Systemic lupus erythematosus (SLE) is an autoimmune disease that involves multiple organ systems. The pathogenic mechanisms that cause SLE remain unclear; however, it is well recognized that the immune balance is disturbed and that this imbalance contributes to the autoimmune symptoms of SLE. Oxidative stress represents an imbalance between the production and manifestation of reactive oxygen species and the ability of the biological system to readily detoxify the reactive intermediates or to repair the resulting damage. In humans, oxidative stress is involved in many diseases, including atherosclerosis, myocardial infarction, and autoimmune diseases. Numerous studies have confirmed that oxidative stress plays an important role in the pathogenesis of SLE. This review mainly focuses on the recent research advances with respect to oxidative stress and regulatory T (Treg)/helper T 17 (Th17) cell dysfunction in the pathogenesis of SLE. Systemic lupus erythematosus (SLE) develops following an encounter by genetically predisposed people to environmental agents, such as ultraviolet light (UV), infections, and cigarette smoke, that cause oxidative stress, but how this oxidative damage modifies the immune system to induce lupus flares remains unknown –3. Activ2 consumption in the peripheral blood lymphocytes (PBL) of SLE patients was increased. Furthermore, SLE PBL consumed more O2 upon in-chamber T cell activation, and N-acetylcysteine diminished O2 consumption [2 consumption that is inhibited by N-acetylcysteine, which may have therapeutic efficacy through reducing oxidative stress in SLE [In T cells from SLE patients and animal models of the disease, the main intracellular antioxidant glutathione is consumed and oxidized , 12. Revsumption . These ds in SLE . A clinis in SLE . Togethe+CD25+ regulatory T cells [+CD25− naïve T cells into CD4+CD25+ regulatory T cells [+Foxp3− T cells into Foxp3+ T cells via transforming growth factor-beta (TGF-β) [Regulatory T (Treg) cells are capable of modulating the function of effector T cells, maintaining immunologic homeostasis, and preventing autoimmunity . Circula T cells , and ind T cells . In addi (TGF-β) , 24. In 2+ flux in lupus T cells [2+ fluxing in T cells contribute to injury in lupus [2+ flux, skews the expression of tyrosine kinases in T cells, and blocks the expression of Foxp3 and the expansion of regulatory cells [Activation of the mammalian target of rapamycin (mTOR) has recently emerged as a key sensor of MHP and mediator of enhanced Ca T cells , 26. mTO T cells , 28. The T cells . Recent in lupus , 31. In in lupus . Additiory cells .+ T cells, whereas, in Treg cell differentiation, the Akt-mTOR axis is a negative regulator of Treg cell [+CD25+Foxp3+ Treg cells [Recently, additional work further demonstrated that mTOR controls the differentiation and functions of Treg cells, suggesting that its activity could be targeted to modulate Treg responses , 29. Spereg cell , 34. Spereg cell , which p2+ flux, has been used safely and effectively to treat lupus [2+ flux is fundamental to understanding the pathogenesis of lupus [Furthermore, mTORC1 activity is high in human and murine Tregs , 33, 36,at lupus , 32, 37.at lupus . The effof lupus , 29.β to augment Foxp3 expression in vitro [+CD25+Foxp3+ Treg cell depletion in APS patients [Another T cell biology regulator is leptin, which is a hormone derived from adipocytes. Leptin is closely related to adipokines, which are associated with oxidative stress and are reported to be overproduced in SLE patients , 40. Prein vitro , 47, 48.in vitro , 49, 50.patients .These data indicate that oxidative stress may contribute to the depletion of Treg cells in SLE patients, suggesting that antioxidation treatment may be an effective method to relieve lupus symptoms via improving Treg cell differentiation. Thus, strategies to enhance the number and function of Treg cells may benefit patients with SLE. Leptin and mTOR may also serve as effective intervention targets to relieve oxidative stress and improve the differentiation of Treg cells in SLE . In addi+ T cells and is characterized by the production of IL-17 [The Th17 lineage is a lineage of effector CD4of IL-17 , 53. Expof IL-17 . Our stuof IL-17 , 54. Envof IL-17 . UV radiof IL-17 , 56. Whiof IL-17 –59. To dof IL-17 . In the of IL-17 . In the of IL-17 . UV radiof IL-17 .β and IL-6 and promotes IL-22 production and enhances Th17 development [The aryl hydrocarbon receptor (AHR) is a ligand-dependent transcription factor best known for mediating the toxicity of dioxin . AHR waselopment . The ligelopment , 65. Intelopment . AHR actelopment , 64. Conelopment . This coelopment . These d2O2), and 3′-methoxy-4′-nitroflavone (MNF) promote the generation of FICZ and thereby give rise to prolonged AHR signaling [ CYP1A1 gene expression and elicits inflammation enlargement [AHR typically resides in the cytoplasm in complex with Hsp90 until the binding of its ligand triggers conformational changes, resulting in an exchange of Hsp90 for the nuclear translocation component ARNT , 67. Theignaling , 71. Intargement . UVB radargement , and UVBargement , 69. ThuBased on the above discussion, we speculate that, in SLE patients, UV exposure can directly induce DNA damage, generation of DNA photoproducts, and apoptotic material, which consequently induces B cell activation and autoantibody production. Furthermore, UV exposure elicits AHR activation and the generation of FICZ, which is a high-affinity ligand for AHR activation and can further promote the differentiation of the Th17 cell population, thereby worsening the severity of lupus . Further+ flux has been a hallmark characteristic in SLE patients and recent studies have demonstrated the involvement of calcium/calmodulin-dependent protein kinase IV (CaMK4) in the pathogenesis of SLE [γt-) mediated Th17 cell differentiation. Importantly, silencing CaMK4 in T cells from patients with SLE inhibited Th17 differentiation. Collectively, these data suggest that CaMK4 inhibition has potential as a therapeutic strategy for Th17-related SLE patients [Enhanced Ca2s of SLE , 73. CaMs of SLE . In addipatients .γt [mTOR is identified as a component of two interacting complexes, mTORC1 and mTORC2, that regulate T cell lineage specification differentiation . Oxidatiγt , 76. Botγt , 35. In +CD25+Foxp3+ Treg cell differentiation in vitro and that baicalin treatment protected lupus-prone MRL/lpr mice against nephritis and improved the Treg cells in vivo [ Antrodia camphorata with antioxidant activities, has been proved to prevent the transformation of mild lupus nephritis into higher-grade nephritis in a murine lupus model [−CD8− T cells, stimulate Foxp3 expression in CD4+CD25+ T cells, and reduce anti-DNA production [Our previous findings demonstrated that baicalin, a compound isolated from a Chinese herb, increased CD4 in vivo , 78. Cur in vivo , 80. The in vivo . Our res in vivo , 83, furus model . Antroquus model . Epigallus model . Epigallus model . A randooduction . AltogetTaken together, these data show that oxidative stress plays a key role in the pathogenesis of SLE. Oxidative stress can induce and aggravate SLE by linking environmental stimulation with immune imbalance. Oxidative stress contributed the Th17/Treg imbalance in SLE patients. Under the normal stage, Th17 cells and Treg cells stay in a dynamic immune balance, while oxidative stress was induced in SLE patients, which can further induce and expand the proinflammatory Th17 cells expansion and inhibit the anti-inflammatory Treg cell differentiation and aggravate autoimmune injuries . Above,


In traditional Chinese medicine (TCM), a combination of kudzu and Chen-Pi is frequently prescribed for relieving colds, fever, bronchitis, and cough. It contains daidzein and hesperetin, selective inhibitors of family 3 (PDE3), and 4 (PDE4) of phosphodiesterases (PDEs), respectively. In passively sensitized human airways, allergen-induced contraction was reported to be inhibited only by the simultaneous inhibition of PDE3 and PDE4, but not by single inhibition of either isozyme. Therefore, we are interested in investigating the interaction between daidzein and hesperetin on their antispasmodic effects in the isolated sensitized and non-sensitized guinea-pig tracheas, to clarify the difference between these two tissues, because effects of TCM prescription on patients with or without allergic asthma are often different. Guinea-pigs were sensitized by subcutaneous injection of ovalbumin (OVA) into legs. After sensitization, the baseline and cumulative OVA-induced contractions of the sensitized trachea were isometrically recorded on a polygraph. In the same way, the histamine (30 μM)-induced tonic contraction of non-sensitized guinea-pig trachea was recorded. The isobole method was used to analyze the antagonism and synergism between daidzein and hesperetin. The isoboles showed antagonism between daidzein and hesperetin on baseline relaxant effect and OVA (100 μg/ml)-induced contraction in the sensitized guinea-pig trachea. In contrast, the isobole showed synergism between daidzein and hesperetin on the relaxant effect of histamine-induced tonic contraction in non-sensitized guinea-pig trachea. These results suggest that the combination of kudzu and Chen-Pi for relieving colds, fever, bronchitis and cough is effective in patients without, but might show little effect in patients with allergic asthma. Pueraria pseudo-hirsuta Tang et Wang, Leguminosae), known as Radix Puerariae. In TCM, kudzu is used against colds, fever, influenza, diarrhea, urticaria, tonsillitis, headache, and spasm (Citrus aurantium L. (Rutaceae), well-known as “Chen-Pi” in TCM, which is used as an expectorant  is about 1% present in the tuber of kudzu  was purchased from Pierce Biotechnology.Daidzein , hesperetin , aminophylline, histamine, indomethacin, nifedipine (purity > 98%) and OVA were purchased from Sigma–Aldrich. Freund’s complete adjuvant (ad libitum. Under a protocol (LAC-94-0091) approved by the Animal Care and Use Committee of Taipei Medical University on December 22, 2005, the guinea-pig tracheas were dissected under anesthesia .Male Hartley guinea pigs (500∼600 g) were purchased from the Animal Center of the Ministry of Science and Technology , and housed in ordinary cages at 22 ± 1°C with a humidity of 50%∼60% under a constant 12/12-h light/dark cycle and provided with food and water 4 1.2, KH2PO4 1.2, CaCl2 2.5, NaHCO3 25, and dextrose 10.1. Indomethacin (3 μM) was added to the Krebs solution to prevent the synthesis of prostaglandins throughout the experiment. The sensitized tissues were incubated with hesperetin, daidzein, combination  or its vehicle (control) for 15 min, and then cumulatively added OVA . The effects of hesperetin, daidzein, combination, or its vehicle on the baseline tension and OVA-induced contractions were expressed as a percentage of 60 mM KCl-induced contraction (100%). In the dark, nifedipine (1 μM) was used as a reference drug. After the non-sensitized tissues were precontracted