ALBERT

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Xing-Wei Xiang, Jin-Xing Xiao, Yu-Fang Zhou, Bin Zheng, Zheng-Shun Wen〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The black seabream (〈em〉Sparus macrocephlus〈/em〉) is an economically pivotal aquaculture species cultured in China and Southeast Asian countries. To understand the molecular immune mechanisms underlying the response to 〈em〉Vibrio parahaemolyticus〈/em〉, a comparative gene transcription analysis were performed with utilized fresh livers of 〈em〉V. parahaemolyticus〈/em〉-immunized 〈em〉Sparus macrocephlus〈/em〉 with a control group through RNA-Seq technology. A total of 256663 contigs were obtained after excluded the low-quality sequences and assembly. The average length of contigs collected from this research is 1066.93 bp. Furthermore, blast analysis indicates 30747 contigs were annotated based on homology with matches in the NT, NR, gene, and string databases. A gene ontology analysis was employed to classify 21598 genes according to three major functional categories: molecular function, cellular component, and biological process. A total of 14470 genes were discovered in 303 KEGG pathways. RSEM and EdgeR were introduced to estimate 3841 genes significantly different expressed (False Discovery Rate〈0.001) which includes 4072 up-regulated genes and 3771 down-regulated genes. A significant enrichment analysis of these differentially expressed genes and isogenes were conducted to reveal the major immune-related pathways which refer to the toll-like receptor, complement, coagulation cascades, and chemokine signaling pathways. In addition, 92175 potential simple sequence repeats (SSRs) and 121912 candidate single nucleotide polymorphisms (SNPs) were detected and identified sequencely in the 〈em〉Sparus macrocephlus〈/em〉 liver transcriptome. This research characterized a gene expression pattern for normal and the 〈em〉V. parahaemolyticus〈/em〉 -immunized 〈em〉Sparus macrocephlus〈/em〉 for the first time and not only sheds new light on the molecular mechanisms underlying the host-〈em〉V. parahaemolyticus〈/em〉 interaction but contribute to facilitate future studies on 〈em〉Sparus macrocephlus〈/em〉 gene expression and functional genomics.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Yi-Hong Chen, Jian-Guo He〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The shrimp aquaculture industry is plagued by disease. Due to the lack of deep understanding of the relationship between innate immune mechanism and environmental adaptation mechanism, it is difficult to prevent and control the diseases of shrimp. The shrimp innate immune system has received much recent attention, and the functions of the humoral immune response and the cellular immune response have been preliminarily characterized. The role of environmental stress in shrimp disease has also been investigated recently, attempting to clarify the interactions among the innate immune response, the environmental stress response, and disease. Both the innate immune response and the environmental stress response have a complex relationship with shrimp diseases. Although these systems are important safeguards, allowing shrimp to adapt to adverse environments and resist infection, some pathogens, such as white spot syndrome virus, hijack these host systems. As shrimp lack an adaptive immune system, immunization therapy cannot be used to prevent and control shrimp disease. However, shrimp diseases can be controlled using ecological techniques. These techniques, which are based on the innate immune response and the environmental stress response, significantly reduce the impact of shrimp diseases. The object of this review is to summarize the recent research on shrimp environmental adaptation mechanisms, innate immune response mechanisms, and the relationship between these systems. We also suggest some directions for future research.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Yinnan Mu, Shimin Zhou, Ning Ding, Jingqun Ao, Xinhua Chen〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Chemokines are a superfamily of structurally related chemotactic cytokines exerting significant roles in regulating cell migration and activation. Currently, five subgroups of fish specific CXC chemokines, named CXCL_F1-CXCL_F5, have been identified in teleost fish. However, understanding of the functions of these fish specific CXC chemokines is still limited. Here, a new member of fish specific CXC chemokines, 〈em〉Lc〈/em〉CXCL_F6, was cloned from large yellow croaker 〈em〉Larimichthys crocea〈/em〉. Its open reading frame (ORF) is 369 nucleotides long, encoding a peptide of 122 amino acids (aa). The deduced 〈em〉Lc〈/em〉CXCL_F6 protein contains a 19-aa signal peptide and a 103-aa mature polypeptide, which has four conserved cysteine residues (C〈sup〉28〈/sup〉, C〈sup〉30〈/sup〉, C〈sup〉56〈/sup〉, and C〈sup〉72〈/sup〉), as found in other known CXC chemokines. Phylogenetic analysis showed 〈em〉Lc〈/em〉CXCL_F6 formed a separate clade with sequences from other fish species, tentatively named CXCL_F6, distinct from the clades formed by fish CXCL_F1-5 and mammalian CXC chemokines. The 〈em〉Lc〈/em〉CXCL_F6 transcripts were constitutively expressed in all examined tissues and significantly up-regulated in the spleen and head kidney tissues by poly (I:C) and 〈em〉Vibrio alginolyticus〈/em〉. Its transcripts were also detected in primary head kidney leukocytes (HKLs), peripheral blood leucocytes (PBLs), and large yellow croaker head kidney (LYCK) cell line, and significantly up-regulated by poly(I:C), lipopolysaccharide (LPS), and peptidoglycan (PGN) in HKLs. Recombinant 〈em〉Lc〈/em〉CXCL_F6 protein (r〈em〉Lc〈/em〉CXCL_F6) could not only chemotactically attract monocytes/macrophages and lymphocytes from PBLs, but also enhance NO release and expression of proinflammatory cytokines (TNF-α, IL-1β, and CXCL8) in monocytes/macrophages. These results indicate that 〈em〉Lc〈/em〉CXCL_F6 plays a role in mediating the inflammatory response.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Jinghua Chen, Lu Zhang, Ning Yang, Mengyu Tian, Qiang Fu, Fenghua Tan, Chao Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Galectins are a family of galactoside-binding proteins with an affinity for β-galactosides, involved in mediating fundamental processes including development, inflammation, cell migration and apoptosis. Galectin-4 is a member of tendem-repeat galectins, plays vital roles in intestinal epithelial barrier. Here, one galectin-4 gene was captured in turbot (〈em〉Sm〈/em〉Lgals4) contains a 1197 bp open reading frame (ORF). In comparison to other species, 〈em〉Sm〈/em〉Lgals4 showed the highest similarity and identity both to large yellow croaker. The genomic structure analysis showed that 〈em〉Sm〈/em〉Lgals4 had conserved exons in the CRD domains compared to other vertebrate species. The syntenic analysis revealed that galectin-4 had the same neighboring genes across all the selected species, which suggested the synteny encompassing galectin-4 region during vertebrate evolution. Subsequently, 〈em〉Sm〈/em〉Lgals4 was widely expressed in all the examined tissues, with the highest expression level in intestine and the lowest expression level in skin. In addition, 〈em〉Sm〈/em〉Lgals4 was significantly down-regulated in intestine following both Gram-negative bacteria 〈em〉Vibrio anguillarum〈/em〉, and Gram-positive bacteria 〈em〉Streptococcus iniae〈/em〉 immersion challenge. Finally, the 〈em〉rSm〈/em〉Lgals4 showed strong binding ability to all the examined microbial ligands. Taken together, our results suggested 〈em〉Sm〈/em〉Lgals4 plays vital roles in fish intestinal immune responses against infection, but the detailed roles of galectin-4 in teleost are still lacking, further studies are needed to be carried out to characterize whether galectin-4 plays similar roles in teleost intestinal immunity.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Yanxiu Mo, Yunpeng Fan, Wen Fu, Wenting Xu, Shujuan Chen, Yuanhui Wen, Shaojun Liu, Liangyue Peng, Yamei Xiao〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Previous research has indicated that the small compound, SP600125, could induce polyploidy of fish cells, and has established a stable tetraploid cell line from diploid fish cells. In order to explore how fish cells maintain homeostasis under SP600125-stress 〈em〉in vitro〈/em〉, this study investigates impacts of SP600125-stress on intracellular pathways, as well as on regulation of the cellular homeostasis feedback in fish cells. Transcriptomes are obtained from the SP600125-treated cells. Compared with unigenes expressed in control group (crucial carp fin cells), a total of 2670 and 1846 unigenes are significantly upregulated and downregulated in these cells, respectively. Differentially expressed genes are found, which are involved in innate defense, inflammatory pathways and cell adhesion molecules-related pathways. The SP600125-stress enhances cell-mediated immunity, characterized by significantly increasing expression of multiple immune genes. These enhanced immune genes include the pro-inflammatory cytokines (IL-1β, TNF-ɑ, IL-6R), the adaptor signal transducers (STAT, IκBɑ), and the integrins (ɑ2β1, ɑMβ2). Furthermore, mitochondria are contributed to the cellular homeostasis regulation upon the SP600125-stress. The results show that acute inflammation is an adaptive and controlled response to the SP600125-stress, which is beneficial for alleviating toxicity by SP600125. They provide a potential way of breeding fish polyploidy induced by SP600125 in the future research.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Maciej Woźny, Kazimierz Obremski, Piotr Hliwa, Piotr Gomułka, Rafał Różyński, Paweł Wojtacha, Maciej Florczyk, Helmut Segner, Paweł Brzuzan〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉To investigate the effects of feed contamination with zearalenone (ZEN) at the current European Commission (EC) guidance value (2 mg⋅kg〈sup〉−1〈/sup〉 feed) on the growth and health of rainbow trout, we performed a long-term feeding trial under aquaculture conditions. It started with the external feeding of the fish larvae, and continued for 96 weeks, at which point the fish had reached market size. To assess the growth of fish and their feeding efficiency throughout this period, the fish were regularly weighed and measured, and their feed consumption was monitored. Additionally, to investigate potential health effects, after 72 weeks of the exposure to ZEN, the fishes' blood was analyzed for major hematological and biochemical indices, and their head kidney, spleen, and liver were examined for morphological, histopathological, cytological, and molecular changes. Finally, to gain insight into the metabolism and distribution of ZEN in fish, the content of free and glucuronidated forms of ZEN and its major metabolites was measured in the intestine, liver, and muscles of the exposed fish. The feed-borne exposure of rainbow trout to ZEN at a dose of 2 mg⋅kg〈sup〉−1〈/sup〉 feed resulted in higher feeding efficiency and growth rate, most probably due to the anabolic properties of the ZEN metabolite. Importantly for the consumers of fish, despite absorption and metabolism of ZEN in the digestive system of the fish that had been exposed for 72 weeks, the residuals of ZEN were not transferred to the fishes’ muscles, which rules out a potential risk to human health related to the consumption of fish meat. However, the increased growth of fish fed with the contaminated feed may come at some cost, as the exposure to ZEN was associated with modulation of key components of the adaptive and innate immune systems. Moreover, the trunk kidney of ZEN-fed fish showed massive inflammation that was likely caused by pathogen infection. These findings raise concerns about fish health under the current recommended EC guidance values.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 130〈/p〉 〈p〉Author(s): Wei Feng, Tong Lei, Yue Wang, Run Feng, Juntao Yuan, Xiyue Shen, Yongguang Wu, Junling Gao, Wenjun Ding, Zhongbing Lu〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Excessive myocardial lipid accumulation is a major feature of diabetic cardiomyopathy (DCM). Although general control nonderepressible 2 (GCN2) has been identified as a sensor of amino acid availability, it also functions as an important regulator of hepatic lipid metabolism. Our previous studies have reported that GCN2 promotes pressure overload or doxorubicin-induced cardiac dysfunction by increasing cardiomyocyte apoptosis and myocardial oxidative stress. However, the impact of GCN2 on the development of DCM remains unclear. In this study, we investigated the effect of GCN2 on DCM in type 1 and type 2 diabetes animal models. After streptozotocin (STZ) or high-fat diet (HFD) plus low-dose STZ treatments, GCN2〈sup〉-/-〈/sup〉 mice developed less cardiac dysfunction, hyperlipidemia, myocardial hypertrophy, fibrosis, lipid accumulation, oxidative stress, inflammation and apoptosis compared with wild-type (WT) mice. In diabetic hearts, GCN2 deficiency attenuated the upregulation of peroxisome proliferator-activated receptor alpha (PPARα) and gamma (PPARγ), the phosphorylation of eIF2α and the induction of activating transcription factor 4 (ATF4) and C/EBP homologous protein (CHOP), as well as the reduction of Bcl-2. Furthermore, we found that knockdown of GCN2 attenuated, whereas overexpression of GCN2 exacerbated, high glucose or palmitic acid-induced cell death, oxidative and endoplasmic reticulum stress and lipid accumulation in H9C2 cells. Collectively, our data provide evidence that GCN2 deficiency protects cardiac function by reducing lipid accumulation, oxidative stress and cell death. Our findings suggest that strategies to inhibit GCN2 activity in the heart may be novel approaches for DCM therapy.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918315594-fx1.jpg" width="219" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 130〈/p〉 〈p〉Author(s): Daria M. Svistunova, Jillian N. Simon, Elzbieta Rembeza, Mark Crabtree, Wyatt W. Yue, Peter L. Oliver, Mattéa J. Finelli〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Protein aggregation, oxidative and nitrosative stress are etiological factors common to all major neurodegenerative disorders. Therefore, identifying proteins that function at the crossroads of these essential pathways may provide novel targets for therapy. Oxidation resistance 1 (Oxr1) is a protein proven to be neuroprotective against oxidative stress, although the molecular mechanisms involved remain unclear. Here, we demonstrate that Oxr1 interacts with the multifunctional protein, peroxiredoxin 2 (Prdx2), a potent antioxidant enzyme highly expressed in the brain that can also act as a molecular chaperone. Using a combination of 〈em〉in vitro〈/em〉 assays and two animal models, we discovered that expression levels of Oxr1 regulate the degree of oligomerization of Prdx2 and also its post-translational modifications (PTMs), specifically suggesting that Oxr1 acts as a functional switch between the antioxidant and chaperone functions of Prdx2. Furthermore, we showed in the Oxr1 knockout mouse that Prdx2 is aberrantly modified by overoxidation and S-nitrosylation in the cerebellum at the presymptomatic stage; this in-turn affected the oligomerization of Prdx2, potentially impeding its normal functions and contributing to the specific cerebellar neurodegeneration in this mouse model.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918312711-fx1.jpg" width="258" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 130〈/p〉 〈p〉Author(s): Hoi-Shan Wong, Bérengère Benoit, Martin D. Brand〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The relative contributions of different mitochondrial and cytosolic sources of superoxide and hydrogen peroxide in cells are not well established because of a lack of suitable quantitative assays. To address this problem using resting C2C12 myoblasts we measured the effects of specific inhibitors that do not affect other pathways on the rate of appearance of hydrogen peroxide in the extracellular medium. We used inhibitors of NADPH oxidases (NOXs), suppressors of site I〈sub〉Q〈/sub〉 electron leak (S1QELs) at mitochondrial Complex I, and suppressors of site III〈sub〉Qo〈/sub〉 electron leak (S3QELs) at mitochondrial Complex III. Around 40% of net cellular hydrogen peroxide release was from NOXs and approximately 45% was from the two mitochondrial sites; 30% from site III〈sub〉Qo〈/sub〉 and 15% from site I〈sub〉Q〈/sub〉. As expected, decreasing cytosolic antioxidant capacity by lowering glutathione levels increased the absolute rates from all sites without changing their proportions, whereas decreasing antioxidant defenses in the mitochondrial matrix increased only the absolute and relative contributions of the two mitochondrial sites. These results show directly that mitochondria are a major contributor to cytosolic hydrogen peroxide in resting C2C12 myoblasts, and provide the first direct evidence of superoxide/hydrogen peroxide production from site I〈sub〉Q〈/sub〉 in unstressed cells.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918315752-fx1.jpg" width="206" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: Available online 31 October 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine〈/p〉 〈p〉Author(s): Brígida R. Pinho, Sara D. Reis, Richard C. Hartley, Michael P. Murphy, Jorge M.A. Oliveira〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Superoxide generation by mitochondria respiratory complexes is a major source of reactive oxygen species (ROS) which are capable of initiating redox signalling and oxidative damage. Current understanding of the role of mitochondrial ROS in health and disease has been limited by the lack of experimental strategies to selectively induce mitochondrial superoxide production. The recently-developed mitochondria-targeted redox cycler MitoParaquat (MitoPQ) overcomes this limitation, and has proven effective 〈em〉in vitro〈/em〉 and in 〈em〉Drosophila〈/em〉. Here we present an 〈em〉in vivo〈/em〉 study of MitoPQ in the vertebrate zebrafish model in the context of Parkinson's disease (PD), and in a human cell model of Huntington's disease (HD). We show that MitoPQ is 100-fold more potent than non-targeted paraquat in both cells and in zebrafish in vivo. Treatment with MitoPQ induced a parkinsonian phenotype in zebrafish larvae, with decreased sensorimotor reflexes, spontaneous movement and brain tyrosine hydroxylase (TH) levels, without detectable effects on heart rate or atrioventricular coordination. Motor phenotypes and TH levels were partly rescued with antioxidant or monoaminergic potentiation strategies. In a HD cell model, MitoPQ promoted mutant huntingtin aggregation without increasing cell death, contrasting with the complex I inhibitor rotenone that increased death in cells expressing either wild-type or mutant huntingtin. These results show that MitoPQ is a valuable tool for cellular and 〈em〉in vivo〈/em〉 studies of the role of mitochondrial superoxide generation in redox biology, and as a trigger or co-stressor to model metabolic and neurodegenerative disease phenotypes.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918313182-fx1.jpg" width="500" alt="Graphical abstract for this article" title=""〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Xin Cai, Chengbin Gao, Huanhuan Song, Ning Yang, Qiang Fu, Fenghua Tan, Chao Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Cathepsin Z (CTSZ) is a lysosomal cysteine protease of the papain superfamily. It participates in the host immune defense via phagocytosis, signal transduction, cell-cell communication, proliferation, and migration of immune cells such as monocytes, macrophages, and dendritic cells. In this study, we reported the identification of 〈em〉Sm〈/em〉CTSZ, a CTSZ homolog from turbot (〈em〉Scophthalmus maximus〈/em〉 L.). 〈em〉Sm〈/em〉CTSZ was 317 residues in length and contains a Pept-C1 domain. In multiple species comparison, 〈em〉Sm〈/em〉CTSZ shared 65–93% overall sequence identities with the CTSZ counterparts from human, rat, and several fish species. In the phylogenetic analysis, 〈em〉Sm〈/em〉CTSZ showed the closest relationship to 〈em〉Cynoglossus semilaevis〈/em〉. The syntenic analysis revealed the similar neighboring genes of CTSZ across all the selected species, which suggested the synteny encompassing CTSZ region during vertebrate evolution. Subsequently, 〈em〉Sm〈/em〉CTSZ was constitutively expressed in various tissues, with the lowest and highest levels in brain and intestine respectively. In addition, 〈em〉Sm〈/em〉CTSZ was significantly up-regulated in intestine following both Gram-negative bacteria 〈em〉Vibrio anguillarum〈/em〉, and Gram-positive bacteria 〈em〉Streptococcus iniae〈/em〉 immersion challenge. Finally, the 〈em〉rSm〈/em〉CTSZ showed strong binding ability to all the examined microbial ligands, and the agglutination effect to different bacteria. Taken together, these results indicated 〈em〉Sm〈/em〉CTSZ could play important roles in mucosal immune response in the event of bacterial infection in teleost. However, the knowledge of CTSZ are still limited in teleost species, further studies should be carried out to better characterize its detailed roles in teleost mucosal immunity.