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1.
Dev Comp Immunol ; 126: 104242, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34450131

RESUMO

Although, in mammals, the Krüppel-like transcription factor 13 (KLF13) plays an essential role in cell proliferation, survival, differentiation, apoptosis, tumorigenesis, immune regulation, and inflammation, its role in penaeid shrimp is unclear. In the current study, we characterized a KLF13 homolog in Penaeus vannamei (PvKLF13), with full-length cDNA of 1677 bp and 1068 bp open reading frame (ORF) encoding a putative protein of 355 amino acids, which contains three ZnF_C2H2 domains. Sequence and phylogenetic analysis revealed that PvKLF13 shares a close evolutionary relationship with KLF13 from invertebrates. Transcript levels of PvKLF13 were ubiquitously expressed in shrimp and induced in hemocytes upon challenge with Vibrio parahaemolyticus, Streptococcus iniae, and white spot syndrome virus (WSSV), suggesting the involvement of PvKLF13 in shrimp immune response to pathogens. Besides, knockdown of PvKLF13 decreased hemocytes apoptosis in terms of increased expression of pro-survival PvBcl-2, but decreased expression of pro-apoptotic PvBax and PvCytochrome C, coupled with high PvCaspase3/7 activity, especially upon V. parahaemolyticus challenge. The findings here indicate the involvement of PvKLF13 in apoptotic cell clearance as an essential part of shrimp innate immune response to pathogens.

2.
Int J Syst Evol Microbiol ; 71(10)2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34623951

RESUMO

A Gram-stain-negative, non-motile, rod-shaped, aerobic bacterium (designated as LMIT005T) was isolated from shrimp ponds in Shantou, China. The new isolate was characterized taxonomically using a polyphasic approach. Based on 16S rRNA gene sequence analysis, strain LMIT005T was found to be affiliated with the family Cyclobacteriaceae of the order Cytophagales while appearing as a distinct lineage. The 16S rRNA gene sequence similarity between strain LMIT005T and Algoriphagus yeomjeoni KCTC 12309T, the closest type strain in the family, was 91.3 %. Strain LMIT005T grew optimally at 25 °C, pH 7 and in the presence of 2.0 % (w/v) NaCl. The DNA G+C content (data from genome sequence) was 40.5 mol%. Compared with reference strain A. yeomjeoni KCTC 12309T, the average nucleotide identity (ANI) of LMIT005T was 70 %. The sole respiratory quinone of LMIT005T was menaquinone (MK-7), and the major fatty acids were summed feature 3 (C16 : 1 ω6c / C16 : 1 ω7c). The polar lipids of strain LMIT005T were mainly composed of phosphatidylethanolamine, phosphatidylcholine, two unidentified amino lipids, two unidentified lipids, one unidentified glycolipid and one unidentified phospholipid. The draft genome of strain LMIT005T comprised 3 089 781 bp (3.09 Mb) nucleotides and 2773 genes. Antimicrobial resistant-related genes (blal, mexA, and mexb) were annotated in the genome of strain LMIT005T, which indicated that it might be able to resist ß-lactam antibiotics. This was further verified by antimicrobial resistant test. Given its distinct genomic, morphological, and physiological differences from previously described type strains, strain LMIT005T is proposed as a representative of a novel genus of the family Cyclobacteriaceae, with the name Penaeicola halotolerans gen. nov., sp. nov. The type strain is LMIT005T (=KCTC 82616T=CICC 25047T).