with histamine (30 μM), daidzein, hesperetin, combination  or their vehicle was cumulatively added to the organ bath. At the end of the experiment without washout, 1 mM of aminophylline was added to standardize the tissues relaxing maximally (100%).Guinea-pigs were subcutaneously injected a mixture of 10% OVA (w/v) and Freund’s complete adjuvant (1:1) into both sides of legs (0.7 ml for each side) on day 1 and 4. On day 25, these sensitized guinea-pigs were sacrificed to study their tracheal relaxations . In the I) < 1, = 1 or >1 represents synergism, zero-interaction or antagonism, respectively , n is the number of experiment. The difference between two values was determined by Student’s t-test. Differences with P < 0.05 were considered statistically significant.All values including drug combination in isobole are given as the mean ± SEM (n = 11). Daidzein , hesperetin (100 μM), or combination  significantly relaxed the baseline when compared to the control. Nifedipine (1 μM), a blocker of calcium channels, significantly relaxed the baseline about 40% of the KCl (60 mM)-induced contraction . Isobole showed antagonism between daidzein and hesperetin on the baseline relaxant effect in the sensitized guinea-pig trachea . Cumulative OVA  concentration-dependently evoked contractions to the maximum of 589 ± 105 mg (n = 11), which was 121.5 ± 6.7% of the KCl (60 mM)-induced contraction. Daidzein (30 and 100 μM), hesperetin (100 μM), or combination  significantly inhibited the contraction when compared to the control . Nifedipine (1 μM) did not affect these contractions (data not shown). Isobole showed antagonism between daidzein and hesperetin on the OVA (100 μg/ml)-induced contraction in the sensitized guinea-pig trachea .The KCl (60 mM)-induced contraction was 715 ± 189 mg (n = 16) in isolated non-sensitized guinea-pig trachea. Daidzein , hesperetin  or combination  significantly and concentration-dependently relaxed the histamine-evoked contractions . Isobole showed synergism between daidzein and hesperetin in the tracheal relaxation .Histamine (30 μM) evoked a tension of 920 ± 50 mg  present throughout the experiment. However, daidzein , hesperetin (100 μM), and combination  significantly relaxed the baseline, suggesting that the nifedipine-insensitive baseline tension is related to PDE3 and PDE4 activities, but other isozymes may be involved  on partially purified PDE3 and PDE4 are similar  significantly relaxed the baseline about 40%, but did not affect cumulative OVA (0.01∼100 μg/ml)-induced contractions, which may involve the release of tachykinins from sensory nerve ending , suggestinvolved . The inh similar . These r similar .Panax ginseng, Lithospermi radix, Glycyrrhizae radix, kudzu, Chen-Pi, etc., is mainly used for relieving infection of upper respiratory tract, bronchitis, pneumonia, and emphysema in children or elderly patients without asthmatic history (In non-sensitized guinea-pig trachea, the isobole showed relaxing synergism between daidzein and hesperetin on histamine-induced tonic contraction. Consistently, the bronchodilatory and anti-inflammatory effects of dual PDE3/4 inhibitors are more obvious when compared with those of either isozyme inhibition . These r history .These findings in sensitized and non-sensitized guinea-pig tracheas suggest that the combination of kudzu and Chen-Pi for relieving colds, fever, bronchitis and cough is effective in patients without, but might show little effect in patients with allergic asthma.C-HS and W-CK conceived and designed the study. T-YC performed the experiments and analyzed the data. C-HS and W-CK wrote the manuscript. All the authors read and aproved the final 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.


Cullin-RING ligases (CRLs), the largest E3 ubiquitin ligase family, promote ubiquitination and degradation of various cellular key regulators involved in a broad array of physiological and pathological processes, including cell cycle progression, signal transduction, transcription, cardiomyopathy, and tumorigenesis. Autophagy, an intracellular catabolic reaction that delivers cytoplasmic components to lysosomes for degradation, is crucial for cellular metabolism and homeostasis. The dysfunction of autophagy has been proved to associate with a variety of human diseases. Recent evidences revealed the emerging roles of CRLs in the regulation of autophagy. In this review, we will focus mainly on recent advances in our understandings of the regulation of autophagy by CRLs and the cross-talk between CRLs and autophagy, two degradation systems. We will also discuss the pathogenesis of human diseases associated with the dysregulation of CRLs and autophagy. Finally, we will discuss current efforts and future perspectives on basic and translational research on CRLs and autophagy. The long-term health of a cell is closely associated with protein quality control which requires a well-regulated balance between protein synthesis and degradation . It is cThe UPS, a clearance system, directs target proteins with their lysine residues and the N-terminal methionine residue covalently attached by ubiquitin molecules, to the 26S proteasome for degradation, leading to the elimination of short-lived, misfolded, and damaged proteins –8. ProteAutophagy is a highly conserved eukaryotic intracellular catabolic degradation process in which cytoplasmic contents, like misfolded proteins and damaged organelles, are engulfed by double-membrane autophagosomes and degraded in lysosomes fused with autophagosomes . In geneAutophagy, or “self-eating”, plays a vital role in the maintenance of cellular homeostasis. The self-digestion maintains critical physiological functions by providing nutrients during fasting and by eliminating the “garbage” in cells, such as aggregated proteins, damaged organelles, and invading pathogens . Such fuGiven the primary role of autophagy in cellular homeostasis, it is not surprising that the whole process is tightly controlled. Including phosphorylation, ubiquitination, and acetylation, multiple types of post-translational modifications have been found in the regulation of autophagy . This reCullin-RING ligases (CRLs), the largest family of E3 ubiquitin ligases, account for ubiquitination of approximately 20 % cellular proteins degraded by UPS . The folGenerally, CRLs consist of four elements: cullins, RING-finger proteins, adaptor proteins, and substrate recognition receptors Fig. . The humAll CRLs share the similar core architecture with a curved cullin protein acting as a molecular scaffold , 29. AmoNeddylation, like ubiquitination, is a process of the attachment of ubiquitin-like molecule NEDD8 to target proteins, involving the successive actions of E1 NEDD8-activating enzyme , which activates NEDD8; E2 NEDD8-conjugating enzyme , which carries the activated NEDD8; and E3 NEDD8 ligase, which recognizes the substrate and catalyzes the transfer of NEDD8 from E2 to the substrate   blocks the interaction of the substrate receptor-adaptor complex with the N-terminus of cullins. However, covalent conjugation of cullin with one NEDD8 molecule removes the inhibitory binding to CAND1 and restores the CRLs in an active conformation –36. In aIn mammalian cells, autophagy consists of several sequential steps: initiation, autophagosome formation, cargo recognition and delivery, autophagosome–lysosome fusion, and cargo degradation followed by recycling of the resulting macromolecules via permeases, all of which are coordinated by different sets of ATGs Fig. . Among tThe activity of the ULK1 complex  is required for the autophagy induction. Under physiological conditions, the activated mTORC1 directly binds to the ULK1 complex and inhibits autophagy by phosphorylating ULK1 on Ser757 and ATG13 , 48. UndThe Beclin-1/Class III PI3K complex, consisting of Beclin-1, Vps34, and Vps15, is essential for the nucleation of phagophore . It cataATG9, a multipass transmembrane protein, is essential for autophagosome formation. ATG9 is localized to the trans-Golgi network (TGN) and late endosomes. Following the induction of autophagy, ATG9 is rearranged from juxtanuclear to peripheral structures, and is then retrieved from the completed autophagosome . Thus, AThe elongation and expansion of the phagophore membrane is controlled by two ubiquitin-like conjugation systems. First, the E1-like enzyme ATG7 and the E2-like enzyme ATG10 mediate the covalent conjugation of ATG12 to ATG5 . The resAutophagy is induced by a range of cellular stresses, including nutrient and energy depletion, ER stress, hypoxia, redox stress, and oncogenic activation . It is rATGs via AKT-FoxO3 signaling [mTOR is the well-established negative regulator of autophagy. It plays a key role in the coordination of cell growth with autophagy in response to physiological and environmental conditions . mTOR, aignaling , 68 , a master regulator of energy metabolism, is a vital positive regulator of autophagy. As a serine/threonine kinase, AMPK is activated when the levels of AMP and ADP in the cells rise owing to various physiological stresses . Upon phTaken together, by integrating both intracellular and extracellular signals, mTOR and AMPK function coordinately in the regulation of autophagy Figs. , 4.More recently, the study on the roles of post-translational modifications in regulation of autophagic flux by affecting the activity, recruitment, and turnover of autophagic components has become an attractive area due to the implications of dysregulated autophagy in multiple diseases . UbiquitFBXL20 targets Vps34, the catalytic subunit of the Beclin-1/Class III PI3K complex, for ubiquitination and proteasome degradation, and plays an important role in DNA damage-induced suppression of autophagy [ZBTB16. Thus, CRL3ZBTB16 controls the initiation of autophagy by regulating the degradation of ATG14L [Ambra1 mediates K63-linked polyubiquitination of Beclin-1 and enhances its association with Vps34 to promote the activation of Vps34, which is required for starvation-induced autophagy [KLHL20 regulates IFN-induced autophagic death by the destruction of DAPK, a serine/threonine protein kinase, which is in charge of the dissociation of Beclin-1 from its Bcl-2 inhibitors by phosphorylating Beclin-1 on Thr119 located at a key position within its BH3 domain required for the interaction with Bcl-2 family members [Several components of autophagy machinery are subjected to CRLs-mediated regulation. For instance, SCFutophagy , 76. Meaf ATG14L . In addif ATG14L –80. Specutophagy . Notablyutophagy . Second,utophagy . Third, utophagy . Under uutophagy , 83. Alt members –86  , 110. CMIn addition to regulating autophagy machinery and upstream regulators, CRLs also control autophagy at the transcriptional level through modification of several key transcription factors.VHL, is partially responsible for MLN4924-induced autophagy [BNIP3 and BNIP3L , both of which disrupt the Bcl-2/Beclin-1 complex, leading to the release of Beclin-1 from Bcl-2 and the subsequent induction of autophagy [NIX/BNIP3 via HIF1α.We already discussed that accumulation of HIF1α, as a well-established substrate of CRL2utophagy . In factutophagy , 113. Inutophagy , 115. ThNIX/BNIP3 [ATG genes, including ATG4B, ATG8, ATG12, Vps34, and Beclin-1, during muscle atrophy [SKP2 [β-TrCP in an IKKβ-dependent manner [Meanwhile, the transcription factor FoxO3 regulates autophagy in skeletal muscle by transactivating IX/BNIP3 . In addi atrophy –118. And atrophy . Given thy [SKP2 , 121. Adt manner . Moreovet manner , 123.