ALBERT

Description: 〈p〉Publication date: March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 86〈/p〉 〈p〉Author(s): Ying-Jun Ning, Si-Ying Chen, Xin-Jiang Lu, Jian-Fei Lu, Jiong Chen〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The glucocorticoid receptor (GR) is an important feedback regulator of the hypothalamic-pituitary-interrenal (HPI) axis. However, there are a limited number of studies focused on host-pathogen interactions in which an association between GR and immune response has been evaluated in monocytes/macrophages (MO/MФ) after being challenged with highly pathogenic bacteria. Here, we cloned the cDNA sequence of the glucocorticoid receptor (PaGR) gene from ayu fish. The PaGR transcript was expressed in all tissues, and changes in expression were observed in immune tissues and MO/MФ after live 〈em〉Vibrio anguillarum〈/em〉 infection. Subsequently, PaGR was expressed and purified to prepare anti-PaGR antibodies. We analyzed the subcellular localization of PaGR. PaGR was expressed not only in the intracellular space but also in the plasma membrane. PaGR activation decreased the expression of pro-inflammatory cytokines and increased the expression of anti-inflammatory cytokines. However, PaGR activation suppressed the phagocytosis activity of 〈em〉V. anguillarum〈/em〉-infected ayu MO/MФ via a non-genomic pathway. Interestingly, PaGR activation could enhance MO/MФ bacterial killing capability and apoptosis. Therefore, PaGR may modulate the immune response in ayu MO/MФ by genomic and non-genomic pathways.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 86〈/p〉 〈p〉Author(s): Xiaoze Guo, Jie Li, Chao Ran, Anran Wang, Mingxu Xie, Yadong Xie, Qianwen Ding, Zhen Zhang, Yalin Yang, Ming Duan, Zhigang Zhou〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In this study, we firstly tested the effects of dietary nucleotides on the disease resistance and innate immunity of zebrafish. Further, we investigated the role of intestinal microbiota in the nucleotides-induced immunostimulatory effect by using a germ-free zebrafish model and microbiota transfer technique. Fish were fed control or nucleotides (NT)-supplemented diets (at 0.05%,0.1%, 0.15% or 0.2%, 〈em〉m〈/em〉/〈em〉m〈/em〉) for 4 weeks, followed by immersion challenge with 〈em〉Aeromonas hydrophila〈/em〉 NJ-1. The results showed that 0.1% NT group enhanced the resistance of zebrafish against 〈em〉A. hydrophila〈/em〉 infection. We further observed that the relative expressions of mucin, claudin16, occlusin1, hepcidin, defensin beta-like, myeloperoxidase (Mpo), and serum amyloid A (Saa) increased in the 0.1% NT group compared with control (〈em〉P〈/em〉 〈 0.05), indicating that dietary nucleotides enhanced the physical barrier and mucosal immunity in the intestine of zebrafish. Moreover, ROS level in the head kidney was significantly increased in NT fed zebrafish versus control (〈em〉P〈/em〉 〈 0.05), indicating enhanced systematic immunity. Furthermore, dietary NT significantly elevated the relative expressions of 〈em〉mpo〈/em〉, 〈em〉saa〈/em〉 and the ROS activity in germ-free zebrafish, while germ-free zebrafish colonized with NT-altered microbiota had no significant difference in the relative expressions of 〈em〉mpo〈/em〉, 〈em〉saa〈/em〉 and the ROS activity compared with the control microbiota-colonized fish, suggesting that the immunostimulatory effect of dietary NT is mediated by direct action of NT and does not involve the microbiota. Consistently, dietary NT can protect germ-free zebrafish from pathogenic infection, whereas germ-free zebrafish colonized with NT microbiota showed no difference in disease resistance compared with control microbiota colonized counterparts. Together, these results indicated that the immunostimulatory and disease protection effect of dietary nucleotides in zebrafish was mediated by direct action of the nucleotides, and does not involve the intestinal microbiota.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 94〈/p〉 〈p〉Author(s): Taotao Zhang, Mimi Zhang, Ting Xu, Shangwu Chen, Anlong Xu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The lamprey is a primitive jawless vertebrate that occupies a critical phylogenetic position, and its larval stage represents the major portion of its life cycle [1]. Lamprey larvae have been proven to be an important model organism for studying numerous biological problems, such as the immune system, due to their unique biological features [2]. In addition, early-stage larvae have never been obtained from the wild [3]; therefore, it is necessary to establish artificial breeding of lampreys in the laboratory. However, during early development, the larvae exhibit susceptibility to saprolegniasis, and the immune responses of lamprey larvae to this infection remain poorly understood. Here, we established a model of fungal infection in lamprey larvae and then used RNA sequencing to investigate the transcript profiles of lamprey larvae and their immune responses to 〈em〉Saprolegnia ferax〈/em〉. Among the profiled molecules, genes involved in pathogen recognition, inflammation, phagocytosis, lysosomal degradation, soluble humoral effectors, and lymphocyte development were significantly upregulated. The results were validated by analysis of several genes by quantitative real-time PCR and whole-mount 〈em〉in situ〈/em〉 hybridization. Finally, we performed a Western blot for VLRs in infected and uninfected lampreys. This work not only provides an animal model for studying fungal infection but also suggests a molecular basis for developing defensive strategies to manage 〈em〉Saprolegnia ferax〈/em〉 infection.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 94〈/p〉 〈p〉Author(s): Ning Wang, Mengting Qin, Xihua Chen, Yang Lu, Xinxin Zhao, Yuhui Wu, Jie Shi, Yitian Li, Rui Zhang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Complement component C3 is well recognized as the central mediator of complement system, whose activation is responsible for the immune surveillance and elimination of non-self-antigens. In this study, C3 gene (HcC3) from a pearl making mussel, 〈em〉Hyriopsis cumingii〈/em〉, was successfully identified. The putative HcC3 possessed the canonical domains and highly conserved functional residues of C3 family members. In phylogenetic analysis, HcC3 was also clustered into C3 subfamily and separated from α2 macroglobulin clade. HcC3 gene was constitutively expressed in a wide range of tissues of pearl mussels, among which the immune-related tissues like hemocytes got highest expression. After allograft surgery of mantle tissues for aquaculture pearl production, the gene expression of HcC3 exhibited a rapid upregulation on day 1, dropped back on day 3, peaked the value on day 7, and restored to the level similar to control samples on day 14 after mantle allograft. The biphasic expression within the two weeks post the surgery suggests the important roles for HcC3 in alloimmune responses and an intricate complement activation mechanism in mollusks during tissue allograft.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 94〈/p〉 〈p〉Author(s): Chuangye Yang, Xiaodong Du, Ruijuan Hao, Qingheng Wang, Yuewen Deng, Ruijiao Sun〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Postoperative care is a critical step of pearl culture that ultimately determines culture success. To determine the effect of dietary vitamin D3 (VD3) levels on immunity and antioxidant capacity of pearl oyster 〈em〉Pinctada fucata martensii〈/em〉 during postoperative care and explore the mechanisms behind this phenomenon, five isonitrogenous and isolipidic experimental diets were formulated by adding different levels of dietary VD3 (0, 500, 1000, 3000, and 10000 IU/kg), and the diets were fed to five experimental groups (EG1, EG2, EG3, EG4, and EG5) in turn and cultured indoors. The control group (CG) was cultured in the natural sea. Pearl oysters that were 1.5 years old were subjected to nucleus insertion. After culturing for 30 days, EG3 exhibited significantly higher survival rates than those in CG and EG5 (P 〈 0.05). Moreover, EG3 exhibited the highest activities of alkaline phosphatase, acid phosphatase, catalase, superoxide dismutase, and lysozyme. However, EG5 achieved the highest activities of glutathione peroxidase. Metabolomics-based profiling of pearl oysters fed with high levels of dietary VD3 (EG5) and optimum levels of dietary VD3 (EG3) revealed 76 significantly differential metabolites (SDMs) (VIP 〉 1 and P 〈 0.05). Pathway analysis indicated that SDMs were involved in 21 pathways. Furthermore, integrated key metabolic pathway analysis suggested that pearl oysters in EG5 regulated the pentose phosphate pathway, glutathione metabolism, sphingolipid metabolism, and arachidonic acid metabolism in response to stress generated from excessive VD3. These findings had significant implications on strengthening the future development and application of VD3 in aquaculture of pearl oyster 〈em〉P. f. martensii〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 93〈/p〉 〈p〉Author(s): Hamed Kolangi Miandare, Ali Taheri Mirghaed, Marjan Hosseini, Nastaran Mazloumi, Ashkan Zargar, Sajad Nazari〈/p〉

Description: 〈p〉Publication date: November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 94〈/p〉 〈p〉Author(s): Ahmed N.F. Neamat-Allah, Essam A. Mahmoud, Yasser Abd El Hakim〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉As recently applicable, there are few studies on the impact of using nano-selenium (nano-Se) on varied fish species. Where nothing reachable focused on its impact on tilapias so, the present analysis evaluated the efficacy of using nano-Se in tilapias on immune response, antioxidant defense compared by conventional Se form. 480 〈em〉O. niloticus〈/em〉 fingerlings were haphazardly grouped firstly into three groups with four replicates of each. The control one (CT) was fed on a basal diet. The second and third one supplemented with 0.7 mg/kg〈sup〉−1〈/sup〉 Se and nano-Se respectively for ten weeks. At the start day of the ninth week, two replicates from each group were injected by 〈em〉Streptococcus iniae〈/em〉 where, the remaining replicates stand without challenge. Enhancement of growth performance measurements were noted in nano-Se compared to Se or CT groups. Existed anemia in 〈em〉S. iniae〈/em〉 tilapias became alleviated by using nano-Se that also, improves the alteration of leucogram induced by challenge. Elevation of aminotransferases, alkaline phosphatase, lactate dehydrogenase (ALT, AST, ALP and LDH) and creatinine in Se and CT challenged replicates that seemed nearly normal by using nano-Se. Usage of nano-Se showed more powerful antioxidant activities than Se. There were an expansion of immunoglobulin M, lysozymes, glutathione peroxidase, nitric oxide, superoxide dismutase and catalase (IgM, LYZ, GPx, NO, SOD, CAT) and their related gene expression in nano-Se with contrast in Se or CT challenged groups. Nile tilapias challenged by 〈em〉S. iniae〈/em〉 disclosed substantial expansion in the percentage of mortality in CT challenged fish (93.33%), followed by the group supplemented with Se (73.33%), whereas the lowermost one at fish supplemented by nano-Se (26.66%). The mortalities have been stopped from the 5〈sup〉th〈/sup〉, 12〈sup〉th〈/sup〉 and 14〈sup〉th〈/sup〉 days in, nano-Se, Se and CT respectively. It can be concluded that using of Se 0.7 mg/kg〈sup〉−1〈/sup〉induce immunosuppressive, antioxidant, liver and kidneys negative impact on tilapias where the same dose from nano-Se was more potent immunomodulating and antioxidant. Also it is attend in counteracting the serious impact induced by 〈em〉S. iniae〈/em〉 challenge.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: October 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 93〈/p〉 〈p〉Author(s): Ping Wang, Zhanying Zhang, Zhongtian Xu, Baoying Guo, Zhi Liao, Pengzhi Qi〈/p〉

Description: 〈p〉Publication date: November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 94〈/p〉 〈p〉Author(s): Lizhu Tao, Xiaoyan Xu, Yuan Fang, Anqi Wang, Fenglin Zhou, Yubang Shen, Jiale Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Grass carp septicemia is a systemic inflammatory response that develops following a bacterial infection. The hyperinflammatory state develops could lead to septic shock and lethality. There is increasing evidence that microRNAs are involved in the regulation of the inflammatory response. In the present study, miR-21 was confirmed to be involved in the inflammatory response following infection with 〈em〉Aeromonas hydrophila〈/em〉 and LPS stimulation. Both 〈em〉jnk〈/em〉 and 〈em〉ccr7〈/em〉 were identified as target gene of miR-21 by overexpression, inhibition, and dual luciferase reporter assays experiments. Meanwhile, miR-21 targets the 〈em〉jnk〈/em〉 and 〈em〉ccr7〈/em〉 to modulate downstream pro-inflammatory factors 〈em〉tnf-α, il-1β, il-6,〈/em〉 and 〈em〉il-12〈/em〉. Our results provide a theoretical basis for exploring the molecular mechanism of grass carp miR-21 regulating inflammation.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 94〈/p〉 〈p〉Author(s): Yunpeng Cao, Na Jin, Mingli Fan, Caiyun Lv, Xiaojun Song, Ping Jin, Fei Ma〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Transforming growth factor-β activated kinase-1 (TAK1) is an important member of the mitogen-activated protein kinase kinase kinase (MAP3K) family, which plays an important role in animal innate immune response. However, the 〈em〉TAK1〈/em〉 gene has yet not been reported in amphioxus to date. Here, we have identified and characterized a 〈em〉TAK1〈/em〉 gene from amphioxus (〈em〉Branchiostoma belcheri〈/em〉) (named as 〈em〉AmphiTAK1〈/em〉) with the full-length cDNA of 3479 bp, including an ORF sequence of 1905 bp, a 5′ UTR of 394 bp and a 3′ UTR of 1180 bp. Moreover, the predicted AmphiTAK1 protein contains STKc_TAK1 domain, TAB1 and TAB2/3 binding domain which are conserved among chordate, and phylogenetic analysis also shows that the AmphiTAK1 is located at the bottom of the chordate, revealing 〈em〉AmphiTAK1〈/em〉 as a new member of the 〈em〉TAK1〈/em〉 gene family. The further qRT-PCR analysis has shown that 〈em〉AmphiTAK1〈/em〉 is widely expressed in six investigated tissues (gonad, gill, hepatic cecum, intestine, muscle and notochord) of 〈em〉Branchiostoma belcheri〈/em〉, with high expression in notochord and gonad, moderate in gill and hepatic cecum. Notably, the expression level of 〈em〉AmphiTAK1〈/em〉 is significantly up-regulated after LPS stimulation. Specially, we also find that AmphiTAK1 protein can interact with AmphiTAB1 by immunoprecipitation assay. These findings reveal that AmphiTAK1 might interact with AmphiTAB1 to involve in innate immune response of 〈em〉Branchiostoma belcheri〈/em〉. Taken together, our present works provide a new insight into evolution and innate immune response mechanism of 〈em〉AmphiTAK1〈/em〉 gene in 〈em〉Branchiostoma belcheri〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 94〈/p〉 〈p〉Author(s): Chao Peng, Dongchang Xie, Chao Zhao, Haidong Xu, Sigang Fan, Lulu Yan, Pengfei Wang, Lihua Qiu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Akirin, which are members of the NF-κB signaling pathway, play critical roles in regulating the expression of antimicrobial peptides. In the present study, the 〈em〉Akirin〈/em〉 gene from 〈em〉Penaeus monodon〈/em〉 was identified from a transcriptome database and designated as 〈em〉PmAkirin〈/em〉. The complete sequence of the 〈em〉PmAkirin〈/em〉 cDNA was 1508 bp, encoding a protein of 213 amino acids, and it showed 99% amino acid identity to the 〈em〉Litopenaeus vannamei〈/em〉 Akirin. Two predicted nuclear localization signals (NLSs) were found, and the amino acid sequence alignments showed that PmAkirin was highly conserved at the N-terminus and C-terminus. 〈em〉PmAkirin〈/em〉 expression was found to be the highest in the hemolymph, followed by the heart, gill, stomach, hepatopancreas, intestine, and muscle. When challenged with 〈em〉Vibrio parahaemolyticus〈/em〉 infection, the 〈em〉PmAkirin〈/em〉 mRNA and three antimicrobial peptides (AMPs: 〈em〉PmALF2, PmALF3,〈/em〉 and 〈em〉PmCrus4〈/em〉) were upregulated. However, another five AMPs (〈em〉PmALF6, PmCrus1, PmPEN3a, PmPEN3b,〈/em〉 and 〈em〉PmPEN5〈/em〉) were downregulated by 〈em〉V. parahaemolyticus〈/em〉 infection. Silencing 〈em〉PmAkirin〈/em〉 by dsRNA significantly decreased the expression of the eight AMPs, which lead to an increase in the blood concentration of 〈em〉V. parahaemolyticus〈/em〉 and higher mortality in the shrimp. In contrast, the overexpression of 〈em〉PmAkirin〈/em〉 significantly increased the expression of the eight AMPs, which led to a reduction in the blood concentration of 〈em〉V. parahaemolyticus〈/em〉 and promoted the survival of the shrimp. Taken together, we concluded that 〈em〉PmAkirin〈/em〉 plays an important role in regulating the expression of AMPs in black tiger shrimp to defend against 〈em〉V. parahaemolyticus〈/em〉 infection.〈/p〉〈/div〉 〈/div〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉To dissect the 〈em〉TBX5〈/em〉 regulatory circuit, we focused on microRNAs (miRNAs) that collectively contribute to make TBX5 a pivotal cardiac regulator. We profiled miRNAs in hearts isolated from wild-type〈em〉, CRE, Tbx5〈/em〉〈sup〉〈em〉lox/〈/em〉+〈/sup〉〈em〉and Tbx5〈/em〉〈sup〉〈em〉del/〈/em〉+〈/sup〉 mice using a Next Generation Sequencing (NGS) approach. TBX5 deficiency in cardiomyocytes increased the expression of the miR-183 cluster family that is controlled by Kruppel-like factor 4, a transcription factor repressed by TBX5. MiR-182-5p, the most highly expressed miRNA of this family, was functionally analyzed in zebrafish. Transient overexpression of miR-182-5p affected heart morphology, calcium handling and the onset of arrhythmias as detected by ECG tracings. Accordingly, several calcium channel proteins identified as putative miR-182-5p targets were downregulated in miR-182-5p overexpressing hearts. In stable zebrafish transgenic lines, we demonstrated that selective miRNA-182-5p upregulation contributes to arrhythmias. Moreover, cardiac-specific down-regulation of miR-182-5p rescued cardiac defects in a zebrafish model of Holt–Oram syndrome. In conclusion, miR-182-5p exerts an evolutionarily conserved role as a TBX5 effector in the onset of cardiac propensity for arrhythmia, and constitutes a relevant target for mediating the relationship between 〈em〉TBX5〈/em〉, arrhythmia and heart development.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Permanently polarized cells have developed transduction mechanisms linking polarity sites with gene regulation in the nucleus. In neurons, one mechanism is based on long-distance retrograde migration of transcription factors (TFs). 〈em〉Aspergillus nidulans〈/em〉 FlbB is the only known fungal TF shown to migrate retrogradely to nuclei from the polarized region of fungal cells known as hyphae. There, FlbB controls developmental transitions by triggering the production of asexual multicellular structures. FlbB dynamics in hyphae is orchestrated by regulators FlbE and FlbD. At least three FlbE domains are involved in the acropetal transport of FlbB, with a final MyoE/actin filament-dependent step from the subapex to the apex. Experiments employing a T2A viral peptide-containing chimera (FlbE::mRFP::T2A::FlbB::GFP) suggest that apical FlbB/FlbE interaction is inhibited to initiate a dynein-dependent FlbB transport to nuclei. FlbD controls the nuclear accumulation of FlbB through a cMyb domain and a C-terminal LxxLL motif. Overall, results elucidate a highly dynamic pattern of FlbB interactions, which enable timely developmental induction. Furthermore, this system establishes a reference for TF-based long-distance signaling in permanently polarized cells.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Non-alcoholic fatty liver disease (NAFLD) is a metabolic liver disease that is thought to be reversible by changing the diet. To examine the impact of dietary changes on progression and cure of NAFLD, we fed mice a high-fat diet (HFD) or high-fructose diet (HFrD) for 9 weeks, followed by an additional 9 weeks, where mice were given normal chow diet. As predicted, the diet-induced NAFLD elicited changes in glucose tolerance, serum cholesterol, and triglyceride levels in both diet groups. Moreover, the diet-induced NAFLD phenotype was reversed, as measured by the recovery of glucose intolerance and high cholesterol levels when mice were given normal chow diet. However, surprisingly, the elevated serum triglyceride levels persisted. Metagenomic analysis revealed dietary-induced changes of microbiome composition, some of which remained altered even after reversing the diet to normal chow, as illustrated by species of the 〈em〉Odoribacter〈/em〉 genus. Genome-wide DNA methylation analysis revealed a “priming effect” through changes in DNA methylation in key liver genes. For example, the lipid-regulating gene 〈em〉Apoa4〈/em〉 remained hypomethylated in both groups even after introduction to normal chow diet. Our results support that dietary change, in part, reverses the NAFLD phenotype. However, some diet-induced effects remain, such as changes in microbiome composition, elevated serum triglyceride levels, and hypomethylation of key liver genes. While the results are correlative in nature, it is tempting to speculate that the dietary-induced changes in microbiome composition may in part contribute to the persistent epigenetic modifications in the liver.