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Zhi Zhou, Zhaoqun Liu, Lingui Wang, Jian Luo, Hailang Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Giant clams are one of the most important animals in coral reef ecosystem, and its growth and reproduction are being threatened by heat stress due to global warming. In the present study, the symbiont density, the crucial enzyme activities and the transcriptome were investigated in the outer mantle of giant clam 〈em〉Tridacna crocea〈/em〉 after the acute exposure of high temperature. The density of symbiotic zooxanthellae decreased significantly during 12–24 h, with the minimum level (7.75 × 10〈sup〉5〈/sup〉 cell cm〈sup〉−2〈/sup〉, 〈em〉p〈/em〉 〈 0.05) at 12 h after heat stress. The activities of superoxide dismutase in the heat stress group was significantly lower than that in the control group at 24 h after heat stress, while no significant change in the activities of catalase was observed during the entire stress process. The activation level of caspase3 began to increase significantly at 12 h (1.22-fold, 〈em〉p〈/em〉 〈 0.05), and reached the highest level at 24 h (1.38-fold, 〈em〉p〈/em〉 〈 0.05) after heat stress. Six paired-end libraries were sequenced in two groups, including the heat stress and control group at 12 h after heat stress. Through the assembling of 187,116,632 paired-end reads with lengths of 2 × 150 bp, a total of 26,676 genes were obtained which derived from giant clam. Bioinformatics analysis revealed 47 significantly upregulated and 88 significantly downregulated genes at 12 h after the treatment. There were 12 overrepresented GO terms for significantly upregulated genes, mostly related to unfolded protein binding and ATP binding, whereas no GO term was overrepresented for significantly downregulated genes. These results collectively suggest high temperature could induce excessive oxidative stress through the repressed antioxidant ability, the apoptosis activated by the unfolded protein response, and further the collapse of the symbiosis between host and symbiont, which has been threatening the growth and reproduction of the giant clam 〈em〉T. crocea〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Zhi-qiang Du, Yue Wang, Hong-yu Ma, Xiu-li Shen, Kai Wang, Jie Du, Xiao-dong Yu, Wen-hong Fang, Xin-cang Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Crustins play important roles in defending against bacteria in the innate immunity system of crustaceans. In present study, we identified a crustin gene in 〈em〉Scylla paramamosain〈/em〉, which was named as 〈em〉SpCrus6〈/em〉. The ORF of 〈em〉Sp〈/em〉Crus6 possessed a signal peptide sequence (SPS) at the N-terminus and a WAP domain at the C-terminus. And there were 5 Proline residues, 5 Glycine and 4 Cysteine residues between SPS and WAP domain in 〈em〉Sp〈/em〉Crus6. These features indicated that 〈em〉Sp〈/em〉Crus6 was a new member of crustin family. The 〈em〉SpCrus6〈/em〉 mRNA transcripts were up-regulated obviously after bacteria or virus challenge. These changes showed that 〈em〉SpCrus6〈/em〉 was involved in the antimicrobial and antiviral responses of 〈em〉Scylla paramamosain〈/em〉. Recombinant 〈em〉Sp〈/em〉Crus6 (r〈em〉Sp〈/em〉Crus6) showed strong inhibitory abilities against Gram-positive bacteria (〈em〉Bacillus megaterium〈/em〉, 〈em〉Staphylococcus aureus〈/em〉, and 〈em〉Bacillus subtilis〈/em〉). But the inhibitory abilities against four Gram-negative bacteria (〈em〉Vibrio parahemolyticus〈/em〉, 〈em〉Vibrio alginolyticus〈/em〉, 〈em〉Vibrio harveyi〈/em〉 and 〈em〉Escherichia coli〈/em〉) and two fungi (〈em〉Pichia pastoris〈/em〉 and 〈em〉Candida albicans〈/em〉) were not strong enough. Besides, r〈em〉Sp〈/em〉Crus6 could strongly bind to two Gram-positive bacteria (〈em〉B〈/em〉. 〈em〉subtilis〈/em〉 and 〈em〉B〈/em〉. 〈em〉megaterium〈/em〉) and three Gram-negative bacteria (〈em〉V〈/em〉. 〈em〉alginolyticus〈/em〉, 〈em〉V〈/em〉. 〈em〉parahemolyticus〈/em〉, and 〈em〉V〈/em〉. 〈em〉harveyi〈/em〉). And the binding levels to 〈em〉S. aureus〈/em〉 and two fungi (〈em〉P〈/em〉. 〈em〉pastoris〈/em〉 and 〈em〉C〈/em〉. 〈em〉albicans〈/em〉) were weak. The polysaccharides binding assays’ results showed r〈em〉Sp〈/em〉Crus6 had superior binding activities to LPS, LTA, PGN and 〈em〉β〈/em〉-glucan. Through agglutinating assays, we found r〈em〉Sp〈/em〉Crus6 could agglutinate well three Gram-positive bacteria (〈em〉S〈/em〉. 〈em〉aureus〈/em〉, 〈em〉B〈/em〉. 〈em〉subtilis〈/em〉 and 〈em〉B〈/em〉. 〈em〉megaterium〈/em〉). And the agglutinating activities to Gram-negative bacteria and fungi were not found. In the aspect of antiviral functions, r〈em〉Sp〈/em〉Crus6 could bind specifically to the recombinant envelop protein 26 (rVP26) of white spot syndrome virus (WSSV) but not to recombinant envelop protein 28 (rVP28), whereas GST protein could not bind to rVP26 or rVP28. Besides, r〈em〉Sp〈/em〉Crus6 could suppress WSSV reproduction to some extent. Taken together, 〈em〉Sp〈/em〉Crus6 was a multifunctional immunity effector in the innate immunity defending response of 〈em〉S〈/em〉. 〈em〉paramamosain〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Yunkun Li, Jiayu Wu, Dong Li, Anqi Huang, Guixian Bu, Fengyan Meng, Fanli Kong, Xiaohan Cao, Xingfa Han, Xiaofu Pan, Wei Fan, Shiyong Yang, Xianyin Zeng, Xiaogang Du〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Schizothorax prenanti〈/em〉 (〈em〉S. prenanti〈/em〉), an important species of economical fish in Southwest China, is susceptible to 〈em〉Aeromonas hydrophila〈/em〉 (Ah). To understand the immune response to Ah, the transcriptome profiling of spleen of 〈em〉S. prenanti〈/em〉 was analyzed after Ah infection. A total of 6, 213 different expression genes (DEGs) were obtained, including 3, 066 up-regulated DEGs and 3, 147 down-regulated DEGs. These DEGs were annotated by KEGG and GO databases, so that the immune-related DEGs (IRDs) can be identified and classified. Then, the interesting IRDs were screened to build heat map, and the reliability of the transcriptome data was validated by qPCR. In order to clarify the mechanism of signal transduction in the anti-bacterial immunity, the signaling pathway initiated by TLRs was predicted. In this pathway, TLR25 and TLR5 mediate the NF-κB and AP-1 signals via MyD88-dependent pathway. Meanwhile, the type I IFN (IFNα/β) induced by IRF1 and IRF3/7 may play an important role in the anti-bacterial immunity. In conclusion, this study preliminarily provides insights into the mechanism of signal transduction after Ah infection in 〈em〉S. prenanti〈/em〉, which contributes to exploring the complex anti-bacterial immunity.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Hongye Jiang, Qing Bian, Weiwei Zeng, Pengli Ren, Hengchang Sun, Zhipeng Lin, Zeli Tang, Xinyi Zhou, Qing Wang, Yingying Wang, Yensheng Wang, Mei X. Wu, Xuerong Li, Xinbing Yu, Yan Huang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Grass carp (〈em〉Ctenopharyngodon idellus〈/em〉) hemorrhagic disease (GCHD), caused by grass carp reovirus (GCRV), has given rise to an enormous loss in grass carp industry during the past years. Up to date, vaccination remained to be the most effective way to protect grass carp from GCHD. Oral vaccination is of major interest due to its advantages of noninvasive, time-saving, and easily-operated. The introduction of oral vaccination has profound impact on aquaculture industry because of its feasibility of extensive application for fish in various size and age. However, the main challenge in developing oral vaccine is that antigens are easily degraded and are easy to induce tolerance. 〈em〉Bacillus subtilis〈/em〉 (〈em〉B. subtilis〈/em〉) spores would be an ideal oral vaccine delivery system for their robust specialty, gene operability, safety and adjuvant property. VP4 protein is the major outer capsid protein encoded by GCRV segment 6 (S6), which plays an important role in viral invasion and replication. In this study, we used 〈em〉B. subtilis〈/em〉 spores as the oral delivery system and successfully constructed the 〈em〉B. subtilis〈/em〉 CotC-VP4 recombinant spores (CotC-VP4 spores) to evaluate its protective efficacy in grass carp. Grass carp orally immunized with CotC-VP4 spores showed a survival rate of 57% and the relative percent survival (RPS) of 47% after the viral challenge. Further, the specific IgM levels in serum and the specific IgZ levels in intestinal mucus were significantly higher in the CotC-VP4 group than those in the Naive group. The immune-related genes including three innate immune-related genes (IL-4/13A, IL-4/13B, CSF1R), four adaptive immune-related genes (BAFF, CD4L, MHC-II, CD8), three inflammation-related genes (IL-1β, TNF-α, TGF-β) and interferon type I (IFN-I) related signaling pathway genes were significantly up-regulated in the CotC-VP4 group. The study demonstrated that the CotC-VP4 spores produced protection in grass carp against GCRV infection, and triggered both innate and adaptive immunity post oral immunization. This work highlighted that 〈em〉Bacillus subtilis〈/em〉 spores were powerful platforms for oral vaccine delivery, and the combination of 〈em〉Bacillus subtilis〈/em〉 spores with GCRV VP4 protein was a promising oral vaccine.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 130〈/p〉 〈p〉Author(s): Alberto Cruz-Bermúdez, Raquel Laza-Briviesca, Ramiro J. Vicente-Blanco, Aránzazu García-Grande, Maria José Coronado, Sara Laine-Menéndez, Cristina Alfaro, Juan Cristobal Sanchez, Fernando Franco, Virginia Calvo, Atocha Romero, Paloma Martin-Acosta, Clara Salas, José Miguel Garcia, Mariano Provencio〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉 〈p〉Lung cancer is a major public health problem due to its high incidence and mortality rate. The altered metabolism in lung cancer is key for the diagnosis and has implications on both, the prognosis and the response to treatments. Although Cancer-associated fibroblasts (CAFs) are one of the major components of the tumor microenvironment, little is known about their role in lung cancer metabolism.〈/p〉 〈p〉We studied tumor biopsies from a cohort of 12 stage IIIA lung adenocarcinoma patients and saw a positive correlation between the grade of fibrosis and the glycolysis phenotype (Low PGC-1α and High GAPDH/MT-CO1 ratio mRNA levels). These results were confirmed and extended to other metabolism-related genes through the 〈em〉in silico〈/em〉 data analysis from 73 stage IIIA lung adenocarcinoma patients available in TCGA. Interestingly, these relationships are not observed with the CAFs marker α-SMA in both cohorts.〈/p〉 〈p〉To characterize the mechanism, 〈em〉in vitro〈/em〉 co-culture studies were carried out using two NSCLC cell lines (A549 and H1299 cells) and two different fibroblast cell lines. Our results confirm that a metabolic reprogramming involving ROS and TGF-β signaling occurs in lung cancer cells and fibroblasts independently of α-SMA induction. Under co-culture conditions, Cancer-Associated fibroblasts increase their glycolytic ability. On the other hand, tumor cells increase their mitochondrial function. Moreover, the differential capability among tumor cells to induce this metabolic shift and also the role of the basal fibroblasts Oxphos Phosphorylation (OXPHOS) function modifying this phenomenon could have implications on both, the diagnosis and prognosis of patients. Further knowledge in the mechanism involved may allow the development of new therapies.〈/p〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918314461-fx1.jpg" width="500" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Guang-hua Wang, Jing-jing Wang, Bin Yue, Xue Du, He-he Du, Min Zhang, Yong-hua Hu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉High-mobility group box 2 (HMGB2) is a non-histone chromosomal protein that involved diverse functions such as transcriptional regulation and innate immune responses in mammalian. In teleost, very limited studies on HMGB2 proteins have been documented. Black rockfish (〈em〉Sebastes schlegelii〈/em〉) is an economic fish species and cultured worldwide. However, the study of black rockfish about immunology is very scarce. In the present study, a HMGB2 homologue gene (〈em〉SsHMGB2〈/em〉) was identified and characterized in black rockfish. The open reading frame of 〈em〉SsHMGB2〈/em〉 is 648 bp, and the deduced amino acid sequence of 〈em〉SsHMGB2〈/em〉 shares 74.4%–91.2% overall sequence identities with the HMGB2 proteins of several fish species. In silico analysis identified several conserved features, including two basic HMG boxes and an acidic C-terminal tail composed of 24 Asp/Glu residues. Expression of 〈em〉SsHMGB2〈/em〉 occurred in multiple tissues and was upregulated during pathogens infection. Recombinant SsHMGB2 (rSsHMGB2) exhibited apparent binding activities against DNA. 〈em〉In vivo〈/em〉 studies showed that the expressions of multiple immune-related genes in head kidney were significantly enhanced when black rockfish were treated with rSsHMGB2. Furthermore, rSsHMGB2 reduced pathogen dissemination and replication in fish kidney and spleen. Taken together, these results suggest that SsHMGB2 possesses apparent immunoregulatory properties and played a role in fighting bacterial infection.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Sai-Wei Chen, Chun-Hung Liu, Shao-Yang Hu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Bacteria-induced diseases are a major cause of mortality in aquaculture. Probiotics have commonly been used to replace antibiotics for prophylactic biocontrol in aquaculture. In the present study, 〈em〉Paenibacillus ehimensis〈/em〉 NPUST1 was isolated from a tilapia culture pond. This probiotic has bacteriocin-like activities against 〈em〉Aeromonas hydrophila〈/em〉 and was characterized by biochemical analysis and 16S rDNA sequencing. The physiochemical properties of a crude extract of the bacteriocin-like substance revealed low pH and high thermal tolerance. The substance exhibited broad-spectrum antimicrobial activity against diverse aquatic pathogens, food spoilage, clinical pathogens, and plant pathogens. The effect of dietary supplementation with 〈em〉P. ehimensis〈/em〉 NPUST1 was evaluated in regard to the growth of Nile tilapia (〈em〉Oreochromis niloticus〈/em〉) and immunity against pathogenic infection. The results showed significantly increased weight gain (WG), feed conversion ratio (FCR), and feed efficiency (FE) in Nile tilapia fed 〈em〉P. ehimensis〈/em〉 NPUST1 for 2 months compared with fish fed a control diet. When challenged with 〈em〉A. hydrophila〈/em〉 and 〈em〉S. iniae,〈/em〉 the fish fed 〈em〉P. ehimensis〈/em〉 NPUST1 also exhibited a higher survival rate than fish fed the control diet. The immune parameters revealed that the 〈em〉P. ehimensis〈/em〉 NPUST1-fed fish had significantly higher phagocytic activity, respiratory burst, and superoxide dismutase (SOD) of the head kidney leukocytes, as well as higher serum lysozyme activity and expression of cytokines TNF-α and IL-1β than the fish fed the control diet. These results indicate that dietary supplementation with 〈em〉P. ehimensis〈/em〉 NPUST1 improved the growth performance, immunity, and disease resistance in Nile tilapia.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Cheng-cai Zheng, Xin-yi Cai, Meng-meng Huang, Idefonce Mkingule, Cong Sun, Shi-Chao Qian, Zhen-ju Wu, Bing-nan Han, Hui Fei〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Japanese eel (〈em〉Anguilla japonica〈/em〉) has become a commercially important fish species all over the world. High-density aquaculture has led to congestion and contributed to bacterial infection outbreaks that have caused high mortality. Therefore a 56-days feeding trial was conducted to determine the effects of dietary 〈em〉Bacillus amyloliquefaciens〈/em〉 (GB-9) and 〈em〉Yarrowia lipolytica〈/em〉 lipase2 (YLL2) on growth performance, digestive enzymes activity, innate immunity and resistance to pathogens of 〈em〉A. japonica〈/em〉. Fish growth performance was significantly affected by dietary YLL2 supplementation but not by GB-9. Fish fed diets with YLL2 at 2.0 g/kg diet in combination of high and low levels of GB-9 (5.0 g/kg and 2.0 g/kg) produced the highest growth. For digestive enzyme, lipase and trypsin activities was promoted by dietary containing YLL2, while amylase activities was increased by dietary containing YLL2, GB-9 single or combination. For innate immunity, the mucus lysozyme activity, leukocytes phagocytosis activity and reactive oxygen species level of skin, peroxidase and lysozyme activity of serum were enhanced in fish fed with GB-9 compared to those in control group (p 〈 0.05). The highest resistance to 〈em〉Vibrio anguillarum〈/em〉 and 〈em〉Aeromonas hydrophila〈/em〉 was determined in fish fed with 5.0 g kg〈sup〉−1〈/sup〉 GB-9 + 2.0 g/kg YLL2. This study demonstrated that GB-9 and YLL2 enhanced non-specific immune defense system of 〈em〉A. japonica〈/em〉, providing them with higher resistance to pathogens. The present results suggested that the combination of these supplements could be considered as potential biological additives for aquaculture farmed fish.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Xianyong Bu, Xuqiu Lian, Yi Wang, Chengzeng Luo, Shengqiang Tao, Yilu Liao, Jiaming Yang, Aijing Chen, Yuhong Yang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The aim of the present study was to investigate effects of dietary yeast culture on immune response related to TLR2-MyD88-NF-kβ signaling pathway, antioxidant capability and disease resistance against 〈em〉Aeromonas hydrophila〈/em〉 for Ussuri catfish (〈em〉Pseudobagrus ussuriensis〈/em〉). A total of 240 Ussuri catfish (mean weight of 7.39 ± 0.32 g) were randomly distributed into four groups that fed diets containing 0 (Y0), 10 (Y1), 20 (Y2) and 30 (Y3) g kg〈sup〉−1〈/sup〉 yeast culture for 8 weeks. The results indicated that dietary 10 g kg〈sup〉−1〈/sup〉 yeast culture supplementation significantly down-regulated mRNA levels of TLR2, MyD88, NF-kβ p65, IL-1β and IL-8 in the liver tissue compared with the control group (〈em〉P〈/em〉 〈 0.05). Simultaneously, serum lysozyme (LZM) activity, respiratory burst activity (RBA) of phagocytes, plasma alkaline phosphatase (AKP) activity and immunoglobulin M (IgM) content were significantly improved in fish fed Y1 diet (〈em〉P〈/em〉 〈 0.05). Fish fed Y1 diet had significantly higher serum alternative complement pathway activity (ACH50) and plasma complement 3 (C3) content than the Y3 group (〈em〉P〈/em〉 〈 0.05). However, no significant differences were observed in plasma acid phosphatase (ACP) activity and complement 4 (C4) content among the groups (〈em〉P〈/em〉 〉 0.05). Fish cumulative mortality rate (CMR) in the Y1 and Y2 groups were significantly lower than that in Y0 and Y3 groups (〈em〉P〈/em〉 〈 0.05), and the lowest CMR was observed in the Y1 group after challenge by 〈em〉A. hydrophila〈/em〉. The highest hepatic superoxide dismutase and glutathione peroxidase activities, total antioxidant capacity and the lowest malondialdehyde content were found in Y1 group, but no significant difference was found in hepatic catalase activity among the groups (〈em〉P〈/em〉 〉 0.05). These results demonstrate that dietary 10 g kg〈sup〉−1〈/sup〉 yeast culture could effectively improve the immunity, antioxidant capability and disease resistance against 〈em〉A. hydrophila〈/em〉 for Ussuri catfish and could down-regulate the mRNA expression levels of pro-inflammatory cytokines modulated by TLR2-MyD88-NF-kβ signaling pathway.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Tao Wang, Xin Wen, Yadong Hu, Xinyu Zhang, Dan Wang, Shaowu Yin〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Copper nanoparticles (Cu NPs) are a new pollutant in aquaculture, representing a hazard to aquatic organisms. We investigated the effects of Cu NPs exposure on oxidative stress, apoptosis and immune response in an economically important model species, 〈em〉Takifugu fasciatus〈/em〉. The juvenile fish were exposed to control, 20 or 100 μg Cu NPs/L for 30 days. The growth of 〈em〉T. fasciatus〈/em〉 was inhibited after Cu NPs exposure. Copper accumulation in liver increased with increasing Cu NPs dose. Oxidative stress indicators [malondialdehyde (MDA), total superoxide dismutase (T-SOD), catalase (CAT) and glutathione (GSH)], apoptosis index and activities of caspases (caspase-3, caspase-9) were all increased with the increase of Cu NPs concentration in liver. With an increase in Cu NPs dose, the activities of succinate dehydrogenase (SDH) and Na〈sup〉+〈/sup〉-K〈sup〉+〈/sup〉-ATPase as well as cytochrome c (Cyt-c) concentration in mitochondria decreased, accompanied by increased Cyt-c concentration in cytosol. Apoptosis-related gene expressions of 〈em〉p53〈/em〉, 〈em〉caspase-3〈/em〉, 〈em〉caspase-9〈/em〉 and 〈em〉Bax〈/em〉 were increased with the increase of Cu NPs dose. However, the opposite result was found in 〈em〉Bcl2〈/em〉 expression. The physiological indicators of immune response [heat shock protein 70 (HSP70), heat shock protein 90 (HSP90), immunoglobulin M (IgM) and lysozyme (LZM)] as well as the mRNA levels of 〈em〉HSP70〈/em〉, 〈em〉HSP90〈/em〉, 〈em〉IgM〈/em〉 and 〈em〉C-LZM〈/em〉 were all increased after Cu NPs exposure〈em〉.〈/em〉 Our results will be helpful in understanding the mechanism of Cu NPs toxicity in 〈em〉T. fasciatus〈/em〉.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S1050464818306867-fx1.jpg" width="278" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Junguo Ma, Xi Chen, Guangyuan Xin, Xiaoyu Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The present study aimed to determine the chronic toxicity of 1-methyl-3-octylimidazolium bromide ([C〈sub〉8〈/sub〉mim]Br) on the silver carp to further reveal the toxicological mechanisms of ionic liquids. Chronic exposure of silver carp to [C〈sub〉8〈/sub〉mim]Br at concentrations of 1.095 and 4.380 mg/L for 60 d was conducted under laboratory conditions. The results revealed that chronic exposure to [C〈sub〉8〈/sub〉mim]Br inhibited the activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) and reduced glutathione (GSH) levels while markedly increasing malondialdehyde (MDA) and protein carbonyl (PC) levels in fish spleen, indicating that [C〈sub〉8〈/sub〉mim]Br treatment induced oxidative stress. Additionally, long-term exposure to [C〈sub〉8〈/sub〉mim]Br markedly upregulated the expressions of nuclear factor-κB (NF-κB), inducible nitric oxide synthase (iNOS), interleukin-1β (IL-1β), IL-6, tumour necrosis factor-α (TNF-α), and interferon-γ (IFN-γ); altered the levels of transforming growth factor-β (TGF-β); and increased the mRNA levels of p38MAPK, c-fos, c-jun, and c-myc, suggesting that long-term exposure to [C〈sub〉8〈/sub〉mim]Br might promote the inflammatory response in fish spleen and that p38MAPK/NF-κB signalling may potentially be involved in this process. Moreover, [C〈sub〉8〈/sub〉mim]Br-exposure altered lysozyme activity and complement 3 (C3) and immunoglobulin M (IgM) content, indicating that chronic [C〈sub〉8〈/sub〉mim]Br exposure also has immunotoxic effects on silver carp. Furthermore, we also found that [C〈sub〉8〈/sub〉mim]Br exposure reduced miR-125b levels, altered miR-143 levels, and upregulated miR-155 and miR-21 levels, suggesting that these miRNAs may be involved in the [C〈sub〉8〈/sub〉mim]Br-induced inflammatory response in fish spleen. In summary, the present study indicates that chronic exposure to [C〈sub〉8〈/sub〉mim]Br induces inflammation in fish spleen and that oxidative stress-mediated p38MAPK/NF-κB signalling and miRNAs may play a key role in this process.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Chaozheng Li, Shaoping Weng, Jianguo He〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉As invertebrates, shrimps rely on multiple innate defense reactions, including humoral immunity and cellular immunity to recognize and eliminate various invaders, such as viruses. White spot syndrome virus (WSSV) causes the most prevalent and devastating viral disease in penaeid shrimps, which are the most widely cultured species in the coastal waters worldwide. In the last couple of decades, studies about WSSV implicate a dual role of the immune system in protecting shrimps against the infection; these studies also explore on the pathogenesis of WSSV infection. Herein, we review our current knowledge of the innate immune responses of shrimps to WSSV, as well as the molecular mechanisms used by this virus to evade host immune responses or actively subvert them for its own benefit. Deciphering the interactions between WSSV and the shrimp host is paramount to understanding the mechanisms that regulate the balance between immune-mediated protection and pathogenesis during viral infection and to the development of a safe and effective WSSV defensive strategy.