β-TrCP-mediated polyubiquitination [β-TrCP, inhibits autophagosome formation by suppressing p62  expression via TCF4 [β-TrCP paradoxically regulates autophagy through repressing LC3B or inducing p62, two key proteins in the process of autophagy , up-regulates LC3B by directly binding to its promoter to facilitate autophagy , 125. ATtination . More imtination . β-catenvia TCF4 . Thus, SIn conclusion, all these studies highlight the importance and complexity of CRLs in the regulation of autophagy. Given that 1) these findings are mostly associated with mTOR pathway, the ULK1 complex, and the Beclin-1/Class III PI3K complex; 2) one specific CRL can target various substrates; 3) the specific substrate is subjected to the regulation of multiple CRLs; 4) autophagy can be regulated at the transcriptional, translational, and post-translational levels, further studies should be directed to elucidate the functional network of CRLs in the whole process of autophagy.FBXL14 and subsequently degraded by the proteasome [Keap1, resulting in the hyperactivation of NRF2, which may contribute to hepatoma development [Accumulating evidences indicate that the active cross-talk exists between UPS and autophagy, two major intracellular clearance systems , 129. Inoteasome . Howeveroteasome . p62 abroteasome , 133; , a synapse-localized F-box protein, was proved to regulate neuronal synaptic tuning via the destruction of RIM1, which is required for synaptic vesicle release . Scrappeocytosis . In lighocytosis  and autoocytosis  in regulZBTB16-mediated ATG14L degradation, leading to the expanded polyQ degradation and the preservation of neuronal functions [Although a causal pathogenetic linkage between CRLs and neural disorders and diseases has not been established, the ubiquitin conjugates and/or inclusion bodies associated with ubiquitin have been discovered to be accumulated in a wide array of chronic neurodegenerative diseases . In addiunctions . Meanwhiunctions , can indunctions . Thus, CAtrogin-1. These findings were confirmed in Atrogin-1 transgenic and knock-out mouse models, strongly indicating its crucial role in the inhibition of cardiac hypertrophy [Atrogin-1 knock-out mice, Atrogin-1 depletion causes cardiomyopathy and premature death resulting from impaired autophagy [Atrogin-1 promotes the ubiquitination and degradation of CHMP2B, which is part of an endosomal sorting complex required for autophagy [Csn8 knock-out (CR-Csn8KO) mouse model also proved the important role of Atrogin-1 in cardiomyocyte necrosis and dilated cardiomyopathy via autophagy impairment caused by down-regulation of Rab7, which is indispensable for autophagosome-lysosome fusion [Atrogin-1, also known as FBXO32, is a skeletal and cardiac muscle-specific F-box protein . Atroginertrophy . Moreoveertrophy . All theutophagy . Mechaniutophagy . Atroginutophagy . In addie fusion . The unde fusion , 158;  CRLs are being validated as promising anti-cancer targets; (2) MLN4924, a small molecule indirect inhibitor of CRLs, which is currently in several Phase I clinical trials against a number of human malignancies, induces protective autophagy; (3) the inhibitors of autophagy significantly sensitize cancer cells, particularly resistant cancer cells, to MLN4924 treatment, future studies focused on CRLs and autophagy would eventually benefit human anti-cancer therapy.Although some associations of CRLs and autophagy have been clarified, many fundamental questions still remain to be addressed: (1) what are other components of autophagy machinery and regulators associated with autophagy as the substrates of CRLs? (2) How does autophagy regulate the activity of CRLs and UPS? (3) Are the findings in cell culture settings consistent to those in physiological settings (knock-out/knock-in mouse models and patient samples)? (4) What is the function of deubiquitination in autophagy? Some recent studies have shown that deubiquitinases, enzymes catalyzing a reverse process for protein ubiquitination, also play a regulatory role in autophagy, such as USP36 and A20 , 160. Th


Further studies are needed to explore new therapeutic strategies aimed at restoring CRBP-1-mediated intracellular retinol trafficking and the meaning of CRBP-1 expression in cancer patients' screening for a more personalized and efficacy retinoid therapy.Retinol and vitamin A derivatives influence cell differentiation, proliferation, and apoptosis and play an important physiologic role in a wide range of biological processes. Retinol is obtained from foods of animal origin. Retinol derivatives are fundamental for vision, while retinoic acid is essential for skin and bone growth. Intracellular retinoid bioavailability is regulated by the presence of specific cytoplasmic retinol and retinoic acid binding proteins (CRBPs and CRABPs). CRBP-1, the most diffuse CRBP isoform, is a small 15 KDa cytosolic protein widely expressed and evolutionarily conserved in many tissues. CRBP-1 acts as chaperone and regulates the uptake, subsequent esterification, and bioavailability of retinol. CRBP-1 plays a major role in wound healing and arterial tissue remodelling processes. In the last years, the role of CRBP-1-related retinoid signalling during cancer progression became object of several studies. CRBP-1 downregulation associates with a more malignant phenotype in breast, ovarian, and nasopharyngeal cancers. Reexpression of CRBP-1 increased retinol sensitivity and reduced viability of ovarian cancer cells  Vitamin A can be acquired from the diet either as preformed vitamin A  or provitamin A carotenoids . Dietary stimulated by retinoic acid 6 (STRA6) mediates vitamin A uptake from RBP   –42. This15; 17)] , 44 and PML-RARA , 42, 45.ic cells , 47. ATRic cells , 48. Recic cells , sumoylaic cells , or autoic cells . High coic cells . Effectiic cells –55. Moreic cells . In vitrll cycle , but notll cycle , 58; ER-ed cells  and theycells [β . Preclincells [β  through  protein . Primary protein . It is pmal glia . Moreovemal glia  and vitamal glia . The commal glia . While smal glia  in breasmal glia , 69, promal glia , 70–74, mal glia , head anmal glia , ovarianmal glia , 78, botmal glia –81, neurmal glia –84, and mal glia , 86. 4-Hmal glia , 87, 88,mal glia –91 and nmal glia , 93 and mal glia , 94. TACmal glia , 96. Synmal glia , 98. A pmal glia , 100. In vitro, CRBP-1 also channels either retinol towards microsomal dehydrogenases that catalyze the oxidation of retinol into RAL or its addressing towards cytosolic dehydrogenases for oxidation into RA. Several microsomal enzymes, termed retinol dehydrogenases types I, II, and III, have high specificity for CRBP-1-bound retinol. Retinal dehydrogenases oxidize CRBP-1-bound retinal [CRBP-1 is a small cytosolic protein of 15 KDa. CRBP-1 is widely expressed and evolutionarily conserved in many tissues and acts as chaperone protein that regulates the uptake and subsequent esterification of retinol and its bioavailability. CRBP-1 gene is located on 3q21 chromosome. CRBP-1 is required for the efficient synthesis of ATRA . CRBP-1  retinal . Retinal retinal . in vitro [α-null mutation, while the squamous keratinizing metaplasia observed in CRBP-1-null mice fed with the vitamin A deficient diet is similar to those observed in old adult mice lacking RAR-γ [It has been shown that a lack or an excess of RA during embryonic development results in congenital abnormalities. As a matter of fact, RA regulates genes that specify body axis pattern and may help to program limb formation in the developing embryo. In mature vertebrates, RA maintains epithelial tissues, contributes to bone remodelling and sustains reproductive processes, such as the oestrous cycle, spermatogenesis, and placental growth . As RA sin vitro . It coulin vitro . As a main vitro . These lin vitro  or to anin vitro , and, nain vitro . The morng RAR-γ . in vivo, but CRBP-1 is downregulated in both the upper and the lower uterine segments during the first and the second trimester of pregnancy [CRBP-1 expression has been studied at histological distinct stages of the rat oestrus cycle. CRBP-1 was detected during the early proliferative phase and after the formation of a mucinous cell layer during proestrus . CRBP-1 regnancy . By the regnancy . CRBP-1 regnancy . The levregnancy .α-smooth muscle actin. Theα-smooth muscle actin is mainly controlled by TGF-β1, a fibrogenic mediator involved in both myofibroblast differentiation and extracellular matrix deposition. Surprisingly, TGF-β1 induces CRBP-1 gene and protein expression in primary cultures of HSC and in portal fibroblasts. The myofibroblast differentiation of HSC is associated with a decrease in total hepatic levels of retinyl palmitate [ in vivo during the evolution of the experimental aortic IT produced by endothelial injury. CRBP-1 expression in cultured SMCs has shown to be regulated by ATRA and retinol. Moreover, the presence of CRBP-1 in cultured SMCs appears to be correlated with the expression of cytokeratin 8 [One of the key events in the wound repair process is the infiltration of fibroblasts into the damaged area where they proliferate and differentiate into myofibroblasts expressing features of SMCs. These fibroblasts present in the granulation tissue originate from subcutaneous tissue fibroblasts . CRBP-1 almitate , the prealmitate . So, it almitate . The patalmitate . The expalmitate . As a maeratin 8 . CRBP-1 eratin 8 . It is ceratin 8 .β-catenin accumulation [Because of the complexity of vitamin A metabolism, the precise role of CRBP-1 in retinoid signaling remains controversial, despite numerous studies conducted over the past three decades on its binding properties, three-dimensional structure, tissue localization, regulation of expression, involvement in retinal metabolism, and null mutation . The funmulation , 133. Thmulation . In a camulation . CRBP-1 mulation . Importamulation . The strmulation . The lowmulation .