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Adult stem cells have a unique capacity to renew themselves and generate differentiated cells that are needed in the body. These cells are recruited and maintained by the surrounding microenvironment, known as the stem cell niche, during organ development. Thus, the stem cell niche is required for proper tissue homeostasis, and its dysregulation is associated with tumorigenesis and tissue degeneration. The identification of niche components and the mechanisms that regulate niche establishment and maintenance, however, are just beginning to be uncovered. Germline stem cells (GSCs) of the 〈em〉Drosophila〈/em〉 ovary provide an excellent model for studying the stem cell niche in vivo because of their well-characterized cell biology and the availability of genetic tools. In this review, we introduce the ovarian GSC niche, and the key signaling pathways for niche precursor segregation, niche specification, and niche extracellular environment establishment and niche maintenance that are involved in regulating niche size during development and adulthood.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Numerous studies have shown that non-coding RNAs play crucial roles in the development and progression of various tumor cells. Plasmacytoma variant translocation 1 (〈em〉PVT1〈/em〉) mainly encodes a long non-coding RNA (lncRNA) and is located on chromosome 8q24.21, which constitutes a fragile site for genetic aberrations. 〈em〉PVT1〈/em〉 is well-known for its interaction with its neighbor 〈em〉MYC〈/em〉, which is a qualified oncogene that plays a vital role in tumorigenesis. In the past several decades, increasing attention has been paid to the interaction mechanism between PVT1 and MYC, which will benefit the clinical treatment and prognosis of patients. In this review, we summarize the coamplification of 〈em〉PVT1〈/em〉 and 〈em〉MYC〈/em〉 in cancer, the positive feedback mechanism, and the latest promoter competition mechanism of 〈em〉PVT1〈/em〉 and 〈em〉MYC〈/em〉, as well as how PVT1 participates in the downstream signaling pathway of c-Myc by regulating key molecules. We also briefly describe the treatment prospects and research directions of PVT1 and MYC.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉CxxC-finger protein 1 (CFP1)-mediated trimethylated histone H3 at lysine-4 (H3K4me3) during oocyte development enables the oocyte genome to establish the competence to generate a new organism. Nevertheless, it remains unclear to which extent this epigenetic modification forms an instructive component of ovarian follicle development. We investigated the ovarian functions using an oocyte-specific 〈em〉Cxxc1〈/em〉 knockout mouse model, in which the H3K4me3 accumulation is downregulated in oocytes of developing follicles. CFP1-dependent H3K4 trimethylation in oocytes was necessary to maintain the expression of key paracrine factors and to facilitate the communication between an oocyte and the surrounding granulosa cells. The distinct gene expression patterns in cumulus cells within preovulatory follicles were disrupted by the 〈em〉Cxxc1〈/em〉 deletion in oocytes. Both follicle growth and ovulation were compromised after CFP1 deletion, because 〈em〉Cxxc1〈/em〉 deletion in oocytes indirectly impaired essential signaling pathways in granulosa cells that mediate the functions of follicle-stimulating hormone and luteinizing hormone. Therefore, CFP1-regulated epigenetic modification of the oocyte genome influences the responses of ovarian follicles to gonadotropin in a cell-nonautonomous manner.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide, with a high mortality rate. Its dismal prognosis is attributed to late diagnosis, high risk of recurrence and drug resistance. To improve the survival of patients with HCC, new approaches are required for early diagnosis, real-time monitoring and effective treatment. Exosomes are small membranous vesicles released by most cells that contain biological molecules and play a great role in intercellular communication under physiological or pathological conditions. In cancer, exosomes from tumor cells or non-tumor cells can be taken up by neighboring or distant target cells, and the cargoes in exosomes are functional to modulate the behaviors of tumors or reshape tumor microenvironment (TME). As essential components, non-coding RNAs (ncRNAs) are selectively enriched in exosomes, and exosomal ncRNAs participate in regulating specific aspects of tumor development, including tumorigenesis, tumor metastasis, angiogenesis, immunomodulation and drug resistance. Besides, dysregulated exosomal ncRNAs have emerged as potential biomarkers, and exosomes can serve as natural vehicles to deliver tumor-suppressed ncRNAs for treatment. In this review, we briefly summarize the biology of exosomes, the functions of exosomal ncRNAs in HCC development and their potential clinical applications, including as biomarkers and therapeutic tools.〈/p〉

Description: 〈p〉Publication date: Available online 2 November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology〈/p〉 〈p〉Author(s): Jingyi Cen, Lei Cui, Yafei Duan, Hua Zhang, Yarou Lin, Jiping Zheng, Songhui Lu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Palytoxins (PLTXs) are a group of complex and poisonous marine natural products that are toxic to marine life and even human beings. In the present study, the oxidative stress and immune response in the hepatopancreas and gills of 〈em〉Litopenaeus vannamei〈/em〉 were assessed for 72 h after injection with PLTX extracts. Chemical and physiological parameters, e.g., the respiratory burst (O〈sub〉2〈/sub〉〈sup〉−〈/sup〉), activities of antioxidant enzymes, oxidative damage to lipids, carbonylation of proteins, and immune gene mRNA expression levels, were analysed. The results showed that the PLTX extract was not fatal to the shrimp but could reduce their mobility. The O〈sub〉2〈/sub〉〈sup〉−〈/sup〉 levels in the gills gradually increased after exposure to PLTX extracts and were significantly higher than those in the control from 6 to 72 h. The malondialdehyde content, lipid peroxidation, protein carbonyl levels, and total antioxidant capacity in the gills all peaked at 12 h. At the same time, the gills were loosely connected, there was a clear disintegration of the epithelial tissue, and the stratum corneum disappeared after 12 h. In addition, compared to those in the control group, the PLTX extract treatment increased the O〈sub〉2〈/sub〉〈sup〉−〈/sup〉 content, malondialdehyde content, lipid peroxidation, and protein carbonyl levels from 12 to 72 h, 24–48 h, 12–24 h, and 12–72 h after injection in the hepatopancreas of the shrimp, respectively. Both the 〈em〉Crustin〈/em〉 and 〈em〉Toll〈/em〉 gene expression levels significantly increased in the hepatopancreas compared to those in the control 6–72 h after injection of the toxin. In parallel, the expression levels of the manganese superoxide dismutase gene gradually decreased from 6 to 48 h and returned to normal levels after 72 h. Interestingly, the total antioxidant capacity also significantly increased compared to that in the control from 6 to 72 h. Our results indicate that although PLTX extracts cause lipid peroxidation and carbonylation of proteins in hepatopancreatic cells, leading to their damage, they did not cause a decrease in the total antioxidant capacity of the hepatopancreas.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 2 November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology〈/p〉 〈p〉Author(s): Qiu-Ning Liu, Ting-Ting Yang, Cheng Wang, Sen-Hao Jiang, Dai-Zhen Zhang, Bo-Ping Tang, Bao-Ming Ge, Jia-Lian Wang, Dong Wang, Li-Shang Dai〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In this study, we identified a fish-specific Toll-like receptor (TLR) in 〈em〉Pelteobagrus fulvidraco〈/em〉, an economically important freshwater fish in China. This TLR, 〈em〉Pf〈/em〉TLR26, was shown to be encoded by a 3084 bp open reading frame (ORF), producing a polypeptide 1027 amino acids in length. The 〈em〉Pf〈/em〉TLR26 protein contains a signal peptide, eight leucine-rich repeat (LRR) domains, two LRR_TYP domains in the extracellular region, and a Toll/interleukin (IL)-1 receptor (TIR) domain in the cytoplasmic region, consistent with the characteristic TLR domain architecture. This predicted 117.1 kDa protein was highly homologous to those of other fish, with phylogenetic analysis revealing the closest relation to TLR26 of 〈em〉Ictalurus punctatus〈/em〉. Real-time quantitative reverse transcription-PCR (qRT-PCR) analysis showed that the 〈em〉Pf〈/em〉TLR26 gene was expressed in all tissues tested, with the highest expression levels seen in the head kidney and blood, and the lowest seen in muscle. 〈em〉Pf〈/em〉TLR26 exhibited significant upregulation in liver, spleen, head kidney, and blood at different time points following challenge with the common TLR agonists lipopolysaccharide (LPS) and polyriboinosinic polyribocytidylic acid (Poly I:C). Taken together, these results suggest that 〈em〉Pf〈/em〉TLR26 may be an important component of the 〈em〉P. fulvidraco〈/em〉 innate immune system, participating in the transduction of TLR signaling under pathogen stimulation.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 94〈/p〉 〈p〉Author(s): Jing Wang, Jia-Jia Du, Biao Jiang, Run-Zhen He, An-Xing Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Short-term feed deprivation or fasting is commonly experienced by aquaculture fish species and may be caused by seasonal variations, production strategies, or diseases. To assess the effects of fasting on the resistance of Nile tilapia to 〈em〉Streptococcus agalactiae〈/em〉 infection, vaccinated and unvaccinated fish were fasted for zero, one, three, and seven days prior to infection. The cortisol levels of both vaccinated and unvaccinated fish first decreased and then increased significantly as fasting time increased. Liver glycogen, triglycerides, and total cholesterol decreased significantly after seven days of fasting, but glucose content did not vary significantly between fish fasted for three and seven days. Hexokinase (HK) and pyruvate kinase (PK) activity levels were lowest after seven days of fasting, while phosphoenolpyruvate carboxykinase (PEPCK) activity levels varied in opposition to those of HK and PK. Serum superoxide dismutase (SOD) and catalase (CAT) activity levels first increased and then decreased as fasting time increased; SOD activity was highest after three days of fasting. Interleukin-1beta (〈em〉IL-1β〈/em〉) and 〈em〉IL-〈/em〉6 mRNA expression levels first increased and then decreased significantly, peaking after three days of fasting. However, suppressor of cytokine signaling-1 (〈em〉SOCS-1〈/em〉) mRNA expression levels were in opposition to those of 〈em〉IL-1β〈/em〉 and 〈em〉IL-6〈/em〉. Specific antibody levels did not vary significantly among unvaccinated fish fasted for different periods. Although specific antibody level first increased and then decreased in the vaccinated fish as fasting duration increased, there were no significant differences in the survival rates of fasted vaccinated fish after challenge with 〈em〉S. agalactiae〈/em〉. The final survival rates of vaccinated fish fasted for zero, one, three, and seven days were 86.67 ± 5.44%, 80.00 ± 3.14%, 88.89 ± 6.28%, and 84.44 ± 8.32%, respectively. Among the unvaccinated fish, the survival rate was highest (35.56 ± 3.14%) in the fish fasted for three days and lowest (6.67 ± 3.14%) in the fish fasted for seven days. Therefore, our results indicated that short-term fasting (three days) prior to an infection might increase the resistance of unvaccinated Nile tilapia to 〈em〉S. agalactiae〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 94〈/p〉 〈p〉Author(s): A. Miccoli, P.R. Saraceni, G. Scapigliati〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉This review describes and summarizes the knowledge on established and experimental vaccines developed against viral and bacterial pathologies affecting the most important farmed marine finfish species present in the Mediterranean area, namely European seabass 〈em〉Dicentrarchus labrax〈/em〉, sea bream 〈em〉Sparus aurata〈/em〉, turbot 〈em〉Psetta maxima〈/em〉 and meagre 〈em〉Argyrosomus regius〈/em〉. The diseases that have been recorded in seabass, sea bream and meagre are caused by bacteria 〈em〉Vibrio anguillarum〈/em〉, 〈em〉Photobacterium damselae〈/em〉, 〈em〉Tenacibaculum maritimum〈/em〉 as well as by viruses such as Viral Encephalopathy and Retinopathy/Viral Nervous Necrosis and Lymphocystic disease. The main pathologies of turbot are instead bacteriosis provoked by 〈em〉Tenacibaculum maritimum〈/em〉, 〈em〉Aeromonas〈/em〉 sp. and 〈em〉Vibrio anguillarum〈/em〉, and virosis by viral hemorrhagic septicaemia virus. Some vaccines have been optimized and are now regularly available for the majority of the above-mentioned pathogens. A measurable immune protection has been conferred principally against 〈em〉Vibrio anguillarum, Photobacterium damselae〈/em〉 sub. 〈em〉piscicida〈/em〉 and VER/VNN.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: December 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 95〈/p〉 〈p〉Author(s): Yue Ma, Yabo Liu, Yanyan Wu, Lei Jia, Xiaohong Liu, Qiyao Wang, Yuanxing Zhang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Surface display can expose foreign antigenic protein on the surface of the vaccine vector, which is promising choice to elicit better immune responses. In this study, we apply this strategy to develop an immunoactivator by using a live attenuated 〈em〉Vibrio harveyi〈/em〉 as an antigenic protein carrier with surface displayed VP28, a major envelope protein of white spot syndrome virus (WSSV), for two major pathogens of 〈em〉Litopenaeus vannamei.〈/em〉 As a result, the immunoactivator showed self-limited growth and attenuation of virulence in shrimp via different inoculation routes either with single-repetitive dose or high dose. Moreover, either intramuscular injection or oral administration of the immunoactivator did not affect growth of shrimp body weight or cause pathologic changes. Additionally, the rapid immunoprotection was induced by the immunoactivator after administration for one week with highly relative percent survival (RPS) more than 90% against both 〈em〉V. harveyi〈/em〉 and WSSV. Until 4 weeks post administration, the immunoactivator still possessed efficient immune effect with no less than 60% RPS for both pathogens. Totally, the attenuated 〈em〉V. harveyi〈/em〉 surface displaying VP28 could be a potential immunoactivator for WSSV and vibriosis control in 〈em〉L. vannamei〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 94〈/p〉 〈p〉Author(s): Yu Zhou, Long-Feng Lu, Xiao-Bing Lu, Shun Li, Yong-An Zhang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Mammalian cyclic GMP-AMP synthase (cGAS) senses double-stranded (ds) DNA in the cytosol to activate the innate antiviral response. In the present study, a cGAS-like gene, namely cGASL, was cloned from grass carp 〈em〉Ctenopharyngodon idellus〈/em〉, and its role as a negative regulator of the IFN response was revealed. Phylogenetic analysis indicated that cGASL was evolutionarily closest to cGAS, but was not a true ortholog of cGAS. Overexpression of cGASL inhibited poly I:C-stimulated grass carp (gc)IFN1pro and ISRE activities. In addition, MITA-, but not TBK1-mediated activation of gcIFN1pro was impaired by cGASL. Co-immunoprecipitation and Western blot experiments indicated that cGASL interacted with MITA and TBK1, resulting in a reduction in the phosphorylation of MITA. Lastly, overexpression of cGASL reduced the transcriptional levels of several IFN-stimulated genes activated by MITA. Collectively, these data suggest that cGASL is a negative regulator of IFN response by targeting MITA in fish.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 94〈/p〉 〈p〉Author(s): Hongjing Zhao, Yu Wang, Xin Yang, Dongxue Fei, Mengyao Mu, Menghao Guo, Hongxian Yu, Mingwei Xing〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉This study was designed to evaluate the effects of zinc on inflammation and tight junction (TJ) in different intestinal regions of common carp under sub-chronic arsenic insult. Fish were exposed to zinc (0, 1 mg/L) and arsenic trioxide (0, 2.83 mg/L) in individual or combination for a month. Inflammatory infiltration and TJ structure changes were displayed by H&E staining and transmission electron microscope. To further explore these changes, biochemical indicator (SOD), gene or protein expressions of inflammatory responses (NF-κB, IL-1β, IL-6 and IL-8) and TJ proteins (Occludin, Claudins and ZOs) were determined. In the anterior intestine, arsenic decreased activity of SOD, mRNA levels of Occludin, Claudins and ZOs, increased mRNA levels of ILs. However, unlike the anterior intestine, arsenic has an upregulation effects of Occludin and Claudin-4 in the mid intestine. These anomalies induced by arsenic, except IL-8, were completely or partially recovered by zinc co-administration. Furthermore, transcription factor (NF-κB) nuclear translocation paralleled with its downstream genes in both intestinal regions. In conclusion, our results unambiguously suggested that under arsenic stress, zinc can partly relieve intestinal inflammation and disruption of tight junction segment-dependently.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S1050464819309532-fx1.jpg" width="275" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: November 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 94〈/p〉 〈p〉Author(s): Yuan Cui, Kai Yin, Yingzheng Gong, Yingying Qu, Honggui Liu, Hongjin Lin〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Atrazine (ATR) causes environmental problems and damages the health of fish and aquatic animals. MicroRNAs (miRNAs) play important roles in immune regulation. However, the immunotoxicity mechanism of ATR in fish lymphocytes and the role of miRNA in this process remain unclear. To further study these mechanisms, spleen lymphocytes were exposed to 20, 40 and 60 μg/ml ATR for 18 h. Fluorescence staining and flow cytometry showed that the number of necrotic lymphocytes increased after ATR exposure. Compared with the control group, the mRNA expression of miR-181–5p was inhibited and the mRNA levels of TNF-α and HK2 were increased after ATR exposure. Additionally, the NF-κB inflammatory pathway and the levels of glycometabolism-related genes were upregulated. These results suggest that ATR induces inflammation and elevates glycometabolism in lymphocytes. We further found that the mRNA levels of receptor-interacting serine-threonine kinase 1 (RIP1), receptor-interacting serine-threonine kinase 3 (RIP3), mixed lineage kinase domain-like pseudokinase (MLKL), cylindromatosis (CYLD) and Fas-Associated protein with Death Domain (FADD) and the protein levels of RIP3 and MLKL in the treatment groups were significantly increased compared to those in control group, suggesting that ATR causes lymphocyte necroptosis. We conclude that miR-181–5p plays a key role in necroptosis in carp lymphocytes exposed to ATR by downregulating the expression of HK and TNF-α, which increases the level of glycometabolism and induces the inflammatory response, respectively.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S1050464819309520-fx1.jpg" width="485" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: February 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 97〈/p〉 〈p〉Author(s): Xu Zhang, Kai Huang, Huan Zhong, Yanqun Ma, Zhongbao Guo, Zhanyang Tang, Junneng Liang, Yongju Luo, Zhijian Su, Liqun Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In this study, the effect of 〈em〉Lycium barbarum〈/em〉 polysaccharides (LBP) on immunological parameters, apoptosis, and growth performance of Nile tilapia (〈em〉Oreochromis niloticus〈/em〉) was investigated. Dietary supplementation with LBP significantly increased complement 3 (C3) activity and promoted interleukin IL-1β gene expression in spleen tissue, significantly reduced apoptosis in spleen tissue, increased the specific growth rate (SGR), relative length gain (LG), and relative weight gain (WG) of Nile tilapia. However, dietary supplementation with LBP did not have a significant effect on serum alkaline phosphatase (AKP), malondialdehyde (MDA), and superoxide dismutase (SOD), blood constituents, apoptosis, or gene expression of IL-1β in liver tissue. Overall, the results showed that dietary supplementation with LBP increased the nonspecific immunity of Nile tilapia and reduced the apoptosis rate to promote growth and development. Thus, LBP has potential for use as a new immunostimulant in aquaculture.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: February 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 97〈/p〉 〈p〉Author(s): Ying Dong, Jingzhuang Zhao, Xiaoyu Chen, Miao Liu, Guangming Ren, Tongyan Lu, Yizhi Shao, Liming Xu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Infectious pancreatic necrosis virus (IPNV) is a common pathogen that causes huge economic losses for the salmonid aquaculture industry. Autophagy plays an important regulatory role in the invasion of pathogenic microorganisms. In this study, we explored the relationship between IPNV infection and autophagy in Chinook salmon embryo (CHSE-214) cells using standard methods. Transmission electron microscopy showed that IPNV infection produced typical structures of autophagosomes in CHSE-214 cells. Transformation of microtubule-associated protein 1 light chain 3 (LC3)-I to LC3-II protein, a marker of autophagy, was observed in IPNV-infected cells using confocal fluorescence microscopy and western blot analysis. Western blotting also showed that expression of the autophagy substrate p62 was significantly decreased in IPNV-infected cells. The influence of autophagy on IPNV multiplication was further clarified with cell culture experiments using autophagy inducer rapamycin and autophagy inhibitor 3-methyladenine. Rapamycin promoted IPNV multiplication at both the nucleic acid and protein levels, which led to higher IPNV yields; 3-methyladenine treatment had the opposite effect. This study has demonstrated that IPNV can induce autophagy, and that autophagy promotes the multiplication of IPNV in CHSE-214 cells.