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Xin Zhang, Jialong Shi, Yulong Sun, Yusuf Jibril Habib, Huiping Yang, Ziping Zhang, Yilei Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In recent years, the abalone aquaculture industry has been threatened by the deteriorating environmental conditions, such as hypoxia and thermal stress in the hot summers. It is necessary to investigate the molecular mechanism in response to these environmental challenges, and subsequently understand the immune defense system. In this study, the transcriptome profiles by RNA-seq of hemocytes from the small abalone 〈em〉Haliotis diversicolor〈/em〉 after exposure to hypoxia, thermal stress, and hypoxia plus thermal stress were established. A total of 103,703,074 clean reads were obtained and 99,774 unigenes were assembled. Of the 99,774 unigenes, 47,154 and 20,455 had homologous sequences in the Nr and Swiss-Prot protein databases, while 16,944 and 10,840 unigenes could be classified by COG or KEGG databases, respectively. RNAseq analysis revealed that the differentially expressed genes (DEGs) after challenges of hypoxia, thermal stress, or hypoxia plus thermal stress were 24,189, 29,165 and 23,665, among which more than 3000 genes involved in at least 230 pathways, including several classical immune-related pathways. The genes and pathways that were involved in immune response to hypoxia/thermal challenges were identified by transcriptome analysis and further validated by quantitative real-time PCR and RNAi technology. The findings in this study can provide information on 〈em〉H. diversicolor〈/em〉 innate immunity to improve the abalone aquaculture industry, and the analysis of the potential immune-related genes in innate immunity signaling pathways and the obtained transcriptome data can provide an invaluable genetic resource for the study of the genome and functional genes.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Thanthrige Thiunuwan Priyathilaka, S.D.N.K. Bathige, Seongdo Lee, Bo-Hye Nam, Jehee Lee〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Toll-like receptors (TLRs) are well-known pattern recognition receptors that play key immunological roles in a diverse range of organisms. In this study, two novel invertebrate TLRs from disk abalone (designated as AbTLR-A and AbTLR-B) were identified and functionally characterized for the first time. AbTLR-A and AbTLR-B comprised the typical TLR domain architecture containing an extracellular leucine-rich repeat domain, transmembrane domain, and Toll/interleukin-1 receptor domain. Expressional analysis revealed that both TLRs were constitutively expressed at all the early embryonic stages of disk abalone analyzed, with the highest level of 〈em〉AbTLR-A〈/em〉 found at the 16-cell stage and 〈em〉AbTLR-B〈/em〉 at the trochophore stage. According to tissue distribution analysis, prominent mRNA expression of 〈em〉AbTLR-A〈/em〉 and 〈em〉AbTLR-B〈/em〉 was detected in the hemocytes and gills, respectively. 〈em〉AbTLR-A〈/em〉 and 〈em〉AbTLR-B〈/em〉 mRNAs were significantly up-regulated in response to Gram-negative 〈em〉Vibrio parahemolyticus〈/em〉, Gram-positive 〈em〉Listeria monocytogenes〈/em〉, and viral hemorrhagic septicemia virus injections in abalone hemocytes and gills. Overexpression of AbTLR-A and AbTLR-B in HEK293T cells directly activated nuclear factor-κB (NF-κB) and activator protein-1 (AP-1) responsive reporters. Neither TLRs showed a high response to pathogen-associated molecular patterns 〈em〉in vitro〈/em〉. Co-expression of AbTLR-A and AbTLR-B with AbMyD88-2 and AbMyD88-X activated NF-κB-responsive reporters in a synergetic manner. These findings demonstrate the involvement of AbTLR-A and AbTLR-B in abalone innate immunity.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Mona Saleh, Gokhlesh Kumar, Abdel-Azeem S. Abdel-Baki, Mohamed A. Dkhil, Mansour El-Matbouli, Saleh Al-Quraishy〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Ichthyophthirius multifiliis〈/em〉, a ciliated protozoan parasite, causes ichthyophthiriasis and leads to considerable economic losses to the aquaculture industry. Understanding the fish immune response and host-parasite interactions could support developing novel strategies for better disease management and control. Fish skin mucus is the first line of defence against infections through the epidermis. Yet, the common carp, 〈em〉Cyprinus carpio〈/em〉, protein-based defence strategies against infection with 〈em〉I. multifiliis〈/em〉 at this barrier remain elusive. The skin mucus proteome of common carp was investigated at 1 day and 9 days post-exposure with 〈em〉I. multifiliis〈/em〉. Using nano-LC ESI MS/MS and statistical analysis, the abundance of 19 immune related and signal transduction proteins was found to be differentially regulated in skin mucus of common carp in response to 〈em〉I. multifiliis〈/em〉. The analysis revealed increased abundance values of epithelial chloride channel protein, galactose-specific lectin nattection, high choriolytic enzyme 1 (nephrosin), lysozyme C, granulin and protein-glutamine gamma-glutamyltransferase 2 in 〈em〉I. multifiliis-〈/em〉exposed carp skin mucus. Multiple lectins and a diverse array of distinct serpins with protease inhibitor activity were identified likely implicated in lectin pathway activation and regulation of proteolysis, indicating that these proteins contribute to the carp innate immune system and the protective properties of skin mucus. The results obtained from this proteomic analysis enables a better understanding of fish host response to parasitic infection and gives insights into the key role skin mucus plays in protecting fish against deleterious effects of 〈em〉I. multifiliis〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Fufa Qu, Jianzhou Tang, Jinting Liao, Bei Chen, Peng Song, Wenjie Luo, Ding Xiong, Tianting Liu, Qianting Gao, Shuangqing Lu, Zhen Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Mitogen-activated protein kinase kinase 6 (MKK6) is an essential component of the p38MAPK signaling pathway, which is involved in the modulation of inflammation, cell apoptosis and survival responses in mammals. However, the function of MKK6s in teleosts is still unclear. In this study, a fish MKK6 homolog (〈em〉Ci〈/em〉MKK6) was first identified from the grass carp (〈em〉Ctenopharyngodon idella〈/em〉), a freshwater fish. 〈em〉Ci〈/em〉MKK6 cDNA encodes a putative protein of 357 amino acids that contains conserved structural characteristics of the MKK6 family, including the S_TKc domain, SVAKT motif and DVD site. The deduced 〈em〉Ci〈/em〉MKK6 protein exhibits high sequence homology with other reported fish MKK6s and shares the closest relationship with MKK6 from 〈em〉Danio rerio〈/em〉. Quantitative real-time PCR (qRT-PCR) analysis revealed that 〈em〉Ci〈/em〉MKK6 mRNA was widely expressed in all tested tissues and stages of embryonic development. Additionally, the transcript levels of 〈em〉Ci〈/em〉MKK6 in the intestine were significantly upregulated in response to bacterial muramyl dipeptide (MDP) and L-Ala-γ-D-Glu-meso-diaminopimelic acid (Tri-DAP) stimulation. Moreover, subcellular localization analysis indicated that 〈em〉Ci〈/em〉MKK6 was distributed in both the cytoplasm and the nucleus of HEK293T cells. Finally, overexpression of 〈em〉Ci〈/em〉MKK6 significantly enhanced the transcriptional activity of the AP-1 reporter gene in HEK293T cells. Overall, these findings may help better clarify the immune function of teleost MKK6s and provide new insight into the immune defense mechanisms of grass carp.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Liting Wu, Shengli Fu, Xiaoxue Yin, Wenna Leng, Zheng Guo, Anli Wang, Jianmin Ye〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Affinity maturation of the antibody response, a process of antibody affinity increasing over response, is one of the key features of the mammalian immune system. However, the process is incompletely understood in teleost, including channel catfish (〈em〉Ictalurus punctaus〈/em〉). In this study, IgM affinity maturation in channel catfish was investigated by estimating the kinetics of antibody affinity using ELISA and ELISPOT assays. Fish were immunized with a T-cell dependent antigen (TNP-KLH), and individual serum IgM antibody titers and affinities, and IgM〈sup〉+〈/sup〉 antibody-secreting cells (ASCs) in peripheral blood were analyzed over a period of 14 weeks. A detectable serum anti-TNP response developed by 2-weeks post-immunization, and the maximal antibody production was observed by 6-weeks post-immunization. The average affinity of anti-TNP serum antibody increased consistently and reached the maximum by 10-weeks post-immunization. The increase of antibody affinity beyond the point of optimal antibody titer revealed that the affinity maturation of IgM antibody response occurred in channel catfish. Dissection of dynamics of individual affinity subpopulations indicated that a significant proportion of low affinity subpopulations appeared at early response, and high affinity subpopulations appeared predominantly at later, resulting in a 100-fold increase in affinity over response. Additional, TNP〈sup〉+〈/sup〉 IgM〈sup〉+〈/sup〉 ASCs was detected by 2-weeks post-immunization and achieved the maximal number by 6-weeks post-immunization. Using an inhibition ELISPOT assay, the findings of a consistent increase in the average affinity of secreted IgM antibody by peripheral blood ASCs, as the immune response progressed, confirmed the occurrence of the affinity maturation. Taken together, the results of this study indicated that affinity maturation occurred in channel catfish following immunization with a TD antigen TNP-KLH.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Weihua Gao, Shuai Li, Qiaoqing Xu, Dashi Zhu, Qin Zhang, Kai Luo, Wenbing Zhang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The CXC chemokine receptors (CXCRs) play critical roles in innate and adaptive immune systems. In this study, six Asian swamp eel (〈em〉Monopterus albus〈/em〉) CXCRs (〈em〉Ma〈/em〉CXCR1–4) were identified and their molecular characterization and expression patterns were analyzed. The open reading frames (ORFs) of 〈em〉Ma〈/em〉CXCR1a, 〈em〉Ma〈/em〉CXCR1b, 〈em〉Ma〈/em〉CXCR2, 〈em〉Ma〈/em〉CXCR3a, 〈em〉Ma〈/em〉CXCR3b, and 〈em〉Ma〈/em〉CXCR4 were 1074 bp (base pairs), 1080 bp, 1125 bp, 1146 bp, 1083 bp, and 1140 bp, and encoded proteins of 357 aa (amino acids), 359 aa, 374 aa, 381 aa, 360 aa, and 379 aa, respectively. All these CXCRs have seven conserved transmembrane domains and four cysteines (with the exception of 〈em〉Ma〈/em〉CXCR3b). Multiple sequence alignment revealed that the 〈em〉Ma〈/em〉CXCRs possess a typical G-protein receptor family 1 signature and a DRY motif. There are also one to four potential N-glycosylation sites in the extracellular regions of the 〈em〉Ma〈/em〉CXCRs, mainly distributed in the N-terminus and extracellular hydrophilic loop (ECL) 2 region. Phylogenetic analysis demonstrated that the 〈em〉Ma〈/em〉CXCRs were clustered together with homologous proteins from other fish. Taken together with the amino acid identity and similarity analysis, these results suggested that the 〈em〉Ma〈/em〉CXCRs are conserved with other homologous genes, in which CXCR4 is more conserved than CXCR1–3. The 〈em〉Ma〈/em〉CXCRs loci showed conserved synteny among teleost fish, and we found that human CXCR1 shares a common ancestor with fish CXCR1a. 〈em〉Ma〈/em〉CXCRs were constitutively expressed in a wide range of tissues (especially in immune-related tissues) with different expression levels, suggesting that the 〈em〉Ma〈/em〉CXCRs have different roles in un-stimulated tissues, and may play vital roles under normal conditions. 〈em〉Ma〈/em〉CXCRs showed different fold changes in the spleen after 〈em〉Aeromonas veronii〈/em〉 and polyinosinic-polycytidylic acid (poly I:C) challenge, which suggested that 〈em〉Ma〈/em〉CXCR1a and 〈em〉Ma〈/em〉CXCR3a have longer antiviral activities compared with their antibacterial functions, and that 〈em〉Ma〈/em〉CXCR1b possesses stronger antiviral than antibacterial activity. 〈em〉Ma〈/em〉CXCR4 may play vital roles during bacterial and viral infection; however, 〈em〉Ma〈/em〉CXCR2 has relatively small effect in antibacterial and antiviral responses. The differential responses of these genes to bacteria and poly I:C implied the differences in the mechanisms of defense against viruses and bacteria.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Rafael Radi, Ana Denicola, Bruce Morgan, Jacek Zielonka〈/p〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): M.A. Rashid, Seonmin Lee, Eunyoung Tak, Jisun Lee, Tae Gyu Choi, Joo-Won Lee, Jae Bum Kim, Jang H. Youn, Insug Kang, Joohun Ha, Sung Soo Kim〈/p〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Dong Liu, Zhiqiang Ma, Shouyin Di, Yang Yang, Jingang Yang, Liqun Xu, Russel J. Reiter, Shubin Qiao, Jiansong Yuan〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Doxorubicin (DOX) is a highly effective anticancer anthracycline drug, but its side effects at the level of the heart has limited its widespread clinical application. Melatonin is a documented potent antioxidant, nontoxic and cardioprotective agent, and it is involved in maintaining mitochondrial homeostasis and function. The present study established acute DOX-induced cardiotoxicity models in both H9c2 cells incubated with 1 μM DOX and C57BL/6 mice treated with DOX (20 mg/kg cumulative dose). Melatonin markedly alleviated the DOX-induced acute cardiac dysfunction and myocardial injury. Both 〈em〉in vivo〈/em〉 and 〈em〉in vitro〈/em〉 studies verified that melatonin inhibited DOX-induced mitochondrial dysfunction and morphological disorders, apoptosis, and oxidative stress 〈em〉via〈/em〉 the activation of AMPK and upregulation of PGC1α with its downstream signaling (NRF1, TFAM and UCP2). These effects were reversed by the use of AMPK siRNA or PGC1α siRNA in H9c2 cells, and were also negated by the cotreatment with AMPK inhibitor Compound C 〈em〉in vivo〈/em〉. Moreover, PGC1α knockdown was without effect on the AMPK phosphorylation induced by melatonin in the DOX treated H9c2 cells. Therefore, AMPK/PGC1α pathway activation may represent a new mechanism for melatonin exerted protection against acute DOX cardiotoxicity through preservation of mitochondrial homeostasis and alleviation of oxidative stress and apoptosis.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918311845-fx1.jpg" width="243" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Lucilla Ribeiro Ávila, Clayson Moura Gomes, Pollyana Guimarães Oliveira, Rodrigo Saar Gomes, Marina Clare Vinaud, Miriam Leandro Dorta, Silvia Reni Bortolin Uliana, Fátima Ribeiro-Dias, Milton Adriano Pelli Oliveira〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Human leishmaniasis caused by 〈em〉Leishmania (Viannia) braziliensis〈/em〉 can be presented as localized cutaneous leishmaniasis (LCL) or mucosal leishmaniasis (ML). Macrophages kill parasites using nitric oxide (NO) and reactive oxygen species (ROS). The aim of this study was to evaluate the ability of parasites obtained from patients with LCL or ML to produce and resist NO or ROS. Promastigotes and amastigotes from LCL or ML isolates produced similar amounts of NO in culture. Promastigotes from ML isolates were more resistant to NO and H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 than LCL parasites in a stationary phase, whereas amastigotes from LCL isolates were more resistant to NO. In addition, in the stationary phase, promastigote isolates from patients with ML expressed more thiol-specific antioxidant protein (TSA) than LCL isolates. Therefore it is suggested that infective promastigotes from ML isolates are more resistant to microbicidal mechanisms in the initial phase of infection. Subsequently, amastigotes lose this resistance. This behavior of ML parasites can decrease the number of parasites capable of stimulating the host immune response shortly after the infection establishment.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918315454-fx1.jpg" width="433" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Yuan Ji, Fang Dai, Bo Zhou〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Higher levels of copper, reduced glutathione (GSH) and reactive oxygen species (ROS) observed in cancer cells than in normal cells, favor the idea of developing copper ionophores as prooxidative anticancer agents (PAAs) to hit the altered redox homeostasis (redox Achilles heel) of cancer cells. In this work, we used salicylaldehyde isonicotinoyl hydrazone (SIH-1) as a basic scaffold to design Cu(II) ionophores with tunable chelation and release of Cu(II) by introducing electron-withdrawing nitro and electron-donating methoxyl groups in the para position to phenolic hydroxyl, or by blocking the phenolic hydroxyl site using methyl. These molecules were used to probe how chelation and release of copper influence their ionophoric role and ability to target redox Achilles heel of cancer cells. Among these molecules, SIH-1 was identified as the most potent Cu(II) ionophore to kill preferentially HepG2 cells over HUVEC cells, and also superior to clioquinol, a copper ionophore evaluated in clinical trials, in terms of its relatively higher cytotoxicity and better selectivity. Higher oxidative potential, despite of lower stability constant, of the Cu(II) complex formed by SIH-1 than by the other molecules, is responsible for its stronger ability in releasing copper by GSH, inducing redox imbalance and triggering mitochondria-mediated apoptosis of HepG2 cells. This work gives useful information on how to design copper ionophores as PAAs for selective killing of cancer cells.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918310931-fx1.jpg" width="354" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Arnaud Tête, Isabelle Gallais, Muhammad Imran, Martine Chevanne, Marie Liamin, Lydie Sparfel, Simon Bucher, Agnès Burel, Normand Podechard, Brice M.R. Appenzeller, Bernard Fromenty, Nathalie Grova, Odile Sergent, Dominique Lagadic-Gossmann〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉We previously demonstrated that co-exposing pre-steatotic hepatocytes to benzo[〈em〉a〈/em〉]pyrene (B[〈em〉a〈/em〉]P), a carcinogenic environmental pollutant, and ethanol, favored cell death. Here, the intracellular mechanisms underlying this toxicity were studied. Steatotic WIF-B9 hepatocytes, obtained by a 48h-supplementation with fatty acids, were then exposed to B[〈em〉a〈/em〉]P/ethanol (10 nM/5 mM, respectively) for 5 days. Nitric oxide (NO) was demonstrated to be a pivotal player in the cell death caused by the co-exposure in steatotic hepatocytes. Indeed, by scavenging NO, CPTIO treatment of co-exposed steatotic cells prevented not only the increase in DNA damage and cell death, but also the decrease in the activity of CYP1, major cytochrome P450s of B[〈em〉a〈/em〉]P metabolism. This would then lead to an elevation of B[〈em〉a〈/em〉]P levels, thus possibly suggesting a long-lasting stimulation of the transcription factor AhR. Besides, as NO can react with superoxide anion to produce peroxynitrite, a highly oxidative compound, the use of FeTPPS to inhibit its formation indicated its participation in DNA damage and cell death, further highlighting the important role of NO. Finally, a possible key role for AhR was pointed out by using its antagonist, CH-223191. Indeed it prevented the elevation of ADH activity, known to participate to the ethanol production of ROS, notably superoxide anion. The transcription factor, NFκB, known to be activated by ROS, was shown to be involved in the increase in iNOS expression. Altogether, these data strongly suggested cooperative mechanistic interactions between B[〈em〉a〈/em〉]P 〈em〉via〈/em〉 AhR and ethanol 〈em〉via〈/em〉 ROS production, to favor cell death in the context of prior steatosis.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918317234-fx1.jpg" width="232" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Lana E. Greene, Richard Lincoln, Gonzalo Cosa〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Lipid peroxidation of polyunsaturated fatty acids in cells may occur via their catalytic autoxidation through peroxyl radicals under oxidative stress conditions. Lipid peroxidation is related to a number of pathologies, and may be invoked in new forms of regulated cell death, yet it may also have beneficial roles in cell signaling cascades. Antioxidants are a natural line of defense against lipid peroxidation, and may accordingly impact the biological outcome associated with the redox chemistry of lipid peroxidation. Critical to unraveling the physiological and pathological role of lipid peroxidation is the development of novel probes with the partition, chemical sensitivity and more importantly, molecular specificity, enabling the spatial and temporal imaging of peroxyl radicals in the lipid membranes of live cells, reporting on the redox status of the cell membrane. This review describes our recent progress to visualize lipid peroxidation in model membrane systems and in live cell studies. Our work portrays the mechanistic insight leading to the development of a highly sensitive probe to monitor lipid peroxyl radicals (LOO〈sup〉•〈/sup〉). It also describes technical aspects including reagents and fluorescence microscopy methodologies to consider in order to achieve the much sought after monitoring of rates of lipid peroxyl radical production in live cell studies, be it under oxidative stress but also under cell homeostasis. This review seeks to bring attention to the study of lipid redox reactions and to lay the groundwork for the adoption of fluorogenic antioxidant probeshancement and maximum intensity recorded in turn provide a benchmark to estimate, when compared to the control BODIPY dye lacking the intramolecular PeT based switch, the overall exte and related fluorescence microscopy methods toward gaining rich spatiotemporal information on lipid peroxidation in live cells.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918301709-fx1.jpg" width="500" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Angus Lindsay, Preston M. McCourt, Peter Karachunski, Dawn A. Lowe, James M. Ervasti〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Generation of superoxide by xanthine oxidase can be stimulated under ischemic and aberrant calcium homeostasis. Because patients and mice with Duchenne muscular dystrophy (DMD) suffer from ischemia and excessive calcium influx, we tested the hypothesis that xanthine oxidase activity is elevated and contributes to disease pathology. Xanthine oxidase activity was measured by urinary isoxanthopterin in DMD patients at rest and in response to exercise. Urinary isoxanthopterin/creatinine was elevated compared to age-matched controls and Becker muscular dystrophy (BMD) patients. Concentrations were also increased after a six minute walk test in ambulatory patients. We also measured urinary isoxanthopterin in wildtype mice and a number of dystrophic mouse models; the DMD mouse model (〈em〉mdx〈/em〉), 〈em〉mdx〈/em〉 mice overexpressing a variety of transgenic miniaturized and chimeric skeletal muscle-specific dystrophins and utrophin and the β-sarcoglycan deficient (〈em〉Scgb〈/em〉〈sup〉〈em〉-/-〈/em〉〈/sup〉) mouse which represents type 2E human limb-girdle muscular dystrophy. 