α [ In vitro studies confirmed the downregulation of RAR-α and RAR-γ mRNA levels in CRBP-1-stable-transfected ovarian cancer cells [α upregulation characterizes other malignancies, such as lung cancer and metastatic melanoma [γ upregulation is reported to be prooncogenic and to support the growth/progression of mammary tumours [It has been documented that CRBP-1 upregulation exerts a direct antitranscriptional effect through the binding of CRBP-1 promoter to RAR-α . In vitrer cells . Abnormamelanoma . Similar tumours . So, CRB tumours , 137–139vector) or CRBP-1 (MTSVCRBP-1), has shown to prevent tumor cells from taking up, storing, and using retinol in vivo [ in vivo data suggest that somatic CRBP-1 loss results in a local deficit in vitamin A storage and metabolism, with important consequences for the affected tissues.Furthermore, CRBP-1 induces a reduced transcription level and activity of PI3K/Akt pathway, in particular, Akt1 . Althoug in vivo . These i in vivo and in vitro, inducing downregulation of survival and proliferative gene pathways and increases retinoid-mediated apoptosis [Reintroduction of CRBP-1 signaling reduced tumorigenicity bothpoptosis , 143. Sopoptosis , 137. Inpoptosis  suggestsVitamin A and derivatives comprise a group of natural and synthetic molecules, which regulate a variety of essential biological processes during normal development, maintained tissue homeostasis, and also mediate protection from diseases. Retinoids have many important and diverse functions throughout the body including roles in vision, regulation of cell proliferation and differentiation, growth of bone tissue, immune function, and activation of tumour suppressor genes. Genomic functions of the retinoids are mediated via their nuclear DNA-binding receptors, RARs, and RXRs, which regulate gene transcription through recruitment of corepressors and coactivators. Natural and synthetic retinoids have been used as potential chemotherapeutic or chemopreventive agents because of their differentiation, antiproliferative, proapoptotic, and antioxidant effects. The function of the CRBP-1 gene in controlling the availability of retinol to cells suggests that its product has special relevance to inhibition of early steps in transformation. CRBP-1 downregulation occurs in breast and ovarian tumors and compromises RAR activity, leading to loss of cellular differentiation and tumor progression. Furthermore, the CpG island hypermethylation of CRBP-1 is responsible for its inactivation in some cancer cell lines such as cervical, larynx, nasopharyngeal, and gastrointestinal carcinomas and lymphoma , 128–130


Pathophysiological conditions such as inflammation, ischemia, infection and tissue injury can all evoke pain, and each is accompanied by local acidosis. Acid sensing ion channels (ASICs) are proton-gated cation channels expressed in both central and peripheral nervous systems. Increasing evidence suggests that ASICs represent essential sensors for tissue acidosis-related pain. This review provides an update on the role of ASICs in pain sensation and discusses their therapeutic potential for pain management. Tissue acidosis occurs in various physiological and pathophysiological states, including inflammation, infection, ischemia, cancer, tissue injury and metabolic stress ,2,3,4. I+ channel/degenerin (ENaC/DEG) family of cation channels [ASICs belong to the voltage-insensitive, amiloride-sensitive epithelial Nachannels ,10. Theychannels . Howeverchannels ,12. The ASIC subunits share the same overall structure with other ENaC/DEG family members, which is characterized by two hydrophobic transmembrane domains, a large cysteine-rich extracellular loop and short intracellular N- and C-termini ,13. A re+, but the homomeric ASIC1a channels are also permeable to Ca2+ [0.5) of homomeric ASIC channels differs: 6.2–6.8 for ASIC1a, 5.1–6.2 for ASIC1b, 4.1–5 for ASIC2a, and 6.2–6.7 for ASIC3 [i.e. proton concentration increase), whereas the time constant of inactivation for ASIC3 remains constant despite changes in pH [Functional ASICs can be formed by homomultimers as well as heteromultimers ,17. Diff to Ca2+ ,21,22. F to Ca2+ ,23. In hor ASIC3 ,25,26. Mor ASIC3 ,20. In aes in pH .Although the exact subunit composition (or subtypes) of ASICs in most neurons remains unclear, almost all ASIC subunits are known to be present in primary sensory neurons ,19,27,28Physiological pain is initiated by high-threshold unmyelinated C or myelinated Aδ primary sensory neurons that feed into noceciptive pathways of the central nervous system ,44. The A), a common inhibitor of ASICs, and nonsteroid anti-inflammatory drugs (NSAIDs) such as declofenac and ibuprofen, which selectively inhibit ASIC1a and ASIC3, respectively [in vivo knockdown of ASIC3 with a specific siRNA, whereas PcTx1, a specific blocker of homomeric ASIC1a channels [Figure 1B).Direct perfusion of acidic solutions or iontophoresis of protons into the skin causes pain in humans ,47,48. TFigure 1ectively . Previouectively . A recenectively . The hypchannels , has no Indeed, inflammation is known to induce a marked increase of the ASIC channel expression in primary sensory neurons; for example, the mRNA level of ASIC3 is increased by up to 15 fold in complete Freund’s adjuvant (CFA) -induced inflammation . The incThe involvement of ASICs in muscle and joint pain associated with inflammation has also been studied ,55,56,57etc. [The role of ASICs in primary inflammatory pain has been investigated recently in gene-targeted mice, and the data are not conclusive. In several studies mentioned above ,54,57,59etc. ,23,39,63Pain is the only sensation that can be evoked from most internal organs. Visceral pain can be considered as part of the defense reactions of the body against harmful stimuli, particularly of those that impinge on the mucosal lining of hollow organs . It has -/- cardiac afferents match the properties of ASIC2a channels, and currents from ASIC2-/- cardiac afferents match the properties of ASIC3 channels [Myocardial ischemia can lead to pain or discomfort within the chest, termed angina pectoris . It is gchannels . These rin situ hybridization experiments reveal that ASIC3 is the most abundant ASIC transcript within thoracolumbar DRG, followed by ASIC2 and ASIC1 [−/− mice were reported to have markedly reduced visceral mechanosensitivity when compared to control animals and ASIC1−/− or ASIC2−/− mice [−/− mice but was fully preserved in ASIC2−/− mice [Luminal acidity is a physiological challenge in the foregut, and acidosis may occur throughout the gastrointestinal (GI) tract as a result of inflammation, ischemia, cancer, microbial activity, or over distension of GI wall . Acid isnd ASIC1 . In addind ASIC1 . The imp−/− mice . Another−/− mice . Interes−/− mice . The dif−/− mice .etc.), pain sensation is generally not detected in the lower respiratory tract and lung parenchyma. Endogenous airway acidification has been documented in various airway inflammatory diseases including asthma and chronic obstructive pulmonary disease [etc.) is known to evoke cardiorespiratory symptoms such as cough, bronchospasm and dyspnea that are at least partially mediated through the activation of bronchopulmonary C-fibers and the subsequent reflex responses [Most sensory inputs arising from airways and lung structures are conducted in vagus nerves and their branches, and the majority of vagal bronchopulmonary sensory nerves are nociceptive C-fibers ,78. Acti disease ,81,82. Aesponses ,84. Receesponses . By usinesponses . Patch cesponses ,86. Chronic pain generally falls into two categories: inflammatory and neuropathic pain ,87. The C). In animal models of neuropathic pain, ASIC3 immunoreactivity in rat DRG neurons is elevated following lumbar disc herniation [ASICs play an essential role in pain sensation not only from the standpoint of their peripheral nociceptive function, but also their involvement in the development of central sensitization and pain hypersensitivity . Spinal Figure 1rniation , and intrniation . In addirniation ,98,99.2+ and Ca2+) and protein kinases (such as PKA and PKC) can modulate certain subunit-containing ASIC channels [Although extracellular proton is the only known activator of ASICs, increasing evidence shows that the proton-gating of ASICs can be modulated by a variety of factors. As we described earlier, the expression of ASICs is upregulated following inflammation and tissue injury ,38,49,95channels . Althougchannels ,111,112.channels ,20. The channels . Therefochannels .in vitro and in vivo preparations [Psalmopoeus cambridgei, potently and specifically inhibits ASIC1a homomultimers, while does not affect ASIC1a-containing heteromultimers [Anthopleura elegantissima is another ASIC blocker that selectively and effectively inhibits homo- and hetero-meric ASIC3 channels [ASICs emerged as a potential therapeutic target for pain treatment when amiloride, a non-selective ASIC blocker, was shown to attenuate the intradermal acid infusion-evoked pain in humans ,48. Locaarations . To dateultimers ,116. APEchannels . Given tchannels .Tissue acidosis occurs in various physiological and pathophysiological states such as inflammation, ischemia, or tissue injury. The local drop in pH is detected by primary sensory neurons, the signals of which feed into nociceptive pathways of central nervous system and produces pain sensation. Studies using ASIC-gene-targeted approaches and ASICs inhibitors in various pain models indicate that ASICs function as the essential sensors of acid-evoked pain and appear as a potential therapeutic target for pain therapy. However, a better understanding of the structure, activation and modulation mechanisms, and development of additional potent and specific ASIC inhibitors suitable for clinical use will be needed for alleviating acid-related pain syndromes.


Flavihumibacter sp. CACIAM 22H1, isolated from Bolonha Lake, Brazil, which will provide further insight into the production of substances of biotechnological interest.Here, we present a draft genome and annotation of  Flavihumibacter, as previously described , Belém, Pará, Brazil.The draft genome sequence obtained for the heterotrophic bacterium After DNA extraction of the cyanobacterial culture, two sequencing runs were performed by GS FLX 454 sequencer (Roche Life Sciences) using a nonpaired library and one sequencing run was carried out on the Illumina MiSeq platform with a paired-end library with 150-bp read length. The raw reads generated after three sequencing runs were processed, resulting in 3,113,618 quality-filtered trimmed reads, followed by coassembly of all datasets using gsAssembler software (Newbler v2.9) with the following parameters: minimum overlap of 20 bp, minimum overlap identity of 80%, heterozygote mode, and extend low-depth overlap options on.MaxBin 2.0 , which eN50 value of 46,868. The genome includes 35 tRNA genes, 2 rRNA genes, 483 pseudo genes, 3 noncoding RNA genes, and 4,271 protein-coding sequences.Genome structural annotation was carried out using the NCBI Prokaryotic Genome Annotation Pipeline . The genFlavihumibacter sp. CACIAM 22H1 possesses 103  of the 107 essential genes used as models by MaxBin 2.0, allowing completeness near 94.4%. Overall, the data presented here should improve the genomic information about this poorly studied genus Flavihumibacter as well as its association with cyanobacteria. Furthermore, it should provide further insight into the production of substances of biotechnological interest.Our draft genome of LUKG00000000. The version described in this paper is version LUKG01000000.This whole-genome shotgun project has been deposited at DDBJ/EMBL/GenBank under the acession no.