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: February 2020〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 97〈/p〉 〈p〉Author(s): Zhongyu Zhang, Gaoyang Liu, Rui Ma, Xiaozhou Qi, Gaoxue Wang, Bin Zhu, Fei Ling〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Aeromonas hydrophila〈/em〉 is a strong gram-negative bacterium that can cause a mass death of grass carp, and result in the huge economic loss. Development of practical vaccines is the best way to control the outbreak of this bacterial disease. In this study, a whole-cell inactivated vaccine was obtained via sonication, and then single-walled carbon nanotubes (SWCNTs) was used to link to the bacterial lysate (BL) for a novel vaccine (SWCNTs-BL). A total of 400 fish were vaccinated with BL and SWCNTs-BL via immersion (5, 10 mg L〈sup〉−1〈/sup〉) or injection (5, 10 μg/fish) before challenge with live 〈em〉A. hydrophila〈/em〉 at the 28 days post immunization (d.p.i.). The results showed that the antibody titer, enzymatic activity, expression of some immune-related genes (especially 〈em〉IgM〈/em〉 and 〈em〉TNF-α〈/em〉) and RPS of fish in the injection groups were significantly increased compared to the control group after 28 d.p.i. For the immersion groups, immunological parameters were increased compared to the control group. Furthermore, the immuno-protective effects of SWCNTs-BL were better than BL. The above results indicated that BL of 〈em〉A. hydrophila〈/em〉 can effectively induce specific immune response of grass carp, and BL linked with functionalized SWCNTs could enhance the protective effect of immersion immunization. Our results may provide a practical vaccine, with a simple production, to fight against bacterial diseases in aquaculture industry.〈/p〉〈/div〉 〈/div〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Invariant natural killer T (iNKT) cells represent a subgroup of innate-like T cells and play an important role in immune responses against certain pathogens. In addition, they have been linked to autoimmunity and antitumor immunity. iNKT cells consist of several subsets with distinct functions; however, the transcriptional networks controlling iNKT subset differentiation are still not fully characterized. Myc-associated zinc-finger-related factor (MAZR, also known as PATZ1) is an essential transcription factor for CD8〈sup〉+〈/sup〉 lineage differentiation of conventional T cells. Here, we show that MAZR plays an important role in iNKT cells. T-cell lineage-specific deletion of MAZR resulted in an iNKT cell-intrinsic defect that led to an increase in iNKT2 cell numbers, concurrent with a reduction in iNKT1 and iNKT17 cells. Consistent with the alteration in the subset distribution, deletion of MAZR also resulted in an increase in the percentage of IL-4-producing cells. Moreover, MAZR-deficient iNKT cells displayed an enhanced expression of Erg2 and ThPOK, key factors for iNKT cell generation and subset differentiation, indicating that MAZR controls iNKT cell development through fine-tuning of their expression levels. Taken together, our study identified MAZR as an essential transcription factor regulating iNKT cell subset differentiation and effector function.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉In the past two decades, transmembrane channel-like (TMC) proteins have attracted a significant amount of research interest, because mutations of 〈em〉Tmc1〈/em〉 lead to hereditary deafness. As evolutionarily conserved membrane proteins, TMC proteins are widely involved in diverse sensorimotor functions of many species, such as hearing, chemosensation, egg laying, and food texture detection. Interestingly, recent structural and physiological studies suggest that TMC channels may share a similar membrane topology with the Ca〈sup〉2+〈/sup〉-activated Cl〈sup〉−〈/sup〉 channel TMEM16 and the mechanically activated OSCA1.2/TMEM63 channel. Namely, these channels form dimers and each subunit consists of ten transmembrane segments. Despite this important structural insight, a key question remains: what is the gating mechanism of TMC channels? The major technical hurdle to answer this question is that the reconstitution of TMC proteins as functional ion channels has been challenging in mammalian heterologous systems. Since TMC channels are conserved across taxa, genetic studies of TMC channels in model organisms such as 〈em〉C. elegans〈/em〉, 〈em〉Drosophila〈/em〉, and zebrafish may provide us critical information on the physiological function and regulation of TMCs. Here, we present a comparative overview on the diverse functions of TMC channels in different species.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉The human gut microbiota, which underpins nutrition and systemic health, is compositionally sensitive to the availability of complex carbohydrates in the diet. The Bacteroidetes comprise a dominant phylum in the human gut microbiota whose members thrive on dietary and endogenous glycans by employing a diversity of highly specific, multi-gene polysaccharide utilization loci (PUL), which encode a variety of carbohydrases, transporters, and sensor/regulators. PULs invariably also encode surface glycan-binding proteins (SGBPs) that play a central role in saccharide capture at the outer membrane. Here, we present combined biophysical, structural, and in vivo characterization of the two SGBPs encoded by the 〈em〉Bacteroides ovatus〈/em〉 mixed-linkage β-glucan utilization locus (MLGUL), thereby elucidating their key roles in the metabolism of this ubiquitous dietary cereal polysaccharide. In particular, molecular insight gained through several crystallographic complexes of SGBP-A and SGBP-B with oligosaccharides reveals that unique shape complementarity of binding platforms underpins specificity for the kinked MLG backbone vis-à-vis linear β-glucans. Reverse-genetic analysis revealed that both the presence and binding ability of the SusD homolog BoSGBP〈sub〉MLG〈/sub〉-A are essential for growth on MLG, whereas the divergent, multi-domain BoSGBP〈sub〉MLG〈/sub〉-B is dispensable but may assist in oligosaccharide scavenging from the environment. The synthesis of these data illuminates the critical role SGBPs play in concert with other MLGUL components, reveals new structure–function relationships among SGBPs, and provides fundamental knowledge to inform future (meta)genomic, biochemical, and microbiological analyses of the human gut microbiota.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Uncontrolled scarring, or fibrosis, can interfere with the normal function of virtually all tissues of the body, ultimately leading to organ failure and death. Fibrotic diseases represent a major cause of death in industrialized countries. Unfortunately, no curative treatments for these conditions are yet available, highlighting the critical need for a better fundamental understanding of molecular mechanisms that may be therapeutically tractable. The ultimate indispensable effector cells responsible for deposition of extracellular matrix proteins that comprise scars are mesenchymal cells, namely fibroblasts and myofibroblasts. In this review, we focus on the biology of these cells and the molecular mechanisms that regulate their pertinent functions. We discuss key pro-fibrotic mediators, signaling pathways, and transcription factors that dictate their activation and persistence. Because of their possible clinical and therapeutic relevance, we also consider potential brakes on mesenchymal cell activation and cellular processes that may facilitate myofibroblast clearance from fibrotic tissue—topics that have in general been understudied.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Astrocytes are the key homeostatic cells in the central nervous system; initiation of reactive astrogliosis contributes to neuroinflammation. Pro-inflammatory cytokine interferon γ (IFNγ) induces the expression of the major histocompatibility complex class II (MHCII) molecules, involved in antigen presentation in reactive astrocytes. The pathway for MHCII delivery to the astrocyte plasma membrane, where MHCII present antigens, is unknown. Rat astrocytes in culture and in organotypic slices were exposed to IFNγ to induce reactive astrogliosis. Astrocytes were probed with optophysiologic tools to investigate subcellular localization of immunolabeled MHCII, and with electrophysiology to characterize interactions of single vesicles with the plasmalemma. In culture and in organotypic slices, IFNγ augmented the astrocytic expression of MHCII, which prominently co-localized with lysosomal marker LAMP1-EGFP, modestly co-localized with Rab7, and did not co-localize with endosomal markers Rab4A, EEA1, and TPC1. MHCII lysosomal localization was corroborated by treatment with the lysosomolytic agent glycyl-〈span〉l〈/span〉-phenylalanine-β-naphthylamide, which reduced the number of MHCII-positive vesicles. The surface presence of MHCII was revealed by immunolabeling of live non-permeabilized cells. In IFNγ-treated astrocytes, an increased fraction of large-diameter exocytotic vesicles (lysosome-like vesicles) with prolonged fusion pore dwell time and larger pore conductance was recorded, whereas the rate of endocytosis was decreased. Stimulation with ATP, which triggers cytosolic calcium signaling, increased the frequency of exocytotic events, whereas the frequency of full endocytosis was further reduced. In IFNγ-treated astrocytes, MHCII-linked antigen surface presentation is mediated by increased lysosomal exocytosis, whereas surface retention of antigens is prolonged by concomitant inhibition of endocytosis.〈/p〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Jianping Fu, Zhiqiang Yi, Hao Cui, Chunhui Song, Miao Yu, Yi Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉IFN-γ is a pleiotropic cytokine with significant roles in antiviral, antitumor and immune regulation. It could be used as an immuno-enhancer to improve fish protectiveness against pathogens. In this study, the prokaryotic expression plasmid pTwin1-N-IFN-γ was constructed to express 〈em〉Cyprinus carpio〈/em〉 (common carp) IFN-γ fused with a chitin binding domain (CBD) and a self-cleavable intein-tag, 〈em〉Synechocystis sp〈/em〉 DnaB. The recombinant protein CBD-DnaB-IFN-γ with the molecular weight of 44.25 kD was successfully expressed in soluble form, and the rIFN-γ (approximate 18.61 kD) was further cleaved and eluted under pH = 7.0 at 25 °C. rIFN-γ could be recognized by western blotting with rabbit anti-grass carp IFN-γ polyclonal antibody. Cytotoxicity studies on EPC cells showed that only 500 ng/ml rIFN-γ had a subtle effect on cells growth and its proliferation rate was reduced to 76.2%. EPC cells incubated with 100 ng/ml rIFN-γ showed significantly higher resistance against SVCV, reducing the TCID〈sub〉50〈/sub〉/ml by more than 800-fold. In vivo studies suggested that intraperitoneal injection of rIFN-γ significantly improved the survival rate of common carps compared with SVCV challenge alone. These results implied that rIFN-γ would act as an immuno-enhancer in carp aquaculture.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Nermeen M. Abu-Elala, Ahmed Samir, Momtaz Wasfy, Magdy Elsayed〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉A vaccine against streptococcosis, lactococcosis and enterococcosis in tilapia was formulated, ME-VAC Aqua Strept, as a polyvalent inactivated vaccine containing 〈em〉Streptococcus agalactiae〈/em〉, 〈em〉S. iniae〈/em〉, 〈em〉Lactococcus garvieae〈/em〉 and 〈em〉Enterococcus faecalis〈/em〉 along with a nano-particulate adjuvant. Use of ME-VAC Aqua Strept by injection or immersion resulted in an improved non-specific and adaptive immunity of broodstock and offspring. Intra-peritoneal vaccination of tilapia broodstock increased the total leukocyte count, phagocytosis, lysozyme activity, antibody titer, number of seeds/vaccinated broodstock, seeds quality and survival rates. Also, immersion mass vaccination of tilapia larvae provided a long period of protection up to three months, with a relative percent of survivability (RPS) not less than 60% at this time. To our knowledge, this vaccine may be the first to offer a combined protection against streptococcosis, lactococcosis and enterococcosis in tilapia. The results support the use of this vaccine as an effective tool for disease control and well-being of fish.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S1050464819301160-fx1.jpg" width="264" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Chong Han, Qiang Li, Qinghua Chen, Guofeng Zhou, Jianrong Huang, Yong Zhang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉〈em〉Mastacembelus armatus〈/em〉, also known as the zigzag eel, is an economically important species of freshwater fish that is very popular with consumers as a high-grade table fish in China. Recently, the wild population of this fish has declined gradually due to overfishing and various types of ecological imbalance. Meanwhile, the aquaculture of this spiny eel has flourished in southern China. To understand the immune response of zigzag eel to 〈em〉Aeromonas veronii〈/em〉, we carried out transcriptome sequencing of zigzag eel spleens after artificial bacterial infection. After assembly, 110,328 unigenes were obtained with 44.42% GC content. A total of 27,098 unigenes were successfully annotated by four public protein databases, namely, Nr, UniProt, KEGG and KOG. Differential expression analysis revealed the existence of 1278 significantly differentially expressed unigenes at 24 h post infection, with 767 unigenes upregulated and 511 unigenes downregulated. After GO and KEGG enrichment analyses, many immune-related GO categories and pathways were significantly enriched. The typical significantly enriched pathways included toll-like receptor signaling pathway, cytokine-cytokine receptor interaction and TNF signaling pathway. In addition, 40,027 microsatellites (SSRs) and 52,716 candidate single nucleotide polymorphisms (SNPs) were identified from the infection and control transcriptome libraries. Overall, this transcriptomic analysis provided valuable information for studying the immune response of zigzag eels against bacterial infection.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Gustavo S. Claudiano, Jefferson Yunis-Aguinaga, Fausto A. Marinho-Neto, Renata L. Miranda, Isabela M. Martins, Fabrizia S. Otani, Antonio V. Mundim, Cleni M. Marzocchi-Machado, Julieta R.E. Moraes, Flávio Ruas de Moraes〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The pathogenesis of sepsis involves complex systems and multiple interrelationships between the host and pathogen producing high mortality rates in various animal species. In this study, hematological disturbances, innate immunity and survival during the septic process in 〈em〉Piaractus mesopotamicus〈/em〉 inoculated with 〈em〉Aeromonas hydrophila〈/em〉 were studied. For this aim, fish blood samples were taken from control and infected groups 1, 3, 6, and 9 h post-inoculation (HPI). Leukogram showed reduction in the number of leukocytes and thrombocytes, followed by cessation of leukocyte chemotaxis 6 HPI and severe morphological changes in leukocytes and erythrocytes. At 3 HPI production of reactive oxygen species increased and at 6 HPI decreased. There was no change in serum lysozyme concentration and lytic activity of the complement system, despite the progressive increase in serum lytic activity and bacterial agglutination. Finally, the changes in clinical signs due to aeromonosis and increasing septicemia resulted in a reduction in survival to 57.14% after 36 HPI. It was possible concluded that these hematological and immune are crucial event in the worsening of sepsis in 〈em〉P. mesopotamicus〈/em〉, and these findings are utility for diagnosing and understanding the pathophysiology sepsis in pacu induced by 〈em〉A. hydrophila〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Fang Li, Limei Xu, Xuan Hui, Wanzhen Huang, Feng Yang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Hemocytes are the major immune cells of crustaceans. New hemocyte production is required throughout the life cycle of these animals to maintain a functional immune system. The mechanism of crustacean hematopoiesis has just begun to be understood and new methods are needed for the investigation of this process. Here we report the directed differentiation of granular cells (GCs) from the hematopoietic tissue (HPT) cells of 〈em〉Cherax quadricarinatus in vitro〈/em〉. We started by providing the cultured HPT cells with different additives to induce possible differentiation. We found that crayfish muscle extract greatly promoted the physical status of the cells and induced the formation of refractile cytoplasmic granules. The transcription of marker genes and the production of functional prophenoloxidase further confirmed the formation of mature GCs. In our experiments, young GCs usually started to develop in ∼2 weeks post induction and over 60% of the cells became mature within 3–4 weeks. This is the first time that the fully differentiation of crustacean hemocytes is accomplished 〈em〉in vitro〈/em〉. It provides a powerful tool for in-depth study of crustacean hematopoiesis.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): M. Tarnawska, M. Augustyniak, P. Łaszczyca, P Migula, I. Irnazarow, M. Krzyżowski, A. Babczyńska〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Pharmaceuticals and household chemicals are important components of municipal sewage. Many of them are biologically active, disrupting not only hormonal regulation of aquatic animals but also, indirectly, disturbing their immunological protection. In the environment, chemicals rarely act as individual substances, but as elements of mixtures. Therefore, the aim of this study was to check whether the acute laboratory exposure of common carp juveniles to a mixture of ibuprofen, sodium dodecyl sulphate (SDS), dimethyl sulfoxide (DMSO) and 17 α-ethynylestradiol in increasing concentrations, modifies the levels of innate immunity (lysozyme, C-reactive protein) as well as general stress (metallothioneins, heat shock proteins HSP70) markers in brain, liver, gills, spleen and mucus. The levels of the markers were measured by an immunodetection technique. Not only do the pharmaceuticals and household chemicals impair immunological reactions of young carp in various tissues but also do that in a concentration-dependent manner in the liver, gills, spleen and mucus. This has a very important implication, since it may result in higher sensitivity of young fish to pathogens due to energy allocation to defence processes. The comparisons of the pattern of stress reactions in the studied organ samples indicated that mucus appeared to be a good, non-invasive material for monitoring of environmental state and fish conditions.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Hongkuan Zhang, Dewei Cheng, Karsoon Tan, Hongxing Liu, Ting Ye, Shengkang Li, Hongyu Ma, Huaiping Zheng〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉As a major intracellular iron storage protein, ferritin plays important roles in iron homeostasis and innate immunity. In this study, two novel 〈em〉ferritin〈/em〉 subunits from noble scallop 〈em〉Chlamys nobilis〈/em〉 (〈em〉CnFer1〈/em〉 and 〈em〉CnFer2〈/em〉) were identified and analyzed. The open reading frame of 〈em〉CnFer1〈/em〉 and 〈em〉CnFer2〈/em〉 was 522 and 519bp long, encoding 173 and 172 amino acids, respectively. Both ferritins contained a putative iron-binding region signature (IBRS). Analysis of putative conserved domains showed the two 〈em〉CnFer〈/em〉 genes contained three key domains of ferritin subunits, a ferroxidase diiron center (E25, Y32, E59, E60, H63, E105, and Q139), an iron ion channel (H116, D129, E132) and a ferrihydrite nucleation center (D58, E59, and E62) that present in M type subunits. A putative iron response element (IRE) was observed at both 〈em〉CnFer〈/em〉 genes in the 5’ UTR. Phylogenetic analysis result suggested that the two genes are cytoplasmic 〈em〉ferritins〈/em〉 and have the closest evolution relationship with 〈em〉ferritins〈/em〉 from 〈em〉Mizuhopecten yessoensis〈/em〉. The two 〈em〉ferritin〈/em〉 genes were wildly expressed in examined tissues and the highest level was found in gill. After 〈em〉V. parahaemolyticus〈/em〉 challenged, both 〈em〉CnFer〈/em〉 genes were significantly up-regulated suggesting that they are important proteins involved in host immune defense. Moreover, under bacterial challenge, the expression levels of both two genes in Golden scallops (rich in carotenoids) were significantly higher than that in Brown scallops (less in carotenoids) which suggesting that carotenoids enhance the immunity in scallops to defense against the bacterial stress.