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[Display omitted] Aqueous Zn-ion battery is expected to become a substitute for Li-ion battery due to its inherent safety, low cost, and environmental friendliness. Dendrite growth and side reaction problems during electroplating lead to its low Coulombic efficiency and unsatisfactory life, which greatly limits its practical application. Here, we propose a dual-salts hybrid electrolyte, which alleviates the above issues by mixing Zn(OTf) 2 to ZnSO 4 solution. Extensive tests and MD simulations have shown that the dual-salts hybrid electrolyte can regulate the solvation structure of Zn2+, facilitating uniform Zn deposition, and inhibiting side reactions and dendrite growth. Hence, the dual-salts hybrid electrolyte exhibits good reversibility in Zn//Zn batteries, which can provide a lifetime of more than 880 h at 1 mA cm−2 and 1 mAh cm−2. Moreover, the average Coulombic efficiency of Zn//Cu cells in hybrid system can reach 98.2% after 520 h, much better than that of 90.7% in pure ZnSO 4 electrolyte and 92.0% in pure Zn(OTf) 2 electrolyte. Benefiting from the fast ion exchange rate and high ion conductivity, Zn-ion hybrid capacitor in hybrid electrolyte also displays excellent stability and capacitive performance. This effective strategy for dual-salts hybrid electrolytes provides a promising direction for designing aqueous electrolytes for Zn-ion batteries. [ABSTRACT FROM AUTHOR]

Polyinosinic-polycytidylic acid (poly I:C) is a synthetic analog of double-stranded RNA (dsRNA) that activates anti-infective innate immunity. The underlying mechanisms are identified as targeting pattern recognition receptors and Th1-inducing. However, whether poly I:C manipulates metabolism to implement this anti-infective function is unknown. Here, GC-MS based metabolomics was used to characterize metabolic profiles induced by different doses of poly I:C. Analysis on the dose-dependent metabolomes shows that elevation of the TCA cycle and malate with the increasing dose of ploy I:C forms the most characteristic feature of the poly I:C stimulation. Exogenous malate activates the TCA cycle and elevates survival of zebrafish infected with Vibrio alginolyticus , which is related to the elevated expression of il-1b, il-6, il-8, tnf-a , and c3b. These results reveal a previously unknown regulation of poly I:C that boosts the TCA cycle to enhance innate immunity against bacterial infection. • Metabolic profiles induced by different doses of poly I:C. was characterized. • Elevated TCA cycle and malate forms the most characteristic feature. • Malate boosts TCA cycle and elevates zebrafish survival post bacterial infection. • This is related to the elevated expression of il-1b, il-6, il-8, tnf-a , and c3b. • These reveal a metabolic modulation of poly I:C against bacterial infection. [ABSTRACT FROM AUTHOR]

Abstract Stearoyl-CoA desaturase 1 (SCD1), a lipogenic enzyme that adds a double bond at the delta 9 position of stearate (C18: 0) and palmitate (C16: 0), has been proven to be important in the development of obesity. Mice with skin-specific deficiency of SCD1 (SKO) display increased whole-body energy expenditure, which is protective against adiposity from a high-fat diet because it improves glucose clearance, insulin sensitivity, and hepatic steatosis. Of note, these mice also display elevated levels of the "pro-inflammatory" plasma interleukin-6 (IL-6). In whole skin of SKO mice, IL-6 mRNA levels are increased, and protein expression is evident in hair follicle cells and in keratinocytes. Recently, the well-known role of IL-6 in causing white adipose tissue lipolysis has been linked to indirectly activating the gluconeogenic enzyme pyruvate carboxylase 1 in the liver, thereby increasing hepatic glucose production. In this study, we suggest that skin-derived IL-6 leads to white adipose tissue lipolysis, which contributes to the lean phenotype of SKO mice without the incidence of meta-inflammation that is associated with IL-6 signaling. Highlights • IL-6 expression is significantly increased in the skin of SKO mice and is also dramatically elevated in blood. • White adipose tissue lipolysis, a well-known effect of IL-6, is evident in SKO mice. • Hepatic PC1 is activated by WAT-derived fatty acids and leads to increased gluconeogenesis in SKO mice. [ABSTRACT FROM AUTHOR]

Alcoholic fatty liver is a threat to human health. It has been long known that abstinence from alcohol is the most effective therapy, other effective therapies are not available for the treatment in humans. Curcumin has a great potential for anti-oxidation and anti-inflammation, but the effect on metabolic reconstruction remains little known. Here we performed metabolomic analysis by gas chromatography/mass spectrometry and explored ethanol pathogenic insight as well as curcumin action pattern. We identified seventy-one metabolites in mouse liver. Carbohydrates and lipids were characteristic categories. Pathway analysis results revealed that ethanol-induced pathways including biosynthesis of unsaturated fatty acids, fatty acid biosynthesis and pentose and glucuronate interconversions were suppressed by curcumin. Additionally, ethanol enhanced galactose metabolism and pentose phosphate pathway. Glyoxylate and dicarboxylate metabolism and pyruvate metabolism were inhibited in mice fed ethanol diet plus curcumin. Stearic acid, oleic acid and linoleic acid were disease biomarkers and therapical biomarkers. These results reflect the landscape of hepatic metabolism regulation. Our findings illustrate ethanol pathological pathway and metabolic mechanism of curcumin therapy. [ABSTRACT FROM AUTHOR]

Malignancies including ovarian cancer (OvCa) are genetically unstable. Genomic integrity is maintained by tumor suppressor p53 and DNA damage response network, which crosstalk to each other via not well characterized mechanisms. In this work, we characterize features of damage-related signals in cultured epithelial OvCa cells and tumor biopsies. We found that endogenous burden of DNA damage in OvCa tissues were ubiquitously accumulated in high-grade malignancies than lower grade of cancer that cannot be obviously explained by disturbed function of in DNA damage response (DDR). In contrast, CHK1 phosphorylation (CHK1-pS345) marking the checkpoint activation in nucleolar compartments are prevalent in high-grade OvCa, coincident to the elevated DNA damage in nucleoplasm. Generation of CHK1-pS345 requires the presence of p53 protein in addition to the well-known activities of ATM/ATR kinases. Apparently, mutant forms of p53 possess higher activity in triggering CHK1 phosphorylation than wild type, implying a potential role of p53 in maintaining rDNA integrity. Loss of p53 function would cause replication stress in nucleoli. Altogether, our study reveals endogenous nucleoli stress in OvCa that is coupled to perturbed function of p53 in DNA repair. [ABSTRACT FROM AUTHOR]

Control of bacterial infection resides in the core of human health and sustainable animal breeding. Vaccines as an economic and efficient immunoprophylaxis have been widely accepted, but mechanisms for vaccines do not fully understand. Information regarding to metabolome in response to vaccines is not available. Here we explore the metabolic features by using GC/MS based metabolic profile and trace metabolic mechanisms in zebrafish ( Dario rerio ) in response to live Edwardsiella tarda vaccine. Pathway enrichment analysis shows that live vaccine activates biosynthesis of unsaturated fatty acids and the TCA cycle and reduces aminoacyl-tRNA biosynthesis, suggesting a metabolic characteristic feature in response to the live vaccine. We further demonstrate that hydroxyl radical is limited during stimulation. Finally, we reveal oleate induces effective protection against E. tarda infection. These results have implications for immunity study that metabolic regulation contributes to immune protection. Our findings enable us to propose novel therapeutic strategies on metabolism against bacterial infections. [ABSTRACT FROM AUTHOR]