Assuntos
Lagoas , Água do Mar , Técnicas de Tipagem Bacteriana , Bacteroidetes/genética , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
3.
mSystems ; : e0091721, 2021 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-34636669

RESUMO

Molting is a crucial lifelong process in the growth, development, and reproduction of crustaceans. In mud crab (Scylla paramamosain), new exoskeleton, gills, and appendages are formed after a molting, which contributes to a 40 to 90% increase in body weight. However, little is currently known about the associations between molting and the dynamic changes of microbiota and physiological characteristics in mud crabs. In this study, the effects of molting on changes of the microbiome, immune response, and digestive enzyme activities in mud crabs were investigated. The results showed dynamic changes in the abundances and community compositions of crab-associated microbiota harboring the gills, subcuticular epidermis, hepatopancreas, midgut, and hemolymph during molting. Renewed microbiota was observed in the gills and midgut of crabs at the postmolt stages, which seems to be related to the formation of a new exoskeleton after the molting. A significant positive correlation between the expression of two antimicrobial peptide (AMP) genes (SpALF5 and SpCrustin) and the relative abundance of two predominant microorganisms (Halomonas and Shewanella) in hemolymph was observed in the whole molt cycle, suggesting that AMPs play a role in modulating hemolymph microbiota. Furthermore, digestive enzymes might play a vital role in the changes of microbiota harboring the hepatopancreas and midgut, which provide suitable conditions for restoring and reconstructing host-microbiome homeostasis during molting. In conclusion, this study confirms that molting affects host-associated microbiota and further sheds light on the effects on the immune response and the digestive systems as well. IMPORTANCE Molting is crucial for crustaceans. In mud crab, its exoskeleton is renewed periodically during molting, and this process is an ideal model to study the effects of host development on its microbiota. Here, multiple approaches were used to investigate the changes in microbial taxa, immune response, and digestive enzyme activity with respect to molting in mud crab. The results found that a renewed microbiota was generated in the gills and midgut of crab after a molt. A significant positive correlation between changes in the relative abundances of microbes (such as Halomonas and Shewanella) and the expression of AMP genes (SpALF5 and SpCrustin) was observed in the hemolymph of crabs during the whole molt cycle, suggesting the modulation of hemolymph microbes by AMPs. Furthermore, the digestive enzymes were found to participate in the regulation of microbiota in hepatopancreas and midgut, consequently providing a suitable condition for the restoration and reconstruction of host-microbiome homeostasis during the molting. This study confirms that molting affects the microbial communities and concomitantly influences the immune and digestive systems in mud crabs. This is also the first time the homeostasis of the host and microbiome, and the associations between molting and physiological characteristics in crustaceans, have been revealed.

4.
Dev Comp Immunol ; 127: 104293, 2021 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-34648768

RESUMO

Arginine metabolism pathway enzymes and products are important modulators of several physiological processes in animals, including immune response. Although some components of the arginine metabolic pathway have been reported in penaeid shrimps, no systematic study has explored all the key pathway enzymes involved in shrimp antimicrobial response. Here, we explored the role of the three key arginine metabolism enzymes (nitric-oxide synthase (NOS), arginase (ARG), agmatinase (AGM)) in Penaeus vannamei antimicrobial immunity. First, P. vannamei homologs of ARG and AGM (PvARG and PvAGM) were cloned and found to be evolutionally conserved with invertebrate counterparts. Transcript levels of PvARG, PvAGM, and PvNOS were ubiquitously expressed in healthy shrimp tissues and induced in hemocytes and hepatopancreas upon challenge with Gram-negative (Vibrio parahaemolyticus) and Gram-positive (Streptoccocus iniae) bacteria, suggesting their involvement in shrimp antimicrobial immune response. Besides, RNA interference knockdown and enzyme activity assay revealed an antagonistic relationship between PvARG/PvAGM and PvNOS, while this relationship was broken upon pathogen stimulation. Interestingly, knockdown of PvNOS increased Vibrio abundance in shrimp hemolymph, whereas knockdown of PvAGR reduced Vibrio abundance. Taken together, our present data shows that homologs of the key arginine metabolism pathway enzymes in penaeid shrimp (PvARG, PvAGM, and PvNOS) work synergistically and/or antagonistically to modulate antibacterial immune response.