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Zhenzhu Sun, Xiaohong Tan, Minglei Xu, Qingying Liu, Huaqun Ye, Cuiyun Zou, Yuanyuan Zhou, Ningning Su, Leling Chen, Anli Wang, Chaoxia Ye〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The study investigated the effects of dietary supplementation with dandelion extracts (DE) on growth performance, feed utilization, body composition, serum biochemical, liver histology, immune-related gene expression and CCl〈sub〉4〈/sub〉 resistance of hybrid grouper 〈em〉(Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀)〈/em〉. A basal diet supplemented with DE at 0% (diet 0%), 0.1% (diet 0.1%), 0.2% (diet 0.2%), 0.4% (diet 0.4%) and 0.8% (diet 0.8%) were fed to hybrid grouper for 8 weeks. The results revealed that dietary DE had not a significant impact on growth performance and feed utilization (〈em〉P〈/em〉 〉 0.05), but it could decrease the percent of crude lipids in whole body and increase the percent of crude protein in muscle (〈em〉P〈/em〉 〈 0.05). Dietary DE increased the mRNA levels of antioxidant enzymes (catalase, glutathione peroxidase and glutathione reductase) and reduced inflammatory factor in the spleen and head-kidney of fish (〈em〉P〈/em〉 〈 0.05), but reduced the expression of the liver antioxidant gene except for glutathione reductase (〈em〉P〈/em〉 〈 0.05). Dietary supplementation with 0.2%–0.4% DE could effectively improve liver health. After injection of CCL〈sub〉4〈/sub〉 by 72 h, fish fed Diet0.2% and Diet0.4% showed regular hepatocyte morphology while fish fed Diet 0%, Diet 0.1% and Diet 0.8% showed hepatocyte damage. Higher survival rate and total blood cell count was observed in fish fed 0.1%–0.4% dietary DE (〈em〉P〈/em〉 〈 0.05). In conclusion, DE could be used as a functional feed additive to enhance liver function of farmed fish. The best level of it should be between 0.2% and 0.4%.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Pengcheng Bao, Xiang Sun, Qin Liu, Yuanxing Zhang, Xiaohong Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In aquaculture, more than one pathogen usually be isolated from the sick fish, creating an urgent need for developing combined vaccines to control fish disease caused by multiple pathogens simultaneously. In our previous work, two live attenuated vaccines against 〈em〉Vibrio anguillarum〈/em〉 and 〈em〉Edwardsiella piscicida〈/em〉 were vaccinated in turbot, exhibiting an efficient protection. However, some immunological processes such as antigenic competition, antigenic cross-reaction and antigen induced suppression during combined vaccination are unknown. In this study, we evaluated the effectiveness of the combined live vaccines and explored the immunological processes after vaccination. We found that the combined two live attenuated vaccines for 〈em〉V. anguillarum〈/em〉 and 〈em〉E. piscicida〈/em〉 induced a stronger immune response without existing antigen competition. Instead, a synergistic effect was observed not only for triggering innate immune response but for stimulation of adaptive immunity. Our study suggested that the two combined live vaccines against 〈em〉V. anguillarum〈/em〉 and 〈em〉E. piscicida〈/em〉 could be used simultaneously in the future.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Chan-Hee Kim, Eun Jeong Kim, Yoon Kwon Nam〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Hepcidin, a cysteine-rich antimicrobial peptide (AMP), plays key roles as a regulatory hormone in iron homeostasis, providing a link between iron metabolism and innate immunity. Unlike many other AMPs displaying a high degree of sequence variability among closely related organisms, hepcidin is highly conserved from teleosts to mammals. However, little is known about the early ancestry of hepcidins in the vertebrate lineage. Here, we first report potential a prototype hepcidin from the Siberian sturgeon 〈em〉Acipenser baerii〈/em〉, a primitive chondrostean species. The 〈em〉A. baerii〈/em〉 hepcidin (〈em〉Ab〈/em〉HAMP) gene showed a tripartite exon-intron organization, which encoded a precursor protein comprised of three structural signatures containing eight cysteine residues, a common structure in vertebrate hepcidin genes and proteins. mRNA expression by iron-overloading and bacterial infection and antibacterial activity revealed that 〈em〉Ab〈/em〉HAMP might play a role in iron metabolism regulator in the liver, and in direct and/or indirect host immune response in the kidney against invading pathogen. Comparison of gene and protein sequences reveled that 〈em〉Ab〈/em〉HAMP possesses intermediate characteristics between tetrapodian and teleostean hepcidins (HAMP1s). Phylogenetically, 〈em〉Ab〈/em〉HAMP had a closer genetic affiliation to tetrapodian orthologs than to teleostean orthologs, suggesting that the structures of this chondrostean hepcidin may closely reflect the structures of an evolutionarily ancestral form that might have evolved into extant hepcidins in tetrapods and teleosts, respectively. Based on the identification of hepcidin from the chondrostean group, the emergence of the common ancestral hepcidin should be traced back to in early Osteichthyes: no later than sarcopterygian (lobe-finned fishes) – actinopterygian (ray-finned fishes) split.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Jixing Feng, Denglai Li, Liming Liu, Yongzheng Tang, Rongbin Du〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In this study, GST Pull-down and mass spectrometry was applied to the precipitation and identification of the small GTP-binding protein (Rab7) interacting protein in hemocyte of 〈em〉Litopenaeus vannamei〈/em〉. According to the search in GenBank with the peptide mass fingerprint, the 45 kDa protein which was pulled down with the GST-tagged Rab7 (GST-Rab7, GTP bound form) was identified to be β-actin with 28% coverage of amino acid sequences. The interaction of Rab7 with β-actin was verified by both GST Pull-down and ELISA 〈em〉in vitro〈/em〉. Meanwhile, confocal microscopic observation showed that Rab7 could be co-localized with β-actin in hemocytes at 12 h post white spot syndrome virus (WSSV) infection (hpi). GST Pull-down and western blotting were used to analyze the cross-interaction between WSSV VP28, Rab7 and β-actin. The results showed that the GST-VP28, His-tagged Rab7 (His-Rab7) and His-β-actin formed a tripartite complex. At 12 hpi, confocal microscopic observation showed that WSSV could be co-localized with Rab7 and β-actin in hemocytes respectively. Furthermore, based on the 〈em〉in vivo〈/em〉 neutralization assay, recombinant His-β-actin accelerated the infection of WSSV, conversely, recombinant His-Rab7 delayed WSSV infection in shrimp. These results suggested the interaction of Rab7 with β-actin and this interaction was involved in WSSV infection.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Yi-Ting Tseng, Ramya Kumar, Han-Ching Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The white Spot Syndrome Virus (WSSV) is a pathogen that causes huge economic losses in the shrimp-farming industry globally. At the WSSV genome replication stage (12 hpi) in WSSV-infected shrimp hemocytes, activation of the PI3K-Akt-mTOR pathway triggers metabolic changes that resemble the Warburg effect. In shrimp, the upstream regulators of this pathway are still unknown, and in the present study, we isolate, characterize and investigate two candidate factors, i.e. the shrimp Ras GTPase isoforms LvRas and LvRap, both of which are upregulated after WSSV infection. dsRNA silencing experiments show that virus replication is significantly reduced when expression of either of these genes is suppressed. Pretreatment with the Ras inhibitor Salirasib further suggests that LvRas, which is a homolog to a commonly overexpressed human oncoprotein, may be involved in regulating the WSSV-induced Warburg effect. We also show that while both the PI3K-Akt-mTOR and Raf-MEK-ERK pathways are activated by WSSV infection, LvRas appears to be involved only in the regulation of the mTOR pathway.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Shunyou Lv, Ya Zhang, Jiaying Zheng, Xiaohong Huang, Youhua Huang, Qiwei Qin〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Tripartite motif (TRIM) proteins have been demonstrated to exhibit critical functions in multiple cellular processes, including development, carcinogenesis, and programmed cell death, and are also widely recognized to be important antiviral restriction factors or modulators of immune and inflammatory signaling pathways. However, in teleosts, additional TRIM members have been identified and their functions remain largely unknown. Here, a novel finTRIM gene from orange spotted grouper (EcfinTRIM82) was cloned and characterized. Sequence analysis indicated that EcfinTRIM82 encoded a 575 amino acid peptide which shared 94% and 82% identity with Asian sea bass (〈em〉Lates calcarifer〈/em〉), and zebrafish (〈em〉Danio rerio〈/em〉) finTRIM82, respectively. EcfinTRIM82 contained three conserved domains, including a RING, B-Box, and SPRY domain. Using fluorescence microscopy, we found that green fluorescence aggregates were observed in the cytoplasm of EcfinTRIM82-EGFP transfected grouper spleen (GS) cells. As the infection proceeded, EcfinTRIM82 transcription was significantly upregulated in Singapore grouper iridovirus (SGIV) or red-spotted grouper nervous necrosis virus (RGNNV) infected GS cells. This suggests that EcfinTRIM82 might be involved in fish virus infection. The 〈em〉in vitro〈/em〉 overexpression of EcfinTRIM82 in GS cells significantly enhanced the replication of SGIV and RGNNV, evidenced by increased expression of viral genes, including the SGIV major capsid protein (MCP), VP19, ICP-18, RGNNV coat protein (CP), and RNA-dependent RNA polymerase (RdRp). Furthermore, the ectopic expression of EcfinTRIM82 significantly decreased the expression of interferon (IFN)-related signaling molecules, including interferon regulatory factor 3 (IRF3), IRF7, interferon stimulated gene 15 (ISG15), ISG56, IFP35, and myxovirus resistance gene (MXI), suggesting that EcfinTRIM82 regulated viral replication via the negative regulation of the host IFN response. In addition, EcfinTRIM82 overexpression substantially decreased the level of proinflammatory cytokine transcription. Furthermore, the ectopic expression of EcfinTRIM82 significantly weakened the melanoma differentiation-associated protein 5 (MDA5), mediator of IRF3 activation (MITA) and mitochondrial antiviral-signaling (MAVS) protein-induced IFN response by detecting the transcription of interferon related cytokines and the promoter activity of IFN. Together, our results demonstrate that finTRIM82 negatively regulates the innate antiviral immune response against grouper virus infection.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Bingzhi Zhang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Three test diets containing three different levels (1, 3, and 5 g kg〈sup〉−1〈/sup〉) of dietary chitosan oligosaccharides (COs) were formulated and used to test the growth performance, body composition, intestine digestive enzymes, antioxidant responses and resistance to 〈em〉Aeromonas hydrophila〈/em〉 of loach 〈em〉Paramisgurnus dabryanus〈/em〉. A basal diet without any COs served as the control. After 60 days of feeding, the growth performance, intestine digestive-enzyme activities, body protein content and total polyunsaturated fatty acids, antioxidant responses, and resistance to 〈em〉A. hydrophila〈/em〉 of loach 〈em〉P. dabryanus〈/em〉 were higher than those of the control when the loach 〈em〉P. dabryanus〈/em〉 was fed with CO-containing diets. The optimum dose of dietary COs required for the maximum growth of loach was 3 g kg〈sup〉−1〈/sup〉 of the diet. Results indicated that dietary COs can improve the growth performance, body composition, intestine digestive enzymes, antioxidant responses, and resistance to 〈em〉A. hydrophila〈/em〉 of loach 〈em〉P. dabryanus〈/em〉 and can thus be used as a diet supplement for them.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Yafei Duan, Yun Wang, Qingsong Liu, Dalin Xiong, Jiasong Zhang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Harmful effects of water pollutants are myriad. Sulfide from water bodies affects the aquatic animals. Intestine barrier function serves as the front-line of animals defense. Our previous study confirmed the toxic effect of sulfide on intestine immune response of 〈em〉Litopenaeus vannamei〈/em〉, but the underlying mechanisms remained elusive. Therefore, in this study, we investigated the transcriptomic and microbiota responses of the 〈em〉L. vannamei〈/em〉 intestine subjected to acute sulfide exposure. Sulfide decreased bacterial richness and altered the intestine microbiota composition. Specifically, sulfide increased the abundances of Bacteroidetes and Actinobacteria, but decreased the abundance of Proteobacteria. At the genus level, sulfide increased typical cellulolytic characteristics bacteria, such as 〈em〉Formosa〈/em〉, 〈em〉Sphingomonas〈/em〉, and 〈em〉Demequina〈/em〉. RNA-seq analysis identified differential expression of 1799 genes (701 up-regulated and 1098 down-regulated) were grouped into 267 pathways. The most enriched pathway ‘amoebiasis’ was related to the intestine mucus homeostasis. A number of immune-related genes associated with antimicrobial, antioxidant, pathogen attachment and recognition, and apoptosis processes in contrasting accessions; they were correlated with the abundance of intestine bacterial at the phylum level. This study provides an insight into the mechanisms associated with molecular and microbiota response and processes involved in adaptation strategies towards sulfide stress.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): A. Anju, C.K. Smitha, K. Preetha, R. Boobal, Philip Rosamma〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Antimicrobial peptides (AMP) are potential alternatives to conventional antibiotics with the prospect to treat infections caused by multidrug resistant bacteria. This is the report of the first arasin sequence from the mud crab, 〈em〉Scylla serrata〈/em〉, designated as Ss-arasin. The complete cDNA sequences of the open reading frame (ORF) is comprised of 198 bp encoding 65 amino acid with a predicted molecular weight of 7 kDa and a predicted isoelectric point of 10.68. The sequence of the N-terminal 24 amino acid residues is indicative of a signal sequence directing the newly synthesize protein toward the secretory pathway. The 41-residue mature peptide is composed of two domains, an N-terminal Gly/Arg-rich domain and a C-terminal cysteine-rich domain. Challenging the mud crab with lipopolysaccharide (LPS) increased expression of Ss-arasin mRNA in haemocytes, reaching the highest level at 6 h, before dropping to basal levels at 24 h. Recombinant rSs-arasin showed antimicrobial activity against three bacterial species 〈em〉Staphylococcus aureus〈/em〉 (40 mM)〈em〉, Pseudomonas aeruginosa〈/em〉 (40 mM) and 〈em〉Escherichia coli〈/em〉 (40 mM) implying significant anti-bacterial action. In addition, recombinant rSs-arasin inhibited human cervical carcinoma (HeLa) and colon carcinoma (HT-29) cell growth. These initial findings are encouraging to further study the structure-activity relationships to optimize these biological functions for future drug development.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): QingJian Liang, MuFei Ou, YingHao Ren, ZeNa Yao, Rui Hu, JieZhen Li, Yuan Liu, Weina Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉SAHH is an enzyme, playing a significant role in the catalyzation of the S-adenosyl homocysteine (SAH) into homocysteine (Hcy) and adenosine (Ado). However, little is known information of the enzyme in crustaceans. In the present study, SAHH cDNA was cloned from 〈em〉Litopenaeus vannamei〈/em〉 (〈em〉LvSAHH〈/em〉). The full length of the 〈em〉LvSAHH〈/em〉 was found, containing a 5′ UTR of 119 bp, an ORF of 1236 bp and a 3′ UTR of 549 bp. The 〈em〉Lv〈/em〉SAHH gene encoded a polypeptide of 411 amino acids with an estimated molecular mass of 45.55 kD and a predicted isoelectronic point (pI) of 5.63. Comparison of the deduced amino acid sequence showed that 〈em〉Lv〈/em〉SAHH has high identity (70 %–82%) with other known species. qRT-PCR analysis revealed that 〈em〉LvSAHH〈/em〉 mRNA was broadly expressed in all of the examined tissues, while the highest expression level was observed in muscle, followed by the expression in stomach, gill, pleopod, hepatopancreas, heart, eye and intestine. Subcellular localization analysis revealed that 〈em〉Lv〈/em〉SAHH was predominantly localized in the cytoplasm and nucleus. 〈em〉LvSAHH〈/em〉 mRNA expression levels in hepatopancreas and gill were significantly up-regulated from 6 to 48 h after 〈em〉V. alginolyticus〈/em〉 injection and reached the highest level (15-fold and 8-fold, 〈em〉p〈/em〉 〈 0.01) at 24 h, respectively. Additionally, the Toll-like receptors (〈em〉TLR〈/em〉) and interleukins-16 (〈em〉IL-16〈/em〉) were detected in hepatopancreas and gill of 〈em〉LvSAHH〈/em〉-knockdown SAHH. 〈em〉LvRack1〈/em〉, 〈em〉LvToll1〈/em〉, 〈em〉LvToll2〈/em〉, 〈em〉LvToll3〈/em〉 and 〈em〉LvIL-16〈/em〉 transcripts were decreased significantly in 〈em〉LvSAHH〈/em〉-knockdown shrimp at 24 h post 〈em〉V. alginolyticus〈/em〉 stimulation in hepatopancreas and gill. But 〈em〉LvToll3〈/em〉 was no significant difference in gill. In summary, these results indicated that 〈em〉Lv〈/em〉SAHH may play a regulatory role in the invertebrate innate immune defense by regulating 〈em〉TLR〈/em〉 and 〈em〉IL-16〈/em〉 expression.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Yeyu Chen, Quan Gong, Mingjiang Song, Jiansheng Lai, Jiahua Sun, Ya Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Antimicrobial peptides (AMPs) play essential roles in the innate immune system to protect against a wide variety of pathogens in aquatic environments. In this study, three very important AMPs, cathelicidin, hepcidin and defensin, were identified in the critically endangered 〈em〉Acipenser dabryanus〈/em〉. The full-length cDNA sequences of these three AMPs were identified from transcriptome sequencing and the rapid amplification of cDNA ends (RACE) technique. Phylogenetic analysis showed that cathelicidin formed a clade with the other members of the cathelicidin family, and similar results were obtained for hepcidin. The 〈em〉A. dabryanus〈/em〉 β-defensin belonged to the fish class 2 β-defensins. A tissue distribution study showed that the three AMP transcripts could be detected constitutively in various tissues. The highest expression levels of cathelicidin and hepcidin were found in the liver, while defensin was primarily expressed in the skin. Bacterial challenge 〈em〉in vivo〈/em〉 revealed significant changes in the gene expression of the three AMPs at both mucosal sites and systemic sites. Striking upregulation of cathelicidin and hepcidin was observed in the skin at 12 h post-challenge, with increases of more than 7000-fold and 1000-fold, respectively, compared to the control, and the expression of defensin mRNA was remarkably elevated in the hindgut (by 230-fold at 6 h post-challenge). Moreover, according to the expression profiles of the AMPs post-challenge, we found that the mucosal immune response occurred earlier than the systemic immune response following bacterial infection. Our results suggest that these three novel AMPs may play important roles in the innate immune system of 〈em〉A. dabryanus〈/em〉 to protect against invading pathogens, especially during the mucosal immune response.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Ronald Lulijwa, Andrea C. Alfaro, Fabrice Merien, Mark Burdass, Tim Young, Jill Meyer, Thao V. Nguyen, Caroline Trembath〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉New Zealand Chinook salmon (〈em〉Oncorhynchus tshawytscha〈/em〉) industry has great potential for growth and expansion. While production is relatively free of health problems, there is limited literature on haematology, and immunological tools to safeguard against possible future health threats. The current study aim was to characterise New Zealand farmed 〈em〉O. tshawytscha〈/em〉 peripheral blood cellular composition, develop a micro-volume method to isolate peripheral blood mononuclear cells (PBMCs) and validate a microcapillary flow cytometry assay kit for PBMC cell count and viability assessment. We used light microscopy to characterise peripheral blood and PBMC cellular composition in combination with a flow cytometer Sysmex XT 2000i Haematology Analyser. ImageJ version 1.52 was used for cell size characterisation of freshly stained blood. The stability of PBMCs stained with the Muse〈sup〉®〈/sup〉 Cell Count and Viability Assay Kit and the Trypan blue assay stains were studied at 4 °C and 21 °C for 60 min; while the Muse〈sup〉®〈/sup〉 Cell Count and Viability Assay Kit was validated against the Trypan blue assay haemocytometer chamber to assess PBMC count and viability. Findings showed that 〈em〉O. tshawytscha〈/em〉 smolt yearlings had total blood cell counts in the range of 1.9–2.7 × 10〈sup〉6〈/sup〉 μL〈sup〉−1〈/sup〉. Differential cell counts revealed five cell types, comprising 97.18% erythrocytes, 2.03% lymphocytes, 0.67% thrombocytes, 0.09% monocytes, and unquantifiable neutrophils. Using micro-volumes of blood and Lymphoprep™, we successfully isolated fish PBMCs. Significantly, stained PBMCs remained stable for up to 45 min at 4 °C and 21 °C; while validation of the Muse〈sup〉®〈/sup〉 protocol showed that this microfluidic instrument delivered more accurate and precise viability results than the haemocytometer. The Muse〈sup〉®〈/sup〉 protocol is rapid, easy to use, has quick calibration steps, and is suitable for field use to facilitate onsite sample processing. These findings pave the way for future assessments of fish health and 〈em〉in vitro〈/em〉 immunological studies in 〈em〉O. tshawytscha〈/em〉.