Abstract: Microglia are resident immune cells of the central nervous system. Their persistent activation in neurodegenerative diseases, traditionally attributed to neuronal dysfunction, may be due to a microglial failure to modulate the release of cytotoxic mediators such as nitric oxide (NO). The persistent activation of microglia with the subsequent release of NO vis-á-vis the accumulation of redox transition metals such as copper (Cu) in neurodegenerative diseases, prompted the hypothesis that copper would alter NO signaling by changing the redox environment of the cell and that, by altering the fate of NO, microglia would adopt a different phenotype. We have used the microglial cell model, BV2, to examine the effects of Cu(I) on NO production and activation as they have been shown to be phenotypically plastic. Our results show that cell viability is not affected by Cu(I) in BV2 microglia and that it has no effect on iNOS mRNA, protein expression and nitrite release. However, when LPS is added to Cu(I)-treated medium, nitrite release is abrogated while iNOS expression is not significantly altered. This effect is Cu(I)-specific and it is not observed with other non-redox metals, suggesting that Cu(I) modulates NO reactivity. Immunofluorescence analysis shows that the M1 (inflammatory) phenotype of BV2 microglia observed in response to LPS, is shifted to an M2 (adaptive) phenotype when Cu(I) is administered in combination with LPS. This same shift is not observed when iNOS function is inhibited by 1400W. In the present study we show that Cu(I) modulates the release of NO to the media, without altering iNOS expression, and produces phenotypic changes in BV2 microglia. [Copyright &y& Elsevier]

Abstract: Nitric oxide and secondary oxides of nitrogen react with unsaturated fatty acids such as linoleic acid to yield oxidized and nitrated products. Fatty acid nitroalkene derivatives, (e.g. nitrolinoleate [LNO2]) are produced by oxidative inflammatory reactions, detected clinically, display potent electrophilic reactivity and induce post-translational protein modifications that mediate adaptive inflammatory signaling responses. LNO2 signaling was examined in lung epithelial cells because the alveolar compartment is a rich site for the transduction of redox and inflammatory reactions. LNO2 did not directly induce Ca2+ influx in cultured lung epithelial cells, but inhibited bradykinin-induced Ca2+ influx in a cGMP-independent manner. In contrast, LNO2 activated MAP kinase (Erk1/2) by a mechanism independent of bradykinin. It was hypothesized that these unique responses were transduced by activation of different protein kinase C isotypes, supported by the observation that LNO2-mediated inhibition of Ca2+ influx was blocked by the non-selective PKC inhibitors chelerythine chloride and calphostin C, but not by the calcium dependent “classic” PKC inhibitor Gö6976. Western blot analysis showed that atypical PKCζ was activated by LNO2 stimulation, with PKCζ and Erk activation also demonstrated in primary culture of human lung type II cells. Addition of pseudotypical PKCζ substrate peptide reversed LNO2-mediated induction of Ca2+ influx and MAP kinase activation. Finally, the electrophilic nature of LNO2 resulted in a novel mode of PKCζ activation, covalent adduction of the enzyme. In summary, LNO2 mediated signaling in lung type II epithelial cells occurs via a unique pathway involving PKCζ. [Copyright &y& Elsevier]

Abstract: Hsp20, referred to as HspB6, is constitutively expressed in various tissues. Specifically, HspB6 is most highly expressed in different types of muscle including vascular, airway, colonic, bladder, and uterine smooth muscle; cardiac muscle; and skeletal muscle. It can be phosphorylated at Ser-16 by both cAMP- and cGMP-dependent protein kinases (PKA/PKG). Recently, Hsp20 and its phosphorylation have been implicated in multiple physiological and pathophysiological processes including smooth muscle relaxation, platelet aggregation, exercise training, myocardial infarction, atherosclerosis, insulin resistance and Alzheimer''s disease. In the heart, key advances have been made in elucidating the significance of Hsp20 in contractile function and cardioprotection over the last decade. This mini-review highlights exciting findings in animal models and human patients, with special emphasis on the potential salutary effects of Hsp20 in heart disease. This article is part of a special issue entitled "Key Signaling Molecules in Hypertrophy and Heart Failure." [Copyright &y& Elsevier]

Abstract: The JAK/STAT signal transduction pathway plays a critical role in host defence against viral and bacterial infections. In the present study, we report cDNA cloning and characterization of the JAK family (mJAK1-3 and mTYK2) and STAT family members (mSTAT1, mSTAT3-6) from the mandarin fish Siniperca chuatsi. To our knowledge, JAK2, TYK2 and STAT6 genes were cloned from fish for the first time. The mJAK family proteins consist of 1112–1177 residues with a FERM domain, an SH2 domain, a pseudokinase domain, and a tyrosine kinase domain. The mSTAT family members contain 716–786 residues with similar architecture, including an N-terminal domain, a coiled coil domain, a DNA binding domain, a linker domain, an SH2 domain, and a transcription activation domain. Multiple sequence alignments of mJAKs/mSTATs and phylogenetic analysis showed that mJAK1 was closed to mTYK2, and mJAK2 was closed to mJAK3. Quantitative real-time PCR results revealed that mJAK/mSTAT family members were expressed in most tissues examined except muscle. In mandarin fish fry cells, the expressions of IRF-1, Mx, SOCS1 and SOCS3 genes were significantly induced by poly(I:C) stimulation, indicating that the mJAK/mSTAT signal pathway is activated by poly(I:C). Furthermore, expressions of all four mJAKs and four mSTATs were all up-regulated after poly(I:C) stimulation, but expression of mSTAT5 was inhibited by poly(I:C). These results suggest that mandarin fish has the JAK/STAT signal transduction pathways similar to those in mammals, and these signalling pathways may play an important role in regulation of antiviral responses in fish. [Copyright &y& Elsevier]

Abstract: Bone marrow mesenchymal stem cells (MSCs) participate in myocardial repair following myocardial infarction. However, their in vivo reparative capability is limited due to lack of their survival in the infarcted myocardium. To overcome this limitation, we genetically engineered male rat MSCs overexpressing CXCR4 in order to maximize the effect of stromal cell-derived factor-1α (SDF-1α) for cell migration and regeneration. MSCs were isolated from adult male rats and cultured. Adenoviral transduction was carried out to over-express either CXCR4/green fluorescent protein (Ad-CXCR4/GFP) or Ad-null/GFP alone (control). Flow cytometry was used to identify and isolate GFP/CXCR4 over-expressing MSCs for transplantation. Female rats were assigned to one of four groups (n =8 each) to receive GFP-transduced male MSCs (2×106) via tail vein injection 3 days after ligation of the left anterior descending (LAD) coronary artery: GFP-transduced MSCs (Ad-null/GFP-MSCs, group 1) or MSCs over-expressing CXCR4/GFP (Ad-CXCR4/GFP-MSCs, group 2), or Ad-CXCR4/GFP-MSCs plus SDF-1α (50 ng/μl) (Ad-CXCR4/GFP-MSCs/SDF-1α, group 3), or Ad-miRNA targeting CXCR4 plus SDF-1α (Ad-miRNA/GFP-MSCs+SDF-1α treatment, group 4). Cardiodynamic data were obtained 4 weeks after induction of regional myocardial infarction (MI) using echocardiography after which hearts were harvested for immunohistochemical studies. The migration of GFP and Y-chromosome positive cells increased significantly in the peri- and infarct areas of groups 2 and 3 compared to control group (p <0.05), or miRNA-CXCR4 group (p <0.01). The number of CXCR4 positive cells in groups 2, 3 was intimately associated with angiogenesis and myogenesis. MSCs engraftment was blocked by pretreatment with miRNA (group 4). Cardiac function was significantly improved in rats receiving MSCs over-expressing CXCR4 alone or with SDF-1α. The up-regulation of matrix metalloproteinases (MMPs) by CXCR4 overexpressing MSCs perhaps facilitated their engraftment in the collagenous tissue of the infarcted area. CXCR4 over-expression led to enhance in vivo mobilization and engraftment of MSCs into ischemic area where these cells promoted neomyoangiogenesis and alleviated early signs of left ventricular remodeling. [Copyright &y& Elsevier]