〈em〉Mdx〈/em〉 and 〈em〉Scgb〈/em〉〈sup〉〈em〉-/-〈/em〉〈/sup〉〈em〉mice〈/em〉 had greater urinary isoxanthopterin/creatinine than wildtype mice while 〈em〉mdx〈/em〉 mice expressing dystrophin or utrophin linking the extracellular matrix to the actin cytoskeleton were not different than wildtype. We also measured higher levels of urinary 〈em〉ortho〈/em〉-tyrosine in humans and mice deficient for dystrophin to confirm elevated oxidative stress. Surprisingly, 〈em〉mdx〈/em〉 had lower xanthine oxidase protein levels and higher mRNA in gastrocnemius muscle compared to wildtype mice, however, the enzymatic activity of skeletal muscle xanthine oxidase was elevated above wildtype and a transgenic rescued 〈em〉mdx〈/em〉 mouse (Dys〈sup〉ΔMTB〈/sup〉-〈em〉mdx〈/em〉). Downhill treadmill running also caused significant increases in 〈em〉mdx〈/em〉 urinary isoxanthopterin that was prevented with the xanthine oxidase inhibitor allopurinol. Similarly, 〈em〉in vitro〈/em〉 eccentric contraction-induced force drop of 〈em〉mdx〈/em〉 muscle was attenuated by the allopurinol metabolite, oxypurinol. Together, our data suggests hyper-activity of xanthine oxidase in DMD, identifies xanthine oxidase activity as a contributing factor in eccentric contraction-induced force drop of dystrophin-deficient skeletal muscle and highlights the potential of isoxanthopterin as a noninvasive biomarker in DMD.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S089158491831311X-fx1.jpg" width="276" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Iva I. Podgorski, Marija Pinterić, Sandra Sobočanec, Marijana Popović Hadžija, Mladen Paradžik, Ana Dekanić, Maja Marinović, Mirna Halasz, Robert Belužić, Grazia Davidović, Andreja Ambriović Ristov, Tihomir Balog〈/p〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): A.H. Falomir-Lockhart, E.E. Villegas-Castagnaso, G. Giovambattista, A. Rogberg-Muñoz〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The development of high-throughput technologies in the last decade produced an exponential increase in the amount of biological data available. The case of redox biology and apoptosis is not an exception, and nowadays there is a need to integrate information from multiple “omics” studies. Therefore, validation of proposed discoveries is essential. However, the study in biological systems of the effect of the massive amounts of sequence variation data generated with next-generation sequencing (NGS) technologies can be a very difficult and expensive process. In this context, the present study aimed to demonstrate the advantages of a computational methodology to systematically analyze the structural and functional effects of protein variants, in order to prioritize further studies. This approach stands out for its easy implementation, low costs and low time consumed. First, the possible impact of mutations on protein structure and function was tested by a combination of tools based on evolutionary and structural information. Next, homology modeling was performed to predict and compare the 3D protein structures of unresolved amino acid sequences obtained from genomic resequencing. This analysis applied to the bovine GSTP1 allowed to determine that some of amino acid substitutions may generate important changes in protein structure and function. Moreover, the haplotype analysis highlighted three structure variants worthwhile studying through 〈em〉in vitro〈/em〉 or 〈em〉in vivo〈/em〉 experiments.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918309407-fx1.jpg" width="494" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Rong Li, Xiaoqi Zhou, Dan Liu, Wei Feng〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Dismutation of superoxide by superoxide dismutase (SOD) generates hydrogen peroxide, which may be reduced to hydroxyl radical. The generated H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 during the catalysis can have an oxidative damage to SOD. Hydrogen peroxide decomposition by catalase (CAT) can help circumvent the problem. Mn-superoxide dismutase (herein referred to as SOD) and CAT are dimeric and tetrameric proteins, respectively. Herein, through intein-mediated in vivo subunit splicing, the C-terminus of the CAT subunit (CAT〈sup〉S〈/sup〉) has been specifically ligated to the N-terminus of the SOD subunit (SOD〈sup〉S〈/sup〉) with a peptide bond. Thus, the splicing product SOD&CAT combines the superoxide anion (•O〈sub〉2〈/sub〉〈sup〉−〈/sup〉) scavenging ability and the ability of decomposing H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉. The in vivo subunit splicing has little effect on the secondary structures of the enzymes as confirmed by circular dichroism (CD) spectra. Fluorescence spectra showed that the splicing product SOD&CAT has a higher stability than SOD. In the splicing product SOD&CAT, the SOD subunits are in close proximity to the CAT subunits, facilitating immediate transfer of H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 between the enzymes and enabling efficient decomposition of H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉. SOD&CAT exhibited a superoxide anion (•O〈sub〉2〈/sub〉〈sup〉−〈/sup〉) scavenging ability 244% higher than that of SOD and 46% higher than that of the mixed enzymes SOD+CAT.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918316058-fx1.jpg" width="428" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): O.G. Lyublinskaya, S.A. Antonov, S.G. Gorokhovtsev, N.A. Pugovkina, Ju.S. Kornienko, Ju.S. Ivanova, A.N. Shatrova, N.D. Aksenov, V.V. Zenin, N.N. Nikolsky〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉HyPer is a genetically encoded fluorogenic sensor for hydrogen peroxide which is generally used for the ratiometric imaging of H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 fluxes in living cells. Here, we demonstrate the advantages of HyPer-based ratiometric flow cytometry assay for H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉, by using K562 and human mesenchymal stem cell lines expressing HyPer. We show that flow cytometry analysis is suitable to detect HyPer response to submicromolar concentrations of extracellularly added H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 that is much lower than concentrations addressed previously in the other HyPer-based assays (such as cell imaging or fluorimetry). Suggested technique is also much more sensitive to hydrogen peroxide than the widespread flow cytometry assay exploiting H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉-reactive dye H〈sub〉2〈/sub〉DCFDA and, contrary to the H〈sub〉2〈/sub〉DCFDA-based assay, can be employed for the kinetic studies of H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 utilization by cells, including measurements of the rate constants of H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 removal. In addition, flow cytometry multi-parameter ratiometric measurements enable rapid and high-throughput detection of endogenously generated H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 in different subpopulations of HyPer-expressing cells. To sum up, HyPer can be used in multi-parameter flow cytometry studies as a highly sensitive indicator of intracellular H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918309493-fx1.jpg" width="405" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Matthew Krilis, Miao Qi, Jian Qi, Jason W.H. Wong, Robyn Guymer, Gerald Liew, Alex P. Hunyor, Michele Madigan, Peter McCluskey, James Weaver, Steven A. Krilis, Bill Giannakopoulos〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉 〈p〉Complement Factor H (CFH) is an important inhibitor of the alternate complement pathway in Bruch's membrane (BM), located between the choriocapillaris and the retinal pigment epithelium. Furthermore dysfunction of its activity as occurs with certain polymorphisms is associated with an increased risk of age related macular degeneration (AMD).〈/p〉 〈p〉The retina is a site of high generation of reactive oxygen species (ROS) and dysfunction of redox homeostasis in this milieu also contributes to AMD pathogenesis. In this study we wanted to explore if CFH exists in distinct redox forms and whether these species have unique protective biological functions. CFH can be reduced by the naturally occurring thioredoxin − 1 in CFH domains 1–4, 17–20. We found a duality of function between the oxidised and reduced forms of CFH. The oxidised form was more efficient in binding to C3b and lipid peroxidation by-products that are known to accumulate in the retinae and activate the alternate complement pathway.〈/p〉 〈p〉Oxidised CFH enhances Factor I mediated cleavage of C3 and C3b whereas the reduced form loses this activity. In the setting of oxidative stress (hydrogen peroxide)-mediated death of human retinal pigment epithelial cells as can occur in AMD, the free thiol form of CFH offers a protective function compared to the oxidised form. We found for the first time using a novel ELISA system we have developed for free thiol CFH, that both redox forms of CFH are found in the human plasma. Furthermore there is a distinct ratio of these redox forms in plasma depending if an individual has early or late AMD, with individuals with early AMD having higher levels of the free thiol form compared to late AMD.〈/p〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918310670-fx1.jpg" width="254" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Weiche Wu, Sisi Wang, Qing Liu, Xinxia Wang, Tizhong Shan, Yizhen Wang〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Dysregulated activation of inflammation is associated with the development and progression of many diseases. Generation of reactive oxygen species (ROS) has been shown to promote an inflammatory response. Cathelicidin peptides not only defend against the invasion of various microbes but also play an important role in regulating immune responses. The objective of this study was to investigate the effects and mechanisms of Cathelicidin-WA (CWA) on the inflammatory response and oxidative stress in macrophages. Our results showed that CWA efficiently attenuated lipopolysaccharide (LPS)-stimulated inflammation and oxidative stress both 〈em〉in vivo〈/em〉 and 〈em〉in vitro〈/em〉. Mechanistically, we found that CWA significantly reduced the LPS-induced nuclear translocation of NF-κB, thus decreasing the production of the pro-inflammatory cytokines TNF-α and IL-6 in macrophages. On the other hand, CWA markedly promoted the nuclear translocation of Nrf2 via the AKT pathway and p38 signaling. This resulted in increased expression of the anti-oxidative genes NQO-1 and HO-1 and alleviated oxidative stress in LPS-stimulated macrophages. Interestingly, the effects of CWA were diminished when AMPK was knocked down. Consistently, we noticed that CWA failed to ameliorate the LPS-induced inflammatory response and oxidative stress in AMPK knockout mice. Furthermore, we discovered that LKB1 was essential for AMPK activation by CWA. These data demonstrated for the first time that CWA attenuated LPS-stimulated inflammation and redox imbalance through regulating LKB1-AMPK signaling. Such knowledge provides new insights into the mechanisms through which Cathelicidin peptides modulate immune responses.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918314047-fx1.jpg" width="249" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Quach Ngoc Tung, Nico Linzner, Vu Van Loi, Haike Antelmann〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉 〈p〉Gram-negative bacteria utilize glutathione (GSH) as their major LMW thiol. However, most Gram-positive bacteria do not encode enzymes for GSH biosynthesis and produce instead alternative LMW thiols, such as bacillithiol (BSH) and mycothiol (MSH). BSH is utilized by 〈em〉Firmicutes〈/em〉 and MSH is the major LMW thiol of 〈em〉Actinomycetes〈/em〉. LMW thiols are required to maintain the reduced state of the cytoplasm, but are also involved in virulence mechanisms in human pathogens, such as 〈em〉Staphylococcus aureus〈/em〉, 〈em〉Mycobacterium tuberculosis〈/em〉, 〈em〉Streptococcus pneumoniae〈/em〉, 〈em〉Salmonella enterica〈/em〉 subsp. Typhimurium and 〈em〉Listeria monocytogenes〈/em〉. Infection conditions often cause perturbations of the intrabacterial redox balance in pathogens, which is further affected under antibiotics treatments. During the last years, novel glutaredoxin-fused roGFP2 biosensors have been engineered in many eukaryotic organisms, including parasites, yeast, plants and human cells for dynamic live-imaging of the GSH redox potential in different compartments. Likewise bacterial roGFP2-based biosensors are now available to measure the dynamic changes in the GSH, BSH and MSH redox potentials in model and pathogenic Gram-negative and Gram-positive bacteria.〈/p〉 〈p〉In this review, we present an overview of novel functions of the bacterial LMW thiols GSH, MSH and BSH in pathogenic bacteria in virulence regulation. Moreover, recent results about the application of genetically encoded redox biosensors are summarized to study the mechanisms of host-pathogen interactions, persistence and antibiotics resistance. In particularly, we highlight recent biosensor results on the redox changes in the intracellular food-borne pathogen 〈em〉Salmonella〈/em〉 Typhimurium as well as in the Gram-positive pathogens 〈em〉S. aureus〈/em〉 and 〈em〉M. tuberculosis〈/em〉 during infection conditions and under antibiotics treatments. These studies established a link between ROS and antibiotics resistance with the intracellular LMW thiol-redox potential. Future applications should be directed to compare the redox potentials among different clinical isolates of these pathogens in relation to their antibiotics resistance and to screen for new ROS-producing drugs as promising strategy to combat antimicrobial resistance.〈/p〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S089158491830073X-fx1.jpg" width="216" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Ludwig K. Groebler, Xiao Suo Wang, Hyun Bo Kim, Anu Shanu, Farjaneh Hossain, Aisling C. McMahon, Paul K. Witting〈/p〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Amanda A. Felizardo, Ivo S. Caldas, Andréa A.S. Mendonça, Reggiani V. Gonçalves, Fernanda L. Tana, Leonardo A. Almeida, Rômulo D. Novaes〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Elderly organisms are more susceptible to infectious diseases. However, the impact of aging on antiparasitic mechanisms, especially the nitric oxide pathway, is poorly understood. Using an integrated 〈em〉in vivo〈/em〉 and 〈em〉in vitro〈/em〉 model, we compared the severity of 〈em〉Trypanosoma cruzi〈/em〉 infection in young and elderly (8 or 72 weeks old) mice. Forty C57BL/6 mice were randomized into four groups: Y-inf, young infected; Yn-inf, young uninfected; A-inf, aged infected; An-inf, aged uninfected. Parasitemia was measured daily, and animals were euthanized after 15 days of infection. 〈em〉Trypanosoma cruzi〈/em〉-induced inflammatory processes were analyzed in blood and heart samples, as well as in bone marrow-derived macrophages (BMDMs) co-cultured with splenocytes isolated from young or elderly mice. Our results indicated upregulated IgG2b and IL-17 production in elderly animals, which was not sufficient to reduce parasitemia, parasitic load and myocarditis to levels observed in young animals. The higher susceptibility of elderly mice to 〈em〉T. cruzi〈/em〉 infection was accompanied by reduced cardiac inducible nitric oxide synthase (iNOS) gene expression, nitric oxide (NO) and IFN-γ levels, as well as an antagonistic upregulation of arginase-1 expression and arginase activity. The same responses were observed when BMDMs co-cultured with splenocytes from elderly mice were stimulated with 〈em〉T. cruzi〈/em〉 antigens. Our findings indicate that elderly mice were more susceptible to 〈em〉T. cruzi〈/em〉 infection, which was potentially related to an attenuated response to antigenic stimulation, inhibition of iNOS gene expression and NO production, and antagonistic upregulation of arginase gene expression and activity, which created favorable conditions for heart parasitism and myocarditis development.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918309286-fx1.jpg" width="275" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 130〈/p〉 〈p〉Author(s): Chengliang Luo, Qiang Yang, Yuancai Liu, Shuanhu Zhou, Jiying Jiang, Russel J. Reiter, Pallab Bhattacharya, Yongchun Cui, Hongwei Yang, He Ma, Jiemin Yao, Sean E. Lawler, Xinmu Zhang, Jianfang Fu, Renato Rozental, Hany Aly, Mark D. Johnson, E. Antonio Chiocca, Xin Wang〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Melatonin is a neurohormone associated with sleep and wakefulness and is mainly produced by the pineal gland. Numerous physiological functions of melatonin have been demonstrated including anti-inflammation, suppressing neoplastic growth, circadian and endocrine rhythm regulation, and its potent antioxidant activity as well as its role in regeneration of various tissues including the nervous system, liver, bone, kidney, bladder, skin, and muscle, among others. In this review, we summarize the recent advances related to the multiple protective roles of melatonin receptor agonists, melatonin and N-acetylserotonin (NAS), in brain injury, liver damage, and bone health. Brain injury, including traumatic brain injury, ischemic stroke, intracerebral hemorrhage, subarachnoid hemorrhage, and newborn perinatal hypoxia-ischemia encephalopathy, is a major cause of mortality and disability. Liver disease causes serious public health problems and various factors including alcohol, chemical pollutants, and drugs induce hepatic damage. Osteoporosis is the most common bone disease in humans. Due in part to an aging population, both the cost of care of fracture patients and the annual fracture rate have increased steadily. Despite the discrepancy in the pathophysiological processes of these disorders, time frames and severity, they may share several common molecular mechanisms. Oxidative stress is considered to be a critical factor in these pathogeneses. We update the current state of knowledge related to the molecular processes, mainly including anti-oxidative stress, anti-apoptosis, autophagy dysfunction, and anti-inflammation as well as other properties of melatonin and NAS. Particularly, the abilities of melatonin and NAS to directly scavenge oxygen-centered radicals and toxic reactive oxygen species, and indirectly act through antioxidant enzymes are disscussed. In this review, we summarize the similarities and differences in the protection provided by melatonin and/or NAS in brain, liver and bone damage. We analyze the involvement of melatonin receptor 1A (MT1), melatonin receptor 1B (MT2), and melatonin receptor 1C (MT3) in the protection of melatonin and/or NAS. Additionally, we evaluate their potential clinical applications. The multiple mechanisms of action and multiple organ-targeted properties of melatonin and NAS may contribute to development of promising therapies for clinical trials.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918309481-fx1.jpg" width="222" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Paul E. Pace, Alexander V. Peskin, Andreas Konigstorfer, Christine J. Jasoni, Christine C. Winterbourn, Mark B. Hampton〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Hydrogen peroxide (H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉) acts as a signaling molecule in cells by oxidising cysteine residues in regulatory proteins such as phosphatases, kinases and transcription factors. It is unclear exactly how many of these proteins are specifically targeted by H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 because they appear too unreactive to be directly oxidised. One proposal is that peroxiredoxins (Prxs) initially react with H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 and then oxidise adjacent proteins via a thiol relay mechanism. The aim of this study was to identify constitutive interaction partners of Prx2 in Jurkat T-lymphoma cells, in which thiol protein oxidation occurs at low micromolar concentrations of H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉. Immunoprecipitation and proximity ligation assays identified a physical interaction between collapsin response mediator protein 2 (CRMP2) and cytoplasmic Prx2. CRMP2 regulates microtubule structure during lymphocyte migration and neuronal development. Exposure of Jurkat cells to low micromolar levels of H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 caused rapid and reversible oxidation of CRMP2, in parallel with Prx2 oxidation, despite purified recombinant CRMP2 protein reacting slowly with H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 (k~1 M〈sup〉−1〈/sup〉s〈sup〉−1〈/sup〉). Lowering Prx expression should inhibit oxidation of proteins oxidised by a relay mechanism, however knockout of Prx2 had no effect on CRMP2 oxidation. CRMP2 also interacted with Prx1, suggesting redundancy in single knockout cells. Prx 1 and 2 double knockout Jurkat cells were not viable. An interaction between Prx2 and CRMP2 was also detected in other human and rodent cells, including primary neurons. However, low concentrations of H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 did not cause CRMP2 oxidation in these cells. This indicates a cell-type specific mechanism for promoting CRMP2 oxidation in Jurkat cells, with insufficient evidence to attribute oxidation to a Prx-dependent redox relay.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918311225-fx1.jpg" width="249" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 130〈/p〉 〈p〉Author(s): Elis Aguiar Morra, Paula Lopes Rodrigues, Itamar Couto Guedes de Jesus, Patrícia Ribeiro Do Val Lima, Renata Andrade Ávila, Tadeu Ériton Caliman Zanardo, Breno Valentim Nogueira, Donald M. Bers, Silvia Guatimosim, Ivanita Stefanon, Rogério Faustino Ribeiro Júnior〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉We previously demonstrated that the loss of female hormones induces cardiac and mitochondrial dysfunction in the female heart. Here, we show the impact of endurance training for twelve weeks, a nonpharmacological therapy against cardiovascular disease caused by ovariectomy and its contribution to cardiac contractility, mitochondrial quality control, bioenergetics and oxidative damage. We found that ovariectomy induced cardiac hypertrophy and dysfunction by decreasing SERCA2 and increasing phospholamban protein expression. Endurance training restored myocardial contractility, SERCA2 levels, increased calcium transient in ovariectomized rats but did not change phospholamban protein expression or cardiac hypertrophy. Additionally, ovariectomy decreased the amount of intermyofibrillar mitochondria and induced mitochondrial fragmentation that were accompanied by decreased levels of mitofusin 1, PGC-1α, NRF-1, total AMPK-α and mitochondrial Tfam. Endurance training prevented all these features except for mitofusin 1. Ovariectomy reduced O〈sub〉2〈/sub〉 consumption, elevated O〈sub〉2〈/sub〉〈sup〉.