5.
Arq Bras Oftalmol ; 2021 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-34586233

RESUMO

PURPOSE: To investigate the incidence, risk factors, and visual outcomes of epiretinal membrane development following rhegmatogenous retinal detachment repair. METHODS: This was a retrospective study of 309 eyes that underwent initial surgery for primary uncomplicated rhegmatogenous retinal detachment. Examinations were conducted preoperatively and then postoperatively at 1, 3, 6, and 12 months. The study patients were categorized into two groups depending on the presence or absence of the epiretinal membrane. RESULTS: The incidence of postoperative epiretinal membrane was 28.5%; 42.7% of these patients had severe epiretinal membrane development and therefore underwent the epiretinal membrane removal. Logistic regression analyses revealed that giant retinal tears (OR: 2.66; 95% CI: 1.045-6.792, p=0.040) and horseshoe tears (OR: 0.534; 95% CI: 0.295-0.967, p=0.039) were the significant predictors of postoperative epiretinal membrane. Triamcinolone acetonide staining was significantly associated with the prevention of epiretinal membrane (p=0.022). A total of 34 patients showed a better or an equal final best-corrected visual acuity; of which 4 eyes were evaluated at the final follow-up visit and exhibited a reduced best-corrected visual acuity. CONCLUSION: Our analysis demonstrated that horseshoe tears and giant retinal tears represent the risk factors for the postoperative epiretinal membrane. Triamcinolone acetonide staining had a significant preventive effect on the postoperative epiretinal membrane. Furthermore, a second round of pars plana vitrectomy, including membrane removal, led to a significant improvement in the final best-corrected visual acuity as per the last follow-up examination, albeit the recovery was limited.

6.
Mol Immunol ; 138: 181-187, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34450346

RESUMO

Prophenoloxidase (proPO)-activating system is a critical innate immune defense in invertebrates. However, the mechanisms involved in regulating the phenoloxidase (PO) activity in shrimp hemolymph remain ill-defined. Our previous studies showed that Penaeus vannamei hemocyanin (HMC) and α2-macroglobulin (α2M), two key regulators of proPO-activating system in plasma, might interact with each other, indicating that this interaction could be implicated in controlling PO activity. Herein, we further confirmed that HMC specifically bind to α2M using Pull down and Far-Western blot analyses. Further studies demonstrated that HMC could directly interact with the receptor binding domain of α2M. In addition, HMC and α2M followed similar expression pattern upon Vibrio parahaemolyticus infection, suggesting the interaction of HMC and α2M might have a role in immune response. Finally, we found that α2M, as a broad-spectrum proteinase inhibitor, suppressed the serum PO activity in vitro, while hemocyanin could partially restore this inhibitory effect. In sum, the present data indicate that HMC interacts with α2M and therefore modulates the PO activity. This finding contributes to better understanding of stable state maintenance of PO activity in shrimp.

7.
J Neurointerv Surg ; 2021 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-34349014

RESUMO

OBJECTIVE: In this review and meta-analysis we sought to compare the efficacy and safety of direct endovascular thrombectomy (EVT) and bridging therapy for intravenous thrombolysis (IVT)-eligible patients with acute ischemic stroke caused by large vessel occlusions (AIS-LVO). METHODS: We searched Medline, Embase, and the Cochrane Library for published randomized clinical trials (RCTs) and observational studies providing outcomes of patients with IVT-eligible AIS-LVO who have undergone EVT with or without IVT. The primary outcome was the proportion of patients achieving a modified Rankin Scale (mRS) score of 0-2 at 90 days. The secondary outcomes included the rates of (1) an excellent outcome defined as an mRS score of 0 or 1 at 90 days, (2) mortality at 90 days, (3) symptomatic intracranial hemorrhage (sICH), (4) any type of intracranial hemorrhage (ICH), (5) successful recanalization, and (6) clot migration. RESULTS: We included three RCTs and six observational studies (4 of which were propensity score-adjusted studies) with a total of 3133 patients. In unadjusted and adjusted analyses, no differences in the rates of mRS scores 0-2, mRS scores 0-1, mortality at 90 days, sICH or successful recanalization were detected between patients with AIS-LVO who underwent direct EVT or bridging therapy. The patients treated with direct EVT had a lower risk ratio for any type of ICH and clot migration than did the patients treated with bridging therapy. CONCLUSION: Compared with bridging therapy, direct EVT may be equally effective and yield a lower rate of ICH and clot migration in patients with AIS. TRAIL REGISTRATION NUMBER: PROSPERO: CRD42021236691.