Abstract: Junctin is one of the components of the ryanodine receptor Ca release channel complex in sarcoplasmic reticulum. To determine the role of acute alteration of junctin protein levels on cardiomyocyte contractility, we used adenoviral-mediated gene transfer techniques in adult rat cardiomyocytes. Acute downregulation of junctin by 40% resulted in significant increases in cell shortening, rate of contraction (+dL/dt), and rate of relaxation (−dL/dt). The alteration of contractile parameters was associated with increased Ca transient peak and accelerated Ca decay. However, all these contractile and Ca kinetic parameters were depressed significantly when junctin levels were upregulated by 60%. Importantly, there were no alterations in other Ca-cycling protein levels when junctin levels were either decreased or increased. These findings suggest that junctin plays a prominent role in cardiomyocyte Ca-cycling and contractility. [Copyright &y& Elsevier]

Abstract: It has been reported that N-methyl-d-aspartate (NMDA) receptor is implicated in drug addiction and antagonists of the NMDA receptor complex can inhibit the development and expression of conditioned place preference (CPP) induced by several addictive drugs, implying that this class of compounds might be considered as candidate for the treatment of substance abuse. To explore this possibility, it is important to evaluate whether the inhibitory effect of NMDA receptor antagonists would be confined to behaviors produced by drugs of abuse only, but not by natural reinforcers. According to the quantitative changes of NMDA receptor subunits, including NR1, NR2A, and NR2B, induced by diverse types of reinforcers, we chose NR2B subunit as the target of research. Experimental results showed that (1) an augmented expression of NR2B subunit was revealed by Western blotting in the nucleus accumbens (NAc) and the hippocampus in rats with CPP induced by morphine, but not by natural rewards such as food, novel environment and social interaction. (2) Ifenprodil, an antagonist highly selective for NR2B subunit of the NMDA receptor, produced a dose-dependent reduction in CPP induced by morphine and novel environment, but not that by food consumption and social interaction. Taking together, these findings suggested that NR2B containing NMDA receptor may be more involved with morphine reward rather than natural rewards, and that antagonism of NR2B may have a potential for the treatment of morphine abuse. [Copyright &y& Elsevier]

In oogamous reproduction of multicellular organisms, a striking phenomenon is the prevailing synchronous development of male germ cells connected by wide intercellular bridges (IBs, 0.1–2 μm), which is well conserved in both animal and plant species ranging from algae to human. In the literature, IBs are believed either to allow genetically segregated haploid spermatids to share diploid gene products after meiosis, or to mediate rapid transfer of some vital signals or nutrients. Although intercellular sharing of gene transcripts has experimental evidences, these hypotheses are still not satisfactory. To explore the unknown roles of IB, we assume that developing male germ cells may be especially sensitive to stochastic gene expression to become heterogeneous. To achieve best gamete quality, such heterogeneity must be eliminated so that relatively uniform gametes with normal functions can be produced. Development within a common syncytium may be the only way for this purpose. The process may require not only the intercellular exchange of a few molecular signals but also the mixing of protoplasm between the connected cells so that they have similar levels/states of mRNAs, proteins and organelles, which can be achieved only through wide IBs. This hypothesis can explain some quite intriguing aspects of male gametogenesis and provide unique predictions that can be tested experimentally. [Copyright &y& Elsevier]

The investigation of factors that regulate expression of CC-chemokines, the important mediators in immune responses and inflammation processes, has an important significance in understanding the immunopathogenesis of liver diseases. We examined the role of interleukin-1β (IL-1β), a multifunctional cytokine, in regulating the expression of macrophage inflammatory protein (MIP)-1β in human hepatocytes (Huh7 and HepG2). IL-1β significantly enhanced MIP-1β expression in these cells at both the mRNA and protein levels. Cytokine-enriched supernatants from monocyte-derived macrophage (MDM) cultures also induced MIP-1β expression. IL-1β is responsible for MDM supernatant-mediated up-regulation of MIP-1β since the antibody to IL-1β abolished MDM supernatant action. Investigation of the mechanism involved in MIP-1β induction by IL-1β showed that IL-1β activated the nuclear factor kappa B (NF-κB) promoter in Huh7 cells. In addition, caffeic acid phenethyl ester (CAPE), a specific inhibitor of the activation of NF-κB, not only abolished IL-1β-mediated NF-κB promoter activation, but also blocked IL-1β-induced MIP-1β expression. These observations suggest that IL-1β-mediated up-regulation of MIP-1β production in the hepatic cells may contribute a critical mechanism for continuous recruitment of inflammatory cell to liver and maintenance of inflammation. [Copyright &y& Elsevier]

[Display omitted] The structural defects and oxygen-containing functional groups of carbon materials as electrode materials for supercapacitors or capacitive deionization devices are critical to their electrochemical performance. The tuning of surface oxygen-containing functional groups and carbon defects during pyrolysis is key to achieve a high performance in ion storage. Herein, quinonyl-dominant defective porous carbon is prepared by a pyrolysis and cross-linking route using lavender stem and potassium acetate as precursor. Benefiting from the presence of abundant defect and surface quinonyl groups, porous carbon shows an ultra-high specific capacitance of 401 F g−1 (1 A g−1) and a high capacitance retention of 63% at a high current density of 100 A g−1 in a KOH solution. Meanwhile, as a capacitive deionization electrode material, it also exhibited a high adsorption capacity of 25.5 mg g−1 in 500 mg L-1 NaCl solution at 1.2 V. Theoretical density functional theory (DFT) calculation demonstrates that surface quinonyl groups and carbon defects can synergistically facilitate the adsorption of K+ and Na+ during the charge/discharge process. This work provides a new perspective for understanding the role of surface oxygen-containing groups and intrinsic defects of porous carbon materials in electrochemical energy storage and desalination applications. [ABSTRACT FROM AUTHOR]

Mechanisms by which vaccines enhance immunity to combat bacterial pathogens are not fully understood. Recently, we have found that live Edwardsiella tarda vaccine enhances ability against the bacterial challenge by metabolic modulation in zebrafish. Here we first explored the metabolic modulation promoted by inactivated E. tarda to eliminate the pathogen. Inactivated E. tarda vaccine modulated a similar metabolome to combat with the pathogen in zebrafish as live E. tarda vaccine did. Specifically, both vaccines promoted biosynthesis of unsaturated fatty acids and the TCA cycle. However, due to relatively higher activated TCA cycle in inactivated vaccine than live vaccine, live vaccine promoted higher abundance of palmitate than inactivated vaccine. Consistently, the protection against E. tarda challenge was palmitate dose-dependent. Live vaccine activated higher expression of IL-1β, IL-8,Cox-2 genes and lower expression of IL-15, IL-21 genes than inactivated vaccine, which is similar to the results stimulated by high and low doses of palmitate, respectively. These findings indicate live and inactivated E. tarda vaccines stimulate differential abundances of palmitate that contribute to differential innate immunities against bacterial infection. Thus, metabolic environment contributes to immune response. Image 1 • Metabolic feature is revealled in zebrafish immunized with dead E. tarda vaccine. • Similar metabolic profile but low palmitate is detected compared with live vaccine. • Palmitate induces immune effective protection in a dose-dependent manner. • High and low palmitate induce altered innate immunity as live and dead vaccines do. • Metabolic environment contributes to immune response. [ABSTRACT FROM AUTHOR]