-〈/sup〉 release and increased Ca〈sup〉2+〈/sup〉-induced mitochondrial permeability transition pore opening in both mitochondrial subpopulations. Ovariectomy also increased NOX-4 protein expression in the heart, reduced mitochondrial Mn-SOD, catalase protein expression and increased protein carbonylation in both mitochondrial subpopulations, which were prevented by endurance training. Taken together, our findings show that endurance training prevented cardiac contractile dysfunction and mitochondrial quality control in ovariectomized rats.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918309705-fx1.jpg" width="423" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Sonu Singh, Akanksha Mishra, Soni Jignesh Mohanbhai, Virendra Tiwari, Rajnish Kumar Chaturvedi, Sukant Khurana, Shubha Shukla〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Wnts and the components of Wnt/β-catenin signaling are widely expressed in midbrain and required to control the fate specification of dopaminergic (DAergic) neurons, a neuronal population that specifically degenerate in Parkinson's disease (PD). Accumulating evidence suggest that mitochondrial dysfunction plays a key role in pathogenesis of PD. Axin-2, a negative regulator of Wnt/β-catenin signaling affects mitochondrial biogenesis and death/birth of new DAergic neurons is not fully explored. We investigated the functional role of Axin-2/Wnt/β-catenin signaling in mitochondrial biogenesis and DAergic neurogenesis in 6-hydroxydopamine (6-OHDA) induced rat model of PD-like phenotypes. We demonstrate that single unilateral injection of 6-OHDA into the medial forebrain bundle (MFB) potentially dysregulates Wnt/β-catenin signaling in substantia nigra pars compacta (SNpc). We used shRNA lentiviruses to genetically knockdown Axin-2 to up-regulate Wnt/β-catenin signaling in SNpc in parkinsonian rats. Genetic knockdown of Axin-2 up-regulates Wnt/β-catenin signaling by destabilizing the β-catenin degradation complex in SNpc in parkinsonian rats. Axin-2 shRNA mediated activation of Wnt/β-catenin signaling improved behavioural functions and protected the nigral DAergic neurons by increasing mitochondrial functionality in parkinsonian rats. Axin-2 shRNA treatment reduced apoptotic signaling, autophagy and ROS generation and improved mitochondrial membrane potential which promotes mitochondrial biogenesis in SNpc in parkinsonian rats. Interestingly, Axin-2 shRNA-mediated up-regulation of Wnt/β-catenin signaling enhanced net DAergic neurogenesis by regulating proneural genes (Nurr-1, Pitx-3, Ngn-2, and NeuroD1) and mitochondrial biogenesis in SNpc in parkinsonian rats. Therefore, our data suggest that pharmacological/genetic manipulation of Wnt signaling that enhances the endogenous regenerative capacity of DAergic neurons may have implication for regenerative approaches in PD.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉A proposed schematic representation of the means by which Promote Mitochondrial Biogenesis and Dopaminergic Neurogenesis by regulating Wnt/β-catenin Signaling in Rat Model of Parkinson's Disease.〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918312176-fx1.jpg" width="314" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Lucas Kich Grun, Nevton da Rosa Teixeira, Lúcia von Mengden, Marco Antônio de Bastiani, Mariana Migliorini Parisi, Rafael Bortolin, Patrícia Lavandoski, Vinícius Pierdoná, Letícia Biscaino Alves, José Cláudio Fonseca Moreira, Cláudio Corá Mottin, Marcus Herbert Jones, Fábio Klamt, Alexandre Vontobel Padoin, Fátima Costa Rodrigues Guma, Florencia María Barbé-Tuana〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Obesity is a prevalent multifactorial chronic disorder characterized by metabolic dysregulation. Sustained pro-oxidative mediators trigger harmful consequences that reflect at systemic level and contribute for the establishment of a premature senescent phenotype associated with macromolecular damage (DNA, protein, and lipids). Telomeres are structures that protect chromosome ends and are associated with a six-protein complex called the shelterin complex and subject to regulation. Under pro-oxidant conditions, telomere attrition and the altered expression of the shelterin proteins are central for the establishment of many pathophysiological conditions such as obesity. Thus, considering that individuals with obesity display a systemic oxidative stress profile that may compromise the telomeres length or its regulation, the aim of this study was to investigate telomere homeostasis in patients with obesity and explore broad/systemic associations with the expression of shelterin genes and the plasma redox state. We performed a cross-sectional study in 39 patients with obesity and 27 eutrophic subjects. Telomere length (T/S ratio) and gene expression of shelterin components were performed in peripheral blood mononuclear cells by qPCR. The oxidative damage (lipid peroxidation and protein carbonylation) and non-enzymatic antioxidant system (total radical-trapping antioxidant potential/reactivity, sulfhydryl and GSH content) were evaluated in plasma. Our results demonstrate that independently of comorbidities, individuals with obesity had significantly shorter telomeres, augmented expression of negative regulators of the shelterin complex, increased lipid peroxidation and higher oxidized protein levels associated with increased non-enzymatic antioxidant defenses. Principal component analysis revealed TRF1 as a major contributor for firstly telomeres shortening. In conclusion, our study is first showing a comprehensive analysis of telomeres in the context of obesity, associated with dysregulation of the shelterin components that was partially explained by TRF1 upregulation that could not be reversed by the observed adaptive non-enzymatic antioxidant response.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918315521-fx1.jpg" width="441" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Marta Sanchez-Marinas, David Gimenez-Zaragoza, Edgar Martin-Ramos, Julia Llanes, José Cansado, Maria Jesús Pujol, Oriol Bachs, Rosa Aligue〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Different studies have demonstrated multiple effects of arsenite on human physiology. However, there are many open questions concerning the mechanism of response to arsenite. 〈em〉Schizosaccharomyces pombe〈/em〉 activates the Sty1 MAPK pathway as a common response to several stress conditions. The specificity of the response is due to the activation of different transcription factors and specific targets such the Cmk2 MAPKAP kinase. We have previously shown that Cmk2 is phosphorylated and activated by the MAPK Sty1 in response to oxidative stress. Here, we report that Cmk2 kinase is specifically necessary to overcome the stress caused by metalloid agents, in particular arsenite. Deletion of 〈em〉cmk2〈/em〉 increases the protein level of various components of the MAPK pathway. Moreover, Cmk2 negatively regulates translation through the Cpc2 kinase: the RACK1 orthologue in fission yeast. RACK1 is a receptor for activated C-kinase. Interestingly, RACK1 is a constituent of the eukaryotic ribosome specifically localized in the head region of the 40 S subunit. Cmk2 controls arsenite response through Cpc2 and it does so through Cpc2 ribosomal function, as observed in genetic analysis using a Cpc2 mutant unable to bind to ribosome. These findings suggest a role for Cmk2 in regulating translation and facilitating adaptation to arsenite stress in the ribosome.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918316101-fx1.jpg" width="287" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Ligia Akemi Kiyuna, Rudá Prestes e Albuquerque, Che-Hong Chen, Daria Mochly-Rosen, Julio Cesar Batista Ferreira〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Mitochondrial dysfunction characterized by impaired bioenergetics, oxidative stress and aldehydic load is a hallmark of heart failure. Recently, different research groups have provided evidence that selective activation of mitochondrial detoxifying systems that counteract excessive accumulation of ROS, RNS and reactive aldehydes is sufficient to stop cardiac degeneration upon chronic stress, such as heart failure. Therefore, pharmacological and non-pharmacological approaches targeting mitochondria detoxification may play a critical role in the prevention or treatment of heart failure. In this review we discuss the most recent findings on the central role of mitochondrial dysfunction, oxidative stress and aldehydic load in heart failure, highlighting the most recent preclinical and clinical studies using mitochondria-targeted molecules and exercise training as effective tools against heart failure.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918311304-fx1.jpg" width="440" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Ling Zhou, Yuetao Yi, Qing Yuan, Jing Zhang, Youjie Li, Pingyu Wang, Maolei Xu, Shuyang Xie〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Previous studies have confirmed that protein tyrosine phosphatase 1B (PTP1B) can promote tumour progression in non-small cell lung cancer (NSCLC). Vanadyl alginate oligosaccharides (VAOS) is a new coordination compounds that possesses a good PTP1B inhibitory activity. However, the potent anticancer efficacy of VAOS in human NSCLC requires further study. In this study, VAOS exhibited effective inhibitory effects in NSCLC both in cultured cells and in a xenograft mouse model. VAOS was further identified to induce NSCLC cell apoptosis through activating protein kinase B (AKT) to elevate intracellular reactive oxygen species (ROS) levels by increasing in oxygen consumption and impairing the ROS-scavenging system. Neither silencing of 〈em〉PTP1B〈/em〉 by siRNA nor transient overexpression of 〈em〉PTP1B〈/em〉 had an effect on the AKT phosphorylation triggered by VAOS, indicating that PTP1B inhibition was not involved in VAOS-induced apoptosis. Through phosphorus colorimetric assay, we demonstrated that VAOS notably inhibited phosphatase and tensin homologue deleted on chromosome 10 (PTEN) dephosphorylation activity, another member of the protein tyrosine phosphatases (PTPases)-upstream factor of AKT. Interestingly, 〈em〉PTEN〈/em〉 knockdown sensitized cells to VAOS, whereas ectopic expression of 〈em〉PTEN〈/em〉 markedly rescued VAOS-mediated lethality. 〈em〉In vivo〈/em〉, VAOS treatment markedly reduced PTEN activity and tumour cell burden with low systemic toxicity. Thus, our data not only provided a new therapeutic drug candidate for NSCLC, but presented new understanding into the pharmacological research of VAOS.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918310815-fx1.jpg" width="252" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Renata Smulik-Izydorczyk, Karolina Dębowska, Jakub Pięta, Radosław Michalski, Andrzej Marcinek, Adam Sikora〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Nitroxyl (〈strong〉HNO)〈/strong〉, which according to the IUPAC recommended nomenclature should be named azanone, is the protonated one-electron reduction product of nitric oxide. Recently, it has gained a considerable attention due to the interesting pharmacological effects of its donors. Although there has been great progress in the understanding of 〈strong〉HNO〈/strong〉 chemistry and chemical biology, it still remains the most elusive reactive nitrogen species, and its selective detection is a real challenge. The development of reliable methodologies for the direct detection of azanone is essential for the understanding of important signaling properties of this reactive intermediate and its pharmacological potential. Over the last decade, there has been considerable progress in the development of low-molecular-weight fluorogenic probes for the detection of 〈strong〉HNO,〈/strong〉 and therefore, in this review, we have focused on the challenges and limitations of and perspectives on nitroxyl detection based on the use of such probes.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S089158491830741X-fx1.jpg" width="498" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Leonel Malacrida, Enrico Gratton〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Fluorescence spectroscopy, coupled with microscopy, opens new frontiers for the study of dynamic processes with high spatio-temporal resolution. The application of phasor plots to FLIM and hyperspectral imaging demonstrate unprecedented capabilities to study complex photophysics at the subcellular level. Using these approaches we studied the effects of an H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 bolus on NIH-3T3 membranes dynamics monitored by LAURDAN fluorescence. Exposure of NIH-3T3 cells to a bolus of H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 modifies the cell membranes and, in particular, the plasma membrane in a complex manner. The LAURDAN results reveal that the peroxide treatment decreases membrane fluidity but surprisingly increases dipolar relaxation around the excited probe. Using the Multidimensional-phasor approach we elucidated the complex photophysics of LAURDAN incorporated into cell membrane after H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 exposure. The results indicate the occurrence of LAURDAN fast-diffusion from gel↔〈em〉l〈/em〉〈sub〉〈em〉d〈/em〉〈/sub〉 phases in membranes exposed to a H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 bolus. An ad hoc hypothesis is presented to interpret the results in the context of H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 oxidative distress/eustress.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918310086-fx1.jpg" width="473" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Sebastián Carballal, Valeria Valez, Damián Alvarez-Paggi, Artak Tovmasyan, Ines Batinic-Haberle, Gerardo Ferrer-Sueta, Daniel H. Murgida, Rafael Radi〈/p〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Juan A. Parga, Ana I. Rodriguez-Perez, Maria Garcia-Garrote, Jannette Rodriguez-Pallares, Jose L. Labandeira-Garcia〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Nuclear factor-E2-related factor 2 (NRF2) is a transcription factor that activates the antioxidant cellular defense in response to oxidative stress, leading to neuroprotective effects in Parkinson's disease (PD) models. We have previously shown that Angiotensin II (AngII) induces an increase in reactive oxygen species (ROS) via AngII receptor type 1 and NADPH oxidase (NOX), which may activate the NRF2 pathway. However, controversial data suggest that AngII induces a decrease in NRF2 signaling leading to an increase in oxidative stress. We analyzed the effect of AngII and the dopaminergic neurotoxin 6-hydroxydopamine (6-OHDA) in culture and in vivo, and examined the effects on the expression of NRF2-related genes. Treatment of neuronal cell lines Mes23.5, N27 and SH-SY5Y with AngII, 6-OHDA or a combination of both increased ROS production and reduced cell viability. Simultaneously, these treatments induced an increase in expression in the NRF2-regulated genes heme oxygenase 1 (〈em〉Hmox1〈/em〉), NAD(P)H quinone dehydrogenase 1 (〈em〉Nqo1〈/em〉) and Kruppel like factor 9 (〈em〉Klf9〈/em〉). Moreover, overexpression of KLF9 transcription factor caused a reduction in the production of ROS induced by treatment with AngII or 6-OHDA and improved the survival of these neuronal cells. Rats treated with AngII, 6-OHDA or a combination of both also showed an increased expression of NRF2 related genes and KLF9. In conclusion, our data indicate that AngII induces a damaging effect in neuronal cells, but also acts as a signaling molecule to activate NRF2 and KLF9 neuroprotective pathways in cellular and animal models of PD.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918312383-fx1.jpg" width="430" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Ying Ma, Zhenyu Huang, Zhaoli Zhou, Xiaoyan He, Ying Wang, Chao Meng, Gang Huang, Ningyuan Fang〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉A bulk of cholesteryl esters accumulation in macrophage foam cells drives the occurrence and development of atherosclerosis. Evidence now shows that autophagy plays key roles in the degradation of intracellular lipid droplets 〈em〉via〈/em〉 autolysosome, and also in the release of intracellular lipids 〈em〉via〈/em〉 cholesterol efflux. In this study, we identified that a mitochondria-targeted antioxidant, Mito-Tempol, has protective effects against cholesteryl esters accumulation by activating autophagy. Mito-Tempol was shown to ameliorate the lipid burden for atherosclerosis, both 〈em〉in vitro〈/em〉 and 〈em〉in vivo.〈/em〉 In the established 〈em〉in vitro〈/em〉 foam cell formation system using oxidized low-density lipoprotein (ox-LDL)-loaded THP-1 macrophages, Mito-Tempol prevented intracellular oxidative stress and attenuated lipid accumulation. Mito-Tempol rescued ox-LDL-impaired autophagic flux, thereby facilitating autophagy-mediated lipid degradation in THP-1 macrophages. Meanwhile, Mito-Tempol also increased the efflux of cholesterol 〈em〉via〈/em〉 autophagy-dependent ABCA1 and ABCG1 up-regulation. The classical autophagy pathway of mTOR may be one of the effector for the autophagy restoration of Mito-Tempol. Our findings give the first insight that cardiovascular system disease may benefits more from the treatment of Mito-Tempol for its impact of reversing atherosclerosis 〈em〉via〈/em〉 autophagy.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918321828-fx1.jpg" width="438" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Emrah Eroglu, Suphachai Charoensin, Helmut Bischof, Jeta Ramadani, Benjamin Gottschalk, Maria R. Depaoli, Markus Waldeck-Weiermair, Wolfgang F. Graier, Roland Malli〈/p〉 〈h5〉Short abstract〈/h5〉 〈div〉〈p〉Over the last decades a broad collection of sophisticated fluorescent protein-based probes was engineered with the aim to specifically monitor nitric oxide (NO), one of the most important signaling molecules in biology. Here we report and discuss the characteristics and fields of applications of currently available genetically encoded fluorescent sensors for the detection of NO and its metabolites in different cell types.〈/p〉〈/div〉 〈div〉 〈h6〉Long abstract〈/h6〉 〈p〉Because of its radical nature and short half-life, real-time imaging of NO on the level of single cells is challenging. Herein we review state-of-the-art genetically encoded fluorescent sensors for NO and its byproducts such as peroxynitrite, nitrite and nitrate. Such probes enable the real-time visualization of NO signals directly or indirectly on the level of single cells and cellular organelles and, hence, extend our understanding of the spatiotemporal dynamics of NO formation, diffusion and degradation. Here, we discuss the significance of NO detection in individual cells and on subcellular level with genetic biosensors. Currently available genetically encoded fluorescent probes for NO and nitrogen species are critically discussed in order to provide insights in the functionality and applicability of these promising tools. As an outlook we provide ideas for novel approaches for the design and application of improved NO probes and fluorescence imaging protocols.〈/p〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918300376-fx1.jpg" width="498" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Michael J. Houghton, Asimina Kerimi, Sarka Tumova, John P. Boyle, Gary Williamson〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Hyperglycemia augments formation of intracellular reactive oxygen species (ROS) with associated mitochondrial damage and increased risk of insulin resistance in type 2 diabetes. We examined whether quercetin could reverse chronic high glucose-induced oxidative stress and mitochondrial dysfunction. Following long-term high glucose treatment, complex I activity was significantly decreased in isolated mitochondria from HepG2 cells. Quercetin dose-dependently recovered complex I activity and lowered cellular ROS generation under both high and normal glucose conditions. Respirometry studies showed that quercetin could counteract the detrimental increase in inner mitochondrial membrane proton leakage resulting from high glucose while it increased oxidative respiration, despite a decrease in electron transfer system (ETS) capacity, and lower non-ETS oxygen consumption. A quercetin-stimulated increase in cellular NAD〈sup〉+〈/sup〉/NADH was evident within 2 h and a two-fold increase in PGC-1α mRNA within 6 h, in both normal and high glucose conditions. A similar pattern was also found for the mRNA expression of the repulsive guidance molecule b (RGMB) and its long non-coding RNA (lncRNA) RGMB-AS1 with quercetin, indicating a potential change of the glycolytic phenotype and suppression of aberrant cellular growth which is characteristic of the HepG2 cells. Direct effects of quercetin on PGC-1α activity were minimal, as quercetin only weakly enhanced PGC-1α binding to PPARα 〈em〉in vitro〈/em〉 at higher concentrations. Our results suggest that quercetin may protect mitochondrial function from high glucose-induced stress by increasing cellular NAD〈sup〉+〈/sup〉/NADH and activation of PGC-1α-mediated pathways. Lower ROS in combination with improved complex I activity and ETS coupling efficiency under conditions of amplified oxidative stress could reinforce mitochondrial integrity and improve redox status, beneficial in certain metabolic diseases.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918313480-fx1.jpg" width="302" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Chun-Yu Chen, Yung-Fong Tsai, Wei-Ju Huang, Shih-Hsin Chang, Tsong-Long Hwang〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Critically ill patients have a high risk of sepsis. Various studies have demonstrated that propofol has anti-inflammatory effects that may benefit critically ill patients who require anesthesia. However, the mechanism and therapeutic effect remain incompletely understood. Our previous data suggest that propofol can act as a formyl peptide receptor 1 (FPR1) antagonist. Here, we hypothesize that propofol mitigates sepsis-induced acute lung injury (ALI) by inhibiting mitochondria-derived 〈em〉N〈/em〉-formyl peptide-mediated neutrophil activation. Oxidative stress caused by activated neutrophils is involved in the pathogenesis of ALI. In human neutrophils, propofol competitively reduced the release of superoxide and associated reactive oxygen species induced by fMMYALF, a human mitochondria-derived 〈em〉N〈/em〉-formyl peptide, suggesting that propofol effectively suppresses neutrophilic oxidative stress. In addition, propofol significantly inhibited fMMYALF-induced elastase release, chemotaxis, calcium mobilization, and phosphorylation of protein kinase B and mitogen-activated protein kinases. These results indicate that propofol suppresses neutrophil activation by blocking the interaction between endogenous 〈em〉N〈/em〉-formyl peptide and its receptor, FPR1, thus inhibiting downstream signaling. Furthermore, propofol alleviated alveolar wall disruption, edematous changes, and neutrophil infiltration in lipopolysaccharide-induced ALI in mice. Noticeably, propofol improved the survival of sepsis mice. This study indicates that the anti-neutrophil effects of propofol may benefit critically ill septic patients.