8.
PLoS Pathog ; 17(8): e1009837, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34379706

RESUMO

It is well known that exosomes could serve as anti-microbial immune factors in animals. However, despite growing evidences have shown that the homeostasis of the hemolymph microbiota was vital for immune regulation in crustaceans, the relationship between exosomes and hemolymph microbiota homeostasis during pathogenic bacteria infection has not been addressed. Here, we reported that exosomes released from Vibrio parahaemolyticus-infected mud crabs (Scylla paramamosain) could help to maintain the homeostasis of hemolymph microbiota and have a protective effect on the mortality of the host during the infection process. We further confirmed that miR-224 was densely packaged in these exosomes, resulting in the suppression of HSP70 and disruption of the HSP70-TRAF6 complex, then the released TRAF6 further interacted with Ecsit to regulate the production of mitochondrial ROS (mROS) and the expression of Anti-lipopolysaccharide factors (ALFs) in recipient hemocytes, which eventually affected hemolymph microbiota homeostasis in response to the pathogenic bacteria infection in mud crab. To the best of our knowledge, this is the first document that reports the role of exosome in the hemolymph microbiota homeostasis modulation during pathogen infection, which reveals the crosstalk between exosomal miRNAs and innate immune response in crustaceans.

9.
Ultrason Sonochem ; 77: 105676, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34315058

RESUMO

In recent years, foodborne diseases caused by Escherichia coli are a major threat to the food industry and consumers. Antimicrobial peptides (AMPs) and ultrasound both have good inhibitory effects on E. coli. In this work, the mechanism of action and synergistic effect of an in silico predicted AMP, designated as TGH2 (AEFLREKLGDKCTDRHV), from the C-terminal sequence of Tegillarca granosa hemoglobin, combined with low-intensity ultrasound was explored. The minimal inhibitory concentration (MIC) of TGH2 on E. coli decreased by 4-fold to 31.25 µg/mL under 0.3 W/cm2 ultrasound treatment, while the time kill curve analysis showed that low-intensity ultrasound combined with peptide TGH2 had an enhanced synergistic bactericidal effect after 0.5 h. The permeability on E. coli cell membrane increased progressively during combined treatment with peptide TGH2 and low-intensity ultrasound, resulting in the leakage of intracellular solutes, as shown by transmission electron microscopy (TEM). Structural analysis using circular dichroism (CD) revealed that peptide TGH2 has an α-helical structure, showing a slight untwisting effect under 0.3 W/cm2 ultrasound treatment for 0.5 h. The findings here provide new insight into the potential application of ultrasound and AMPs combination in food preservation.


Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Escherichia coli/efeitos dos fármacos , Peptídeos/química , Peptídeos/farmacologia , Ondas Ultrassônicas , Sequência de Aminoácidos , Cinética , Testes de Sensibilidade Microbiana
10.
Eur Radiol ; 2021 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-34327578