Wear particles liberated from the surface of prostheses are considered to be main reason for osteoclast bone resorption and that extensive osteoclastogenesis leads to peri-implant osteolysis and subsequent prosthetic loosening. The aim of this study was to assess the effect of rifampin on osteoclastogenesis and titanium (Ti) particle-induced osteolysis. The Ti particle-induced osteolysis mouse calvarial model and bone marrow-derived macrophages (BMMs) were used. Rifampin, at dose of 10 or 50 mg/kg/day, was respectively given intraperitoneally for 14 days in vivo . The calvariae were removed and processed for Further histological analysis. In vitro, osteoclasts were generated from mouse BMMs with receptor activator of nuclear factor-κB ligand (RANKL) and the macrophage colony stimulating factor. Rifampin at different concentrations was added to the medium. The cell viability, tartrate-resistant acid phosphatase (TRAP) staining, TRAP activity and resorption on bone slices were analysis. Osteoclast-specific genes and RANKL-induced MAPKs signaling were tested for further study of the mechanism. Rifampin inhibited Ti-induced osteolysis and osteoclastogenesis in vivo . In vitro data indicated that rifampin suppressed osteoclast differentiation and bone resorption in a dose-dependent manner. Moreover, rifampin significantly reduced the expression of osteoclast-specific markers, including TRAP, cathepsin K, V-ATPase d2, V-ATPase a3, c-Fos, and nuclear factor of activated T cells (NFAT) c1. Further investigation revealed that rifampin inhibited osteoclast formation by specifically abrogating RANKL-induced p38 and NF-κB signaling. Rifampin had significant potential for the treatment of particle-induced peri-implant osteolysis and other diseases caused by excessive osteoclast formation and function. [ABSTRACT FROM AUTHOR]

Highlights: [•] Dietary supplementation with tocopherol increases the concentration in the lung. [•] Tocopherol blocks the bleomycin-dependent increase of NO metabolites. [•] Tocopherol reduces bleomycin-stimulated inflammation. [•] Bleomycin-dependent loss of lung mechanical function due is reduced by tocopherol. [Copyright &y& Elsevier]

Abstract: The NF-κB/IκBα pathway plays an important role in the regulation of immune and inflammatory responses. IκBα is an inhibitory molecule that sequesters transcription activator NF-κB dimer in the cytoplasm of unstimulated cells. Here, we isolated the full-length cDNAs of the mandarin fish (Siniperca chuatsi) alpha inhibitor of NF-κB (ScIκBα) and p65 NF-κB (Scp65). Multiple sequence alignments showed that the amino acid sequences of both ScIκBα and Scp65 contain conserved domains similar to those of mammalian counterparts. Protein pull-down and coimmunoprecipitation assays showed that ScIκBα directly bound with Scp65. Real-time quantitative PCR analysis showed that ScIκBα mRNA was constitutive in all mandarin fish tissues detected. After challenge with infectious spleen and kidney necrosis virus (ISKNV), the mRNA level of ScIκBα was decreased nearly 6 fold in the spleen. This result suggests that the NF-κB/IκBα pathway in mandarin fish may play a role in the immune response against ISKNV. [Copyright &y& Elsevier]

Abstract: Glutamate receptors are known to be densely distributed in the forebrain rewarding circuits, and glutamatergic transmission is actively involved in the regulation of rewarding and reinstating effects of drugs of abuse. Here we investigated the possible involvement of the N-methyl-d-aspartate (NMDA) receptors in the reinstatement of extinguished morphine conditioned place preference (CPP) in rats. We found that previously extinguished morphine (3 mg/kg, i.p.) CPP was markedly reinstated by a priming injection of morphine (2 mg/kg, i.p.) or an acute environmental stressor (forced swim for 10 min), but not by the stress induced by a 24-h food deprivation. Parallel with this, protein levels of the NMDA receptor 2B subunit (NR2B) were elevated in the nucleus accumbens (NAc) and the hippocampus, but not the prefrontal cortex, of reinstated rats. Systemic administration of an NR2B selective antagonist ifenprodil (1, 3, 10 mg/kg, i.p.) attenuated the reinstatement induced by a priming morphine injection, although not by the forced swim. Ifenprodil (2.0 μg/rat) directly injected into the NAc shell or the CA1 region of the dorsal hippocampus produced a similar effect. These results indicate that the NR2B-containing NMDA receptors in the NAc and the dorsal hippocampus play a significant role in mediating the reinstatement of rewarding responses to morphine. [Copyright &y& Elsevier]

Tiger frog virus (TFV) belongs to the genus Ranavirus (family Iridoviridae) and causes significant harm in cultured frogs, resulting in substantial losses in ecological and economic field in Southern China. Attachment is the first step in viral life cycle, which is dependent on the interactions of virions with extracellular matrix (ECM) components. Studying this process will help in understanding virus infection and controlling viral diseases. In this study, the roles of primary ECM components in TFV attachment were investigated. The results on the kinetics of virus attachment showed TFV successful attachment to the cell surface as a relatively rapid process after TFV was used to inoculate cells for 10 min at 4 °C. Western blot and quantitative PCR analyses results showed that soluble fibronectin, collagen IV, laminin, or hyaluronic acid treatment with TFV caused no significant effect on virus attachment. Soluble heparin, heparan sulfate and chondroitin sulfate A/B could inhibit TFV attachment in a dose-dependent manner. Enzymic digestion by cell surface heparin/heparan sulfate using heparinase I, II, and III could significantly prevent TFV attachment, suggesting that heparan sulfate plays an important role in TFV attachment. Furthermore, the binding assays of heparin–agarose beads and virion showed that TFV virions specifically bound with heparin in a dose-dependent manner. Given that heparin is a structural analogue of heparan sulfate, the above results suggest that heparan sulfate might serve as an attachment factor of TFV infection. Our work would be beneficial to understand the mechanisms of TFV attachment and the interactions of TFV with cellular receptor(s). • Kinetics of virus attachment showed that TFV attachment is a relatively rapid process. • Extracellular matrix glycosaminoglycans play an important role in TFV attachment. • Heparan sulfate may be an attachment factor for TFV infection. [ABSTRACT FROM AUTHOR]

• The AMO with hollow flower-like was prepared by nanosheets self-assembled. • Controllable anchoring nanoparticles on the AMO by hydrophobic driving mechanism. • The TEOOS and PVP are selected as effective hydrophobic modified reagents. • The prepared catalyst presents excellent catalytic reaction performance. • A possible mechanism illustrates dehydrogenative homo-coupling of pyridines. In this study, we synthesized a series of Ni-based catalysts with different CeO 2 contents (0, 1, 2, 5 and 10 wt%) anchoring on the Al-Mg oxide spheres (AMO) with three-dimensional hollow flower-like structure. The triethoxyoctylsilane (TEOOS) and polyvinylpyrrolidone (PVP) were selected as effective modified reagents to introduce hydrophobic functionality on the surface of AMO and Ni-CeO 2 nanoparticles, respectively, for controllable anchoring Ni-CeO 2 nanoparticles on AMO by hydrophobic interaction driving mechanism. The effects of CeO 2 content on the texture, dispersion of Ni species, the reducibility of Ni2+, surface electronic states and acidity were investigated in detail. The Ni-5CeO 2 /AMO was found to achieve the highest 2,2′-bipyridine selectivity of 90.6% and maximum turnover number (TON) of 8611, specifically. Moreover, the Ni-5CeO 2 /AMO exhibits superior stability, maintaining 89% of 2,2′-bipyridine yield after 6 runs, due to the mitigated nickel leaching and promotion of the desorption of 2,2′-bipyridine. Furthermore, with the help of DFT calculations, a possible reaction mechanism was discussed and illustrated. [ABSTRACT FROM AUTHOR]