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S1050464819301330-fx1.jpg" width="461" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): C. Laltlanmawia, Ratan Kumar Saha, Himadri Saha, Pradyut Biswas〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The present study was conducted to elucidate the possible effect of a combination of 〈em〉Withania somnifera〈/em〉 (Ashwagandha) root extract and dietary 〈span〉l〈/span〉-ascorbic acid (vitamin C) on the immune responses and survivability in 〈em〉Labeo rohita〈/em〉 fingerlings against low pH and waterborne iron toxicity. Four diets were prepared containing 0% (control), 0.01% (T1), 0.1% (T2) and 1.0% (T3) of a combination of 〈em〉Withania somnifera〈/em〉 (Ashwagandha) root extract and dietary 〈span〉l〈/span〉-ascorbic acid (vitamin C) at the rate of 1:1 (w/w). Fishes were fed for 60 days to find out the best dose and duration of feeding by examining different haematological (haemoglobin, haematocrit, total RBC count, total WBC count, MCV, MCH and MCHC), immunological (NBT, lysozyme and total immunoglobulin activity) and biochemical (glucose, total protein, ALP, SGPT and SGOT level) parameters on 7, 15, 30 and 60 days of feeding. The best response was observed in fish fed with diet T3 within 15 days. The result showed a significant (p 〈 0.05) increase in haemoglobin, haematocrit, total RBC count, total WBC count, NBT, lysozyme activity, total immunoglobulin and total protein whereas a significant (p 〈 0.05) decrease in glucose, ALP, SGPT and SGOT level compared to control. Challenged study against waterborne iron (6.02 mg l〈sup〉−1〈/sup〉) and low pH (4.2–4.5) after feeding the fish for 15 days with diet T3 showed significantly (P 〈 0.05) higher protection than control. The study concludes that inclusion of 1.0% 〈em〉Withania somnifera〈/em〉 (Ashwagandha) root extract and dietary 〈span〉l〈/span〉-ascorbic acid (vitamin C) combination in diet have a stimulatory effect on immune response and reduces the effect of multiple stresses (i.e., low pH and waterborne iron toxicity) in 〈em〉L. rohita〈/em〉 fingerlings.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Konda Subramanian Shalini, Omita Yengkhom, Parasuraman Aiya Subramani, R. Dinakaran Michael〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The polysaccharide fraction (PF) isolated from the hemiparasitic mistletoe, 〈em〉Dendrophthoe falcata〈/em〉 (L.f.) Ettingsh (DF) leaves was tested for its immunostimulatory properties in 〈em〉Oreochromis niloticus〈/em〉 (Linn.). Different groups of experimental fish were fed for 1, 2 or 3 weeks with three different doses [low (0.01%), mid (0.1%), or high (1%)] of 〈em〉D. falcata〈/em〉 polysaccharide fraction (DFPF) - supplemented diet. After every feeding regimen, the fish were assessed for non-specific immunological parameters, immune related gene expression and disease protection. The DFPF treated groups showed significant (P 〈 0.05) enhancement of non-specific immune parameters. Significant (P 〈 0.05) upregulation of lysozyme and TNF-α gene expression was observed in DFPF treated groups. In pathogen challenge studies using 〈em〉Aeromonas hydrophila〈/em〉, the DFPF treated groups displayed significant (P 〈 0.05) decrease in percentage mortality and the consequent increase in relative percent survival (RPS). Supplementation of 1% DFPF in the feed for a week conferred the maximum protection against the virulent pathogen challenge, recording a RPS of 100. These results suggest that DFPF has the potential to be used as an immunostimulating feed additive in finfish aquaculture.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): G.I. Godahewa, N.C.N. Perera, Hyukjae Kwon, Jehee Lee〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The fourth member of the typical 2-cysteine peroxiredoxin (Prx4) is a well-known antioxidant enzyme, which reduces different peroxides in their catalytic process. The present study reports the identification of the rockfish 〈em〉Sebastes schlegelii〈/em〉 Prx4 (SsPrx4) at a genomic level, as well as the characterization of its structural and functional features. SsPrx4 harbors a complete ORF of 786 bp encoding a polypeptide (29 kDa) of 262 amino acids (aa) with an isoelectric point of 6.2. Thioredoxin 2 domain was prominent in the SsPrx4 sequence, which has a signal peptide (31 bp) at the N-terminus. Hence, the SsPrx4 may be functionally active in the cytoplasm of rockfish cells. Moreover, two VCP motifs and three catalytic triad residues (〈sup〉112〈/sup〉T, 〈sup〉115〈/sup〉C, 〈sup〉191〈/sup〉R) were identified in the SsPrx4 protein sequence. A peroxidatic cysteine (〈sup〉115〈/sup〉C〈sub〉P〈/sub〉) and resolving cysteines (〈sup〉236〈/sup〉C〈sub〉R〈/sub〉) were detected at the VCP motifs. The rockfish Prx4 genome consists of seven exons, which are similar to the architecture of other Prx4 orthologs. The deduced amino acid sequence of SsPrx4 shares a relatively high amino acid sequence identity (91.6%) and close evolutionary relationship with 〈em〉Miichthys miiuy〈/em〉 and 〈em〉Stegastes partitus〈/em〉 Prx4. The potential for scavenging extracellular H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉 was evidenced by the purified recombinant SsPrx4 protein (rSsPrx4) 〈em〉in vitro〈/em〉 system. Moreover, rSsPrx4 may protect the plasmid DNA in a metal-catalyzed oxidation system and catalyze the reduction of an insulin disulfide bond. Quantitative real-time PCR revealed that SsPrx4 mRNA was ubiquitously expressed in fourteen different tissues, with the highest expression observed in the liver followed by the ovary, and kidney tissues. Transcriptional modulations were observed in liver and spleen tissues of rockfish after injecting them with bacterial stimuli, including 〈em〉Streptococcus iniae〈/em〉, LPS, and a viral mimic of poly I:C. Together, the results suggest that SsPrx4 may play an important role in both the antioxidant and innate immune defense of black rockfish. These findings provide structural and functional insights into the SsPrx4 of the teleost.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Bartolomeo Gorgoglione, Nick G.H. Taylor, Jason W. Holland, Stephen W. Feist, Christopher J. Secombes〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Simultaneous and sequential infections often occur in wild and farming environments. Despite growing awareness, co-infection studies are still very limited, mainly to a few well-established human models. European salmonids are susceptible to both Proliferative Kidney Disease (PKD), an endemic emergent disease caused by the myxozoan parasite 〈em〉Tetracapsuloides bryosalmonae〈/em〉, and Viral Haemorrhagic Septicaemia (VHS), an OIE notifiable listed disease caused by the 〈em〉Piscine Novirhabdovirus〈/em〉. No information is available as to how their immune system reacts when interacting with heterogeneous infections. A chronic (PKD) + acute (VHS) sequential co-infection model was established to assess if the responses elicited in co-infected fish are modulated, when compared to fish with single infections. Macro- and microscopic lesions were assessed after the challenge, and infection status confirmed by RT-qPCR analysis, enabling the identification of singly-infected and co-infected fish. A typical histophlogosis associated with histozoic extrasporogonic 〈em〉T. bryosalmonae〈/em〉 was detected together with acute inflammation, haemorrhaging and necrosis due to the viral infection. The host immune response was measured in terms of key marker genes expression in kidney tissues. During 〈em〉T. bryosalmonae〈/em〉/VHSV-Ia co-infection, modulation of pro-inflammatory and antimicrobial peptide genes was strongly influenced by the viral infection, with a protracted inflammatory status, perhaps representing a negative side effect in these fish. Earlier activation of the cellular and humoral responses was detected in co-infected fish, with a more pronounced upregulation of Th1 and antiviral marker genes. These results reveal that some brown trout immune responses are enhanced or prolonged during PKD/VHS co-infection, relative to single infection.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Xiudan Wang, Xiaoli Zhao, Chunyu Yan, Zhihao Jia, Zhao Lv, Cuiping Ma, Mengqiang Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉A variety of combinations of leucine-rich repeat (LRR) and immunoglobulin-like (Ig) domains have been found and discovered in invertebrates and vertebrates, but the functions remain largely unexplored. In the present study, a novel LRR and Ig domain-containing protein (LRRIG), CgLRRIG-3, was identified and characterized from oyster 〈em〉Crassostrea gigas〈/em〉. It contained two typical LRR motifs, a LRRNT motif and an Ig domain and PSI-BALST and phylogeny analysis revealed that the sequence of CgLRRIG-3 was most related with leucine-rich repeat neuronal 1 proteins from vertebrate. Its mRNA transcripts were constitutively expressed in muscle, gill, hepatopancreas, mantle, gonad and hemocytes with the highest level in hepatopancreas. The mRNA expression level of CgLRRIG-3 in hemocytes could respond to the stimulations of variety PAMPs including lipopolysaccharide (LPS), peptidoglycan (PGN), glucan (GLU) and polyinosinic-polycytidylic acid (poly I:C). The recombinant proteins exhibited a wide PAMP binding repertoire to four typical PAMPs and could significantly induce the expression of CgTNF-1 and CgIL17-5 as well as increase phagocytosis in primary cultured oyster hemocytes. In hepatopancreas, CgLRRIG-3 was mainly distributed in the basolateral membrane of digestive tubule and the hemocoel sinusoid between the digestive tubules. And in hemocytes, the positive signal was mainly distributed in a special group of granulocytes. These results collectively indicated that CgLRRIG-3 could not only function as an immune effector.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Dinglong Yang, Yijing Han, Lizhu Chen, Yongliang Liu, Ruiwen Cao, Qing Wang, Jianmin Zhao〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In the present study, a scavenger receptor class B type I (designed as RpSR-BI) was cloned and characterized from manila clam 〈em〉Ruditapes philippinarum〈/em〉. The full-length cDNA of RpSR-BI was of 2000 bp, containing an open reading frame (ORF) of 1515 bp. Multiple alignments and phylogenetic analysis strongly suggested that RpSR-BI was a member of the scavenger receptors family. The mRNA transcript of RpSR-BI was constitutively expressed in all tested tissues, and mainly expressed in hepatopancreas and hemocytes. Generally, 〈em〉Vibrio anguillarum〈/em〉 or 〈em〉Micrococcus luteus〈/em〉 challenge induced the expression of RpSR-BI transcripts in hemocytes of manila clams. Recombinant protein of RpSR-BI (rRpSR-BI) could bind lipopolysaccharides, peptidoglycan and glucan, but not chitin 〈em〉in vitro〈/em〉. Coinciding with the PAMPs binding assay, a broad agglutination spectrum was displayed by rRpSR-BI including Gram-positive bacteria and Gram-negative bacteria. Moreover, rRpSR-BI could enhance the phagocytosis and chemotaxis of hemocytes. These results showed that RpSR-BI functioned as a pattern recognition receptor (PRR) with distinct recognition spectrum, and also as an opsonin involved in the innate immune response of 〈em〉R. philippinarum〈/em〉.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Xinzhan Meng, Yubang Shen, Shentong Wang, Xiaoyan Xu, Yunfei Dang, Meng Zhang, Lisen Li, Jiahua Zhang, Rongquan Wang, Jiale Li〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Complement is traditionally recognized as part of the innate immune system, defending the host against the invasion of foreign pathogens. In complement system, C3 (complement component 3) is a central component. Therefore, research into C3 can help us better understand the functions of fish complement system. In this study, we detected the grass carp C3 (〈em〉gcC3〈/em〉) mRNA expression in all sample tissues from healthy grass carp, which was highest in the liver, followed by the heart and the spleen, and lowest in the muscle, head kidney, trunk kidney, blood and intestine. After infection with 〈em〉Aeromonas hydrophila〈/em〉, 〈em〉gcC3〈/em〉 mRNA expression levels were significantly upregulated in the gill, liver, spleen, intestine, trunk kidney and head kidney. Interestingly, C3 protein levels were downregulated and subsequently upregulated in the liver and serum. Histologically, C3 protein at 24 h pi was over expressed in necrotic liver sites, and the liver index (LI) at this point was significantly higher than that of the control. These findings are indicated that C3 plays an important role in the immune response of grass carp after 〈em〉A. hydrophila〈/em〉 infection, and C3 protein may play an assistant role in repairing liver tissues from 〈em〉A. hydrophila〈/em〉 injury.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Hyun Ju Lim, Najib Abdellaoui, Ki Hong Kim〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Rhabdoviral G protein-based DNA vaccines have been recognized as a useful way to protect cultured fish from rhabdoviral diseases. In Korea, viral hemorrhagic septicemia virus (VHSV) genotype IVa has been the primary culprit of high mortalities of cultured olive flounder (〈em〉Paralichthys olivaceus〈/em〉). In this study, we inserted a miR-155-expressing cassette into the VHSV's G protein-based DNA vaccine, and analyzed the effects of miR-155 on the antiviral activity and on the vaccine efficacy in olive flounder. Olive flounder fingerlings were intramuscularly (i.m.) immunized with 10 μg/fish (1st experiment) or 1 μg/fish (2nd experiment) of DNA vaccine plasmids. However, there were no significant differences in mortalities and serum neutralization titers between fish immunized with 1 μg and 10 μg plasmids/fish, suggesting that i.m. injection with 1 μg plasmids/fish would be enough to induce effective adaptive immune responses in olive flounder fingerlings. In survival rates, as fish immunized with just G protein expressing plasmids showed no or too low mortalities, the adjuvant effect of miR-155 was not discernible. Also, in the serum neutralization activities, although G gene or G gene plus miR-155 expressing DNA vaccines induced significantly higher activities than control vaccines (PBS and vacant vector), no significant differences were found between G gene alone and G gene plus miR-155 expressing DNA vaccines. In the serum virucidal activity, fish immunized with G gene plus miR-155 expressing DNA vaccine showed significantly higher activity against hirame rhabdovirus (HIRRV) at 3 days post-immunization (d.p.i.) compared to other groups, suggesting that miR-155 produced from the vector can enhance innate immune responses in olive flounder. The significantly enhanced serum virucidal activities against VHSV especially at 28 d.p.i. in the groups immunized with G gene alone and G gene plus miR-155 expressing DNA vaccines reflect the increased antibodies against G protein, which could activate the classical complement pathway and subsequent viral inactivation. As the available information on the DNA vaccines in olive flounder is not sufficient, more diverse researches on the protective efficacy of DNA vaccines are needed to make more practical use of DNA vaccines in olive flounder farms.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Qing Wang, Min Yang, Chen Li, Shaowen Wang, Yuxin Wang, Fangmei Lin, Leyun Zheng, Yepin Yu, Qiwei Qin〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Chemokine receptors are a superfamily of seven-transmembrane domain G-coupled receptors and have important roles in immune surveillance, inflammation, and development. In previous studies, a series of CXCRs in grouper (〈em〉Epinephelus coioides〈/em〉) was identified; however, the function of CXCR in viral infection has not been studied. To better understand the effect of the CXCR family on the fish immune response, full-length CXCR1a was cloned, and its immune response to Singapore grouper iridovirus (SGIV) was investigated. Grouper CXCR1a shared a seven-transmembrane (7-TM) region and a G protein-coupled receptor (GPCR) family 1 that contained a triaa stretch (DRY motif). Phylogenetic analysis indicated that CXCR1a showed the nearest relationship to 〈em〉Takifugu rubripes〈/em〉, followed by other fish, bird and mammal species. Fluorescence microscopy revealed that CXCR1a was expressed predominantly in the cytoplasm. Overexpression of CXCR1a in grouper cells significantly inhibited the replication of SGIV, demonstrating that CXCR1a delayed the occurrence of cytopathic effects (CPE) induced by SGIV infection and inhibited viral gene transcription. Furthermore, our results also showed that CXCR1a overexpression significantly increased the expression of interferon-related cytokines and activated ISRE and IFN promoter activities. Taken together, the results demonstrated that CXCR1a might have an antiviral function against SGIV infection.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Qiang Wang, Jing Li, Hongmei Guo〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The 〈em〉Solen strictus〈/em〉 (Gould, 1861) has been recognized as an important marine economic bivalve in Eastern and Southeast Asia. To gain a better understanding of the 〈em〉S. strictus〈/em〉 immune system and its related genes in response to bacterial infections, we performed a comparative gene transcription analysis from 〈em〉S. strictus〈/em〉 with 〈em〉Vibrio anguillarum〈/em〉 through RNA-Seq technology, meanwhile the differentially expressed genes (DEGs) were investigated. After assembly, a total of 195,774 transcripts with an average length of 996 bp were obtained. Total 153,038 unigenes were annotated in the nr, Swiss-Prot, KEGG, COG, KOG, GO and Pfam databases, and 56,597 unigenes (36.98%) were annotated in at least one database. After bacterial challenge, there were 1588 significant differentially expressed genes (DEGs) between the challenged and control groups, including 999 up-regulated and 589 down-regulated genes. All the DEGs were classified into three gene ontology categories, and allocated to 225 KEGG pathways. Immune-related genes were detected from immune system pathways among the top 20 enriched pathways, such as Toll-like receptor signaling, RIG-I-like receptor signaling and NOD-like receptor signaling pathway. In addition, 56,079 potential simple sequence repeats (SSRs) and 1,031,521 candidate single nucleotide polymorphisms (SNPs) were detected and identified in the 〈em〉S. strictus〈/em〉 transcriptome. Results of the present study will provide valuable theoretical resources for future genetic and genomic research on 〈em〉S. strictus〈/em〉. The research results will be helpful for improving the efficiency and quality of artificial breeding, establishing genetic linkage map, and enhancing health management for this species.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Se-Young Cho, Rachael A. Protzman, Yeong O. Kim, Bipin Vaidya, Myung-Joo Oh, Joseph Kwon, Duwoon Kim〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Seasonal temperature has a major influence on the infectivity of pathogens and the host immune system. Viral hemorrhagic septicemia virus (VHSV) is one such pathogen that only causes the mortality of fish at low temperatures. This study aims to discover the host defense mechanism and pathway for resistance to VHSV at higher temperatures. We first observed the VHSV infection patterns at low and higher temperatures in fathead minnow (FHM) cells (20 °C and 28 °C) and zebrafish (15 °C and 25 °C). In comparison to the 20 °C infection, FHM cells infected at 28 °C showed decreased apoptosis, increased cell viability, and reduced VHSV N gene expression. In zebrafish, infection at 25 °C caused no mortality and significantly reduced the N gene copy number in comparison to infection at 15 °C. To explore the antiviral infection mechanisms induced by high temperature 〈em〉in vitro〈/em〉 and 〈em〉in vivo〈/em〉, the changes in the proteomic profile were measured through UPLC-MS〈sup〉E〈/sup〉 analysis. ACADL, PTPN6, TLR1, F7, A2M, and GLI2 were selected as high temperature-specific biomarkers in the FHM cell proteome; and MYH9, HPX, ANTXR1, APOA1, HBZ, and MYH7 were selected in zebrafish. Increased immune response, anticoagulation effects, and the formation of lymphocytes from hematopoietic stem cells were analyzed as functions that were commonly induced by high temperature 〈em〉in vitro〈/em〉 and 〈em〉in vivo〈/em〉. Among these biomarkers, GLI2 was predicted as an upstream regulator. When treated with GANT58, a GLI-specific inhibitor, cell viability was further reduced due to GLI2 inhibition during VHSV infection at varying temperatures in FHM cells, and the mortality in zebrafish was induced earlier at the low temperature. Overall, this study discovered a new mechanism for VHSV infection 〈em〉in vitro〈/em〉 and 〈em〉in vivo〈/em〉 that is regulated by GLI2 protein.