S-transferases (GSTs) are multifunctional enzymes that are involved in the metabolism of endogenous and exogenous compounds and are related to insecticide resistance. The purpose of this study was to provide new information on the molecular characteristics and the positive selection of locust GSTs. Based on the transcriptome database, we sequenced 28 cytosolic GSTs and 4 microsomal GSTs from the migratory locust (Locusta migratoria). We assigned the 28 cytosolic GSTs into 6 classes—sigma, epsilon, delta, theta, omega and zeta, and the 4 microsomal GSTs into 2 subclasses—insect and MGST3. The tissue- and stage-expression patterns of the GSTs differed at the mRNA level. Further, the substrate specificities and kinetic constants of the cytosolic GSTs differed markedly at the protein level. The results of likelihood ratio tests provided strong evidence for positive selection in the delta class. The result of Bayes Empirical Bayes analysis identified 4 amino acid sites in the delta class as positive selection sites. These sites were located on the protein surface. Our findings will facilitate the elucidation of the molecular characteristics and evolutionary aspects of insect GST superfamily.Glutathione  Locusta migratoria) is an important agricultural pest in East and South Asia and the Pacific Region. The locust has developed insecticide resistance because of frequent applications of insecticides. Increased glutathione S-transferase  activity has been reported to be one of the main reasons for the resistance in L. migratoriaThe migratory locust  and microsomal GSTs. The latter are also known as membrane-associated proteins in eicosanoid and glutathione metabolism (MAPEG).The GSTs constitute a superfamily of multifunctional proteins found in most aerobic prokaryotes and eukaryotes Anopheles gambiae play a role in response to oxidative stress, by catalyzing the dehydrochlorination of DDT to yield DDE, thereby conferring resistance to DDT Sarcoptes scabiei is related to permethrin resistance 2 to PGD2Based on the sequence identity, substrate specificity, and immunological properties, insect cytosolic GSTs are assigned into at least 7 distinct classes, namely delta, epsilon, sigma, omega, theta, zeta, and unknown 4S, and FLAP subclasses), class 3 (MGST3 subclass), Synechocystis class, E. coli class, and bacteria class 2 to PGE2 under basal and inflammatory conditions 4 to form LTC4, and the reduction of 5S-hydroperoxy-6t,8c,11c,14c-eicosatetraenoic acid (5-HPETE) to 5S-hydroxy- eicosatetraenoic acid (5-HETE) in the leukotriene synthesis pathway Microsomal GSTs are classified into 6 classes, namely class 1 , class 2 . We heterologously expressed 25 of these GST genes by using E. coli expression system We purified the GSTs by using NTA-Ni2+ affinity chromatography. Finally, we identified positive selection sites of the GSTs by using the branch model, site model, and Bayes empirical Bayes (BEB) methods, and labeled these sites by using 3-dimensional homology models. Our findings will facilitate the elucidation of the molecular characteristics and evolutionary history of insect GSTs.In the present study, we sequenced and classified 32 full-length GST sequences from RevertAid H minus reverse transcriptase was purchased from Fermentas . Fastpfu DNA polymerase and the pEASY-Blunt zero cloning kit were obtained from Transgen Biotech Co. Ltd. . The SMARTer RACE cDNA amplification kit and the His60 Ni Superflow resin were purchased from Clontech . The substrates of GSTs, 2,4-dinitrochlorobenzene (CDNB), 2,4-dinitrofluorobenzene (FDNB), 3,4- dichloronitrobenzene (DCNB), 4-chloro-7-nitrobenzofurazan (NBD-Cl), 4-nitrobenzyl chloride (pNBC), 4-nitrophenethyl bromide (pNPB), and reduced glutathione (GSH) were from Sigma . All other reagents used were of the highest grade commercially available.Total RNA was extracted by using TRIzol , and mRNA was subsequently isolated by using the PolyATtract mRNA isolation systems. The cDNA was synthesized by using the SMARTer RACE cDNA amplification kit. RACE PCR was performed by using Fastpfu DNA polymerase, according to the manufacturer's instructions. The PCR product was subcloned into the pEASY-Blunt zero vector and sequenced by Sangon Biotech . The phylogenetic tree was constructed based on full-length DNA sequences, by using the neighbor-joining algorithm with mega 5.0 software.L. migratoria. For the stage-specific expression profile analysis, we collected whole bodies of L. migratoria individuals at 8 different developmental stages, including the egg (5 days and 10 days), and the 1st, 2nd, 3rd, 4th, and 5th-instar nymphs and adults. We extracted total RNA from these samples, and used 4 µg total RNA to synthesize cDNA, by using oligo-d(T) and RevertAid H minus reverse transcriptase. The primers used for RT-qPCR analysis are shown in β-actin as a reference gene. The experiment was repeated with 3 biological replicates, each with 2 technical replicates.All data were analyzed by the one-way ANOVA, LSD test.For the tissue-specific expression profile analysis, we dissected 12 different tissues, including the foregut, midgut, hindgut, gastric caecum, Malpighian tubules, cuticle, spermary, ovary, trachea, fat bodies, antenna, and muscles (from legs and back) from adult individuals of E. coli BL21 (DE3). The recombinant GSTs were induced by using 1 mM isopropyl-β-d-thiogalactopyranoside (IPTG) at 25°C, and purified by using NTA-Ni2+-affinity chromatography. The enzymatic activities were measured spectrophotometrically, as described previously The open reading frames of the GSTs were subcloned into the pET28a vector, and the recombinant vectors were used to transform http://blast.ncbi.nlm.nih.gov/). Then the program searched against Protein Database (PDB) of known protein structures, and then the X-ray structure of Delta GST (3EIN) from Drosophila melanogaster were selected as template structure. Finally, the homology model of LmGSTD1 was generated by MODELLER 9v12 through comparative protein structure prediction The sequence of LmGSTD1 as a target sequence was submitted to BLASTP program L. migratoria were detected based on the transcriptome, and were identified and sequenced by using RACE PCR.Based on the transcriptome of gratoria , we idengratoria , Table 1LmGSTe2 and LmGSTe3, the cytosolic GSTs of L. migratoria had an acidic theoretical pI, and a relatively high molecular mass (22.1–27.0 kDa) (L. migratoria GSTs. The sigma GSTs were most numerous (10 members), and were followed by the delta and epsilon GSTs , the omega and theta GSTs , and the zeta class (1 member). The former unclassified GST (LmGSTu1) was reassigned with high confidence into the delta class (LmGSTd6). Furthermore, 9 of the 10 sigma GSTs contained a class-specific Tyr9 residue in the G-site; the omega GSTs had an ancestral Cys9 residue; and the zeta, theta, and delta GSTs contained a conserved Ser in the G-site (With the exception of 7.0 kDa) . To enha7.0 kDa) . Based o7.0 kDa) . It was e G-site .LmGSTm1, LmGSTm2, and LmGSTm3) were classified into class 1 (insect subclass), and LmGSTm4 was assigned into class 3 (MGST3 subclass). To the best of our knowledge, our study is the first to report an insect GST  as a member of the MGST3 subclass.On the other hand, the 4 microsomal GSTs had an alkaline theoretical pI and a relatively low molecular mass  were widely expressed in ≥6 tissues (P<0.05). In comparison, 12 GSTs , had relatively narrow tissue-expression ranges (P<0.05). Moreover, LmGSTt1 was specifically located in the muscles and spermary (P<0.05), whereas LmGSTz1, LmGSTm1, and LmGSTm4 were mainly expressed in the trachea, spermary, and muscles, respectively (P<0.05).Our RT-qPCR analyses showed that the tissue-expression patterns of GSTs could be divided into 3 types, including wide-expression, partial-expression, and specific-expression . We obseLmGSTd1, LmGSTd3, LmGSTd5, LmGSTd7, LmGSTe2, LmGSTe3, LmGSTe4, LmGSTe5, LmGSTs3, LmGSTs8, LmGSTo1, and LmGSTz1) showed relatively high expression in the egg stage (P<0.05). Among these, 7 GSTs  showed a high expression level in the early egg stage (LmGSTo1) was primarily expressed in the late egg stage, and 3 GSTs  were expressed in the early and late egg stages (P<0.05). The remaining GSTs were mainly expressed in the nymphal and adult stages. The different mRNA expression patterns within each GST class may suggest different roles of these genes in different stages and tissues.The results of stage-dependent expression pattern analysis revealed that the GSTs could be classified into 2 groups, including the egg stage-expression group, and the nymph and adult stages-expression group . We obsegg stage  (P<0.05)2+-NTA column. The exceptions were 4 microsomal GSTs, 1 sigma GST (LmGSTs6) and 2 epsilon GSTs (LmGSTe2 and LmGSTe3). However, the reason for the no-expression of these genes is unclear.The locust GSTs were selected for protein expression and purification. Most of the GSTs were successfully purified by using a NiIn order to explain the possible relationships between the catalytic activity and substrate diversity, we determined the substrate specificity of the recombinant locust GSTs toward 6 electrophilic substrates, including CDNB, FDNB, DCNB, NDB-Cl, pNPC, and pNPB . We obseKmCDNB varied <70-fold among the GST classes . By contrast, the catalytic efficiency (kcat/Km) of CDNB differed markedly among the GST classes . On the other hand, the apparent KmGSH varied <50-fold among the GST classes . By contrast, the catalytic efficiency (kcat/Km) of GSH differed markedly among the GST classes .We determined the kinetic constants of the 25 GSTs, by using CDNB and GSH as substrates . The appThe observed variations of enzyme catalytic properties not only reflect environmental changes in the GSTs active site, but also embody the diversity of the physiological substrates. This may partially explain why insects possess so many different GSTs.A, ωB and ωC were significantly greater than the background ratio ω0  were located in the N-domain of GSTs.Our present results focused on whether the specific sites have undergone positive selection. Therefore, we chose 5 models of CODEML to detect positive selection sites. We conducted 2 LRTs, which compared M1a with M2a, and M7 with M8, to test whether each GST class had evolved under positive selection . The oneN- and C-domain residues.The results of protein 3-dimensional-structure modeling suggested that the overall structures of the delta GSTs, which are involved in positive selection pressure, were relatively conserved, especially regarding the β-sheet and α-helix . TherefoIn the present study, we identified and sequenced cDNAs putatively encoding 28 cytosolic GSTs and 4 microsomal GSTs based on the transcriptome database. Our phylogenetic analysis revealed that the 28 cytosolic GSTs belonging to 6 different cytosolic classes , and the 4 microsomal GSTs belonging to 2 classes—class 1 (insect subclass) and class 3 (MGST3 subclass). The sigma class (10 members), delta class (7 members), epsilon class (5 members), and omega class (3 members) have apparently undergone gene expansion. However, it remains unclear whether these genes play different roles in different tissues and at different developmental stages. To explore these matters, we determined the gene expression patterns at the mRNA level, and the enzymatic properties at the protein level.We observed 12 GSTs highly expressed during the egg stage. Among the GSTs that were highly expressed in the egg stage, some showed a high expression level in the early egg stage, others were primarily expressed in the late egg stage, and others were expressed in both the early and late stages . BecauseOncorhynchus kisutch) are located in the olfactory rosettes and are critical to maintaining olfaction in vivo. In the present study, we observed differences in the substrate specificities and steady kinetic constants of locust GSTs. The architecture of the functional region may have been altered by amino acid substitutions. The G-site residues of the N-terminal domain are known to be relatively conserved; on the other hand, the H-site residues of the C-terminal domain are relatively flexible, thereby contributing to adaptation to a wider spectrum of toxic reagents. Classic studies in the literature have paid more attention to the obvious functional changes caused by amino acid replacements in critical regions N-acetyl-l-ornithine transcarbamylase and N-succinyl-l-ornithine transcarbamylase E. coli isocitrate dehydrogenase systematically inverted coenzyme specificity Tissue-specific expressed GSTs may play a vital part in the respective organs or tissues through their detoxification activity. For instance, the pi, rho, and omega GSTs of coho salmon , the hydrophobic ligand binding site (H-site) and the positive selective sites are labeled with green, red and blue square, respectively. The residues located at the interface of the N- and C-termianl domain and dimer interface are label with orange and pink square, respectively.