In mouse, there are four stearoyl-CoA desaturase isoforms (SCD1-4) that catalyze the synthesis of monounsaturated fatty acids. Previously, we have shown that mice harboring a whole body deletion of the SCD1 isoform (SCD1KO) are protected from diet and genetically induced adiposity. Here, we report that global deletion of the SCD2 isoform (SCD2KO) provides a similar protective effect against the onset of both high-fat diet (HFD) and high-carbohydrate diet (HCD) induced adiposity. After 10 weeks of HFD feeding or 6 weeks of HCD feeding, SCD2KO mice failed to gain weight and had decreased fat mass. On HFD, SCD2KO mice remained glucose and insulin tolerant. Lastly, the markers for energy expenditure, UCP1 and PGC-1α, were increased in the brown adipose tissue of HFD fed SCD2KO mice. • Stearoyl-CoA desaturase-2 deficiency protects against diet-induced weight gain and adiposity. • Stearoyl-CoA desaturase-2 deficiency improves glucose and insulin tolerance. • Stearoyl-CoA desaturase-2 deficiency alters brain fatty acid composition. [ABSTRACT FROM AUTHOR]

The mandarin fish Siniperca chuatsi is a cultured freshwater fish species that is popular in China because of its high market value. With the development of high-density cultural mode in mandarin fish, viral diseases such as Infectious spleen and kidney necrosis virus (ISKNV) are becoming increasingly serious. Stimulator of interferon genes (STING) is a central component in the innate immune response to cytosolic DNA and RNA derived from different pathogens. However, the roles of STING in innate immune response of mandarin fish remain unknown. In the present study, S. chuatsi STING (sc STING)-mediated host immune response against ISKNV infection was investigated. Sc STING transcription level increased remarkably in response to ISKNV infection, LPS, PMA, or poly (I:C) stimulation in mandarin fish fry (MFF-1) cells. Immunofluorescence results showed that sc STING localized majorly in the endoplasmic reticulum. sc STING overexpression remarkably increased the expression levels of scIFN-h , scMx , scISG15 , scPKR , scViperin , scIL-1β , scIL-18 , and scTNF-α genes. IFN-β-luciferase report assay results showed that the relative expressions of luciferin were remarkably increased in MFF-1 cells. Site mutation of serine (S) on C-terminus of sc STING showed that both S388 and S396 were important for mediated signaling. Furthermore, sc STING overexpression inhibited ISKNV infection, and knockdown of sc STING promoted ISKNV infection, indicating that sc STING could suppress ISKNV infection in MFF-1 cells. These observations suggested that the sc STING played an important role in innate immune against ISKNV infection. Our work would help elucidate the roles of teleost fish STING in innate immunity. • sc STING can evoke interferon and inflammatory immune responses. • The Ser388 and Ser396 sites are crucial for sc STING-mediated signaling. • sc STING mediates host antiviral immune response against ISKNV infection. [ABSTRACT FROM AUTHOR]

Transition metals are increasingly attracting interest in electrocatalysts for use in water decomposition due to their excellent catalytic activity and stability. Simultaneously, metal-organic frameworks with designable metal ion centers and organic ligands are the promising precursors for the one-step synthesis of metal encapsulated in carbon composites for alkaline hydrogen evolution reaction (HER). Herein, we report the successful construction of Co-CoO/ZnFe 2 O 4 encapsulated in carbon nanowires (Co-CoO/ZnFe 2 O 4 @CNWs) by annealing as-synthesized nanowire Co/Zn/Fe-MOF at 400 °C in N 2. This structure provides the rich defect sites and active centers, and the synergy of Co, CoO and ZnFe 2 O 4 lead to efficient hydrogen evolution when the composite is used as a catalytic electrode for HER in 1.0 M KOH. The catalyst shows a low initial overpotential (97 mV) and a small Tafel slope (138 mV dec−1), and the overpotential at 10 mA cm−2 is only 226 mV. In addition, this composite material exhibits excellent long-term durability even after 1000 cycles. It is expected that it is a potential alternative catalyst for rational utilization in the field of electrolytic water decomposition. [ABSTRACT FROM AUTHOR]

Mandarin fish (Siniperca chuatsi) is a significant cultured species with high added value in China. With the expansion of farming, diseases of mandarin fish such as Infectious spleen and kidney necrosis virus (ISKNV) diseases are becoming more and more serious. Human endogenous retrovirus subfamily H long terminal repeat associating protein 2 (HHLA2) is a type 1 transmembrane molecule with three extracellular Ig domains (IgV-IgC-IgV) and plays important roles in the T cell proliferation and tumorigenesis. The HHLA2-homologues have not been found in virus. In this study, a viral HHLA2 protein encoded by ISKNV ORF069L was identified and the virulence of the deleted ORF069L reconstruction ISKNV strain (Δ ORF069L) was investigated. ISKNV ORF069L gene was predicted to encode a 222-amino acids peptide. The bioinformation analysis revealed that ISKNV ORF069L contained an Ig HHLA2 domain and was homologous to vertebrate B7-CD28 family proteins. The recombinant virus strain of Δ ORF069L was constructed by homologous recombination technology. The virus titer and growth curves between ISKNV wild type (WT) and Δ ORF069L on cellular level showed no significant differences indicating that the ORF069L did not influence the ISKNV replication. The expression levels of immune-related genes (Mx1, IL-1β, IL-8, TNF-a and IgM) were increased in fish infected with Δ ORF069L , compared to those in fish infected with ISKNV WT. Furthermore, the lethality caused by Δ ORF069L declined by 40% compared with ISKNV WT, indicating that ORF069L was a virulence gene of ISKNV. Most importantly, the protection rate was nearly 100% for fish immunized with Δ ORF069L strain. Those results suggested that Δ ORF069L could be developed as a potential attenuated vaccine against ISKNV. Our work will be beneficial to promote the development of gene deletion attenuated vaccines for ISKNV disease. • A viral HHLA2 protein encoded by ISKNV ORF069L was identified. • A recombinant virus of ISKNV ORF069L deletion (Δ ORF069L) was constructed. • ISKNV ORF069L was an important virulence gene. • The protection rate was nearly 100% for fish immunized with Δ ORF069L strain. • Δ ORF069L could be developed as an attenuated vaccine against ISKNV. [ABSTRACT FROM AUTHOR]