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S1050464819301111-fx1.jpg" width="376" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Shijun Chen, Zhenxiao Zhuang, Peng Yin, Xu Chen, Yanmei Zhang, Lixia Tian, Jin Niu, Yongjian Liu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉A 58-day feeding trial was conducted to evaluate the effects of dietary 〈em〉myo〈/em〉-inositol (MI) supplementation on growth performance, haematological parameters, hepatopancreas histopathology and antioxidant status of 〈em〉Litopenaeus vannamei〈/em〉 fed with oxidized fish oil (OFO). Control diet contained fresh fish oil (FFO) without MI supplementation. The other four diets contained two oxidation levels of OFO (peroxide value: 133.2 and 268.7 meq kg〈sup〉−1〈/sup〉) with or without 200 mg MI kg〈sup〉−1〈/sup〉 diets (MI0+L, MI0+H, MI200 + L and MI200 + H). Results showed that OFO-supplemented groups (without MI supplementation) showed better growth performance and lower whole-body inositol content when opposed to control group. MI supplementation significantly improved whole-body inositol content in high-oxidized fish oil (HOFO) groups, and also reduced whole-body lipid in low-oxidized fish oil (LOFO) groups. Moreover, Supplementation of OFO and MI markedly hit the fatty acid profile of muscle. HOFO caused severe histopathological changes in hepatopancreas of shrimp, which slightly alleviated by MI supplementation. MI supplementation also grew the total protein (TP) content and alkaline phosphatase (AKP) activity and decreased the activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) of serum in OFO-supplemented groups. Ingestion of OFO increased levels of lipid peroxidation and protein oxidation in serum or hepatopancreas, which partly ameliorated by MI supplementation. Activities of antioxidant enzymes exhibited different expression patterns because of OFO and MI. In addition, HOFO markedly increased mRNA expression levels of antioxidant genes including ferritin (FT), thioredoxin (Trx), GPX, glutathione S-transferase (GST) and catalase (CAT) and decreased peroxiredoxin (Prx) expression, in which expression of GPX and Prx were increased owing to MI supplementation. Therefore, it suggested that dietary OFO stimulated growth performance, but also induced oxidative stress and caused impairment to hepatopancreas in 〈em〉L. vannamei〈/em〉. The negative impact brought about by OFO was partially mitigated by dietary MI supplementation.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Yanfei Cao, Shuai Yang, Chuzhang Feng, Waner Zhan, Zhe Zheng, Qingheng Wang, Yuewen Deng, Yu Jiao, Xiaodong Du〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Interleukin-17 (IL-17) is a proinflammatory cytokine that plays an important role in immune responses. In this study, we identified 57 IL-17 genes from the genomes of six marine invertebrates, including 〈em〉Pinctada fucata martensii〈/em〉, 〈em〉Crassostrea gigas〈/em〉, 〈em〉Lottia gigantea〈/em〉, 〈em〉Capitella teleta〈/em〉, 〈em〉Mizuhopecten yessoensis〈/em〉, and 〈em〉Mytilus galloprovincialis〈/em〉. Phylogenetic analysis showed that all invertebrate IL-17 genes were clustered into one group, implying that invertebrate IL-17 evolved from one common ancestral gene. From the extron–intron analysis, we found many intronless IL-17 genes in mollusks, which may be caused by retroposition. Tissue and development transcriptomic analysis showed that the expression of PmIL-17 was tissue and developmental stage-specific. Moreover, we cloned the full length of the IL-17-2 gene from 〈em〉P. f. martensii〈/em〉 (PmIL-17-2) and explored its function in the immune response. The full-length cDNA of PmIL-17-2 is 719 bp, containing an open reading frame of 564 bp, a 5′ -untranslated region (UTR) of 31 bp, and a 3′ -UTR of 124 bp with a 30 bp poly (A) tail. PmIL-17-2 had a strong response to lipopolysaccharide (LPS), indicating that the PmIL-17-2 participates in innate immune responses. In situ hybridization of hemocytes showed that PmIL-17-2 was mainly produced by granulosa cells, and the number of the stained granulosa increased after LPS stimulation. These results lay the foundation for the research of IL-17 family in marine invertebrates.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Mariana Rangel Pilotto, Samuel Milanez, Renato Teixeira Moreira, Rafael Diego Rosa, Luciane Maria Perazzolo〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Herein, we evaluated the immunomodulatory and the antiviral protective properties of a cyanobacteria-enriched diet on the immune responses of the Pacific white shrimp 〈em〉Litopenaeus vannamei〈/em〉 challenged with the White spot syndrome virus (WSSV). Shrimp were fed with an 〈em〉Arthrospira platensis〈/em〉 supplemented feed during 20 days, and its effects were examined by evaluating well-known standardized shrimp immune parameters (total hemocyte counts, total protein concentration, phenoloxidase activity, and serum agglutination titer). Additionally, we assessed the expression of crucial genes involved in both hemolymph- and gut-based immunities related to the shrimp capacity to circumvent viral and microbial infections. Dietary supplementation improved shrimp survival rates after challenge with a median lethal dose of WSSV. From all immune parameters tested, only the serum agglutination titer was higher in treated animals. On the other hand, the expression of some representative marker genes from different immune response pathways was only modulated in the midgut and not in the circulating hemocytes, suggesting that this feed supplementation can be used as an attractive strategy to enhance immunity in shrimp gut. Altogether, our results evidence the immunomodulatory properties of 〈em〉A. platensis〈/em〉 supplemented feed in shrimp humoral and intestinal defenses and highlight the potential use of cyanobacteria-based immunostimulants in shrimp farming for protection against infectious diseases.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Xiaoxue Yin, Yanjian Yang, Kailiang Han, Liting Wu, Hairong Wu, Xia Bian, Xiufang Wei, Zheng Guo, Liangliang Mu, Jianmin Ye〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Transferrin (TF), an iron-binding multifunctional protein, could participate in the iron-withholding strategy, an effective antimicrobial defense mechanism in innate immunity, and is involved in host defense against pathogenic infection. In this study, a TF homologue (OnTF) was purified from serum of Nile tilapia (〈em〉Oreochromis niloticus〈/em〉) through a two-step affinity chromatography, and characterized its antibacterial function and the role in inflammatory response. The identification by mass spectrometry showed that peptide sequence of the purified OnTF was highly consistent with its amino acids sequence, containing two conserved iron binding lobes: N-lobe and C-lobe. The native OnTF was able to bond iron ions, and possessed capability to inhibit the growth of both bacterial pathogens (〈em〉Streptococcus agalactiae〈/em〉 and 〈em〉Aeromonas hydrophila〈/em〉) 〈em〉in vitro〈/em〉. Upon infections of 〈em〉S. agalactiae〈/em〉 and 〈em〉A. hydrophila〈/em〉, the expression of OnTF protein was significantly up-regulated 〈em〉in vivo〈/em〉 and 〈em〉in vitro〈/em〉. In addition, the OnTF participated in the regulation of inflammation, migration, and enhancement of phagocytosis and respiratory burst activity in head kidney macrophages/monocytes. Taken together, the results of this study indicated that OnTF is likely to involve in innate immunity to play a role in host defense against bacterial infection in Nile tilapia.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Moonika H. Marana, Dagoberto Sepúlveda, Defang Chen, Azmi Al-Jubury, Rzgar M. Jaafar, Per W. Kania, Niels Henrik Henriksen, Bjørn Krossøy, Inger Dalsgaard, Niels Lorenzen, Kurt Buchmann〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉 〈p〉Mariculture in Denmark is based on production of rainbow trout grown two years in fresh water followed by one growth season in sea cages. Although the majority of rainbow trout are vaccinated against the most serious bacterial pathogens – 〈em〉Aeromonas salmonicida〈/em〉 subsp. 〈em〉salmonicida〈/em〉, 〈em〉Vibrio anguillarum〈/em〉 and 〈em〉Yersinia ruckeri〈/em〉, by the use of commercially available vaccines, disease outbreaks requiring treatment with antibiotics still occur.〈/p〉 〈p〉The present study tested the potential of a new experimental multicomponent vaccine that is based on local bacterial strains, isolated from rainbow trout in Danish waters, and thus custom-designed for Danish rainbow trout mariculture. The vaccination with the multicomponent vaccine resulted in protection against three relevant bacterial diseases (yersiniosis, furunculosis, vibriosis) under experimental conditions. We showed that i.p. injection of the vaccine induced specific antibody responses in trout against the different bacterial antigens and regulated expression of genes encoding SAA, C3, IL-1β, IL-6, IL-8, IgD and MHCII.〈/p〉 〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Euan R.O. Allan, Liping Yang, Jacob A. Tennessen, Michael S. Blouin〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Freshwater snails are obligate intermediate hosts for numerous parasitic trematodes, most notably schistosomes. Schistosomiasis is a devastating human and veterinary illness, which is primarily controlled by limiting the transmission of these parasites from their intermediate snail hosts. Understanding how this transmission occurs, as well as the basic immunobiology of these snails may be important for controlling this disease in the future. Allelic variation in the Guadeloupe resistance complex (GRC) of 〈em〉Biomphalaria glabrata〈/em〉 partially determines their susceptibility to parasitic infection, and can influence the microbiome diversity and microbial defenses in the hemolymph of these snails. In the present study, we examine the most abundant proteins present in the hemolymph of snails that are resistant or susceptible to schistosomes, as determined by their GRC genotype. Using proteomic analysis, we found that snails with different GRC genotypes have differentially abundant hemolymph proteins that are not explained by differences in transcription. There are 13 revealed hemolymph proteins that differ significantly between resistant and susceptible genotypes, nearly 40% of which are involved in immune responses. These findings build on the mounting evidence that genes in the GRC region have multiple physiological roles, and likely contribute more extensively to the general immune response than previously believed. These data also raise the intriguing possibility that the GRC region controls resistance to schistosomes, not directly, but indirectly via its effects on the snail's proteome and potentially its microbiome.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: Available online 6 March 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology〈/p〉 〈p〉Author(s): Hourong Liu, Yuan Liu, Chengwen Song, Junhao Ning, Zhaoxia Cui〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Clip domain serine proteases (cSPs), a family of multifunctional proteins, play a crucial role in innate immune system. Here, we report the functional characterization of two clip domain serine proteases (PtcSP1 and PtcSP3) from the swimming crab 〈em〉Portunus trituberculatus〈/em〉. The recombinant N-terminal clip domains and the C-terminal SP-like domains of PtcSP1 and PtcSP3 were expressed in 〈em〉Escherichia coli〈/em〉 system, and assayed for various biological functions: protease activity, antimicrobial activity, bacterial clearance and microbial-binding activity. The recombinant SP-like domains of PtcSP1 and PtcSP3 exhibited trypsin-like protease activity, while their recombinant clip domains showed strong antibacterial activity and could bind to bacteria and yeast, suggesting the potential roles of PtcSP1 and PtcSP3 in immune defense and pattern recognition. Unlike PtcSP3, PtcSP1 revealed the opsonic activity as shown by a higher bacterial clearance rate of 〈em〉Vibrio alginolyticus〈/em〉 coated with the combination of the recombinant clip domain and SP-like domain of PtcSP1 as compared with 〈em〉V. alginolyticus〈/em〉 only. Knockdown of PtcSP1 or PtcSP3 by RNA interference resulted in a significant decrease of total phenoloxidase (PO) activity in crab, suggesting that PtcSP1 and PtcSP3 are involved in the proPO system. In addition, suppression of PtcSP1 or PtcSP3 changed the expression of PtALFs and complement-like components. All these findings suggest that PtcSP1 and PtcSP3 are multifunctional immune molecules and perform different protective functions in crab defense.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Cai-xia Lei, Jing-jing Tian, Wen Zhang, Yu-ping Li, Hong Ji, Er-meng Yu, Wang-bao Gong, Zhi-fei Li, Kai Zhang, Guang-jun Wang, De-guang Yu, Jun Xie〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Lipid droplets (LDs) are increasingly being recognized as important immune modulators in mammals, in additional to their function of lipid ester deposition. However, the role of LDs in fish immunity remains poorly understood. In this study, the function of LDs in the innate immune response of 〈em〉Ctenopharyngodon idella〈/em〉 kidney (CIK) cells, which are the equivalent of myeloid cells in vertebrates, was investigated. LD number and TG content significantly increased in the CIK cells following exposure to lipopolysaccharide (LPS), peptidoglycan (PGN), and polyriboinosinic-polyribocytidylic acid (Poly [I: C]) for 24 h, accompanied by increases in the relative expression of several innate immune genes. However, fatty acid compositions of the triglycerides were not changed after treatment with these three pathogenic mimics. LPS, PGN, and Poly (I: C) did not alter the relative expressions of lipogenic (〈em〉FAS〈/em〉, 〈em〉SCD〈/em〉, and 〈em〉DGAT〈/em〉) and lipid catabolic (〈em〉PPARα〈/em〉, 〈em〉ATGL〈/em〉, and 〈em〉CPT-1〈/em〉) genes. However, these treatments did increase the mRNA levels of lipid transportation genes (〈em〉FATP/CD36〈/em〉, 〈em〉ACSL1〈/em〉, and 〈em〉ACSL4〈/em〉), and also decreased the non-esterified fatty acid level in the medium. To further explore the role of LDs in the immune response, CIK cells were incubated with different concentrations (0, 100, 200, 300, 400, 500 μM) of exogenous lipid mix (LM; oleic acid [OA]:linoleic acid [LA]:linolenic acid [LNA] = 2:1:1), and were then transferred to a lipid-free medium and incubated for 24 h. LD size and number increased with the increase in lipid levels, and this was accompanied by increased expression of innate immune genes, including 〈em〉MyD88〈/em〉, 〈em〉IRF3,〈/em〉 and 〈em〉IL-1β〈/em〉, which were expressed at their highest levels in 300 μM exogenous lipid mix. Interestingly, after incubating with different fatty acids (LM, OA, LA, LNA, arachidonic acid [ARA], and docosahexaenoic acid [DHA]; 300 μM), ARA and DHA were more potent in inducing LD formation and innate immune gene expression in the CIK cells. Finally, atglistatin, an ATGL inhibitor, effectively attenuated the expression of most genes upregulated by ARA or DHA, suggesting that lipolysis may be involved in the regulation of immune genes at the transcriptional level. Overall, the findings of this study demonstrate that LDs are functional organelles that could act as modulators in the innate immune response of CIK cells. Additionally, long-chain polyunsaturated fatty acid enriched LDs play a unique role in regulating this process.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: April 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 87〈/p〉 〈p〉Author(s): Ghasem Ashouri, Nasrollah Mahboobi Soofiani, Seyed Hossein Hoseinifar, Seyed Amir Hossein Jalali, Vahid Morshedi, Hien Van Doan, Mansour Torfi Mozanzadeh〈/p〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Dinglong Yang, Yijing Han, Yongliang Liu, Ruiwen Cao, Qing Wang, Zhijun Dong, Hui Liu, Xiaoli Zhang, Qianqian Zhang, Jianmin Zhao〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Peptidoglycan recognition proteins (PGRPs) are important pattern recognition receptors in the innate immune system of invertebrates. In the study, a short PGRP (designed as RpPGRP) was identified and characterized from the manila clam 〈em〉Ruditapes philippinarum〈/em〉. The open reading frame of RpPGRP encoded a polypeptide of 249-amino acids with a calculated molecular mass of 27.2 kDa and an isoelectric point of 6.62. Multiple alignments and phylogenetic analysis strongly suggested that RpPGRP was a new member of the PGRP superfamily. In non-stimulated clams, RpPGRP exhibited different tissue expression pattern, and highly expressed in hepatopancreas and hemocytes. Expression of RpPGRP transcripts was significantly up-regulated in hemocytes of clams post 〈em〉Vibrio anguillarum〈/em〉 or 〈em〉Micrococcus luteus〈/em〉 challenge. The recombinant RpPGRP (rRpPGRP) exhibited high affinity to PGN, LPS and zymosan in a concentration-dependent manner. With a broad spectrum of bacterial binding activities, rRpPGRP exhibited strong agglutination activity to 〈em〉Escherichia coli〈/em〉, 〈em〉Vibrio splendidus〈/em〉, 〈em〉V. anguillarum〈/em〉 and 〈em〉M. luteus〈/em〉. Furthermore, rRpPGRP exhibited Zn〈sup〉2+〈/sup〉-dependent amidase activity and catalyzed the degradation of insoluble PGN. Especially, rRpPGRP exhibited significant antibacterial activity against 〈em〉E. coli〈/em〉 and 〈em〉M. luteus〈/em〉. Moreover, the biofilm formation of 〈em〉E. coli〈/em〉 could be inhibited after rRpPGRP incubation in the presence of Zn〈sup〉2+〈/sup〉. This inhibitory effect of rRpPGRP might attribute to its amide bactericidal activity. Taken together, rRpPGRP played important roles in PGRP-mediated immune defense mechanisms, especially by recognizing antigens and eliminating bacteria.〈/p〉〈/div〉 〈/div〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Mitochondria are essential components of eukaryotic cells, carrying out critical physiological processes that include energy production and calcium buffering. Consequently, mitochondrial dysfunction is associated with a range of human diseases. Fundamental to their function is the ability to transition through fission and fusion states, which is regulated by several GTPases. Here, we have developed new methods for the non-subjective quantification of mitochondrial morphology in muscle and neuronal cells of 〈em〉Caenorhabditis elegans〈/em〉. Using these techniques, we uncover surprising tissue-specific differences in mitochondrial morphology when fusion or fission proteins are absent. From ultrastructural analysis, we reveal a novel role for the fusion protein FZO-1/mitofusin 2 in regulating the structure of the inner mitochondrial membrane. Moreover, we have determined the influence of the individual mitochondrial fission (DRP-1/DRP1) and fusion (FZO-1/mitofusin 1,2; EAT-3/OPA1) proteins on animal behaviour and lifespan. We show that loss of these mitochondrial fusion or fission regulators induced age-dependent and progressive deficits in animal movement, as well as in muscle and neuronal function. Our results reveal that disruption of fusion induces more profound defects than lack of fission on animal behaviour and tissue function, and imply that while fusion is required throughout life, fission is more important later in life likely to combat ageing-associated stressors. Furthermore, our data demonstrate that mitochondrial function is not strictly dependent on morphology, with no correlation found between morphological changes and behavioural defects. Surprisingly, we find that disruption of either mitochondrial fission or fusion significantly reduces median lifespan, but maximal lifespan is unchanged, demonstrating that mitochondrial dynamics play an important role in limiting variance in longevity across isogenic populations. Overall, our study provides important new insights into the central role of mitochondrial dynamics in maintaining organismal health.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Here, we review melanocyte development and how the embryonic melanoblast, although specified to become a melanocyte, is prone to cellular plasticity and is not fully committed to the melanocyte lineage. Even fully differentiated and pigment-producing melanocytes do not always have a stable phenotype. The gradual lineage restriction of neural crest cells toward the melanocyte lineage is determined by both cell-intrinsic and extracellular signals in which differentiation and pathfinding ability reciprocally influence each other. These signals are leveraged by subtle differences in timing and axial positioning. The most extensively studied migration route is the dorsolateral path between the dermomyotome and the prospective epidermis, restricted to melanoblasts. In addition, the embryonic origin of the skin dermis through which neural crest derivatives migrate may also affect the segregation between melanogenic and neurogenic cells in embryos. It is widely accepted that, irrespective of the model organism studied, the immediate precursor of both melanoblast and neurogenic populations is a glial-melanogenic bipotent progenitor. Upon exposure to different conditions, melanoblasts may differentiate into other neural crest-derived lineages such as neuronal cells and vice versa. Key factors that regulate melanoblast migration and patterning will regulate melanocyte homeostasis during different stages of hair cycling in postnatal hair follicles.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Cellular protein quality control (PQC) plays a significant role in the maintenance of cellular homeostasis. Failure of PQC mechanism may lead to various neurodegenerative diseases due to accumulation of aberrant proteins. To avoid such fatal neuronal conditions PQC employs autophagy and ubiquitin proteasome system (UPS) to degrade misfolded proteins. Few quality control (QC) E3 ubiquitin ligases interplay an important role to specifically recognize misfolded proteins for their intracellular degradation. Leucine-rich repeat and sterile alpha motif-containing 1 (LRSAM1) is a really interesting new gene (RING) class protein that possesses E3 ubiquitin ligase activity with promising applications in PQC. LRSAM1 is also known as RING finger leucine repeat rich (RIFLE) or TSG 101-associated ligase (TAL). LRSAM1 has various cellular functions as it modulates the protein aggregation, endosomal sorting machinery and virus egress from the cells. Thus, this makes LRSAM1 interesting to study not only in protein conformational disorders such as neurodegeneration but also in immunological and other cancerous disorders. Furthermore, LRSAM1 interacts with both cellular protein degradation machineries and hence it can participate in maintenance of overall cellular proteostasis. Still, more research work on the quality control molecular functions of LRSAM1 is needed to comprehend its roles in various protein aggregatory diseases. Earlier findings suggest that in a mouse model of Charcot–Marie–Tooth (CMT) disease, lack of LRSAM1 functions sensitizes peripheral axons to degeneration. It has been observed that in CMT the patients retain dominant and recessive mutations of LRSAM1 gene, which encodes most likely a defective protein. However, still the comprehensive molecular pathomechanism of LRSAM1 in neuronal functions and neurodegenerative diseases is not known. The current article systematically represents the molecular functions, nature and detailed characterization of LRSAM1 E3 ubiquitin ligase. Here, we review emerging molecular mechanisms of LRSAM1 linked with neurobiological functions, with a clear focus on the mechanism of neurodegeneration and also on other diseases. Better understanding of LRSAM1 neurobiological and intracellular functions may contribute to develop promising novel therapeutic approaches, which can also propose new lines of molecular beneficial targets for various neurodegenerative diseases.