(PDF)Click here for additional data file.S2 FigurePrediction of transmembrane segments of microsomal GSTs from L. migratoria.A: LmGSTm1, B: LmGSTm2,C: LmGSTm3,D: LmGSTm4.(TIF)Click here for additional data file.S3 FigureSuperimposed homology structure of delta class GSTs.N-domain, C-domain and link region are colored yellow, red and green, respectively.(TIF)Click here for additional data file.S4 FigurePhylogenetic analysis of delta GSTs from L.migtatoria. The bootstrap neighbor-joining tree was generated using MEGA from ClustalW alignments. Branch numbers represent bootstrap values (1000 replicates). Branch A, branch B and branch C are labeled with A, B, C(TIF)Click here for additional data file.S1 TablePrimer sequence for Real-time PCR analysis.(DOC)Click here for additional data file.S2 TableComparison of GST gene number from L. migratoria and five insects.(DOC)Click here for additional data file.S1 DataAnalysis of L. migratoria transcriptome.(PDF)Click here for additional data file.S2 DataSequences of L. migratoria GSTs.(DOC)Click here for additional data file.S3 DataSequences of cytosolic GSTs used for phylogenetic analysis.(DOC)Click here for additional data file.S4 DataSequences of microsomal GSTs used for phylogenetic analysis.(DOC)Click here for additional data file.S5 DataOutput file from PAML.(RAR)Click here for additional data file.


TRAPPC11 orthologue showed hepatomegaly with steatosis and defects in visual system development. In humans, TRAPPC11 mutations have been reported in only three families showing limb-girdle muscular dystrophy (LGMD) or myopathy with movement disorders and intellectual disability.Transport protein particle (TRAPP) is a multiprotein complex involved in endoplasmic reticulum-to-Golgi trafficking. Zebrafish with a mutation in the We screened muscular dystrophy genes using next-generation sequencing and performed associated molecular and biochemical analyses in a patient with fatty liver and cataract in addition to infantile-onset muscle weakness.TRAPPC11 mutations. Muscle pathology demonstrated typical dystrophic changes and liver biopsy revealed steatosis. The patient carried compound heterozygous mutations of a previously reported missense and a novel splice-site mutation. The splice-site change produced two aberrantly-spliced transcripts that were both predicted to result in translational frameshift and truncated proteins. Full-length TRAPPC11 protein was undetectable on immunoblotting.We identified the first Asian patient with TRAPPC11-opathy as the patient showed the following: (1) congenital muscular dystrophy phenotype rather than LGMD; (2) steatosis and infantile-onset cataract, both not observed in previously reported patients; but (3) no ataxia or abnormal movement, clearly indicating that TRAPPC11 plays a physiological role in multiple tissues in human.This report widens the phenotype of  TRAPPC11 orthologue is characterized by hepatomegaly with steatosis, thereby named foie gras mutant, and by defects in visual system development [TRAPPC11 mutations, describing one Syrian family with limb-girdle muscular dystrophy (LGMD) phenotype, which was labeled LGMD2S, and two families of Hutterite ancestry with myopathy phenotype, movement disorders and intellectual disability [TRAPPC11 gene who developed congenital muscular dystrophy (CMD) phenotype with prominent fatty liver and infantile-onset cataract, further broadening the clinical phenotype of TRAPPC11-opathy.Transport protein particle (TRAPP) is a multiprotein complex involved in endoplasmic reticulum (ER)-to-Golgi trafficking and possibly other membrane trafficking steps , 2. Oligelopment , 5. In hsability . In addiThe currently 8-year-old Han Chinese girl residing in Taiwan, born to non-consanguineous parents, was found unable to stand up at age 1 year. Timeline of the developmental milestones before then was not recalled by the parents. She started walking independently at age 1 year and 6 months and readily fell down. At age 2 years, speech delay was noticed as she could not speak any significant single word though she could understand and follow simple orders. Bilateral cataracts were also found at the same age. Her birth history was uneventful, and there was no relevant family history. However, rehabilitation did not show marked improvement in her speech and motor functions. At age 3 years and 6 months, high levels of transaminases were found immediately prior to cataract surgery. Subsequently high creatine kinase (CK) level was identified resulting in a referral to a pediatric myologist. Physical examination showed mild lordosis, positive Gowers’ sign with waddling gait, and decreased deep tendon reflexes, as well as hepatomegaly. Neither ataxia nor abnormal movement was observed. Blood biochemistry indicated that the levels of AST , ALT , and serum CK  were markedly elevated. Further assessment identified borderline cognitive function  = 82–87; motor DQ = 67). Brain magnetic resonance imaging (MRI) did not show cerebral or cerebellar atrophy, and there was no structural abnormality except for slightly reduced periventricular white matter volume with angular-shaped ventricles Fig. . On muscDue to persistent high levels of AST/ALT and hepatomegaly, liver biopsy was performed and revealed excessive lipid accumulation in hepatic cells, suggestive of steatohepatitis Fig. . Muscle AGRN, ALG13, ANO5, B3GALNT2, B3GNT1, CAPN3, CAV3, CHKB, COL12A1, COL6A1, COL6A2, COL6A3, DAG1, DES, DMD, DNAJB6, DOK7, DOLK, DPAGT1, DPM1, DPM2, DPM3, DYSF, EMD, FAT1, FHL1, FKRP, FKTN, FLNC, GFPT1, GMPPB, ISPD, ITGA7, KLHL9, LAMA2, LARGE, LMNA, MEGF10, MICU1, MYOT, PLEC, POMGNT1, POMGNT2, POMT1, POMT2, PTRF, SGCA, SGCB, SGCD, SGCG, POMK, SMCHD1, STIM1, SYNE1, SYNE2, TCAP, TMEM43, TMEM5, TNPO3, TRAPPC11, TRIM32) was performed on genomic DNA extracted from blood lymphocytes of the proband. Multiplex primer pools were designed using Ion AmpliSeq™ Designer software (Life Technologies). This custom gene panel covers 96.8 % of the coding sequence region of these genes. Enrichment of exonic sequences was performed with Ion AmpliSeq™ Library Kit 2.0 (Thermo Scientific) and sequenced on an IonPGM™ (Thermo Scientific) according to the manufacturer’s protocol. Compound heterozygous mutations of c.2938G > T (p.Gly980Arg) and c.661-1G > T in TRAPPC11 (NM_021942.5) were identified. Subsequent Sanger sequencing of genomic DNA was performed in both the proband and her parents to confirm the detected variants  to allow detection of all mRNA products by sequencing with fluorescent dideoxy chain terminators  on an ABI 3130 sequencing instrument (Applied Biosystems). Two additional aberrantly spliced transcripts were identified, and both were predicted to result in translational frameshift (Fig. In order to identify the cause of this syndrome, targeted next-generation sequencing covering reported muscular dystrophy related genes (nts Fig. . This reift Fig. , c. In bift Fig. .Fig. 3MoWe have demonstrated that the novel splice-site mutation c.661-1G > T results in two different aberrant transcripts, predicted to produce two truncated proteins. The absence of a full-length TRAPPC11 protein by Western blot analysis suggests the possibility that Gly980Arg mutation may destabilize the protein, which was also shown in the previous report describing Gly980Arg in a homozygous manner, or may enhance its degradation [TRAPPC11 mutations presented with two groups of clinical manifestations: one with more prominent muscular and skeletal symptoms and the other with microcephaly, hyperkinetic movements, ataxia, and intellectual disability, apparently reflecting the difference of the two genotypes, Gly980Arg and Ala372_Ser429del. Three patients in one family carried homozygous Gly980Arg, and five affected members from two unrelated families had homozygous Ala372_Ser429del. No notable interfamiliar difference of phenotype was observed between the two families with Ala372_Ser429del mutation. Noteworthily, only one patient with Gly980Arg mutation was reported to develop mild cataracts after school age. Clinical manifestation of the patient in the present study is different from the previously reported patients in several respects. First, the patient in the present study presented with steatosis and very early-onset cataract, similar to what was seen in the foie gras mutant in zebrafish. Second, the patient developed a CMD phenotype rather than the LGMD seen in the previous study. Finally, the patient does not display choreiform movement, ataxia, nor any skeletal abnormality. These differences may partly be explained by a more deleterious effect of the splice-site mutation on the TRAPPC11 protein which we expect to lead to complete loss of function, compared to Gly980Arg or a 58 amino acid in-frame deletion in homozygosity in the previous study. Table TRAPPC11 mutations.In the previous study, the affected individuals with TRAPPC11 mutations [TRAPPC11-associated disease.It is noteworthy that, although the patient presented here did not have microcephaly, abnormal involuntary movements, nor cerebral atrophy, which were previously reported in the patients with utations , her brautations . Howeverutations , 10. RegTRAPPC11-opathy. Although the detailed mechanism causing intracellular lipid storage in liver is still unknown, the phenotype of the patient in this study clearly indicates that TRAPPC11 plays a physiological role in multiple tissues in humans including the liver, muscle, eye, brain, and bone. This may be due to impairment of TRAPPC11 functions in multiple membrane-trafficking pathways or other processes.Collectively, this study widens the phenotype of Written informed consent was obtained from the patient’s mother for publication of this Case Report and any accompanying images. A copy of the written consent is available to Editors of this journal on request.