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918311675-fx1.jpg" width="218" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Carolina Prolo, Natalia Rios, Lucia Piacenza, María Noel Álvarez, Rafael Radi〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In the last two decades, there has been a significant advance in understanding the biochemistry of peroxynitrite, an endogenously-produced oxidant and nucleophile. Its relevance as a mediator in several pathologic states and the aging process together with its transient character and low steady-state concentration, motivated the development of a variety of techniques for its unambiguous detection and estimation. Among these, fluorescence and chemiluminescence approaches have represented important tools with enhanced sensitivity but usual limited specificity. In this review, we analyze selected examples of molecular probes that permit the detection of peroxynitrite by fluorescence and chemiluminescence, disclosing their mechanism of reaction with either peroxynitrite or peroxynitrite-derived radicals. Indeed, probes have been divided into 1) redox probes that yield products by a free radical mechanism, and 2) electrophilic probes that evolve to products secondary to the nucleophilic attack by peroxynitrite. Overall, boronate-based compounds are emerging as preferred probes for the sensitive and specific detection and quantitation. Moreover, novel strategies involving genetically-modified fluorescent proteins with the incorporation of unnatural amino acids have been recently described as peroxynitrite sensors. This review analyzes the most commonly used fluorescence and chemiluminescence approaches for peroxynitrite detection and provides some guidelines for appropriate experimental design and data interpretation, including how to estimate peroxynitrite formation rates in cells.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918300728-fx1.jpg" width="290" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Jacek Zielonka, Balaraman Kalyanaraman〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Reactive oxygen species (ROS) have been implicated in both pathogenic cellular damage events and physiological cellular redox signaling and regulation. To unravel the biological role of ROS, it is very important to be able to detect and identify the species involved. In this review, we introduce the reader to the methods of detection of ROS using luminescent (fluorescent, chemiluminescent, and bioluminescent) probes and discuss typical limitations of those probes. We review the most widely used probes, state-of-the-art assays, and the new, promising approaches for rigorous detection and identification of superoxide radical anion, hydrogen peroxide, and peroxynitrite. The combination of real-time monitoring of the dynamics of ROS in cells and the identification of the specific products formed from the probes will reveal the role of specific types of ROS in cellular function and dysfunction. Understanding the molecular mechanisms involving ROS may help with the development of new therapeutics for several diseases involving dysregulated cellular redox status.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918301357-fx1.jpg" width="285" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Jing Hodes, Patric Sielaff, Hendrik Metz, Daniela Kessler-Becker, Thomas Gassenmeier, Reinhard H.H. Neubert〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉 〈p〉The control of bleaching reaction is important in hair bleaching and laundry detergents to ensure quality of the final product. A better understanding of the reaction mechanisms is needed to minimize product failures. 〈sup〉31〈/sup〉P NMR-spectroscopy-based spin trap technique was employed to detect and quantify the free radical species that were generated in different bleaching solutions. These solutions contained the key actives in an alkaline hair colorant/bleaching product, an ammonium salt and hydrogen peroxide at pH = 10. Generally, the main radical species detected in hair oxidative coloring or bleaching processes, were hydroperoxyl/superoxide radicals HO〈sub〉2〈/sub〉·/O〈sub〉2〈/sub〉〈sup〉〈strong〉.-〈/strong〉〈/sup〉, amino radicals ·NH〈sub〉2〈/sub〉 and hydroxyl radicals ·OH. Their amounts showed a variation based on the chemical composition of the bleaching systems and the metal ion content. The generation of free radicals from reactions between transition metal ions, such as copper, and hydrogen peroxide at pH = 10 was evaluated. In the absence of chelating agents, the copper ions generated a significant level of hydroxyl radicals in a Fenton-like reaction with hydrogen peroxide at pH = 10. Besides that, an increase in copper ion content led to an increase of amino radical ·NH〈sub〉2〈/sub〉, whereas the concentration of superoxide radical O〈sub〉2〈/sub〉〈sup〉·〈strong〉-〈/strong〉〈/sup〉 decreased which was not yet well reported in the previous literature.〈/p〉 〈p〉The effect of chelating agents like ethylenediaminetetraacetic acid (EDTA), tetrasodium-iminodisuccinate (IDS), a mixture of basic amino acids and dicarboxylic acid on free radical formation was investigated in the presence of binary Cu〈sup〉2+〈/sup〉-Ca〈sup〉2+〈/sup〉 bleaching systems. As expected, in the binary Cu〈sup〉2+〈/sup〉-Ca〈sup〉2+〈/sup〉 ion system EDTA did not suppress hydroxyl radical formation effectively, but the mixture containing sodium succinate, lysine and arginine reduced hydroxyl radical formation, whereas IDS (nearly) completely inhibited hydroxyl radical formation. The results indicated that each bleaching solution has its characteristic performance and damage profile. Whereas the reactivity can be controlled by the usage of chelating agents.〈/p〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918310050-fx1.jpg" width="357" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 130〈/p〉 〈p〉Author(s): Yu-Ting Cheng, Jer-An Lin, Jhih-Jia Jhang, Gow-Chin Yen〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Oxidative stress contributes to the progression of non-steroidal anti-inflammatory drug (NSAID)-induced gastrointestinal (GI) cell apoptosis. In our previous study, we reported that nuclear factor erythroid 2-related factor 2 (Nrf2) plays a protective role against ketoprofen-induced GI mucosal oxidative injury. Recent reports suggest that Nrf2 could exhibit antioxidative and antiapoptosis responses through up-regulation of DJ-1 (PARK7). In the current study, we proposed that induction of DJ-1 expression by protocatechuic acid (PCA) might provide a potential therapeutic approach for treating oxidative stress-associated GI ulcer diseases. The results indicated that PCA increased mRNA expression of glutathione peroxidase and heme oxygenase-1 through up-regulation of DJ-1 followed by Nrf2 translocation. Furthermore, PCA protected Int-407 cells against ketoprofen-induced oxidative stress by regulating the DJ-1, PI3K, and mTOR pathways. Pretreatment with PCA inhibited mitochondrial ROS generation, up-regulated the mitochondrial membrane potential, and down-regulated pro-apoptotic Bax as well as downstream caspase-8, caspase-9, and caspase-3 activity, and reversed impaired DJ-1 and anti-apoptotic Bcl-2 protein expression in Int-407 cells induced by ketoprofen. Similar to the 〈em〉in vitro〈/em〉 results, SD rats treated with PCA before administration of ketoprofen exhibited decreased caspase-3 protein expression as well as oxidative damage, and impairment of the antioxidant system and DJ-1 protein expression in the GI mucosa were reversed. The administration of lansoprazole, a type of proton pump inhibitor (PPI), strongly inhibited ketoprofen-induced GI mucosal injuries via up-regulation of DJ-1, indicating that DJ-1 is essential for the dietary antioxidant- and PPI drug-mediated mechanism of ulcer therapy. These results suggest that DJ-1 could be a novel target for protection against ketoprofen-induced GI ulcers due to its antioxidant and anti-apoptosis characteristics.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉The protective effects of PCA on ketoprofen-induced GI oxidative injury〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918321816-fx1.jpg" width="268" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Antonella Tramutola, Giulia Abate, Chiara Lanzillotta, Francesca Triani, Eugenio Barone, Federica Iavarone, Federica Vincenzoni, Massimo Castagnola, Mariagrazia Marziano, Maurizio Memo, Emirena Garrafa, D. Allan Butterfield, Marzia Perluigi, Fabio Di Domenico, Daniela Uberti〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Alzheimer's disease (AD) is a progressive form of dementia characterized by increased production of amyloid-β plaques and hyperphosphorylated tau protein, mitochondrial dysfunction, elevated oxidative stress, reduced protein clearance, among other. Several studies showed systemic modifications of immune and inflammatory systems due, in part, to decreased levels of CD3〈sup〉+〈/sup〉 lymphocytes in peripheral blood in AD. Considering that oxidative stress, both in the brain and in the periphery, can influence the activation and differentiation of T-cells, we investigated the 3-nitrotyrosine (3-NT) proteome of blood T-cells derived from AD patients compared to non-demented (ND) subjects by using a proteomic approach. 3-NT is a formal protein oxidation and index of nitrosative stress. We identified ten proteins showing increasing levels of 3-NT in CD3〈sup〉+〈/sup〉 T-cells from AD patients compared with ND subjects. These proteins are involved in energy metabolism, cytoskeletal structure, intracellular signaling, protein folding and turnover, and antioxidant response and provide new insights into the molecular mechanism that impact reduced T-cell differentiation in AD. Our results highlight the role of peripheral oxidative stress in T-cells related to immune-senescence during AD pathology focusing on the specific targets of protein nitration that conceivably can be suitable to further therapies. Further, our data demonstrate common targets of protein nitration between the brain and the periphery, supporting their significance as disease biomarkers.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918313078-fx1.jpg" width="402" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Maria Moßhammer, Verena Schrameyer, Peter Ø. Jensen, Klaus Koren, Michael Kühl〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉There is a strong need for techniques that can quantify the important reactive oxygen species hydrogen peroxide (H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉) in complex media and 〈em〉in vivo〈/em〉. We combined chemiluminescence-based H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 measurements on a commercially available flow injection analysis (FIA) system with sampling of the analyte using microdialysis probes (MDPs), typically used for measurements in tissue. This allows minimally invasive, quantitative measurements of extracellular H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 concentration and dynamics utilizing the chemiluminescent reaction of H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 with acridinium ester. By coupling MDPs to the FIA system, measurements are no longer limited to filtered, liquid samples with low viscosity, as sampling via a MDP is based on a dynamic exchange through a permeable membrane with a specific cut-off. This allows continuous monitoring of dynamic changes in H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 concentrations, alleviates potential pH effects on the measurements, and allows for flexible application in different media and systems. We give a detailed description of the novel experimental setup and its measuring characteristics along with examples of application in different media and organisms to highlight its broad applicability, but also to discuss current limitations and challenges. The combined FIA-MDP approach for H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 quantification was used in different biological systems ranging from marine biology, using the model organism 〈em〉Exaiptasia pallida〈/em〉 (light stress induced H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 release up to ~ 2.7 µM), over biomedical applications quantifying enzyme dynamics (glucose oxidase in a glucose solution producing up to ~ 60 µM H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 and the subsequent addition of catalase to monitor the H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 degradation process) and the ability of bacteria to modify their direct environment by regulating H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 concentrations in their surrounding media. This was shown by the bacteria 〈em〉Pseudomonas aeruginosa〈/em〉 degrading ~ 18 µM background H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 in LB-broth. We also discuss advantages and current limitations of the FIA-MDP system, including a discussion of potential cross-sensitivity and interfering chemical species.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918309523-fx1.jpg" width="498" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 130〈/p〉 〈p〉Author(s): Asokan Devarajan, Namakkal S. Rajasekaran, Claire Valburg, Ekambaram Ganapathy, Snehal Bindra, William A. Freije〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Intrauterine growth restriction (IUGR) leads to adult obesity, cardiovascular disease, and non-alcoholic fatty liver disease/steatohepatitis. Animal models have shown that combined intrauterine and early postnatal calorie restriction (IPCR) ameliorates these sequelae in adult life. The mechanism by which IPCR protects against adult onset disease is not understood. Autophagy, a lysosomal degradative process, recycles cellular constituents and eliminates damaged organelles, proteins, and oxidants. In this study, we hypothesized that IPCR could regulate autophagy in the liver of male rat offspring. At birth (d1) of male IUGR rat offspring and on day 21 (p21) of life, IPCR male rat offspring had a profound decrease in hepatic autophagy in all three stages of development: initiation, elongation, and maturation. However, upon receiving a normal diet ad-lib throughout adulthood, aged IPCR rats (day 450 of life (p450)), had increased hepatic autophagy, in direct contrast to what was seen in early life. The decreased autophagy at d21 led to the accumulation of ubiquitinated proteins and lipid oxidative products, whereas the increased autophagy in late life had the opposite effect. Oxidized lipids were unchanged at d1 by IUGR treatment indicating that decreased autophagy precedes oxidative stress in early life. When cellular signaling pathways regulating autophagy were examined, the 5′ adenosine monophosphate-activated protein kinase pathway (AMPK), and not endoplasmic stress pathways, was found to be altered, suggesting that autophagy is regulated through AMPK signaling pathway in IPCR rats. Taken together, this study reveals that the perinatal nutritional status establishes a nutritionally sensitive memory that enhances hepatic autophagy in late life, a process that perhaps acts as a protective mechanism to limited nutrition.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918311742-fx1.jpg" width="500" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Jun Zhang, Bin Wang, Hong Wang, Hui He, Qiong Wu, Xia Qin, Xi Yang, Linmu Chen, Ge Xu, Zhiyi Yuan, Qiying Yi, Zhen Zou, Chao Yu〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Copper oxide nanoparticles (CuONPs) have been widely used in the industrial and pharmaceutical fields; however, their toxicity profile is deeply concerning. Currently, nanomaterials-induced toxicity in the cardiovascular system is receiving increased attention. Our previous toxicological study found that lysosomal deposition of CuONPs triggered vascular endothelial cell death, indicating that the involvement of autophagic dysfunction was crucial for CuONPs-induced toxicity in human umbilical vein endothelial cells (HUVECs). In the current study, we investigated the detailed mechanism underlying the autophagic dysfunction induced by CuONPs. We demonstrated that CuONPs exposure caused accumulation of superoxide anions, which likely resulted from mitochondrial dysfunctions. MnTBAP, a superoxide anions scavenger, alleviated CuONPs-induced HUVECs death, indicating that excessive superoxide anions were directly related to the CuONPs cytotoxicity in HUVECs. Interestingly, we found that mitophagy (a protective mechanism for clearance of damaged mitochondria and excessive superoxide anions) was initiated but failed to be cleared in CuONPs-treated cells, resulting in the accumulation of damaged mitochondria. Inhibition of mitophagy through Atg5 knockout or blocking of mitochondria fission with Mdivi-1 significantly aggravated CuONPs-induced superoxide anions accumulation and cell death, suggesting that mitophagy is a protective mechanism against CuONPs cytotoxicity in HUVECs. In summary, we demonstrate that superoxide anions (originating from damaged mitochondria) are involved in CuONPs-associated toxicity and that impaired mitophagic flux aggravates the accumulation of excessive superoxide anions, which leads to HUVECs death. Our findings indicate that there are crucial roles for superoxide anions and mitophagy in CuONPs-induced toxicity in vascular endothelial cells.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918311183-fx1.jpg" width="225" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Matías N. Möller, Ana Denicola〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Oxygen and nitric oxide are small hydrophobic molecules that usually need to diffuse a considerable distance to accomplish their biological functions and necessarily need to traverse several lipid membranes. Different methods have been used to study the diffusion of these molecules in membranes and herein we focus in the quenching of fluorescence of pyrenes inserted in the membrane. The pyrene derivatives have long fluorescence lifetimes (around 200 ns) that make them very sensitive to fluorescence quenching by nitric oxide, oxygen and other paramagnetic species. Results show that the apparent diffusion coefficients in membranes are similar to those in water, indicating that diffusion of these molecules in membranes is not considerably limited by the lipids. This high apparent diffusion in membranes is a consequence of both a favorable partition of these molecules in the hydrophobic interior of membranes and a high diffusion coefficient. Altering the composition of the membrane results in slight changes in diffusion, indicating that in most cases the lipid membranes will not hinder the passage of oxygen or nitric oxide. The diffusion of nitric oxide in the lipid core of low density lipoprotein is also very high, supporting its role as an antioxidant. In contrast to the high permeability of membranes to nitric oxide and oxygen, the permeability to other reactive species such as hydrogen peroxide and peroxynitrous acid is nearly five orders of magnitude lower.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918307305-fx1.jpg" width="361" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Alexander I. Kostyuk, Anastasiya S. Panova, Dmitry S. Bilan, Vsevolod V. Belousov〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉A wide variety of genetically encoded fluorescent biosensors are available to date. Some of them have already contributed significantly to our understanding of biological processes occurring at cellular and organismal levels. Using such an approach, outstanding success has been achieved in the field of redox biology. The probes allowed researchers to observe, for the first time, the dynamics of important redox parameters in vivo during embryogenesis, aging, the inflammatory response, the pathogenesis of various diseases, and many other processes. Given the differences in the readout and spectra of the probes, they can be used in multiparameter imaging in which several processes are monitored simultaneously in the cell. Intracellular processes form an extensive network of interactions. For example, redox changes are often accompanied by changes in many other biochemical reactions related to cellular metabolism and signaling. Therefore, multiparameter imaging can provide important information concerning the temporal and spatial relationship of various signaling and metabolic processes. In this review, we will describe the main types of genetically encoded biosensors, the most frequently used readout, and their use in multiplexed imaging mode.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918301680-fx1.jpg" width="306" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Lisa Traeger, Inka Gallitz, Rohit Sekhri, Nicole Bäumer, Tanja Kuhlmann, Claudia Kemming, Michael Holtkamp, Jennifer-Christin Müller, Uwe Karst, Francois Canonne-Hergaux, Martina U. Muckenthaler, Donald B. Bloch, Andrea Olschewski, Thomas B. Bartnikas, Andrea U. Steinbicker〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The bone morphogenetic protein (BMP) type I receptors ALK2 and ALK3 are essential for expression of hepcidin, a key iron regulatory hormone. In mice, hepatocyte-specific 〈em〉Alk2〈/em〉 deficiency leads to moderate iron overload with periportal liver iron accumulation, while hepatocyte-specific 〈em〉Alk3〈/em〉 deficiency leads to severe iron overload with centrilobular liver iron accumulation and a more marked reduction of basal hepcidin levels. The objective of this study was to investigate whether the two receptors have additive roles in hepcidin regulation. Iron overload in mice with hepatocyte-specific 〈em〉Alk2〈/em〉 and 〈em〉Alk3 (Alk2/3)〈/em〉 deficiency was characterized and compared to hepatocyte-specific 〈em〉Alk3〈/em〉 deficient mice. Co-immunoprecipitation studies were performed to detect the formation of ALK2 and ALK3 homodimer and heterodimer complexes 〈em〉in vitro〈/em〉 in the presence and absence of ligands. The iron overload phenotype of hepatocyte-specific 〈em〉Alk2/3〈/em〉-deficient mice was more severe than that of hepatocyte-specific 〈em〉Alk3〈/em〉-deficient mice. 