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Huntington’s disease (HD) is a hereditary and fatal disease causing profound neurodegeneration. Deficits in cerebral energy and neurotransmitter metabolism have been suggested to play a central role in the neuronal dysfunction and death associated with HD. The branched-chain amino acids (BCAAs), leucine, isoleucine and valine, are important for cerebral nitrogen homeostasis, neurotransmitter recycling and can be utilized as energy substrates in the tricarboxylic acid (TCA) cycle. Reduced levels of BCAAs in HD have been validated by several reports. However, it is still unknown how cerebral BCAA metabolism is regulated in HD. Here we investigate the metabolism of leucine and isoleucine in the R6/2 mouse model of HD. Acutely isolated cerebral cortical and striatal slices of control and R6/2 mice were incubated in media containing 〈sup〉15〈/sup〉N- or 〈sup〉13〈/sup〉C-labeled leucine or isoleucine and slice extracts were analyzed by gas chromatography–mass spectrometry (GC–MS) to determine isotopic enrichment of derived metabolites. Elevated BCAA transamination was found from incubations with [〈sup〉15〈/sup〉N]leucine and [〈sup〉15〈/sup〉N]isoleucine, in both cerebral cortical and striatal slices of R6/2 mice compared to controls. Metabolism of [U-〈sup〉13〈/sup〉C]leucine and [U-〈sup〉13〈/sup〉C]isoleucine, entering oxidative metabolism as acetyl CoA, was maintained in R6/2 mice. However, metabolism of [U-〈sup〉13〈/sup〉C]isoleucine, entering the TCA cycle as succinyl CoA, was elevated in both cerebral cortical and striatal slices of R6/2 mice, suggesting enhanced metabolic flux via this anaplerotic pathway. To support the metabolic studies, expression of enzymes in the BCAA metabolic pathway was assessed from a proteomic resource. Several enzymes related to BCAA metabolism were found to exhibit augmented expression in the R6/2 brain, particularly related to isoleucine metabolism, suggesting an increase in the BCAA metabolic machinery. Our results show that the capacity for cerebral BCAA metabolism, predominantly of isoleucine, is amplified in the R6/2 brain and indicates that perturbations in cerebral BCAA homeostasis could have functional consequences for HD pathology.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉The transcription factor Ptf1a is a crucial helix–loop–helix (bHLH) protein selectively expressed in the pancreas, retina, spinal cord, brain, and enteric nervous system. Ptf1a is preferably assembled into a transcription trimeric complex PTF1 with an E protein and Rbpj (or Rbpjl). In pancreatic development, Ptf1a is indispensable in controlling the expansion of multipotent progenitor cells as well as the specification and maintenance of the acinar cells. In neural tissues, Ptf1a is transiently expressed in the post-mitotic cells and specifies the inhibitory neuronal cell fates, mostly mediated by downstream genes such as Tfap2a/b and Prdm13. Mutations in the coding and non-coding regulatory sequences resulting in Ptf1a gain- or loss-of-function are associated with genetic diseases such as pancreatic and cerebellar agenesis in the rodent and human. Surprisingly, Ptf1a alone is sufficient to reprogram mouse or human fibroblasts into tripotential neural stem cells. Its pleiotropic functions in many biological processes remain to be deciphered in the future.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Receptor tyrosine kinases (RTKs) play key roles in various aspects of cell biology, including cell-to-cell communication, proliferation and differentiation, survival, and tissue homeostasis, and have been implicated in various diseases including cancer and neurodevelopmental disorders. Ligand-activated RTKs recruit adapter proteins through a phosphotyrosine (p-Tyr) motif that is present on the RTK and a p-Tyr-binding domain, like the Src homology 2 (SH2) domain found in adapter proteins. Notably, numerous combinations of RTK/adapter combinations exist, making it challenging to compare receptor activities in standardised assays. In cell-based assays, a regulated adapter recruitment can be investigated using genetically encoded protein–protein interaction detection methods, such as the split TEV biosensor assay. Here, we applied the split TEV technique to robustly monitor the dynamic recruitment of both naturally occurring full-length adapters and artificial adapters, which are formed of clustered SH2 domains. The applicability of this approach was tested for RTKs from various subfamilies including the epidermal growth factor (ERBB) family, the insulin receptor (INSR) family, and the hepatocyte growth factor receptor (HGFR) family. Best signal-to-noise ratios of ligand-activated RTK receptor activation was obtained when clustered SH2 domains derived from GRB2 were used as adapters. The sensitivity and robustness of the RTK recruitment assays were validated in dose-dependent inhibition assays using the ERBB family-selective antagonists lapatinib and WZ4002. The RTK split TEV recruitment assays also qualify for high-throughput screening approaches, suggesting that the artificial adapter may be used as universal adapter in cell-based profiling assays within pharmacological intervention studies.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Embryonic stem cells (ESCs) are immortal stem cells that own multi-lineage differentiation potential. ESCs are commonly derived from the inner cell mass (ICM) of pre-implantation embryos. Due to their tremendous developmental capacity and unlimited self-renewal, ESCs have diverse biomedical applications. Different culture media have been developed to procure and maintain ESCs in a state of naïve pluripotency, and to preserve a stable genome and epigenome during serial passaging. Chromatin modifications such as DNA methylation and histone modifications along with microRNA activity and different signaling pathways dynamically contribute to the regulation of the ESC gene regulatory network (GRN). Such modifications undergo remarkable changes in different ESC media and determine the quality and developmental potential of ESCs. In this review, we discuss the current approaches for derivation and maintenance of ESCs, and examine how differences in culture media impact on the characteristics of pluripotency via modulation of GRN during the course of ICM outgrowth into ESCs. We also summarize the current hypotheses concerning the origin of ESCs and provide a perspective about the relationship of these cells to their in vivo counterparts (early embryonic cells around the time of implantation). Finally, we discuss generation of ESCs from human embryos and domesticated animals, and offer suggestions to further advance this fascinating field.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Cancers of the reproductive organs have a strong association with mitochondrial defects, and a deeper understanding of the role of this organelle in preneoplastic–neoplastic changes is important to determine the appropriate therapeutic intervention. Mitochondria are involved in events during cancer development, including metabolic and oxidative status, acquisition of metastatic potential, resistance to chemotherapy, apoptosis, and others. Because of their origin from melatonin-producing bacteria, mitochondria are speculated to produce melatonin and its derivatives at high levels; in addition, exogenously administered melatonin accumulates in the mitochondria against a concentration gradient. Melatonin is transported into tumor cell by GLUT/SLC2A and/or by the PEPT1/2 transporters, and plays beneficial roles in mitochondrial homeostasis, such as influencing oxidative phosphorylation and electron flux, ATP synthesis, bioenergetics, calcium influx, and mitochondrial permeability transition pore. Moreover, melatonin promotes mitochondrial homeostasis by regulating nuclear DNA and mtDNA transcriptional activities. This review focuses on the main functions of melatonin on mitochondrial processes, and reviews from a mechanistic standpoint, how mitochondrial crosstalk evolved in ovarian, endometrial, cervical, breast, and prostate cancers relative to melatonin’s known actions. We put emphasis on signaling pathways whereby melatonin interferes within cancer-cell mitochondria after its administration. Depending on subtype and intratumor metabolic heterogeneity, melatonin seems to be helpful in promoting apoptosis, anti-proliferation, pro-oxidation, metabolic shifting, inhibiting neovasculogenesis and controlling inflammation, and restoration of chemosensitivity. This results in attenuation of development, progression, and metastatic potential of reproductive cancers, in addition to lowering the risk of recurrence and improving the life quality of patients.〈/p〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Xuebing Bai, Yanna Guo, Yonghong Shi, Jinxing Lin, Imran Tarique, Xindong Wang, Waseem Ali Vistro, Yufei Huang, Hong Chen, Abdul Haseeb, Ping Yang, Qiusheng Chen〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Intercellular communication of gut epithelial cells is critical to gut mucosal homeostasis. Exosomes are important intercellular mediators in communication between cell to cell. Although many literature focus on the immunologic roles in the gut by the exosomes, the biological process of exosomes in the absorptive cells remains unknown. Uncovering the distribution, classification and formation process of multivesicular bodies (MVBs) and their exosomes in the absorptive cells of the zebrafish gut, is urgently needed to establish a platform for immunological research of fish gut exosomes. The expression levels of CD63 and TSG101 were different among the three segments of the gut, and they were enriched at the apex of the mid gut villi. The characteristics of MVBs and their exosomes in the absorptive cells were further revealed by transmission electron microscopy (TEM). Early endosomes (ee) were mainly present in the apical and basal cytoplasm of absorptive cells. Late endosomes (le) were mostly distributed with the supranuclear part of these cells. “Heterogeneous” MVBs were detected underlying the apical membranes of absorptive cells. Many exosomes with some MVB-like structures occurred in the lumen, indicating that the release process was mainly through apical secretion. Various MVBs with exosomes and the endosome-heterogeneous MVB-exosome complex existed widely in the mid gut absorptive cells, concluding that zebrafish as a potential model for in vivo MVBs and their exosomes research. All the results were summarized in a schematic diagram illustrating the morphological characteristics of gut MVBs and their exosomes in zebrafish.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Ishaaq Saputra, Ravi Fotedar, Sanjay K. Gupta, Muhammad A.B. Siddik, Md Javed Foysal〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉A two phased feeding trial was conducted to evaluate the effects of alternative protein sources on the immunophysiological responses of marron. During the phase I, marron were fed with five alternative protein supplemented diets for 90 days, while in phase II, the same marron were exposed to elevated temperature (30 °C) and their immunophysiological responses were investigated post exposure. Five isoproteic (crude protein 30%) and isoenergetic diets were prepared by containing fishmeal, poultry by-product meal, feather meal, lupin meal, and meat and bone meal as the main protein source. A hundred and fifty juvenile marron (〈em〉Cherax cainii〈/em〉) of the average weight 9.09 ± 0.21 g were randomly distributed into 15 tanks (three replicates per feeding treatments). In the Phase I, general immune response parameters, such as, total haemocyte count (THC), proportion of hyaline cells, neutral red retention time (NRRT), phagocytic rate (PR), heamolymph bacteraemia, and condition indices of marron were investigated. The highest (P 〈 0.05) THC among dietary protein sources was obtained in marron fed with PbM at the end of experiment. Marron fed with FeM protein sources resulted in the highest survival rate followed by PbM fed group. Longer microvilli length (3.83 ± 0.18 μm) was demonstrated in marron fed with PbM diet. Diets containing FM and PbM protein sources revealed significantly (P 〈 0.05) lower number of microvilli/group than diets containing FeM and LM. The results demonstrated that different dietary protein sources in the marron diets did not detect significant (P 〉 0.05) change of the condition indices throughout the experiment period, however highest Hiw and Hid was recorded in marron fed with PBM at day 45. The PR of marron fed dietary protein from PbM did not change significantly after temperature exposure. Increased NRRT, PR and haemolymph bacteraemia was observed with dietary feeding of FM at the end of the trial. However, results revealed that PbM could be an alternative protein source for culture of marron as reflected in terms of increased THC, longer microvillus length and improved susceptibility to high temperature exposure. Overall, result could serve as useful baseline data in developing cost effective potential diets for marron aquaculture.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Xiaowen Xu, Meifeng Li, Dongming Li, Zeyin Jiang, Changxin Liu, Xiao Shi, Chuxin Wu, Xingxing Chen, Gang Lin, Chengyu Hu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉SAMHD1 is an innate immunity restriction factor that inhibits virus infection through IRF3-mediated antiviral and apoptotic responses. Fish SAMHD1 shares some similar properties with those in mammals. In this study, a 〈em〉SAMHD1〈/em〉 orthologue from grass carp (〈em〉Ctenopharyngodon idellus〈/em〉) was cloned and characterized. The full-length cDNA of 〈em〉CiSAMHD1〈/em〉 is 2792 bp with an ORF of 1884 bp encoding a polypeptide of 627 amino acids. Multiple alignments showed that SAMHD1 is highly conserved among different species. Phylogenetic tree analysis revealed that 〈em〉CiSAMHD1〈/em〉 shared a high degree of homology with 〈em〉Sinocyclocheilus rhinocerous SAMHD1〈/em〉. Expression analysis indicated that 〈em〉CiSAMHD1〈/em〉 was widely expressed in all tissues tested including the brain, eyes, spleen, gill, intestine, liver, heart and kidney. It was significantly up-regulated in spleen, liver and intestines after treatment with poly I:C. Also, 〈em〉CiSAMHD1〈/em〉 can be induced following stimulation with recombinant IFN in CIK cells. The promoter sequence of 〈em〉CiSAMHD1〈/em〉 was identified to explore the mechanism underlying the transcriptional regulation of 〈em〉CiSAMHD1〈/em〉. The promoter sequence of 〈em〉CiSAMHD1〈/em〉 (1370 bp) consists of IRF1, IRF3, IRF9 and p65 binding elements. Gel mobility shift assay also showed that IRF1, IRF3, IRF9 and p65 prokaryotic proteins can separately interact with 〈em〉CiSAMHD1〈/em〉 promoter. Dual luciferase assay and q-PCR suggested that the promoter of 〈em〉CiSAMHD1〈/em〉 can be activated by the overexpression of 〈em〉Ci〈/em〉IRF3 and 〈em〉Ci〈/em〉IRF9, but cannot be triggered by 〈em〉Ci〈/em〉IRF1 and 〈em〉Ci〈/em〉p65. In contrast, knockdown of 〈em〉Ci〈/em〉IRF3 or 〈em〉Ci〈/em〉IRF9 inhibits the transcription of 〈em〉CiSAMHD1〈/em〉. Intriguingly, CCK assay suggested that 〈em〉Ci〈/em〉SAMHD1 decreased cell viability. TUNEL apoptosis assay and Hoechst 33258 staining assay indicated that apoptosis is induced by the overexpression of 〈em〉Ci〈/em〉SAMHD1. Crystal violet staining, detection of two GCRV genes (vp3 and vp5) and viral titration showed that 〈em〉Ci〈/em〉SAMHD1 can suppress the proliferation of grass carp reovirus (GCRV) in CIK cells.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Cuiyun Zou, Ningning Su, Jinhui Wu, Minglei Xu, Zhenzhu Sun, Qingying Liu, Leling Chen, Yuanyuan Zhou, Anli Wang, Chaoxia Ye〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉In this study, two experiments were performed to explore the effect of Radix Bupleuri extracts (RBE) on growth, lipid deposition and metabolism and immune response of hybrid grouper (〈em〉Epinephelus lanceolatus♂〈/em〉 × 〈em〉Epinephelus fuscoguttatus♀〈/em〉) using in vitro and in vivo models. In vitro, we used 2 ml/L 20% lipid emulsion (LE)-induced steatosis in hybrid grouper primary hepatocytes, then RBE (200, 400 and 800 μg/ml) was added to the hepatocytes after (post-treatment) the incubation with 20% LE (2 ml/L) in the culture medium. We found that RBE markedly increased cell viability, which were consistent with hepatocytes morphological structure examination and lipid metabolism and immune related genes study. The above result suggested that RBE has a protective effect on this model of hepatocytes damage. In vivo, five graded levels of RBE at 0, 200, 400, 800 and 1600 mg/kg diet were supplemented to a basal diet with 15% lipid levels (high lipid), and fed to a total of 300 hybrid grouper with an average initial weight of 25.58 ± 0.05 g for 8 weeks. Growth performance, liver histology, plasma biochemical parameters, and expression of genes involved in lipid metabolism and immune-related were measured. The study indicated that dietary RBE significantly improved growth performance and feed utilization and reduced hepatosomatic index. Dietary supplementation with 200–800 mg/kg RBE diets effectively decreased serum ALP, ALT, AST and LDH contents in fish. Furthermore, adipogenesis relative mRNA levels of DGAT2, G6PD, ME1 and DGKα in fish fed 200–400 mg/kg RBE diets were lower (〈em〉P〈/em〉 〈 0.05) than in those fed RBE0 diets, while dietary supplementation with 200–800 mg/kg RBE diets up-regulated lipolysis-related genes (CPT1, LPL and PPARα) expression in the liver of hybrid grouper. Moreover, dietary RBE down-regulated the expression of apoptosis-related genes (caspase-9), up-regulated the expression of antioxidant genes (CAT) and immune-related genes (MHC2, IKKα and TGF-β1). Thus, our data suggest that RBE suppressed lipid accumulation and enhanced immune capability in hybrid grouper both in vitro and in vivo. These results offer new insight into RBE as a hepatoprotective in fish.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Meimei Zhang, Yingxue Qin, Lixing Huang, Qingpi Yan, Leilei Mao, Xiaojin Xu, Suyun Wang, Mengmeng Zhang, Liwei Chen〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Several bacteria have been defined as extracellular pathogens; however, in recent years, it has been confirmed that they have the ability to survive and escape the attack of host phagocytes, thus causing further infection. Previous studies have shown that 〈em〉Aeromonas hydrophila〈/em〉 could survive in fish macrophages; however, the mechanism remains unknown. In this study, 〈em〉sodA〈/em〉 and 〈em〉sodB〈/em〉 of the strain 〈em〉A. hydrophila〈/em〉 B11 were stable silenced by shRNA. The survival rates of intracellular 〈em〉sodA-〈/em〉RNAi and 〈em〉sodB-〈/em〉RNAi decreased by 91.8% and 74.9% and the immune escape rates decreased by about 32% and 92% respectively. At the same time, reactive oxygen species (ROS) in fish macrophages that phagocytosed 〈em〉sodA〈/em〉-RNAi and 〈em〉sodB〈/em〉-RNAi increased by 40% and 32.6%, respectively, compared to those of macrophages that phagocytosed the wild-type strain. Compared to 〈em〉sodA,〈/em〉 the expression of 〈em〉sodB〈/em〉 predominates in 〈em〉A. hydrophila〈/em〉 without oxidative stress; however, when exposed to oxidative stress, the magnitude of up-regulation of 〈em〉sodA〈/em〉 expression is significantly higher than that of 〈em〉sodB〈/em〉. With increased of methyl viologen concentration, the survival rates of 〈em〉sodA〈/em〉-RNAi and 〈em〉sodB〈/em〉-RNAi were significantly decreased. The expressions of 〈em〉sodA〈/em〉 and s〈em〉odB〈/em〉 did not affect the growth of 〈em〉A. hydrophila〈/em〉 without oxidative stress, but the inhibition of 〈em〉sodA〈/em〉 and 〈em〉sodB〈/em〉 expression led to a slight decrease in bacterial growth under oxidative stress. These results indicated that (1) 〈em〉sodA〈/em〉 and 〈em〉sodB〈/em〉 play an important role in the process of bacterial resistance to ROS damage in host phagocytic cells, allowing them to survive or even escape fish macrophages; (2) the 〈em〉sodB〈/em〉 expression was dominant in 〈em〉A. hydrophila〈/em〉 without oxidative stress, the 〈em〉sodA〈/em〉 expression was up-regulated more significantly under oxidative stress, and 〈em〉sodA〈/em〉 and s〈em〉odB〈/em〉 contributed equally to the process of bacterial resistance to ROS; (3) 〈em〉sodA〈/em〉 and 〈em〉sodB〈/em〉 complement each other and cooperate in the process of intracellular survival of bacteria to protect against ROS damage.