Mandarin fish (Siniperca chuatsi) is a universally farmed fish species in China and has a large farming scale and economic value. With the high-density cultural mode in mandarin fish, viral diseases, such as infectious spleen and kidney necrosis virus (ISKNV) and Siniperca chuatsi rhabdovirus (SCRV), have increased loss, which has seriously restricted the development of aquaculture. Y-Box binding protein 1 (YB-1) is a member of cold shock protein family that regulates multiple cellular processes. The roles of mammalian YB-1 protein in environmental stress and innate immunity have been studied well, but its roles in teleost fishes remain unknown. In the present study, the characteristic of S. chuatsi YB-1 (sc YB-1) and its roles in cold stress and virus infection were investigated. The sc YB-1 obtained an 1541 bp cDNA that contains a 903 bp open reading frame encoding a protein of 300 amino acids. Tissue distribution results showed that the sc YB-1 is a ubiquitously expressed gene found among tissues from mandarin fish. Overexpression of sc YB-1 can increase the expression levels of cold shock-responsive genes, such as scHsc70a , scHsc70b , and scp53. Furthermore, the role of sc YB-1 in innate immunity was also investigated in mandarin fish fry (MFF-1) cells. The expression level of sc YB-1 was significant change in response to poly (I:C), poly (dG:dC), PMA, ISKNV, or SCRV stimulation. The overexpression of sc YB-1 can significantly increase the expression levels of NF-κB-responsive genes, including scIL-8, scTNF-α , and scIFN-h. The NF-κB-luciferase report assay results showed that the relative expression of luciferin was significantly increased in the cells overexpressed with sc YB-1 compared with those in cells overexpressed with control plasmid. These results indicate that sc YB-1 can induce the NF-κB signaling pathway in MFF-1 cells. Overexpressed sc YB-1 can downregulate the expression of ISKNV viral major capsid protein (mcp) gene but upregulates the expression of SCRV mcp gene. Moreover, knockdown of sc YB-1 using siRNA can upregulate the expression of ISKNV mcp gene but downregulates the expression of SCRV mcp gene. These results indicate that sc YB-1 suppresses ISKNV infection while enhancing SCRV infection. The above observations suggest that sc YB-1 is involved in cold stress and virus infection. Our study will provide an insight into the roles of teleost fish YB-1 protein in stress response and innate immunity. • S. chuatsi YB-1 (sc YB-1) is involved in the cold stress response in mandarin fish. • sc YB-1 can regulate the NF-κB signaling pathway in MFF-1 cells. • sc YB-1 suppresses ISKNV infection but enhances SCRV infection. [ABSTRACT FROM AUTHOR]

Ranaviruses belong to the family Iridoviridae , and have become a serious threat to both farmed and natural populations of fish and amphibians. Previous reports showed that ranaviruses could encode viral Bcl-2 family-like proteins (vBcl-2), which play a critical role in the regulation of cell apoptosis. However, the mechanism of ranaviruses vBcl-2 interactions with host protein in mediating apoptosis remains unknown. Tiger frog virus (TFV) belonging to the genus Ranavirus has been isolated from infected tadpoles of Rana tigrina rugulosa , and it causes a high mortality rate among tiger frog tadpoles cultured in southern China. This study elucidated the molecular mechanism underlying the interaction of TFV ORF104R with the VDAC2 protein to regulate cell apoptosis. TFV ORF104R is highly similar to other ranaviruses vBcl-2 and host Mcl-1 proteins, indicating that TFV ORF104R is a postulate vBcl-2 protein. Transcription and protein expression levels showed that TFV orf104r was a late viral gene. Western blot results suggested that TFV ORF104R was a viral structural protein. Subcellular localization analysis indicated that TFV ORF104R was predominantly colocalized with the mitochondria. Overexpressed TFV ORF104R could suppress the release of cytochrome C and the activities of caspase-9 and caspase-3. These results indicated that TFV ORF104R might play an important role in anti-apoptosis. Furthermore, the interaction between TFV ORF104R and VDAC2 was detected by co-immunoprecipitation in vitro. The above observations suggest that the molecular mechanism of TFV-regulated anti-apoptosis is through the interaction of TFV ORF104R with the VDAC2 protein. Our study provided a mechanistic basis for the ranaviruses vBcl-2-mediated inhibition of apoptosis and improved the understanding on how TFV subverts host defense mechanisms in vivo. • TFV ORF104R was a late viral protein as viral structural protein of virion particles. • TFV ORF104R interacted with cellular VDAC2 protein. • TFV ORF104R inhibited cell apoptosis though intrinsic pathway. [ABSTRACT FROM AUTHOR]

Mandarin fish (Siniperca chuatsi) is a popular cultured freshwater fish species due to its high market value in China. With increasing density of breeding, mandarin fish is often cultured under low environmental oxygen concentrations (hypoxia). In this study, the relative expression levels of hypoxia response element (HRE)-luciferase reporter and the HIF signaling pathway downstream genes (scldha , scvegf , and scglut-1) were significantly increased by hypoxic stress, thereby indicating that mandarin fish has an HIF signaling pathway. The mandarin fish HIF-1α (sc HIF-1α) was also characterized. Multiple sequence alignments showed that sc HIF-1α presented similar architectures to other known vertebrates. Subcellular localization analysis showed that sc HIF-1α was mainly located in the nucleus of the mandarin fish fry-1 (MFF-1) cells. The role of sc HIF-1α in the regulation of the HIF signaling pathway was confirmed. Overexpression of sc HIF-1α could induce the HIF signaling pathway, whereas knockdown of sc HIF-1α inhibited the activity of the HIF-1 signaling pathway. Tissue distribution analysis showed that schif-1α was significantly highly expressed in the blood, heart, and liver, which indicated that the main function of sc HIF-1α was closely related to the circulatory system. Furthermore, sc HIF-1α expression was significantly induced by poly I:C, poly dG:dC or PMA, thereby indicating that sc HIF-1α was involved in the immune response. HIF-1α plays an important role in pathogen infections in mammals, but its role in fish is rarely investigated. Overexpression of sc HIF-1α could inhibit MRV and SCRV infections, whereas knockdown of sc HIF-1α could promote such infections. Those results suggested that sc HIF-1α played an important role in fish virus infection. Our study will help understand the hypoxia associated with the outbreaks of aquatic viral disease. • The sc HIF signaling pathway exists in MFF-1 cells. • The expression of sc HIF-1α was up-regulated in response to the immune challenge. • sc HIF-1α could inhibit MRV and SCRV infections. [ABSTRACT FROM AUTHOR]

Many desired phenotypes for producing cellulosic biofuels are often observed in industrial Saccharomyces cerevisiae strains. However, many industrial yeast strains are polyploid and have low spore viability, making it difficult to use these strains for metabolic engineering applications. We selected the polyploid industrial strain S. cerevisiae ATCC 4124 exhibiting rapid glucose fermentation capability, high ethanol productivity, strong heat and inhibitor tolerance in order to construct an optimal yeast strain for producing cellulosic ethanol. Here, we focused on developing a general approach and high-throughput screening method to isolate stable haploid segregants derived from a polyploid parent, such as triploid ATCC 4124 with a poor spore viability. Specifically, we deleted the HO genes, performed random sporulation, and screened the resulting segregants based on growth rate, mating type, and ploidy. Only one stable haploid derivative (4124-S60) was isolated, while 14 other segregants with a stable mating type were aneuploid. The 4124-S60 strain inherited only a subset of desirable traits present in the parent strain, same as other aneuploids, suggesting that glucose fermentation and specific ethanol productivity are likely to be genetically complex traits and/or they might depend on ploidy. Nonetheless, the 4124-60 strain did inherit the ability to tolerate fermentation inhibitors. When additional genetic perturbations known to improve xylose fermentation were introduced into the 4124-60 strain, the resulting engineered strain (IIK1) was able to ferment a Miscanthus hydrolysate better than a previously engineered laboratory strain (SR8), built by making the same genetic changes. However, the IIK1 strain showed higher glycerol and xylitol yields than the SR8 strain. In order to decrease glycerol and xylitol production, an NADH-dependent acetate reduction pathway was introduced into the IIK1 strain. By consuming 2.4 g/L of acetate, the resulting strain (IIK1A) exhibited a 14% higher ethanol yield and 46% lower byproduct yield than the IIK1 strain from anaerobic fermentation of the Miscanthus hydrolysate. Our results demonstrate that industrial yeast strains can be engineered via haploid isolation. The isolated haploid strain (4124-S60) can be used for metabolic engineering to produce fuels and chemicals. [ABSTRACT FROM AUTHOR]