RESUMO

OBJECTIVES: To assess whether dual-energy computed tomography (DECT), using conventional computed tomography or magnetic resonance imaging as a reference standard, is sufficiently accurate to differentiate intracerebral hemorrhage from contrast extravasation after endovascular thrombectomy for acute ischemic stroke. METHODS: On January 20, 2021, we searched the PubMed Medline, Embase, Web of Science, and Cochrane Library databases. QUADAS-2 was used to assess the risk of bias and applicability. Meta-analyses were performed using a bivariate random-effects model. To explore sources of heterogeneity, meta-regression analyses were performed. Deeks' funnel plot asymmetry test was used to assess publication bias. RESULTS: A total of 7 studies (269 patients, 269 focal areas) were included. The pooled mean sensitivity, specificity, and accuracy of DECT in identifying intracerebral hemorrhage from contrast extravasation after mechanical thrombectomy for acute ischemic stroke were 0.77 (95% confidence interval (CI) 0.29 to 0.96), 1 (95% CI 0.86 to 1), and 0.99 (95% CI 0.98 to 1), respectively. This evidence was of moderate certainty due to the risk of bias. Higgin's I-squared for study heterogeneity was observed for the pooled sensitivity (I2 = 78.88%) and pooled specificity (I2 = 82.12%). Moreover, Deeks' funnel plot asymmetry test revealed no publication bias (p = 0.38). CONCLUSION: DECT shows excellent accuracy and specificity in differentiating intracerebral hemorrhage from contrast extravasation after endovascular thrombectomy for acute ischemic stroke. Nevertheless, there was substantial and moderate heterogeneity among the studies. Future large-scale, prospective cohort studies are warranted to validate our findings. KEY POINTS: • Dual-energy computed tomography shows excellent accuracy and specificity in differentiating intracerebral hemorrhage from contrast extravasation after endovascular thrombectomy for acute ischemic stroke. • Via meta-regression analysis, we found various possible covariates, including the publication date, image analysis, index test time, time of follow-up imaging, and reference standard judgment, that had an important effect on the heterogeneity. • There were no concerns regarding applicability in any of the included studies.

11.
Dev Comp Immunol ; 125: 104147, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34111502

RESUMO

In the Notch signaling pathway in vertebrates and invertebrates, the ligand Delta plays crucial roles in cell proliferation, differentiation, and immunity. Although the Notch signaling pathway has recently been implicated in the immune defense of Penaeus vannamei, the association of Delta with this immune response remains unclear. Here, we cloned and characterized the Delta homolog in P. vannamei (designated as PvDelta). PvDelta has a 2493 bp open reading frame (ORF) encoding a putative protein of 830 amino acids. Bioinformatics analysis revealed that PvDelta contains an N-terminal signal peptide, a conserved Notch ligand (MNNL) domain, a Delta/Serrate/Lag-2 segment, 9 epidermal growth factors segments, a transmembrane domain, and shares high homology with other Delta family members. Transcripts of PvDelta were detected in all shrimp tissues tested and were induced by Vibrio parahaemolyticus, white spot syndrome virus (WSSV), and lipopolysaccharide (LPS), indicating its involvement in shrimp immune response. Moreover, after PvDelta knockdown followed by LPS stimulation, the expression of Notch signaling pathway genes (i.e., PvNotch, PvCSL, and PvHey) was downregulated. Finally, shrimp depleted of PvDelta showed a lower survival rate in response to V. parahaemolyticus challenge. In sum, our data reveal that PvDelta is involved in the innate immunity of shrimp by positively modulating the Notch signaling pathway.

12.
Front Immunol ; 12: 697397, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34122458

RESUMO

Shrimp aquaculture is an essential economic venture globally, but the industry faces numerous challenges, especially pathogenic infections. As invertebrates, shrimp rely mainly on their innate immune system for protection. An increasing number of studies have shown that ubiquitination plays a vital role in the innate immune response to microbial pathogens. As an important form of posttranslational modification (PTM), both hosts and pathogens have exploited ubiquitination and the ubiquitin system as an immune response strategy to outwit the other. This short review brings together recent findings on ubiquitination and how this PTM plays a critical role in immune modulation in penaeid shrimps. Key findings inferred from other species would help guide further studies on ubiquitination as an immune response strategy in shrimp-pathogen interactions.