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Daniel Gonzalez-Silvera, Francisco A. Guardiola, Cristóbal Espinosa, Elena Chaves-Pozo, M. Ángeles Esteban, Alberto Cuesta〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Viral necrosis virus (NNV) or nodavirus causes fish viral encephalopathy and retinopathy worldwide. In some cases, mortalities in aquaculture industry can reach up to 100%, some species being especially sensitive as is the case of European sea bass (〈em〉Dicentrarchus labrax〈/em〉), one of the main cultured species in the Mediterranean, with the consequent economical loses. Development of new vaccines against NNV is in the spotlight though few researches have focused in European sea bass. In this study we have generated a recombinant NNV (rNNV) vaccine produced in 〈em〉Escherichia coli〈/em〉 expressing the capsid protein and administered it to European sea bass juveniles by two different routes (intraperitoneal and oral). The last being considered non-stressful and desired for fish farming of small fish, which in fact are the most affected by NNV. Oral vaccine was composed of feed pellets containing the recombinant whole bacteria, and injected vaccine was composed of recombinant bacteria previously lysed. Our results revealed production of specific anti-NNV IgM following the two vaccination procedures, levels that were further increased in orally-vaccinated group after challenge with NNV. Genes related to interferon (IFN), T-cell and immunoglobulin markers were scarcely regulated in head-kidney (HK), gut or brain. Vaccination by either route elicited a relative survival response of 100% after NNV challenge. To our knowledge, this is the first report of a recombinant vaccine followed by no purification steps which resulted in a complete protection in European sea bass when challenged with NNV.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Chao Pei, Xiaoying Sun, Yi Zhang, Li Li, Yan Gao, Li Wang, Xianghui Kong〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The Polymeric Immunoglobulin Receptor (〈em〉pIgR〈/em〉) gene has been proved to play an important role in transporting polymeric immunoglobulin (Ig) in the mucosal tissues of mammals. 〈em〉pIgR〈/em〉 gene also exists in teleost, but the genetic diversity and functions of this gene still need to be further explored. We obtained seven grass carp pIgR splicing transcripts, a full-length pIgR (〈em〉CipIgR-1〈/em〉) and six truncated variants (〈em〉CipIgR-2〈/em〉 to 〈em〉CipIgR-7〈/em〉). The full-length pIgR contained two immunoglobulin-like domains (ILD), a transmembrane domain (TMD) and a cytoplasmic domain (CyD). The 〈em〉CipIgR-2〈/em〉 lacked a small part in CyD, and 〈em〉CipIgR-3〈/em〉 lost TMD and CyD. Partial cDNA sequences of the other four grass carp pIgR variants (〈em〉CipIgR-4〈/em〉 to 〈em〉CipIgR-7〈/em〉) were also cloned. The total expression levels of 〈em〉CipIgR〈/em〉 and its variants in different tissues were detected by real-time quantitative PCR. The highest expression was found in the intestine, followed by the spleen and the skin. The function of the two extracellular ILDs of 〈em〉CipIgR〈/em〉 was investigated based on its combining capacity with grass carp immunoglobulin M (IgM) and aquatic pathogenic bacteria. The cDNA sequences of two ILDs were cloned and expressed in 〈em〉Escherichia coli〈/em〉 BL21 (DE3). Recombinant ILDs protein was purified and incubated with different bacteria respectively. Results of Western blot showed the recombinant protein could combine 〈em〉Bacillus subtilis〈/em〉, 〈em〉Vibrio parahaemolyticus〈/em〉, and 〈em〉Escherichia coli〈/em〉. In addition, binding activity of rILDs with grass carp IgM was detected. Collectively, these results indicated that multiple variants of 〈em〉pIgR〈/em〉 gene in grass carp might be involved in the antibacterial immunity.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Jiabao Hu, Qijun Le, Yajun Wang, Na Yu, Xiaohuan Cao, Siwen Kuang, Man Zhang, Weiwei Gu, Yibo Sun, Yang Yang, Shanliang Xu, Xiaojun Yan〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Formaldehyde can effectively control ectoparasites in silver pomfret (〈em〉Pampus argenteus〈/em〉). However, there is limited information on the effects of formaldehyde treatment at a molecular level in fishes. In the present study, transcriptome profiling was conducted to investigate the effects of formaldehyde treatment (80 mg/L, bath for 1 h every day for three consecutive days) on the liver and kidney tissues of silver pomfret. A total of 617959982 clean reads were obtained and assembled into 265760 unigenes with an N50 length of 1507 bp, and the assembled unigenes were all annotated by alignment with public databases. A total of 2204 differentially expressed genes (DEGs) were detected in the liver and kidney tissues, and they included 7 detoxification- related genes and 9 immune-related genes, such as 〈em〉CYP450〈/em〉, 〈em〉GST〈/em〉, 〈em〉MHC I & II〈/em〉, and 〈em〉CCR〈/em〉. In addition, 1440 DEGs were mapped to terms in the GO database, and 1064 DEGs were mapped to the KEGG database. The expression of 4 detoxification-related genes and 6 immune-related genes in three days formaldehyde treatment were analyzed using RT-qPCR, and the antioxidant enzyme levels were also determined. The results indicate differential expression of detoxification- and immune-related genes during the three days formaldehyde treatment. Our data could provide a reference for the treatment of parasites to avoid high mortality and help in understanding the molecular activity in fishes after formaldehyde exposure.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Mohamed Faisal, Isaac F. Standish, Mary Ann Vogelbein, Elena V. Millard, Stephen L. Kaattari〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉This study reports the development of a monoclonal antibody (designated 3B10) against the muskellunge (〈em〉Esox masquinongy〈/em〉) IgM. The 3B10 monoclonal antibody (mAb) belongs to the IgG3 kappa isotype. Western blotting demonstrated that 3B10 mAb reacted primarily to muskellunge IgM heavy chain. 3B10 also reacted strongly with the IgM heavy chain of other esocids, including the northern pike (〈em〉Esox lucius〈/em〉), tiger muskellunge (〈em〉E. masquinongy x E. lucius〈/em〉), and, to a much lesser extent, the chain pickerel (〈em〉E. niger〈/em〉). The 3B10 mAb did not bind to IgM from 10 other fish species resident in the Great Lakes basin. Using the 3B10 mAb, it was possible to determine the muskellunge Ig ability to bind to antigens. Using trinitrophenyl hapten conjugated to keyhole limpet hemocyanin (TNP-KLH) as the eliciting antigen, muskellunge Ig subclasses exhibited a range of affinities with log aK values 5.56–6.25 that is considered intermediate compared to other fish species. 3B10 mAb was used to develop and evaluate an indirect ELISA for the detection and quantitation of circulating antibodies against the viral hemorrhagic septicemia virus genotype IVb (VHSV-IVb). Using the newly optimized assay, anti-VHSV-IVb antibodies were detected in sera of VHSV-IVb vaccinated muskellunge as well as from those of wild muskellunge sampled from an endemic waterbody. In addition to its use in immunoassays, the developed 3B10 mAb will enable future investigation aiming at deciphering immune mechanism of this important fish species to pathogens.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Md Tawheed Hasan, Won Je Jang, Bong-Joo Lee, Kang Woong Kim, Sang Woo Hur, Sang Gu Lim, Sungchul C. Bai, In-Soo Kong〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Investigations were carried out to evaluate and quantify the effects of dietary supplementation with heat-killed (HK) 〈em〉Bacillus〈/em〉 sp. SJ-10 (BSJ-10) probiotic (1 × 10〈sup〉8〈/sup〉 CFU g〈sup〉−1〈/sup〉) on the growth and immunity of olive flounder (〈em〉Paralichthys olivaceus〈/em〉). Flounder (averagely 9.64 g) were divided into two groups, and fed control and HK BSJ-10 (HKBSJ-10)-inoculated diets for 8 weeks. Investigations were carried out on growth and feed utilizations, innate immunity, serum biochemical parameters, microvilli length, and pro- and anti-inflammatory cytokine gene (tumor necrosis factor [TNF]-α, interleukin [IL]-1β, IL-6, and IL-10) transcriptions. Compared to control, HKBSJ-10 diet significantly (〈em〉P〈/em〉 〈 0.05) enhanced weight gain and protein efficiency ratio, 1.17 and 1.11 folds respectively. Humoral innate immune parameters, lysozyme and superoxide dismutase in treatment group were also elevated by 1.34 and 1.16 folds. Similarly, an increased (〈em〉P〈/em〉 〈 0.05) relative expressions of TNF-α, IL-1β, IL-6 were recorded in liver (2.71, 3.38, and 4.12 folds respectively), and gill (2.08, 1.98, and 1.81 folds respectively) than that of controls. Moreover, after challenge with 〈em〉Streptococcus iniae〈/em〉 (1 × 10〈sup〉8〈/sup〉 CFU mL〈sup〉−1〈/sup〉), the HKBSJ-10-fed group exhibited significantly higher protection (〈em〉P〈/em〉 〈 0.05) against streptococcosis compared to controls, validating the observed changes in immune parameters and induction on the cytokine-encoding genes. Therefore, HKBSJ-10 increases growth, modulates innate immune parameters, and protects olive flounders against streptococcosis.〈/p〉〈/div〉 〈/div〉

Description: 〈p〉Publication date: May 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 88〈/p〉 〈p〉Author(s): Ziqiang Liu, Yao Liu, Yaqi Gu, Lili Gao, Ao Li, Dongwu Liu, Cuijie Kang, Qiuxiang Pang, Xiaoqian Wang, Qiang Han, Hairui Yu〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉Opioid neuropeptides are developed early in the course of a long evolutionary process. As the endogenous messengers of immune system, opioid neuropeptides participate in regulating immune response. In this study, the mechanism that Met-enkephalin (M-ENK) inhibits ROS production through Wnt/β-catenin signaling was investigated in the ZF4 cells of zebrafish. ZF4 cells were exposed to 0, 10, 20, 40, 80, and 160 μM Met-enkephalin (M-ENK) for 24 h, and the cell viability was detected with 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. The cell viability was significantly increased by 10, 20, 40, 80, and 160 μM M-ENK. After ZF4 cells were exposed to 0, 20, 40, and 80 μM M-ENK for 24 h, the mRNA expression of Wnt10b, β-catenin, and CCAAT/enhancer binding protein α (C/EBPα) was significantly increased by 40 and 80 μM M-ENK. However, the mRNA and protein expression of GSK-3β was significantly decreased by 40 and 80 μM M-ENK. The protein expression of β-catenin was significantly induced by 40 and 80 μM M-ENK, while the protein expression of p-β-catenin was significantly decreased by 20, 40, and 80 μM M-ENK. In addition, the mRNA expression of CAT, SOD, and GSH-PX was significantly increased by 40 and 80 μM M-ENK. The levels of H〈sub〉2〈/sub〉O〈sub〉2〈/sub〉, ·OH, and O〈sub〉2〈/sub〉〈sup〉·-〈/sup〉 were significantly decreased, but the activity of CAT, SOD, and GSH-PX was significantly increased by 40 and 80 μM M-ENK. The fluorescence intensity of reactive oxygen species (ROS) was decreased, and that of mitochondrial membrane potential (MMP) was increased with the increase of M-ENK concentration in ZF4 cells. The results showed that M-ENK could induce Wnt/β-catenin signaling, which further inhibited ROS production through the induction of C/EBPα, MMP, and the activities of antioxidant enzymes.〈/p〉〈/div〉 〈/div〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉In addition to their modulation through de novo expression and degradation, surface levels of chemokine receptors are tuned by their ligand-dependent recycling to the plasma membrane, which ensures that engaged receptors become rapidly available for further rounds of signaling. Dysregulation of this process contributes to the pathogenesis of chronic lymphocytic leukemia (CLL) by enhancing surface expression of chemokine receptors, thereby favoring leukemic cell accumulation in the protective niche of lymphoid organs. In this review, we summarize our current understanding of the process of chemokine receptor recycling, focusing on the impact of its dysregulation in CLL.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Rab18 is one of the small number of conserved Rab proteins which have been traced to the last eukaryotic common ancestor. It is found in organisms ranging from humans to trypanosomes, and localizes to multiple organelles, including most notably endoplasmic reticulum and lipid droplets. In humans, absence of Rab18 leads to a severe illness known as Warburg-Micro syndrome. Despite this evidence that Rab18 is essential, its role in cells remains mysterious. However, recent studies identifying effectors and interactors of Rab18, are now shedding light on its mechanism of action, suggesting functions related to organelle tethering and to autophagy. In this review, we examine the variety of roles proposed for Rab18 with a focus on new evidence giving insights into the molecular mechanisms it utilizes. Based on this summary of our current understanding, we identify priority areas for further research.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Oxygen constitutes a vital element for the survival of every single cell in multicellular aerobic organisms like mammals. A complex homeostatic oxygen-sensing system has evolved in these organisms, including detectors and effectors, to guarantee a proper supply of the element to every cell. The carotid body represents the most important peripheral arterial chemoreceptor organ in mammals and informs about hypoxemic situations to the effectors at the brainstem cardiorespiratory centers. To optimize organismal adaptation to maintained hypoxemic situations, the carotid body has evolved containing a niche of adult tissue-specific stem cells with the capacity to differentiate into both neuronal and vascular cell types in response to hypoxia. These neurogenic and angiogenic processes are finely regulated by the niche and by hypoxia itself. Our recent data on the cellular and molecular mechanisms underlying the functioning of this niche might help to comprehend a variety of different diseases coursing with carotid body failure, and might also improve our capacity to use these stem cells for the treatment of neurological disease. Herein, we review those data about the recent characterization of the carotid body niche, focusing on the study of the phenotype and behavior of multipotent stem cells within the organ, comparing them with other well-documented neural stem cells within the adult nervous system.〈/p〉

Description: 〈p〉Publication date: September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 92〈/p〉 〈p〉Author(s): Lanfen Fan, Lei Wang, Zhenlu Wang〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉To understand the homeostasis mechanism of crustacean hepatopancreas to cold stress, iTRAQ proteomics based on the genome database of 〈em〉Litopenaeus vannamei〈/em〉 (〈em〉L. vannamei〈/em〉) was applied to investigate proteins changes and variety of the hepatopancreas during cold stress stage in this study. A total of 4062 distinct proteins were identified, 137 differentially expressed proteins (DEPs) including 62 differentially up-regulated proteins (DUPs) and 75 differentially down-regulated proteins (DDPs) were identified in G1 (18 °C) compared with CK (28 °C), 359 DEPs including 131 DUPs and 228 DDPs were identified in G2 (13 °C for 24 h) compared with CK. Based on bioinformatics analysis, the cold tolerance of 〈em〉L. vannamei〈/em〉 might be related to energy metabolism such as amino acid, carbohydrate, lipid, and oxidative phosphorylation. Moreover, shrimp immunity was declined during cold stress stage. However, 〈em〉L. vannamei〈/em〉 could cope with cold stress by enhancing the production of ATP and UFA. Notably, arginine kinase, heat shock proteins, and histones may act as positive regulators in 〈em〉L. vannamei〈/em〉 under cold stress. Ten randomly selected proteins were used for validation using qRT-PCR and the expressions on the transcription level for most of the genes were similar to the results of iTRAQ. These results indicated that 〈em〉L. vannamei〈/em〉 can maintain the organism homeostasis by a series of orderly regulatory process during cold stress. Furthermore, the results can provide guidance for shrimp farming.〈/p〉〈/div〉 〈/div〉 〈h5〉Graphical abstract〈/h5〉 〈div〉〈p〉〈figure〉〈img src="https://ars.els-cdn.com/content/image/1-s2.0-S1050464819306849-fx1.jpg" width="488" alt="Image 1" title="Image 1"〉〈/figure〉〈/p〉〈/div〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Disintegrin and metalloproteinases (ADAMs) 10 and 17 can release the extracellular part of a variety of membrane-bound proteins via ectodomain shedding important for many biological functions. So far, substrate identification focused exclusively on membrane-anchored ADAM10 and ADAM17. However, besides known shedding of ADAM10, we identified ADAM8 as a protease capable of releasing the ADAM17 ectodomain. Therefore, we investigated whether the soluble ectodomains of ADAM10/17 (sADAM10/17) exhibit an altered substrate spectrum compared to their membrane-bound counterparts. A mass spectrometry-based N-terminomics approach identified 134 protein cleavage events in total and 45 common substrates for sADAM10/17 within the secretome of murine cardiomyocytes. Analysis of these cleavage sites confirmed previously identified amino acid preferences. Further in vitro studies verified fibronectin, cystatin C, sN-cadherin, PCPE-1 as well as sAPP as direct substrates of sADAM10 and/or sADAM17. Overall, we present the first degradome study for sADAM10/17, thereby introducing a new mode of proteolytic activity within the protease web.〈/p〉

Description: 〈h3〉Abstract〈/h3〉 〈p〉Fibrillin microfibrils are ubiquitous elements of extracellular matrix assemblies that play crucial roles in regulating the bioavailability of growth factors of the transforming growth factor beta superfamily. Recently, several “a disintegrin and metalloproteinase with thrombospondin motifs” (ADAMTS) proteins were shown to regulate fibrillin microfibril function. Among them, 〈em〉ADAMTS17〈/em〉 is the causative gene of Weill-Marchesani syndrome (WMS) and Weill-Marchesani-like syndrome, of which common symptoms are ectopia lentis and short stature. 〈em〉ADAMTS17〈/em〉 has also been linked to height variation in humans; however, the molecular mechanisms whereby ADAMTS17 regulates skeletal growth remain unknown. Here, we generated 〈em〉Adamts17-/-〈/em〉 mice to examine the role of Adamts17 in skeletogenesis. 〈em〉Adamts17〈/em〉-/- mice recapitulated WMS, showing shorter long bones, brachydactyly, and thick skin. The hypertrophic zone of the growth plate in 〈em〉Adamts17〈/em〉-/- mice was shortened, with enhanced fibrillin-2 deposition, suggesting increased incorporation of fibrillin-2 into microfibrils. Comprehensive gene expression analysis of growth plates using laser microdissection and RNA sequencing indicated alteration of the bone morphogenetic protein (BMP) signaling pathway after Adamts17 knockout. Consistent with this, phospho-Smad1 levels were downregulated in the hypertrophic zone of the growth plate and in 〈em〉Adamts17〈/em〉-/- primary chondrocytes. Delayed terminal differentiation of 〈em〉Adamts17〈/em〉-/- chondrocytes, observed both in primary chondrocyte and primordial metatarsal cultures, and was prevented by BMP treatment. Our data indicated that Adamts17 is involved in skeletal formation by modulating BMP-Smad1/5/8 pathway, possibly through inhibiting the incorporation of fibrillin-2 into microfibrils. Our findings will contribute to further understanding of disease mechanisms and will facilitate the development of therapeutic interventions for WMS.〈/p〉

Description: 〈p〉Publication date: September 2019〈/p〉 〈p〉〈b〉Source:〈/b〉 Fish & Shellfish Immunology, Volume 92〈/p〉 〈p〉Author(s): Guilan Di, Yanfei Li, Xianliang Zhao, Ning Wang, Jingqiang Fu, Min Li, Miaoqin Huang, Weiwei You, Xianghui Kong, Caihuan Ke〈/p〉 〈div xml:lang="en"〉 〈h5〉Abstract〈/h5〉 〈div〉〈p〉The haemocytes of the ivory shell, 〈em〉Babylonia areolata〈/em〉 are classified by morphologic observation into the following types: hyalinocytes (H) and granulocytes (G). Haemocytes comprise diverse cell types with morphological and functional heterogene and play indispensable roles in immunological homeostasis of invertebrates. In the present study, two types of haemocytes were morphologically identified and separated as H and G by Percoll density gradient centrifugation. The differentially expressed proteins were investigated between H and G using mass spectrometry. The results showed that total quantitative proteins between H and G samples were 1644, the number of up-regulated proteins in G was 215, and the number of down-regulated proteins in G was 378. Among them, cathepsin, p38 MAPK, toll-interacting protein-like and beta-adrenergic receptor kinase 2-like were up-regulated in G; alpha-2-macroglobulin-like protein, C-type lectin, galectin-2-1, galectin-3, β-1,3-glucan-binding protein, ferritin, mega-hemocyanin, mucin-17-like, mucin-5AC-like and catalytic subunit of protein kinase A were down-regulated in G. The results showed that the most significantly enriched KEGG pathways were the pathways related to ribosome, phagosome, endocytosis, carbon metabolism, protein processing in endoplasmic reticulum and oxidative phosphorylation. For phagosome and endocytosis pathway, the number of down-regulation proteins in G was more than that of up-regulation proteins. For lysosome pathway, the number of up-regulation proteins in G was more than that of down-regulation proteins. These results suggested that two sub-population haemocytes perform the different immune functions in 〈em〉B. areolata〈/em〉.〈/p〉〈/div〉 〈/div〉