Autophagy is an evolutionarily conserved biological process in all eukaryotes for the degradation of intracellular components for nutrient recycling. Autophagy is known to be involved in responses to low nitrogen stress in Arabidopsis . Foxtail millet has strong abiotic stress resistance to both low nutrient and drought stress. However, to date, there have only been a few genes reported to be related with abiotic stress resistance in foxtail millet. In this study, we identified an autophagy-related gene, SiATG8a, from foxtail millet. SiATG8a is mainly expressed in stems and its expression was dramatically induced by drought stress and nitrogen starvation treatments. SiATG8a was localized in the membrane and cytoplasm of foxtail millet. Overexpression of SiATG8a in Arabidopsis conferred tolerance to both nitrogen starvation and to drought stress. Under nitrogen starvation conditions, the SiATG8a transgenic plants had larger root and leaf areas and accumulated more total nitrogen than wild-type plants. The transgenic plants had lower total protein concentrations than did the WT plants. Under drought stress, the SiATG8a transgenic plants had higher survival rates, chlorophyll content, and proline content, but had lower MDA content than wild type plants. Taken together, our results represent the first identified case where overexpression of autophagy related gene can simultaneously improve plant resistance to low nitrogen and drought stresses. These findings implicate plant autophagy in plant stress responses to low nitrogen and drought and should be helpful in efforts to improve stresses resistance to nitrogen starvation and drought of crops by genetic transformation. [ABSTRACT FROM AUTHOR]

GLP-1 can help to overcome problems of liver cells metabolism, not only pancreatic cell. But the explicit mechanism of this effect remains unclear. In recent years, microRNAs have received the attention of researchers and some microRNAs have important implications for diabetes. The mitochondrial protective gene PGC-1α is also closely related to diabetes, and UCP2 is related to anti-mitochondrial oxidative stress, but the mechanism of action of these genes is unclear. In this study, we used HepG2 cell line and used the cell counting kit (CCK) to measure the cell viability with GLP-1(7–36) and/or glucotoxicity. To investigate alterations in gene expression resulting from incubation with GLP-1 (7–36) or hyperglycaemia, the RNA expression levels of miR-23a, PGC-1α, Bak, Bax and UCP2 were quantified using real-time PCR. The protein levels of PGC-1α were determined by western blot. The role of miR-23a in the regulation of PGC-1α was further assessed through cell transfection to downregulate of miR-23a expression. In this study, the viability of HepG2 hepatocytes was decreased under hyperglycaemia, but incubation with 10 nmol/L GLP-1 (7–36) amide for 24 h significantly increased the viability and decreased the mRNA expression levels of Bax and Bak. Incubation with GLP-1(7–36) amide for 24 h attenuated the RNA expression of miR-23a and increased the mRNA and protein expression of PGC-1α. Inhibition of miR-23a expression by cell transfection led to increases in the mRNA and protein expression of PGC-1α. In addition, the mRNA expression of UCP2 increased after incubation with GLP-1(7–36) for 24 h. In conclusion, GLP-1 induced increased expression of mitochondrial protective gene PGC-1α by downregulating miR-23a to inhibit hepatocyte apoptosis and also enhanced UCP2 to reduce apoptosis. [ABSTRACT FROM AUTHOR]

The reparative reaction is considered to be important during the occurrence of collapse in the femoral head with osteonecrosis (ONFH), but little is known about the long-term reparative process. The aim of this study was to determine and analyze the altered microRNA expression profile in the reparative interface of ONFH, and further validate the expression of the involved genes in the predicted pathways. Microarray analysis was performed comparing the reparative interface of patients with ONFH and normal tissue of patients with fresh femoral neck fracture (FNF) and partly validated by real-time PCR. Potential target genes of differentially expressed miRNAs were predicted by TargetScan and miRanda, and the target genes were used for further bioinformatics analysis such as Gene Ontology and Pathway assay. The filtered miRNAs and genes in the predict pathways were further examined by real-time PCR in another 6 independent ONFH patients. Among the 2578 miRNAs identified, 17 were consistently differentially expressed, 12 of which are up-regulated and 5 down-regulated. GO classification showed that the predicted target genes of these miRNAs are involved in signal transduction, cell differentiation, methylation, cell growth and apoptosis. The Kyoto Encyclopedia of Genes and Genomes (KEGG) classification indicated that these genes play a role in angiogenesis and Wnt signaling pathways. The expression of miR-34a and miR-146a and genes in the predict pathways were significantly up-regulated. This study presented a global view of miRNA expression in the reparative interface of osteonecrosis. In addition, our data provided novel and robust information for further researches in the pathogenesis and molecular events of ONFH. [ABSTRACT FROM AUTHOR]

Nicaraven, a hydroxyl radical-specific scavenger has been demonstrated to attenuate radiation injury in hematopoietic stem cells with 5 Gy γ-ray exposures. We explored the effect and related mechanisms of nicaraven for protecting radiation injury induced by sequential exposures to a relatively lower dose γ-ray. C57BL/6 mice were given nicaraven or placebo within 30 min before exposure to 50 mGy γ-ray daily for 30 days in sequences (cumulative dose of 1.5 Gy). Mice were victimized 24 h after the last radiation exposure, and the number, function and oxidative stress of hematopoietic stem cells were quantitatively estimated. We also compared the gene expression in these purified stem cells from mice received nicaraven and placebo treatment. Nicaraven increased the number of c-kit + stem/progenitor cells in bone marrow and peripheral blood, with a recovery rate around 60–90% of age-matched non-irradiated healthy mice. The potency of colony forming from hematopoietic stem/progenitor cells as indicator of function was completely protected with nicaraven treatment. Furthermore, nicaraven treatment changed the expression of many genes associated to DNA repair, inflammatory response, and immunomodulation in c-kit + stem/progenitor cells. Nicaraven effectively protected against damages of hematopoietic stem/progenitor cells induced by sequential exposures to a relatively low dose radiation, via complex mechanisms. [ABSTRACT FROM AUTHOR]

Exosomes, nano-vesicles naturally released from living cells, have been well recognized to play critical roles in mediating cell-to-cell communication. Given that diabetic hearts exhibit insufficient angiogenesis, it is significant to test whether diabetic cardiomyocyte-derived exosomes possess any capacity in regulating angiogenesis. In this study, we first observed that both proliferation and migration of mouse cardiac endothelial cells (MCECs) were inhibited when co-cultured with cardiomyocytes isolated from adult Goto-Kakizaki (GK) rats, a commonly used animal model of type 2 diabetes. However, GK-myocyte-mediated anti-angiogenic effects were negated upon addition of GW4869, an inhibitor of exosome formation/release, into the co-cultures. Next, exosomes were purified from the myocyte culture supernatants by differential centrifugation. While exosomes derived from GK myocytes (GK-exosomes) displayed similar size and molecular markers (CD63 and CD81) to those originated from the control Wistar rat myocytes (WT-exosomes), their regulatory role in angiogenesis is opposite. We observed that the MCEC proliferation, migration and tube-like formation were inhibited by GK-exosomes, but were promoted by WT-exosomes. Mechanistically, we found that GK-exosomes encapsulated higher levels of miR-320 and lower levels of miR-126 compared to WT-exosomes. Furthermore, GK-exosomes were effectively taken up by MCECs and delivered miR-320. In addition, transportation of miR-320 from myocytes to MCECs could be blocked by GW4869. Importantly, the exosomal miR-320 functionally down-regulated its target genes (IGF-1, Hsp20 and Ets2) in recipient MCECs, and overexpression of miR-320 inhibited MCEC migration and tube formation. GK exosome-mediated inhibitory effects on angiogenesis were removed by knockdown of miR-320. Together, these data indicate that cardiomyocytes exert an anti-angiogenic function in type 2 diabetic rats through exosomal transfer of miR-320 into endothelial cells. Thus, our study provides a novel mechanism underlying diabetes mellitus-induced myocardial vascular deficiency which may be caused by secretion of anti-angiogenic exosomes from cardiomyocyes. [ABSTRACT FROM AUTHOR]