Bacteroidetes, Actinobacteria, Proteobacteria, Firmicutes, Fusobacteria, and TM7. The dominant genera included Capnocytophaga, Prevotella, Actinomyces, Corynebacterium, Neisseria, Streptococcus, Rothia, and Leptotrichia. β diversity analysis showed that the plaque microbial community structure was similar in all groups. Using LEfSe analysis, 25 differentially abundant taxa were identified as potential biomarkers. Key genera (27) that potentially contributed to the differential distributions of plaque microbiota between groups were identified by PLS-DA analysis. Finally, co-occurrence network analysis and function predictions were performed. Treatment strategies directed toward modulating microbial interactions and their functional output should be further developed.Dental caries has a polymicrobial etiology within the complex oral microbial ecosystem. However, the overall diversity and structure of supragingival plaque microbiota in adult dental health and caries are not well understood. Here, 160 supragingival plaque samples from patients with dental health and different severities of dental caries were collected for bacterial genomic DNA extraction, pyrosequencing by amplification of the 16S rDNA V1–V3 hypervariable regions, and bioinformatic analysis. High-quality sequences  clustered into 10,365 operational taxonomic units , representing 453 independent species belonging to 122 genera, 66 families, 34 orders, 21 classes, and 12 phyla. All groups shared 7522 OTUs, indicating the presence of a core plaque microbiome. α diversity analysis showed that the microbial diversity in healthy plaques exceeded that of dental caries, with the diversity decreasing gradually with the severity of caries. The dominant phyla of plaque microbiota included  The microorganisms that live in and on the human body are estimated to outnumber human cells by at least an order of magnitude. Human beings are regarded as “superorganisms” whose metabolism represents an amalgamation of microbial and human attributes  index. Furthermore, we aimed to identify bacterial shifts related with dental health and caries, and to investigate the presence of a core plaque microbiome. Our findings expand the understanding of bacterial ecology in dental caries and provide directions for prevention and treatment strategies of dental caries.This study was performed in accordance with the recommendations of the Ethics Committee of the Ninth People's Hospital of Shanghai Jiao Tong University. All subjects gave written informed consent in accordance with the Declaration of Helsinki.A total of 160 subjects were recruited for this study, including 131 patients with dental caries from the Department of Endodontics  and 29 dental healthy volunteers from the College of Stomatology (Shanghai Jiao Tong University). Inclusion criteria of subjects were as follows: (1) No-caries (2) Low-caries (3) Moderate-caries .(4) High-caries , with minor modifications. Briefly, the sampling site was isolated with cotton rolls and dried with a gentle air stream to avoid saliva contamination. Then, a sterile Gracey curette was used to collect a pooled supragingival plaque from the buccal surface of the maxillary first molar or, if necessary, from its adjacent teeth or contralateral teeth, and to avoid gingival bleeding. Each pooled plaque sample was immediately released from the curette by agitation into a sterile, labeled Eppendorf tube containing 1 mL thioglycollate medium, which was placed on ice. Within 4 h of collection, all samples were transported on ice to our laboratory for DNA extraction.Sampling was performed 2 h after eating in the morning. Supragingival plaque samples were collected from each subject according to the Manual of Procedures for Human Microbiome Project , following the manufacturer's instructions, and stored at −40°C prior to further analysis. The quantity and quality of extracted DNAs were measured using a NanoDrop ND-1000 spectrophotometer  and agarose gel electrophoresis, respectively. The results showed that the A260:A280 ratios were 1.8–2.0 and that the DNA concentrations were 20–150 ng/μL, indicating that the genomic DNA extracted was ideal and met the requirements for subsequent sequencing.PCR amplification of the bacterial 16S rDNA V1–V3 region was performed using the forward primer 8F (5′-AGAGTTTGATCCTGGCTCAG-3′) and the reverse primer 533R (5′-TTACCGCGGCTGCTGGCAC-3′)  pipeline was employed to process the pyrosequencing data, as previously described   test. The taxonomy compositions and abundances were visualized using MEGAN (version 4.70.4) and GraPhlAn software (version 0.9.7) , alpha diversity indexes, including the Chao1, ACE, Shannon, Inverse Simpson, Good's coverage, and Simpson's evenness indexes were calculated at 97% identity    was used to detect differentially abundant genera in the four groups for biomarker discovery using the online Galaxy workflow framework . After data trimming and quality filtering, 2,261,700 high-quality sequences  were acquired, with an average of 14,136 sequences per sample , the Inverse Simpson diversity index was significantly different between groups MC and HC , demonstrating the higher bacterial diversity of healthy dental plaques compared to dental caries, with the diversity decreasing gradually with the severity of caries. Good's coverage estimator for each group was over 95%, indicating that the current sequencing depth was sufficient to saturate the bacterial diversity of dental plaques. In addition, Simpson's evenness index indicated that the bacterial-community distribution in the plaque samples was very uneven, which was also observed in the rank-abundance curve which had a high slope and a long tail comprised of low-abundance OTUs , Actinobacteria (28.6%), Proteobacteria (14.6%), Firmicutes (11.3%), Fusobacteria (5.8%), and TM7 (3.6%), together accounting for 99% of the total sequences. The 6 most rare phyla were Spirochaetes, SR1, Synergistetes, GN02, Tenericutes, and Chloroflexi. The most prevalent genera were Capnocytophaga (18.1%), Prevotella (12.8%), Actinomyces (12.3%), Corynebacterium (9.8%), Neisseria (7.0%), Streptococcus (6.3%), Rothia (3.7%), Leptotrichia (3.7%), TM7_[G-1] (3.1%), Porphyromonas (2.8%), Lautropia, (2.8%), Fusobacterium (2.1%), Selenomonas (2.1%), Veillonella (1.7%), Actinobaculum (1.7%), Campylobacter (1.2%), and Propionibacterium (1.0%), together comprising 92.3% of the total sequences. The compositions in taxa of the microbial communities according to the tested sample groupings are provided in Figure Neisseria, Streptococcus, Lautropia, Campylobacter, Ottowia, Cardiobacterium, and Aggregatibacter were relatively abundant, with higher abundances of Lautropia, Cardiobacterium, and Aggregatibacter observed in the NC group and higher abundances of Neisseria and Campylobacter observed in the LC group. Nevertheless, the Streptococcus and Ottowia genera exhibited similar abundances in each group. These significant differences were further tested by LEfSe and PLS-DA analysis. A taxonomy tree visualized by GraPhlAn was used to quickly find the dominant taxa from the complex microbial data. As shown in Figure Firmicutes, Bacteroidetes , Actinobacteria  and Proteobacteria.The bacterial distribution was characterized in terms of the relative taxonomic abundances. A total of 12 phyla, 21 classes, 34 orders, 66 families, 122 genera and 453 species were detected in the supragingival plaque samples. Figure P = 0.462. UPGMA hierarchical clustering analysis also revealed that the samples did not form well-separated clusters corresponding to the four groups, indicating similarity in the bacterial community structures . Since LEfSe was a strict tool, we also generated a PLS-DA model to identify more genera that were differentially distributed among groups. The key genera with VIP > 1 were considered to be important contributors to the model. As shown in Table Cardiobacterium, Corynebacterium, Lachnospiraceae_[G-3], Lachnoanaerobaculum, Aggregatibacter, Eubacterium_[XI][G-7], GN02_[G-1], Fusobacterium, Clostridiales_[F-2][G-1], and Tannerella, which were identified as health-related bacteria. The other 17 genera were significantly more abundant in HC, MC and LC groups, identified as caries-related bacteria. Furthermore, two genera were identified with a VIP > 2, namely the Cardiobacterium and Corynebacterium genera.There were significant differences in the community compositions among the four groups. As shown in Figure Capnocytophaga , Prevotella (13.5%), Actinomyces (13.0%), Corynebacterium (8.9%), Streptococcus (6.6%), and Neisseria (6.4%).To examine the existence of an identifiable common core microbiome , Selenomonas–Prevotella (0.76), and Treponema–Prevotella (0.75) pairs.Co-occurrence analysis was used to discern relationships among the plaque microbiota at the genus level by calculating C-scores. As shown in Table a Figure , 14 geneTo predict bacterial functions of members of the supragingival plaque community, PICRUSt analysis was performed based on the 16S rRNA composition data of each sample Figure . A totalOchrobactrum, Centipeda, Lachnoanaerobaculum, GN02_[G-1], Moryella, Bacteroides, Megasphaera, Rhizobium, Filifactor, and Dietzia. The V1–V3 region has been used in several studies , indicating that the bacterial diversity of healthy dental plaques was higher than that in caries. This finding was in agreement with results from previous studies  or have been identified at significantly higher levels in dental caries   were also identified as health-related genera. Cardiobacterium is a member of the group of HACEK bacteria , which are part of the normal oropharyngeal flora and have common characteristics that easily lead to endocardial infections , and showed no significant difference among the groups. In Aas's study, S. mutans was not detected in 10% of dental caries using checkerboard hybridization , should be further developed.Conceived and designed the experiments: JL and ZH. Performed the experiments: CX and SR. Analyzed the data: CX and ZH. Contributed reagents/materials/analysis tools: CX and SR. Manuscript preparation: CX. Manuscript revisions: JL, ZH, CX, and SR.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.