〈em〉In vitro〈/em〉 co-immunoprecipitation studies in Huh7 cells showed that ALK3 can homodimerize in absence of BMP2 or BMP6. In contrast, ALK2 did not homodimerize in either the presence or absence of BMP ligands. However, ALK2 did form heterodimers with ALK3 in the presence of BMP2 or BMP6. ALK3-ALK3 and ALK2-ALK3 receptor complexes induced hepcidin expression in Huh7 cells. Our data indicate that: (I) ALK2 and ALK3 have additive functions 〈em〉in vivo,〈/em〉 as 〈em〉Alk2/3〈/em〉 deficiency leads to a greater degree of iron overload than 〈em〉Alk3〈/em〉 deficiency; (II) ALK3, but not ALK2, undergoes ligand-independent homodimerization; (III) the formation of ALK2-ALK3 heterodimers is ligand-dependent and (IV) both receptor complexes functionally induce hepcidin expression 〈em〉in vitro〈/em〉.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918316083-fx1.jpg" width="230" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Simona Gargiulo, Daniela Rossin, Gabriella Testa, Paola Gamba, Erica Staurenghi, Fiorella Biasi, Giuseppe Poli, Gabriella Leonarduzzi〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Atherosclerosis is currently understood to be mainly the consequence of a complicated inflammatory process at the different stages of plaque development. Among the several inflammatory molecules involved, up-regulation of the functional cyclooxygenase 2/membrane-bound prostaglandin E synthase 1 (COX-2/mPGES-1) axis plays a key role in plaque development. Excessive production of oxidized lipids, following low-density lipoprotein (LDL) oxidation, is a characteristic feature of atherosclerosis. Among the oxidized lipids of LDLs, the oxysterol 27-hydroxycholesterol (27-OH) and the aldehyde 4-hydroxynonenal (HNE) substantially accumulate in the atherosclerotic plaque, contributing to its progression and instability through a variety of processes. This study shows that 27-OH and HNE promote up-regulation of both the inducible enzymes COX-2 and mPGES-1, leading to increased production of prostaglandin (PG) E〈sub〉2〈/sub〉 and inducible nitric oxide synthase, and the subsequent release of nitric oxide in human promonocytic U937 cells. The study also examined the potential involvement of the functionally coupled COX-2/mPGES-1 in enhancing the production of certain pro-inflammatory cytokines and of matrix metalloproteinase 9 by U937 cells. This enhancement is presumably due to the induction of PGE〈sub〉2〈/sub〉 synthesis, as a result of the up-regulation of the COX-2/mPGES-1, stimulated by the two oxidized lipids, 27-OH and HNE. Induction of PGE〈sub〉2〈/sub〉 synthesis might thus be a mechanism of plaque instability and eventual rupture, contributing to matrix metalloproteinase production by activated macrophages.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918310062-fx1.jpg" width="436" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Benjamin S. Rayner, Yunjia Zhang, Bronwyn E. Brown, Leila Reyes, Victoria C. Cogger, Clare L. Hawkins〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The infiltration of activated leukocytes, including macrophages, at sites of inflammation and the formation and presence of hypochlorous acid (HOCl) are interlinked hallmarks of many debilitating disease processes, including atherosclerosis, arthritis, neurological and renal disease, diabetes and obesity. The production of extracellular traps by activated leukocytes in response to a range of inflammatory stimuli is increasingly recognised as an important process within a range of disease settings. We show that exposure of human monocyte-derived macrophages to pathophysiological levels of HOCl results in the dose-dependent extrusion of DNA and histones into the cellular supernatant, consistent with extracellular trap formation. Concurrent with, but independent of these findings, macrophage exposure to HOCl also resulted in an immediate and sustained cytosolic accumulation of Ca〈sup〉2+〈/sup〉, culminating in the increased production of cytokines and chemokines. Polarisation of the macrophages prior to HOCl exposure revealed a greater propensity for inflammatory M1 macrophages to produce extracellular traps, whereas alternatively-activated M2 macrophages were less susceptible to HOCl insult. M1 macrophages also produced extracellular traps on exposure to phorbol myristate acetate (PMA), interleukin-8 (IL-8) and tumour necrosis factor α (TNFα). Taken together, these data indicate a potential role for macrophages in mediating extracellular trap formation, which may be relevant in pathological conditions characterised by chronic inflammation or excessive HOCl formation.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918315223-fx1.jpg" width="450" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Yuyang Zhou, Min Lin, Jie Wang, Fan Chen, Feiyang Li, Wenge Chen, Longyu Han, Chiahung Wang, Jianming Chen, Jing-Wei Shao, Lee Jia〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉S〈/em〉-nitrosocaptopril (CapNO) possesses dual capacities of both Captopril and an NO donor with enhanced efficacy and reduced side effects. CapNO crystals are difficult to make due to its unstable S-NO bond. Here, we report a novel stable 〈em〉S〈/em〉-nitrosocaptopril monohydrate (CapNO·H〈sub〉2〈/sub〉O) that is stabilized by intermolecular five-membered structure, where one H of H〈sub〉2〈/sub〉O forms a hydrogen bond with O〈sup〉-〈/sup〉 of the stable resonance zwitterion Cap–S〈sup〉+〈/sup〉=N–O〈sup〉-〈/sup〉, and the O in H〈sub〉2〈/sub〉O forms the dipole-dipole interaction with S〈sup〉+〈/sup〉 through two unpaired electrons. With the chelation and common ion effect, we synthesized and characterized CapNO·H〈sub〉2〈/sub〉O that is stable at 4 °C for 180 days and thereafter without significant degradation. Compared to Captopril, CapNO showed direct vasorelaxation and beneficial effect on PAH rats, and could be self-assembled in rat stomach when Captopril and NaNO〈sub〉2〈/sub〉 were given separately. This novel CapNO·H〈sub〉2〈/sub〉O with low entropy paves an avenue for its clinical trials and commercialization.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S089158491831075X-fx1.jpg" width="370" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Kok Leong Chong, Benjamin A. Chalmers, Jason K. Cullen, Amandeep Kaur, Jacek L. Kolanowski, Benjamin J. Morrow, Kathryn E. Fairfull-Smith, Martin J. Lavin, Nigel L. Barnett, Elizabeth J. New, Michael P. Murphy, Steven E. Bottle〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Here we describe new fluorescent probes based on fluorescein and rhodamine that provide reversible, real-time insight into cellular redox status. The new probes incorporate bio-imaging relevant fluorophores derived from fluorescein and rhodamine linked with stable nitroxide radicals such that they cannot be cleaved, either spontaneously or enzymatically by cellular processes. Overall fluorescence emission is determined by reversible reduction and oxidation, hence the steady state emission intensity reflects the balance between redox potentials of critical redox couples within the cell. The permanent positive charge on the rhodamine-based probes leads to their rapid localisation within mitochondria in cells. Reduction and oxidation also leads to marked changes in the fluorophore lifetime, enabling monitoring by fluorescence lifetime imaging microscopy. Finally, we demonstrate that administration of a methyl ester version of the rhodamine-based probe can be used at concentrations as low as 5 nM to generate a readily detected response to redox stress within cells as analysed by flow cytometry.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918301126-fx1.jpg" width="434" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Anibal E. Vercesi, Roger F. Castilho, Alicia J. Kowaltowski, Helena C.F. de Oliveira, Nadja C. de Souza-Pinto, Tiago R. Figueira, Estela N.B. Busanello〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Mitochondria possess a Ca〈sup〉2+〈/sup〉 transport system composed of separate Ca〈sup〉2+〈/sup〉 influx and efflux pathways. Intramitochondrial Ca〈sup〉2+〈/sup〉 concentrations regulate oxidative phosphorylation, required for cell function and survival, and mitochondrial redox balance, that participates in a myriad of signaling and damaging pathways. The interaction between Ca〈sup〉2+〈/sup〉 accumulation and redox imbalance regulates opening and closing of a highly regulated inner membrane pore, the membrane permeability transition pore (PTP). In this review, we discuss the regulation of the PTP by mitochondrial oxidants, reactive nitrogen species, and the interactions between these species and other PTP inducers. In addition, we discuss the involvement of mitochondrial redox imbalance and PTP in metabolic conditions such as atherogenesis, diabetes, obesity and in mtDNA stability.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918309742-fx1.jpg" width="262" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Fang Lei, Wen Wang, Yating Fu, Ji Wang, Yu Zheng〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Cigarette smoke (CS) exposure negatively affects neurodevelopment. We established a CS exposure rat model to determine how maternal CS exposure induces oxidative stress and mitochondrial dysfunction in parafacial respiratory group (pFRG) essential to central chemoreceptive regulation of normal breathing. Pregnant rats were exposed to CS during gestational days 1–20, and the offspring were studied on postnatal day 2. Our data showed that maternal CS exposure resulted in elevated accumulation of ROS, which left a footprint on DNA and lipid with increases in 8-hydroxy-2′-deoxyguanosine and malondialdehyde contents. Furthermore, maternal CS exposure induced decreases in manganese superoxide dismutase, catalase and glutathione reductase activities as well as reduction in glutathione content in pFRG in the offspring. Moreover, maternal exposure to CS led to mitochondrial ultrastructure changes, mitochondrial swelling, reduction in ATP generation, loss of mitochondrial membrane potential and increase in mitochondrial DNA copy number. These findings suggest that maternal exposure to CS alters normal development of pFRG that is critical for normal respiratory control.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918310712-fx1.jpg" width="500" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Hirofumi Sakurai, Yuko Morishima, Yukio Ishii, Kazufumi Yoshida, Masayuki Nakajima, Yoshiya Tsunoda, Shih-Yuan Hayashi, Takumi Kiwamoto, Yosuke Matsuno, Mio Kawaguchi, Masayuki Yamamoto, Nobuyuki Hizawa〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Oxidative stress induced by cigarette smoke and other environmental pollutants contributes to refractory asthma. To better understand the role of smoking in asthma, we investigated the effects of cigarette smoke on allergic airway responses in mice and examined expression of nuclear factor-E2-related factor-2 (Nrf2) and its downstream factors, because Nrf2 is known to play a pivotal role in antioxidant responses. OVA-sensitized and challenged BALB/c mice were exposed to cigarette smoke and then treated with dexamethasone, sulforaphane (an activator of Nrf2), or their combination. Upon exposure to cigarette smoke, Nrf2 and associated transcripts were upregulated in response to oxidative stress, and asthmatic responses were steroid resistant. In OVA-sensitized and challenged mice exposed to cigarette smoke and treated with sulforaphane, Nrf2-mediated antioxidant responses were upregulated to a greater extent, and steroid sensitivity of asthmatic responses was restored. Moreover, the expression and activity of histone deacetylase 2 (HDAC2), a key regulator of steroid responsiveness, was reduced in mice exposed to cigarette smoke, but restored by sulforaphane treatment. No effects of sulforaphane were observed in Nrf2-deficient mice. These findings indicate that cigarette smoke induces steroid unresponsiveness in asthmatic airways, and that sulforaphane restores steroid sensitivity via upregulation of Nrf2 and enhancement of HDAC2 expression and activity. Thus, Nrf2 may serve as a potential molecular target for cigarette smoke-related refractory asthma resistant to steroid therapy.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918317520-fx1.jpg" width="272" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Feng-Yang Wang, Ke-Bin Huang, Hai-Wen Feng, Zhen-Feng Chen, You-Nian Liu, Hong Liang〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Agents with multiple modes of tumor cell death can be effective chemotherapeutic drugs. One example of a bimodal chemotherapeutic approach is an agent that can induce both apoptosis and autophagic death. Thus far, no clinical anticancer drug has been shown to simultaneously induce both these pathways. Mono-functional platinum complexes are potent anticancer drug candidates which act through mechanisms distinct from cisplatin. Here, we describe the synthesis and characterize of two mono-functional platinum complexes containing 8-substituted quinoline derivatives as ligands. In comparison to cisplatin, n-Mon-Pt-1 exhibited a greater in vitro cytotoxicity, was more effective in resistant cells and elicited a better anticancer effect. Mechanistic experiments indicate that n-Mon-Pt-1 mainly accumulates in mitochondria, and stimulates significant TrxR inhibition, ROS release and an ER stress response, ultimately resulting in a simultaneous induction of apoptosis and autophagy. Importantly, compared to cisplatin, n-Mon-Pt-1 exhibits lower acute toxicity and better anticancer activity in a murine tumor model.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918317064-fx1.jpg" width="500" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): Adam J. Trewin, Brandon J. Berry, Alicia Y. Wei, Laura L. Bahr, Thomas H. Foster, Andrew P. Wojtovich〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Oxidants play an important role in the cell and are involved in many redox processes. Oxidant concentrations are maintained through coordinated production and removal systems. The dysregulation of oxidant homeostasis is a hallmark of many disease pathologies. The local oxidant microdomain is crucial for the initiation of many redox signaling events; however, methods to control oxidant product are limited. Some fluorescent proteins, including GFP, TagRFP, KillerRed, miniSOG, and their derivatives, generate oxidants in response to light. These genetically-encoded photosensitizers produce singlet oxygen and superoxide upon illumination and offer spatial and temporal control over oxidant production. In this review, we will examine the photosensitization properties of fluorescent proteins and their application to redox biology. Emerging concepts of selective oxidant species production via photosensitization and the impact of light on biological systems are discussed.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918300455-fx1.jpg" width="500" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Shijun Ma, Afu Fu, Sierin Lim, Geraldine Giap Ying Chiew, Kathy Qian Luo〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Circulation of cancer cells in the bloodstream is a vital step for distant metastasis, during which cancer cells are exposed to hemodynamic shear stress (SS). The actions of SS on tumor cells are complicated and not fully understood. We previously reported that fluidic SS was able to promote migration of breast cancer cells by elevating the cellular ROS level. In this study, we further investigated the mechanisms regulating SS-promoted cell migration and identified the role of MnSOD in the related pathway. We found that SS could enhance tumor cell adhesion to extracellular matrix and endothelial monolayer, and MnSOD also regulated this process. Briefly, SS stimulates the generation of mitochondrial superoxide in tumor cells. MnSOD then converts superoxide into hydrogen peroxide, which activates ERK1/2 to promote tumor cell migration and activates FAK to promote tumor cell adhesion. Combining our previous and present studies, we present experimental evidence on the pro-metastatic effects of hemodynamic SS and reveal the underlying mechanism. Our findings provide new insights into the nature of cancer metastasis and the understanding of tumor cell responses to external stresses and have valuable implications for cancer therapy development.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918309729-fx1.jpg" width="340" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Jonatas E. Nogueira, Patricia Passaglia, Clarissa M.D. Mota, Bruna M. Santos, Marcelo E. Batalhão, Evelin C. Carnio, Luiz G.S. Branco〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Physical exercise induces inflammatory and oxidative markers production in the skeletal muscle and this process is under the control of both endogenous and exogenous modulators. Recently, molecular hydrogen (H〈sub〉2〈/sub〉) has been described as a therapeutic gas able to reduced oxidative stress in a number of conditions. However, nothing is known about its putative role in the inflammatory and oxidative status during a session of acute physical exercise in sedentary rats. Therefore, we tested the hypothesis that H〈sub〉2〈/sub〉 attenuates both inflammation and oxidative stress induced by acute physical exercise. Rats ran at 80% of their maximum running velocity on a closed treadmill inhaling either the H〈sub〉2〈/sub〉 gas (2% H〈sub〉2〈/sub〉, 21% O〈sub〉2〈/sub〉, balanced with N〈sub〉2〈/sub〉) or the control gas (0% H〈sub〉2〈/sub〉, 21% O〈sub〉2〈/sub〉, balanced with N〈sub〉2〈/sub〉) and were euthanized immediately or 3 h after exercise. We assessed plasma levels of inflammatory cytokines [tumor necrosis factor-α (TNF-α), interleukin (IL)−1β and IL-6] and oxidative markers [superoxide dismutase (SOD), thiobarbituric acid reactive species (TBARS) and nitrite/nitrate (NOx)]. In addition, we evaluated the phosphorylation status of intracellular signaling proteins [glycogen synthase kinase type 3 (GSK3α/β) and the cAMP responsive element binding protein (CREB)] that modulate several processes in the skeletal muscle during exercise, including changes in exercise-induced reactive oxygen species (ROS) production. As expected, physical exercise increased virtually all the analyzed parameters. In the running rats, H〈sub〉2〈/sub〉 blunted exercise-induced plasma inflammatory cytokines (TNF-α and IL-6) surges. Regarding the oxidative stress markers, H〈sub〉2〈/sub〉 caused further increases in exercise-induced SOD activity and attenuated the exercise-induced increases in TBARS 3 h after exercise. Moreover, GSK3α/β phosphorylation was not affected by exercise or H〈sub〉2〈/sub〉 inhalation. Otherwise, exercise caused an increased CREB phosphorylation which was attenuated by H〈sub〉2〈/sub〉. These data are consistent with the notion that H〈sub〉2〈/sub〉 plays a key role in decreasing exercise-induced inflammation, oxidative stress, and cellular stress.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918311432-fx1.jpg" width="346" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Laura Kofoed Kjaer, Jens Oellgaard, Trine Henriksen, Peter Gaede, Oluf Pedersen, Henrik Enghusen Poulsen〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉 〈h6〉Objective〈/h6〉 〈p〉The RNA oxidation product, 8-oxo-7,8-dihydroguanosine (8-oxoGuo), has been associated with mortality in patients with type 2 diabetes (T2D). However, the identification and the potential effect of approved treatments decreasing urine 8-oxoGuo level remain unraveled. In the Steno-2 study intensified multifactorial treatment compared with conventional multifactorial treatment reduced mortality in T2D patients with microalbuminuria by 45%. We assessed association between 8-oxoGuo at advanced baseline and total mortality with up to 19.9 years follow-up and from end of intervention to end of follow-up up to (up to 13.9 years).〈/p〉 〈/div〉 〈div〉 〈h6〉Materials and methods〈/h6〉 〈p〉In 1993, 160 T2D patients with microalbuminuria were included in the Steno-2 trial. Urine samples from baseline were not available, but samples were available from 155 patients (97%) in 1995 (advanced baseline) and from 125 patients (96%) in 2001 (end of intervention). Hazard ratios (HR) for log2-transformed 8-oxoGuo and dichotomized (cut-off at median; low vs. high RNA oxidation) were estimated using Cox regressions.〈/p〉 〈/div〉 〈div〉 〈h6〉Results〈/h6〉 〈p〉During follow-up of 19.9 years after advanced baseline, 89 died and no association between 8-oxoGuo and mortality was found (p = 0.40). From the end of 7.8 years of intervention and during remaining 13.9 years of observation, 61 died and doubling the urine 8-oxoGuo level was associated with mortality with a HR 3.08 (95% CI [1.86 −5.12]; p 〈 0.001) after multiple adjustments. Patients with low 8-oxoGuo in the intensified-treatment had the lowest risk of dying compared with high 8-oxoGuo in the conventional-treatment both from advanced baseline onwards, adjusted HR 0.40 (95% CI [0.21 −0.75]; p = 0.004), and from end of intervention onwards, adjusted HR 0.28 (95% CI [0.13 −0.61]; p = 0.001).〈/p〉 〈/div〉 〈div〉 〈h6〉Conclusions〈/h6〉 〈p〉In T2D patients with microalbuminuria, high levels of urine 8-oxoGuo after 7.8 years of multifactorial intervention was associated with higher mortality during 13.9 years of post-trial follow-up. Patients with low 8-oxoGuo in the intensified treatment group had the lowest risk of dying.〈/p〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉Cell clipart image adapted from http://www.servier.com Powerpointimage-bank (Servier Medical Art by Servier) which is licensed under a Creative Commons Attribution 3.0 Unported License 〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918313169-fx1.jpg" width="347" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Yue Deng, Beichen Jiang, Carolyn L. Rankin, Kazuhito Toyo-oka, Mark L. Richter, Julie A. Maupin-Furlow, Jackob Moskovitz〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The methionine sulfoxide reductase (Msr) system is known for its function in reducing protein-methionine sulfoxide to methionine. Recently, we showed that one member of the Msr system, MsrA, is involved in the ubiquitination-like process in 〈em〉Archaea〈/em〉. Here, the mammalian MsrA is demonstrated to mediate the ubiquitination of the 14-3-3 zeta protein and to promote the binding of 14-3-3 proteins to alpha synuclein in brain. MsrA was also found to enhance the ubiquitination and phosphorylation of Ser129 of alpha synuclein in brain. Furthermore, we demonstrate that, similarly to the archaeal MsrA, the mammalian MsrA can compete for capturing ubiquitin using the same active site it contains for methionine sulfoxide binding. Based on our previous observations showing that MsrA knockout mice have elevated expression levels of dopamine and 14-3-3 zeta and our current data, we propose that MsrA-dependent 14-3-3 zeta ubiquitination affects the regulation of alpha synuclein degradation and dopamine synthesis in the brain.