Abstract: Depressed Ca-handling in cardiomyocytes is frequently attributed to impaired sarcoplasmic reticulum (SR) function in human and experimental heart failure. Phospholamban (PLN) is a key regulator of SR and cardiac function, and PLN mutations in humans have been associated with dilated cardiomyopathy (DCM). We previously reported the deletion of the highly conserved amino acid residue arginine 14 (nucleic acids 39, 40 and 41) in DCM patients. This basic amino acid is important in maintaining the upstream consensus sequence for PKA phosphorylation of Ser 16 in PLN. To assess the function of this mutant PLN, we introduced the PLN-R14Del in cardiac myocytes of the PLN null mouse. Transgenic lines expressing mutant PLN-R14Del at similar protein levels to wild types exhibited no inhibition of the initial rates of oxalate-facilitated SR Ca uptake compared to PLN-knockouts (PLN-KO). The contractile parameters and Ca-kinetics also remained highly stimulated in PLN-R14Del cardiomyocytes, similar to PLN-KO, and isoproterenol did not further stimulate these hyper-contractile basal parameters. Consistent with the lack of inhibition on SR Ca-transport and contractility, confocal microscopy indicated that the PLN-R14Del failed to co-localize with SERCA2a. Moreover, PLN-R14Del did not co-immunoprecipitate with SERCA2a (as did WT-PLN), but rather co-immunoprecipitated with the sarcolemmal Na/K-ATPase (NKA) and stimulated NKA activity. In addition, studies in HEK cells indicated significant fluorescence resonance energy transfer between PLN-R14Del-YFP and NKAα1-CFP, but not with the NKA regulator phospholemman. Despite the enhanced cardiac function in PLN-R14Del hearts (as in PLN-knockouts), there was cardiac hypertrophy (unlike PLN-KO) coupled with activation of Akt and the MAPK pathways. Thus, human PLN-R14Del is misrouted to the sarcolemma, in the absence of endogenous PLN, and alters NKA activity, leading to cardiac remodeling. [Copyright &y& Elsevier]

Abstract: Among the identified microRNAs (miRs) thus far, ~50% of mammalian miRs are clustered in the genome and transcribed as polycistronic primary transcripts. However, whether clustered miRs mediate non-redundant and cooperative functions remains poorly understood. In this study, we first identified activation of the promoter of miR-144/451 by GATA-4, a critical transcription factor in the heart. Next, we observed that ectopic expression of miR-144 and -451 individually augmented cardiomyocyte survival, which was further improved by overexpression of miR-144/451, compared to control cells in response to simulated ischemia/reperfusion. In contrast, knockdown of endogenous miR-144 and -451 revealed opposite effects. Using luciferase reporter assay and western blot analysis, we also validated that both miR-144 and miR-451 target CUG triplet repeat-binding protein 2 (CUGBP2), a ubiquitously expressed RNA-binding protein, known to interact with COX-2 3′UTR and inhibit its translation. Accordingly, protein levels of CUGBP2 were greatly reduced and COX-2 activity was markedly increased in miR-144-, miR-451-, and miR-144/451-overexpressing cardiomyocytes, compared to GFP cells. Furthermore, inhibition of COX-2 activity by either NS-398 or DUP-697 partially offset protective effects of the miR-144/451 cluster. Together, these data indicate that both partners of the miR-144/451 cluster confer protection against simulated I/R-induced cardiomyocyte death via targeting CUGBP2–COX-2 pathway, at least in part. Thus, both miR-144 and miR-451 may represent new therapeutic agents for the treatment of ischemic heart disease. [ABSTRACT FROM AUTHOR]

Abstract: A gene manipulated cell patch using a homologous peritoneum substrate was developed and applied after myocardial infarction to repair scarred myocardium. We genetically engineered male rat mesenchymal stem cells (MSC) using adenoviral transduction to over-express CXCR4/green fluorescent protein (GFP) (MSCCXCR4) or MSCNull or siRNA targeting CXCR4 (MSCsiRNA). Gene expression was studied by real-time quantitative PCR (qPCR) and enzyme-linked immunosorbent assay (ELISA). Cells were cultured on excised peritoneum for 9 days. Two weeks after left anterior descending (LAD) coronary artery ligation in female hearts, the peritoneum patch was applied over the scarred myocardium, cell side down. Efficacy of engraftment was determined by presence of GFP positive cells. One month after cell implantation, echocardiography was performed and hearts were harvested for histological analysis. Left ventricle (LV) fibrosis, LV anterior wall thickness (AWT) and blood vessel density at the margins of the graft were measured. There was significant up-regulation of the chemokines in the MSCCXCR4 group cultured under normoxic conditions when compared to the MSCNull group and a further increase was observed after exposure to hypoxia. One month after cell transplantation with the peritoneum patch, substantial numbers of GFP-positive cells were observed in and around the infarcted myocardium in MSCCXCR4 group. LV AWT, LV fibrosis and LV function were significantly improved in the MSCCXCR4 group as compared to these same variables in the MSCNull control. These salutary effects were absent in MSCsiRNA group. The gene manipulated MSC-seeded peritoneum patch promotes tissue nutrition (angiogenesis), reduces myocardial remodeling, and enhances heart function after myocardial infarction. [Copyright &y& Elsevier]

Abstract: The occurrence of cardiovascular dysfunction in sepsis is associated with a significantly increased mortality rate of 70% to 90% compared with 20% in septic patients without cardiovascular impairment. Thus, rectification or blockade of myocardial depressant factors should partly ameliorate sepsis progression. Heat shock protein 20 (Hsp20) has been shown to enhance myocardial contractile function and protect against doxorubicin-induced cardiotoxicity. To investigate the possible role of Hsp20 in sepsis-mediated cardiac injury, we first examined the expression profiles of five major Hsps in response to lipopolysaccharide (LPS) challenge, and observed that only the expression of Hsp20 was downregulated in LPS-treated myocardium, suggesting that this decrease might be one of the mechanisms contributing to LPS-induced cardiovascular defects. Further studies using loss-of-function and gain-of-function approaches in adult rat cardiomyocytes verified that reduced Hsp20 levels were indeed correlated with the impaired contractile function. In fact, overexpression of Hsp20 significantly enhanced cardiomyocyte contractility upon LPS treatment. Moreover, after administration of LPS (25 μg/g) in vivo, Hsp20 transgenic mice (10-fold overexpression) displayed: 1) an improvement in myocardial function; 2) reduced the degree of cardiac apoptosis; and 3) decreased NF-κB activity, accompanied with reduced myocardial cytokines IL-1β and TNF-α production, compared to the LPS-treated non-transgenic littermate controls. Thus, the increases in Hsp20 levels can protect against LPS-induced cardiac apoptosis and dysfunction, associated with inhibition of NF-κB activity, suggesting that Hsp20 may be a new therapeutic agent for the treatment of sepsis. [Copyright &y& Elsevier]