13.
Mol Neurobiol ; 58(7): 3362-3373, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33694060

RESUMO

Glioma is the most common type of solid tumour affecting the central nervous system, and the survival rate of patients with glioma is low. However, the mechanism associated with glioma progression remains unclear. Growing evidence suggests that lncRNAs play essential roles in the initiation and progression of tumours, including gliomas. In the present study, we identified and verified the expression of the novel lncRNA RP11-279C4.1 by analyzing the TANRIC database and performing qRT-PCR assays, the results of which revealed its upregulation in glioma tissues and cell lines. The results of multiple functional experiments demonstrated that RP11-279C4.1 knockdown inhibited glioma malignant phenotypes, including cell proliferation, migration, invasion and cell self-renew ability in vitro. In addition, RP11-279C4.1 downregulation suppressed tumour growth in vivo. Mechanistically, RP11-279C4.1 induced CBX3 activation via competitively sponging miR-1273g-3p, and rescue assay results confirmed the importance of the RP11-279C4.1/miR-1273g-3p/CBX3 axis. Overall, the results of our present study demonstrated that RP11-279C4.1 functions as an oncogene that promotes tumour progression by modulating the miR-1273g-3p/CBX3 axis in glioma, suggesting that RP11-279C4.1 may be a novel therapeutic target for glioma.

14.
Cell Biol Toxicol ; 2021 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-33630204

RESUMO

Current cancer treatment regimens such as chemotherapy and traditional chemical drugs have adverse side effects including the appearance of drug-resistant tumor cells. For these reasons, it is imperative to find novel therapeutic agents that overcome these factors. To this end, we explored a cationic antimicrobial peptide derived from Litopenaeus vannamei hemocyanin (designated LvHemB1) that induces cancer cell death, but sparing normal cells. LvHemB1 inhibits the proliferation of human cervical (HeLa), esophageal (EC109), hepatocellular (HepG2), and bladder (EJ) cancer cell lines, but had no significant effect on normal liver cell lines (T-antigen-immortalized human liver epithelial (THLE-3) cells). In addition to its antiproliferative effects, LvHemB1 induced apoptosis, by permeating cells and targeting mitochondrial voltage-dependent anion channel 1 (VDAC1). Colocalization studies revealed the localization of LvHemB1 in mitochondria, while molecular docking and pull-down analyses confirmed LvHemB1-VDAC1 interaction. Moreover, LvHemB1 causes loss in mitochondrial membrane potential and increases levels of reactive oxygen species (ROS) and apoptotic proteins (caspase-9, caspase-3, and Bax (Bcl-2-associated X)), which results in mitochondrial-mediated apoptosis. Thus, peptide LvHemB1 has the potential of being used as an anticancer agent due to its antiproliferation effect and targeting to VDAC1 to cause mitochondrial dysfunction in cancer cells, as well as its ability to induce apoptosis by increasing ROS levels, and the expression of proapoptotic proteins.

16.
Virulence ; 12(1): 481-492, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-33487106

RESUMO

Acute hepatopancreatic necrosis disease (AHPND) is a major debilitating disease that causes massive shrimp death resulting in substantial economic losses in shrimp aquaculture. The Pir toxin proteins secreted by a unique strain of Vibrio parahaemolyticus play an essential role in the pathogenesis of AHPND. At present, most studies on the effects of Pir toxin proteins in shrimp focus on digestive tissues or organs such as hepatopancreas, stomach, etc., with none on the immune organs. In the present study, two recombinant Pir toxin proteins (rPirA and rPirB) of V. parahaemolyticus were expressed with rPirB shown to enter shrimp hemocytes. Employing pull-down and LC-MS/MS analysis, GST-rPirB was found to interact with 13 proteins in hemocytes, including histone H3 and histone H4 and among which histone H4 had the highest protein score. Further analysis using GST pull-down and Far-Western blot analysis revealed that rPirB could interact with histone H4. In addition, using the purified nucleosome protein from Drosophila S2 cells, it was found that PirB protein could specifically bind to histones. When flow cytometry was applied, it was observed that the interaction between PirB and histones in shrimp hemocytes induces apoptosis, which results in the dephosphorylation of Serine 10 in histone H3. Collectively, the current study shows that in addition to its effect on the digestive tract of shrimp, the PirB toxin protein interacts with histones to affect the phosphorylation of histone H3-S10, thereby inducing apoptosis.