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918310414-fx1.jpg" width="285" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: 20 November 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 128〈/p〉 〈p〉Author(s): 〈/p〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Xia Li, Junzhou Wu, Xiaoying Zhang, Wei Chen〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Malignant melanoma is a highly metastatic and life-threatening cancer. Reactive oxygen species (ROS) play important roles in cancer initiation and progression including metastasis. It has been reported that the oxidative stress spontaneously generated in circulating melanoma cells was able to suppress distant metastasis 〈em〉in vivo〈/em〉. However, little is known regarding the effects and mechanism of glutathione reductase (GR) inhibition-induced oxidative stress in regulation of melanoma metastasis. Here, we demonstrate that GR inhibition generates oxidative stress and suppresses lung metastasis and subcutaneous growth of melanoma 〈em〉in vivo〈/em〉. In addition, inhibitory effects by GR activity reduction were observed on cell proliferation, colony formation, cell adhesion, migration and invasion in melanoma cells 〈em〉in vitro〈/em〉. GR inhibition-induced oxidative stress was also found to block epithelial-to-mesenchymal transition (EMT) by decreasing the expression of Vimentin, ERK1/2, transcription factor Snail and increasing the expression of E-cadherin. In addition, actin rearrangement, a key element involved in cell motility, was also affected by GR-mediated oxidative stress possibly through protein 〈em〉S〈/em〉-glutathionylation on actin. In conclusion, this study identifies GR as an effective regulator of oxidative stress that affects the multistep processes of metastasis in melanoma cells, and it becomes a potential target for melanoma therapy.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918313194-fx1.jpg" width="450" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Yuxin Shua, Nanfei Yang, Yunchen Bei, Pingping Shen〈/p〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Akira Sakurada, Koji Miyanishi, Shingo Tanaka, Masanori Sato, Hiroki Sakamoto, Yutaka Kawano, Kohichi Takada, Yusaku Nakabeppu, Masayoshi Kobune, Junji Kato〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉 〈h6〉Background & aims〈/h6〉 〈p〉The role of base excision repair genes in human hepatocarcinogenesis has not yet been explored. Here, we investigated relationships between variants of these genes and the risk of developing hepatocellular carcinoma (HCC).〈/p〉 〈/div〉 〈div〉 〈h6〉Methods〈/h6〉 〈p〉Nineteen tagging SNPs in base excision repair genes (including 〈em〉MUTYH〈/em〉, 〈em〉OGG1〈/em〉 and 〈em〉MTH1〈/em〉) were genotyped using iPLEX assays; one significant SNP was found and confirmed in Japanese patients with chronic hepatitis C (CHC) (n = 38 HCC and 55 controls). The effects of modifying the intronic variants were determined by luciferase assays. MUTYH-null mice were used to examine the involvement of oxidative stress and DNA repair enzymes in hepatocarcinogenesis.〈/p〉 〈/div〉 〈div〉 〈h6〉Results〈/h6〉 〈p〉Significant associations were found for a single intron SNP (rs3219487) in the 〈em〉MUTYH〈/em〉 gene. The risk of developing HCC in patients with A/A or G/A genotypes was higher than in those with the G/G genotype (OR = 9.27, 95% CI = 2.39 −32.1, P = 0.0005). 〈em〉MUTYH〈/em〉 mRNA levels in both peripheral mononuclear cells were significantly lower in G/A or A/A genotyped subjects (P = 0.0157 and 0.0108, respectively). We found that −2000 in the 〈em〉MUTYH〈/em〉 promoter region is involved in enhanced expression of 〈em〉MUTYH〈/em〉 by insertion of a major allele sequence of rs3219487. Liver tumors were observed in MUTYH-null mice after 12 months´ high iron diet, but no tumors developed when dietary anti-oxidant (N-Acetyl-〈span〉L〈/span〉-cysteine) was also provided.〈/p〉 〈/div〉 〈div〉 〈h6〉Conclusions〈/h6〉 〈p〉CHC patients with the rs3219487 adenine allele had a significantly increased risk of developing HCC. MUTYH-null mice with iron-associated oxidative stress were susceptible to development of liver tumors unless prevented by dietary anti-oxidants.〈/p〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918312164-fx1.jpg" width="420" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Jason P. Eiserich, Sean P. Ott, Tamara Kadir, Brian M. Morrissey, Keri A. Hayakawa, Michele A. La Merrill, Carroll E. Cross〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉 〈h6〉Rationale〈/h6〉 〈p〉Cystic fibrosis (CF) patients are known to produce cyanide (CN〈sup〉-〈/sup〉) although challenges exist in determinations of total levels, the precise bioactive levels, and specificity of its production by CF microflora, especially 〈em〉P. aeruginosa〈/em〉. Our objective was to measure total CN〈sup〉-〈/sup〉 levels in CF sputa by a simple and novel technique in 〈em〉P. aeruginosa〈/em〉 positive and negative adult patients, to review respiratory tract (RT) mechanisms for the production and degradation of CN〈sup〉-〈/sup〉, and to interrogate sputa for post-translational protein modification by CN〈sup〉-〈/sup〉 metabolites.〈/p〉 〈/div〉 〈div〉 〈h6〉Methods〈/h6〉 〈p〉Sputa CN〈sup〉-〈/sup〉 concentrations were determined by using a commercially available CN〈sup〉-〈/sup〉 electrode, measuring levels before and after addition of cobinamide, a compound with extremely high affinity for CN〈sup〉-〈/sup〉. Detection of protein carbamoylation was measured by Western blot.〈/p〉 〈/div〉 〈div〉 〈h6〉Measurements and main results〈/h6〉 〈p〉The commercial CN〈sup〉-〈/sup〉 electrode was found to overestimate CN〈sup〉-〈/sup〉 levels in CF sputum in a highly variable manner; cobinamide addition rectified this analytical issue. Although 〈em〉P. aeruginosa〈/em〉 positive patients tended to have higher total CN〈sup〉-〈/sup〉 values, no significant differences in CN〈sup〉-〈/sup〉 levels were found between positive and negative sputa. The inflammatory oxidant hypochlorous acid (HOCl) was shown to rapidly decompose CN〈sup〉-〈/sup〉, forming cyanogen chloride (CNCl) and the carbamoylating species cyanate (NCO〈sup〉-〈/sup〉). Carbamoylated proteins were found in CF sputa, analogous to reported findings in asthma.〈/p〉 〈/div〉 〈div〉 〈h6〉Conclusions〈/h6〉 〈p〉Our studies indicate that CN〈sup〉-〈/sup〉 is a transient species in the inflamed CF airway due to multiple biosynthetic and metabolic processes. Stable metabolites of CN〈sup〉-〈/sup〉, such as cyanate, or carbamoylated proteins, may be suitable biomarkers of overall CN〈sup〉-〈/sup〉 production in CF airways.〈/p〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918314060-fx1.jpg" width="500" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 130〈/p〉 〈p〉Author(s): Max A. Thorwald, Jose A. Godoy-Lugo, Gema J. Rodriguez, Marco Antonio Rodriguez, Mostofa Jamal, Hiroshi Kinoshita, Daisuke Nakano, Akira Nishiyama, Henry J. Forman, Rudy M. Ortiz〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Diabetic hearts are susceptible to damage from inappropriate activation of the renin angiotensin system (RAS) and hyperglycemic events both of which contribute to increased oxidant production. Prolonged elevation of oxidants impairs mitochondrial enzyme function, further contributing to metabolic derangement. Nuclear factor erythriod-2-related factor 2 (Nrf2) induces antioxidant genes including those for glutathione (GSH) synthesis following translocation to the nucleus. We hypothesized that an acute elevation in glucose impairs Nrf2-related gene expression in diabetic hearts, while AT1 antagonism would aid in Nrf2-mediated antioxidant production and energy replenishment. We used four groups (n = 6–8/group) of 25-week-old rats: 1) LETO (lean strain-control), 2) type II diabetic OLETF, 3) OLETF + angiotensin receptor blocker (ARB; 10 mg olmesartan/kg/d × 8 wks), and 4) ARBM (4 weeks on ARB, 4 weeks off) to study the effects of acutely elevated glucose on cardiac mitochondrial function and Nrf2 signaling in the diabetic heart. Animals were gavaged with a glucose bolus (2 g/kg) and groups were dissected at T0, T180, and T360 minutes. Nrf2 mRNA was 32% lower in OLETF rats compared to LETO and remained suppressed in response to glucose. LETO Nrf2 mRNA increased 25% at T360 in response to glucose while no changes were observed in diabetic hearts. GCLC and GCLM mRNA decreased in diabetic hearts 33% and 44% respectively and remained suppressed in response to glucose while ARB treatment increased GCLM transcripts 90% at T180. These data illustrate that during T2DM and in response to glucose, cardiac Nrf2′s adaptive response to environmental stressors such as glucose is impaired in diabetic hearts and that ARB treatment may aid Nrf2′s impaired dynamic response.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918312814-fx1.jpg" width="446" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Ji Eun Choi, Seo-Hee Heo, Myung Ju Kim, Woo-Hyun Chung〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉A genetic analysis of synthetic lethal interactions in yeast revealed that the mutation of 〈em〉SOD1〈/em〉, encoding an antioxidant enzyme that scavenges superoxide anion radical, impaired the growth of a set of mutants defective in homologous recombination (HR) pathway. Hence, SOD1 inhibition has been proposed as a promising approach for the selective killing of HR-deficient cancer cells. However, we show that the deletion of 〈em〉RAD51〈/em〉 and 〈em〉SOD1〈/em〉 is not synthetic lethal but displays considerably slow growth and synergistic sensitivity to both reactive oxygen species (ROS)- and DNA double-strand break (DSB)-generating drugs in the budding yeast 〈em〉Saccharomyces cerevisiae〈/em〉. The function of Sod1 in regard to Rad51 is dependent on Ccs1, a copper chaperone for Sod1. Sod1 deficiency aggravates genomic instability in conjunction with the absence of Rad51 by inducing DSBs and an elevated mutation frequency. Inversely, lack of Rad51 causes a Sod1 deficiency-derived increase of intracellular ROS levels. Taken together, our results indicate that there is a significant and specific crosstalk between two major cellular damage response pathways, ROS signaling and DSB repair, for cell survival.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918311110-fx1.jpg" width="285" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Bojana Božić, Jelena Korać, Dalibor M. Stanković, Marina Stanić, Mima Romanović, Jelena Bogdanović Pristov, Snežana Spasić, Ana Popović-Bijelić, Ivan Spasojević, Milica Bajčetić〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉An increase in the copper pool in body fluids has been related to a number of pathological conditions, including infections. Copper ions may affect antibiotics via the formation of coordination bonds and/or redox reactions. Herein, we analyzed the interactions of Cu〈sup〉2+〈/sup〉 with eight β-lactam antibiotics using UV–Vis spectrophotometry, EPR spectroscopy, and electrochemical methods. Penicillin G did not show any detectable interactions with Cu〈sup〉2+〈/sup〉. Ampicillin, amoxicillin and cephalexin formed stable colored complexes with octahedral coordination environment of Cu〈sup〉2+〈/sup〉 with tetragonal distortion, and primary amine group as the site of coordinate bond formation. These β-lactams increased the solubility of Cu〈sup〉2+〈/sup〉 in the phosphate buffer. Ceftazidime and Cu〈sup〉2+〈/sup〉 formed a complex with a similar geometry and gave rise to an organic radical. Ceftriaxone-Cu〈sup〉2+〈/sup〉 complex appears to exhibit different geometry. All complexes showed 1:1 stoichiometry. Cefaclor reduced Cu〈sup〉2+〈/sup〉 to Cu〈sup〉1+〈/sup〉 that further reacted with molecular oxygen to produce hydrogen peroxide. Finally, meropenem underwent degradation in the presence of copper. The analysis of activity against 〈em〉Escherichia coli〈/em〉 and 〈em〉Staphylococcus aureus〈/em〉 showed that the effects of meropenem, amoxicillin, ampicillin, and ceftriaxone were significantly hindered in the presence of copper ions. The interactions with copper ions should be taken into account regarding the problem of antibiotic resistance and in the selection of the most efficient antimicrobial therapy for patients with altered copper homeostasis.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918311213-fx1.jpg" width="405" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): 〈/p〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Zhen-Zhou Yang, Ying-Ting Yu, Hong-Ru Lin, De-Chun Liao, Xiang-Huan Cui, Hong-Bing Wang〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Lonicera japonica〈/em〉 (LJ) is widely used as the local medicine to improve body and prevent ills in China, but mechanisms of its healthy beneficial effects remain largely unclear. Here, we evaluated the anti-aging and healthspan promoting activities of 75% ethanol extract of LJ (LJ-E) in the animal model 〈em〉Caenorhabditis elegans〈/em〉. Our results showed that LJ-E (500 μg/mL) treatment enhanced the mean lifespan of worms by over 21.87% and significantly improved age-associated physiological functions in 〈em〉C. elegans〈/em〉. The 500 μg/mL concentration of LJ-E enhanced the survival rates under oxidative and thermal stresses, and decreased reactive oxygen species (ROS) levels and fat accumulation in the worms. Gene-specific mutant studies showed that LJ-E-mediated lifespan extension was dependent on 〈em〉mev-1〈/em〉, 〈em〉daf-2〈/em〉, 〈em〉daf-16〈/em〉, and 〈em〉hsf-1〈/em〉, but not 〈em〉eat-2〈/em〉 genes. LJ-E could upregulate stress-inducible genes, viz., 〈em〉hsp-16.2〈/em〉, 〈em〉sod-3〈/em〉 and 〈em〉mtl-1〈/em〉. Moreover, we found that the D1086.10 protein interacted with superoxide dismutase (SOD)–3 by functional protein association networks analysis according to RNA-sequencing results. It was confirmed that 〈em〉D1086.10〈/em〉 was needed to promote longevity, and positively regulated expression of 〈em〉sod-3〈/em〉 by using 〈em〉D1086.10〈/em〉 mutants. Furthermore, LJ-E significantly delayed amyloid β-protein induced paralysis in CL4176 strain. Given the important role of autophagy in aging and protein homeostasis, we observed that LJ-E could remarkably increase the mRNA expression of autophagy gene 〈em〉bec-1〈/em〉 in CL4176 strain, and decrease expression of autophagy substrate p62 protein by more than 40.0% in BC12921 strain. Finally, we found that combination composed of three major compounds (54 μg/mL chlorogenic acid, 15 μg/mL 1,5-dicaffeoylquinic acid and 7.5 μg/mL 1,3-dicaffeoylquinic acid) of 500 μg/mL LJ-E could significantly delay paralysis in CL4176 worms caused by Aβ toxicity, comparable to that of LJ-E. Overall, our study may have important implications in using 〈em〉Lonicera japonica〈/em〉 to promote healthy aging and have a potency to design therapeutics for age-related diseases.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918313959-fx1.jpg" width="386" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): Hong-Na Mu, Quan Li, Jing-Yu Fan, Chun-Shui Pan, Yu-Ying Liu, Li Yan, Kai Sun, Bai-He Hu, Dan-Dan Huang, Xin-Rong Zhao, Xin Chang, Chuan-She Wang, Shu-Ya He, Ke He, Bao-Xue Yang, Jing-Yan Han〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The transplanted liver inevitably suffers from ischemia reperfusion (I/R) injury, which represents a key issue in clinical transplantation determining early outcome and long-term graft survival. A solution is needed to deal with this insult. This study was undertaken to explore the effect of Caffeic acid (CA), a naturally occurring antioxidant, on I/R injury of grafted liver and the mechanisms involved. Male Sprague–Dawley rats underwent orthotopic liver transplantation (LT) in the absence or presence of CA administration. In vitro, HL7702 cells were subjected to hypoxia/reoxygenation. LT led to apparent hepatic I/R injury, manifested by deteriorated liver function, microcirculatory disturbance and increased apoptosis, along with increased PDIA3 expression and nicotinamide adenosine dinucleotide phosphate (NADPH) oxidase activity, and membrane translocation of NADPH oxidase subunits. Treatment with CA attenuated the above alterations. siRNA/shRNA-mediated knockdown of PDIA3 in HL7702 cells and rats played the same role as CA not only in inhibiting ROS production and NADPH oxidase activity, but also in alleviating hepatocytes injury. CA protects transplanted livers from injury, which is likely attributed to its protection of oxidative damage by interfering in PDIA3-dependent activation of NADPH oxidase.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918315843-fx1.jpg" width="219" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: December 2018〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 129〈/p〉 〈p〉Author(s): I. Ramírez-Camacho, F. Correa, M. El Hafidi, A. Silva-Palacios, M. Ostolga-Chavarría, M. Esparza-Perusquía, S. Olvera-Sánchez, O. Flores-Herrera, C. Zazueta〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉 〈p〉Electron leakage from dysfunctional respiratory chain and consequent superoxide formation leads to mitochondrial and cell injury during ischemia and reperfusion (IR). In this work we evaluate if the supramolecular assembly of the respiratory complexes into supercomplexes (SCs) is associated with preserved energy efficiency and diminished oxidative stress in post-ischemic hearts treated with the antioxidant N-acetylcysteine (NAC) and the cardioprotective maneuver of Postconditioning (PostC). Hemodynamic variables, infarct size, oxidative stress markers, oxygen consumption and the activity/stability of SCs were compared between groups. We found that mitochondrial oxygen consumption and the activity of respiratory complexes are preserved in mitochondria from reperfused hearts treated with both NAC and PostC. Both treatments contribute to recover the activity of individual complexes. NAC reduced oxidative stress and maintained SCs assemblies containing Complex I, Complex III, Complex IV and the adapter protein SCAFI more effectively than PostC. On the other hand, the activities of CI, CIII and CIV associated to SCs assemblies were preserved by this maneuver, suggesting that the activation of other cardioprotective mechanisms besides oxidative stress contention might participate in maintaining the activity of the mitochondrial respiratory complexes in such superstructures.〈/p〉 〈p〉We conclude that both the monomeric and the SCs assembly of the respiratory chain contribute to the in vivo functionality of the mitochondria. However, although the ROS-induced damage and the consequent increased production of ROS affect the assembly of SCs, other levels of regulation as those induced by PostC, might participate in maintaining the activity of the respiratory complexes in such superstructures.〈/p〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918317441-fx1.jpg" width="211" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Free Radical Biology and Medicine, Volume 130〈/p〉 〈p〉Author(s): Eleonora Cremonini, Cesar G. Fraga, Patricia I. Oteiza〈/p〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Emerging evidence supports a beneficial action of the flavan-3-ol (–)-epicatechin (EC) on insulin sensitivity and potential impact on the development/progression of type 2 diabetes (T2D). In humans, supplementation with EC-rich foods, extracts, and pure EC improves insulin sensitivity and glucose tolerance in normal weight, overweight, obese and T2D individuals. These effects of EC are also observed in rodent models of diet-induced obesity and T2D. The events involved in the development of insulin resistance and T2D are multiple and interrelated. EC has been shown to inhibit inflammation, oxidative and endoplasmic reticulum stress, to modulate mitochondrial biogenesis and function, and to regulate events in the gastrointestinal tract and the pancreas that impact glucose homeostasis. A downregulation of oxidant production, particularly through direct inhibition or suppression of NADPH oxidase expression, and of redox sensitive signals (NF-κB, JNK1/2) that inhibit the insulin pathway, appear to be central to the beneficial actions of EC on insulin sensitivity. Overall, EC seems to have a positive role in the regulation of glucose homeostasis, however definitive answers on its importance for the management of T2D will depend on further clinical and mechanistic studies.〈/p〉〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S0891584918310876-fx1.jpg" width="262" alt="fx1" title="fx1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: January 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 84〈/p〉 〈p〉Author(s): Ye Yuan, Min Jin, Jia Xiong, Qicun Zhou〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The experiment was conducted to evaluate the effects of different dietary dosage forms of copper supplementation on growth performance, hematological characteristics, antioxidant capacity, immune responses and gene expressions related to innate immune of juvenile Pacific white shrimp 〈em〉Litopenaeus vannamei〈/em〉. Three isonitrogenous and isolipidic diets were formulated to contain three dosage forms of copper: copper sulfate (Diet I-Cu), copper sulfate + copper amino acid complex (1: 1, Diet M-Cu) and copper amino acid complex from Availa〈sup〉®〈/sup〉Cu100 (Diet Availa-Cu), respectively. 360 Pacific white shrimp juveniles (initial weight 1.86 ± 0.03 g) were randomly allocated in 12 tanks corresponding to quadruplicate tanks of the three dietary treatments, and the 8-week feeding trail was conducted. The results indicated that percent weight gain (PWG) and specific growth rate (SGR) in shrimp fed M-Cu diet were significantly higher than that fed I-Cu diet. Survival, feed efficiency (FE), protein efficiency ratio (PER) of shrimp were not significantly different between all treatment groups. High contents of total protein (TP) and glucose (GLU) were found in shrimp fed the diet containing M-Cu, whereas contents of cholesterol (CHOL) and triacylglycerol (TAG) in shrimp fed M-Cu diet were significantly lower than that in I-Cu diet group. In hemolymph, shrimp fed M-Cu diet had high activities of phenoloxidase (PO), alkaline phosphatase (ALP) and acid phosphatase (ACP). While, Cu/Zn superoxide dismutase (Cu/Zn SOD), ceruloplasmin (CP) and lysozyme (LZM) in hemolymph were not significantly affected by different dietary dosage forms of copper. High activities of Cu/Zn SOD, ALP, ACP and LZM in hepatopancreas were observed in shrimp fed M-Cu diet. Shrimp fed diet supplemented with Availa-Cu showed a significantly higher gene expression levels of Cu/Zn 〈em〉sod〈/em〉, 〈em〉alp〈/em〉, 〈em〉acp〈/em〉 and 〈em〉lzm〈/em〉 in hepatopancreas than that fed I-Cu diet. This study indicated that copper amino acid complex was more effective than copper sulfate to improve growth performance and enhance antioxidant ability and innate immune system.〈/p〉〈/div〉 〈/div〉