The following review will discuss the various actions of CN in glial cells, with particular emphasis on astrocytes, and consider the implications for neurologic dysfunction arising with aging, injury, and/or neurodegenerative disease.Similar to peripheral immune/inflammatory cells, neuroglial cells appear to rely on calcineurin (CN) signaling pathways to regulate cytokine production and cellular activation. Several studies suggest that harmful immune/inflammatory responses may be the most impactful consequence of aberrant CN activity in glial cells. However, newly identified roles for CN in glutamate uptake, gap junction regulation, Ca The CN holoenzyme is a heterodimeric protein containing a 60 kDa catalytic subunit (CN A) and a 19 kDa regulatory subunit (CN B). Multiple isoforms of both subunits have been identified and are expressed differentially throughout brain and most other tissues  is a Caview see ). CN A cfidelity ,5. Withifidelity .in situ hybridization studies on healthy adult rat brain performed in the mid-1980s to early 1990s reported high levels of CN in neurons of the striatum, hippocampus, amygdala, and neocortex [Immunohistochemical and eocortex -9, with eocortex ,7. Many eocortex ,11. Sinceocortex . In the eocortex ,13. Neveeocortex -17. The Though primarily localized to neurons in healthy nervous tissue, CN may also be strongly expressed in glia during aging, injury, and/or disease. Greater numbers of CN-positive astrocytes have been reported in the hippocampus of aged, β-amyloid (Aβ)-bearing transgenic amyloid precursor protein/presenilin 1 (APP/PS1) mice, particularly in the immediate vicinity of extracellular Aβ deposits . A simil+, glutamate, and other neurotransmitters [Astrocytes are an abundant and diverse subtype of glia. As a critical component of the neurovascular unit, astrocytes ensheath most microvessels via specialized end-foot processes, which help to maintain the integrity of the blood-brain barrier and promote osmotic balance (for review see ). Many asmitters ,24. Redismitters . By enshsmitters ,26,27. Tsmitters .+ and glutamate) from the extracellular milieu, which could, in turn, lead to further synaptic dysfunction, neuronal damage, and neuroinflammation [Along with activated microglia, astrocyte activation is widely accepted as a hallmark of neuroinflammation, though the functional phenotype of activated astrocytes remains somewhat elusive. Protective roles for activated astrocytes, particularly after acute injury, have been demonstrated by many studies . Howeveammation ,28,33,34ammation -37 suggeWe have hypothesized that CN is a critical mechanism for triggering phenotype switching  and neuroinflammatory signaling inherent to astrocytes during neural damage and dysfunction ,18,33. IIn astrocytes, CN appears to represent a fundamental link between morphological changes and immune/inflammatory signaling. Forced expression of activated CN in primary mixed neuron/glia cultures was sufficient to cause an increase in the width of astrocyte somata and processes , while iThrough its actions on NFAT- and NFκB-dependent transcriptional regulation, CN appears ideally suited to drive the self-perpetuating “cytokine cycles” implicated in chronic neuroinflammation ,55. Inde2+ elevations in myoblasts, while NFATs 1 and 2 are selectively activated by the same kind of stimulation in myotubes [., NFATs 2 and 4) have also been detected in astrocytes and microglia at the mRNA and protein levels [Even the presence of different NFAT isoforms can dramatically alter the functional impact of elevated CN activity. A striking example of this is found in muscle tissue where NFAT4 is selectively activated by Camyotubes . In skelmyotubes . The idemyotubes ,19,66-68myotubes , we havemyotubes . In contmyotubes -71), sugn levels -74. Seven levels ,68,73, tn levels . Other gIn summary, the available literature suggests that CN plays a major role in shaping the neuroinflammatory phenotype of astrocytes. The impact of CN on neuroinflammation appears to be complex and may depend upon the co-activation of other intracellular signaling pathways and/or the association of CN with multiple transcription factors, including different NFAT isoforms, NFκB, AP1, FOXO3, and others. These differential CN interactions likely do not simply promote a global phenotypic change but instead may selectively affect specific glial functions , which energetically couple glutamate uptake to the import of Na+ ions. The EAAT2 isoform (or Glt 1) is expressed predominantly in astrocytes and plays a lead role in glutamate clearance from many brain regions [One of the most important functions of astrocytes is the removal of potentially toxic factors from the extracellular milieu, including K regions . Loss of regions ,75,76).  regions ,77-83. T regions , suggest regions -86. ThesChanges in EAATs appear to be strongly linked to the activated astrocyte phenotype. Immunohistochemical analyses of postmortem human brain sections revealed an inverse correlation between EAAT2 and GFAP expression levels . Moreove2+ and inositol trisphosphate (IP3), also allow groups of astrocytes to respond in synchrony to highly localized extracellular signaling factors, which, in turn, can influence the plasticity of local synaptic ensembles and/or modulate the tone of the local vascular network [Astrocytes are extensively coupled to one another and, to a lesser degree, to other cell types via gap junctions (GJ). The GJ channel consists of a hexamer of specialized proteins called connexins (Cx), which are localized to the cell membrane and directly apposed to a similar Cx hexamer localized to the plasma membrane of another cell . The cha network . In addi network .+ and glutamate across GJ-coupled astrocyte networks during high levels of neuronal activity is one of the fundamental mechanisms for preserving high fidelity interneuronal communication and for preventing excitotoxicity [The diffusion of Ktoxicity . Transgetoxicity -97. On ttoxicity .2+ mobilizing agents ionomycin and phorbol ester [Similar to EAATs, astrocytic GJs are highly sensitive to extracellular factors linked to neuroinflammation and exhibit altered expression and/or function in a variety of different neurologic disorders and diseases ,99. Multol ester . ParalleDespite these observations, the functional impact of the CN/Cx43 interaction is not presently clear. Using a dye-coupling approach, CN inhibitors were initially shown to prevent the decoupling of GJs in astrocytes following a hypoxic insult . Later s2+ dysregulation including elevated expression of a variety of Ca2+ channels and Ca2+ regulated proteins, as well as higher intracellular Ca2+ levels and/or more frequent Ca2+ oscillations [2+ homeostasis. At the transcriptional level, CN helps drive the expression of many key Ca2+ signaling mediators in multiple cell types [2+ channels, intracellular Ca2+ release channels, and Ca2+-dependent enzymes [2+ transients evoked by Aβ [2+-induced Ca2+ release, including IP3 receptor 1 and metabotropic glutamate receptor 5 [2+ channels, which may, in turn, disrupt cellular activity and viability [2+ channels appear to be present at high levels in activated astrocytes following acute injury [2+ channel blocker nifedipine [2+ channel activator Bayk8644 [2+ channel interactions may play a critical role in promoting Ca2+ dysregulation in activated astrocytes (Figure Similar to neurons, activated astrocytes in models of aging, injury, and disease exhibit numerous signs of Callations -107. Whill types -111. At ed by Aβ  and to sceptor 5 . Other piability -116. Thoe injury . This difedipine , while oIn AD, CN expression and activity levels are directly correlated to increasing levels of the Aβ peptide . Aβ depo2+ levels. Similarly, our group showed that inhibition of astrocytic NFAT activity reduces soluble Aβ levels and plaque load in the hippocampus of APP/PS1 mice [Interestingly, APP/PS1 mice treated with FK506 exhibit reductions in Aβ plaque load relative to vehicle-treated APP mice , suggestPS1 mice  in paralPS1 mice . Nonethe2+ homeostasis in neurons and/or erode neuronal viability leading to greater Aβ levels [In addition to directly influencing Aβ production in astrocytes, it is perhaps just as likely that astrocytic CN indirectly influences Aβ production/metabolism in neurons. Indeed, numerous “pro-inflammatory” factors released by glial cells, many of which are sensitive to CN activity, have been shown to stimulate neuronal APP and/or Aβ production, e.g., see -131. Simβ levels -136. Whe2+ transients, lower extracellular glutamate levels, reduced neuronal excitability, and less excitotoxic neuronal death following inhibition of astrocytic CN/NFATs as discussed in preceding sections.Numerous studies have shown that commercial CN inhibitors dampen glial activation, impart neuroprotection, and/or improve neurologic function in animal models of aging, injury, and disease ,12. SureIn addition to directly suppressing NFAT activity, overexpression of VIVIT may help to divert CN activity to other substrates that are more protective in nature. For instance, Fernandez et al.  reportedWe are just beginning to understand the contribution of CN to glial function. While it seems clear that CN is intimately involved in neuroinflammation, much work is still needed to fully assess the specific interactions between CN-dependent and -independent transcription factors, and how these interactions regulate specific inflammatory phenotypes. While this review has focused primarily on the role of CN in astrocytes, other work showing similar roles of CN in microglia -58,74 un


Our aim was to assess whether perioperative and postoperative optimal 7-day nutrition support could modulate the inflammatory status and clinical outcome of severe malnourished patients with surgery for gastrointestinal neoplasia.A prospective randomized study of 64 patients with gastrointestinal neoplasia, severe malnourished BMI <18.5, albumin level <3 g/dl, BW loss >10%, NRS >3, scheduled for surgery, allocated into two groups. Group A: 32 patients, minimal enteral nutrition in the postoperative period according to tolerance, medium 500 kcal/day. Group B: 32 patients received optimal parenteral nutrition support  3 days before surgery and continued for at least 4 days postoperatively. We measured CRP, fibrinogen, IL-6, TNF, albumin level preoperative and at 96 hours, the incidence of complications, and the length of ICU stay.There was a significant decrease in the values of CRP, IL-6, TNF, albumin at 96 hours in group B. No difference in fibrinogen. A significantly lower rate of complications and a shorter time of ICU stay were observed in group B. See Figures Perioperative optimal nutrition support for at least 7 days could modulate the inflammatory status and clinical outcome of severe malnourished surgical neoplasic patients.