. All authors read and approved the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.