Assuntos
Apoptose/genética , Toxinas Bacterianas/genética , Toxinas Bacterianas/metabolismo , Hemócitos/patologia , Histonas/metabolismo , Vibrio parahaemolyticus/química , Animais , Aquicultura , Drosophila/citologia , Hepatopâncreas/patologia , Penaeidae/citologia , Penaeidae/microbiologia , Fosforilação , Proteínas Recombinantes/genética , Vibrio parahaemolyticus/patogenicidade
17.
Virus Res ; 291: 198218, 2021 01 02.
Artigo em Inglês | MEDLINE | ID: mdl-33152380

RESUMO

Singapore grouper iridovirus (SGIV) is a large double-stranded DNA virus that is a major threat to grouper aquaculture. The pathogenesis of SGIV is not well understood so far. Previous studies have revealed that ICP18, an immediate early protein encoded by SGIV ORF086R gene, promotes viral replication by regulating cell proliferation and virus assembly. In the present study, the potential functions of ICP18 were further explored by probing into its interactors using a proximity-dependent BioID method. Since our in-house grouper embryonic cells (a natural host cell of SGIV) could not be efficiently transfected with the plasmid DNA, and the grouper genome data for mass spectrometry-based protein identification is not currently available, we chosen a non-permissive cell (HEK293 T) as a substitute for this study. A total of 112 cellular proteins that potentially bind to ICP18 were identified by mass spectrometry analysis. Homology analysis showed that among these identified proteins, 110 candidate ICP18-interactors had homologous proteins in zebrafish (a host of SGIV), and shared high sequence identity. Further analysis revealed that the identified ICP18-interacting proteins modulate various cellular processes such as cell cycle and cell adhesion. In addition, the interaction between ICP18 and its candidate interactor, i.e., cyclin-dependent kinase1 (CDK1), was confirmed using Co-immunoprecipitation (Co-IP) and Pull-down assays. Collectively, our present data provides additional insight into the biological functions of ICP18 during viral infection, which could help in further unraveling the pathogenesis of SGIV.


Assuntos
Bass/virologia , Iridovirus/metabolismo , Proteínas Virais/metabolismo , Animais , Adesão Celular , Ciclo Celular , Quinases Ciclina-Dependentes/metabolismo , Doenças dos Peixes/virologia , Células HEK293 , Humanos , Iridovirus/química , Iridovirus/classificação , Iridovirus/genética , Espectrometria de Massas/métodos , Domínios e Motivos de Interação entre Proteínas , Singapura , Proteínas Virais/genética , Replicação Viral
18.
J Proteomics ; 232: 104074, 2021 02 10.
Artigo em Inglês | MEDLINE | ID: mdl-33309928

RESUMO

Notch signaling pathway is a highly evolutionary conserved signaling pathway, which modulates many biological processes such as cell differentiation, tissue development and immune response. Our previous study revealed that Litopenaeus vannamei Notch (LvNotch) was involved in immune response by regulating reactive oxygen species (ROS) production in hemocytes. However, the immune regulatory networks mediated by LvNotch remain unclear in shrimp. In this study, 21 proteins that potentially interact with LvNotch were identified by GST pull-down and liquid chromatography tandem mass spectrometry (LC-MS/MS) analyses. Among these proteins, COP9 signalosome complex subunit 1 (CSN1) was chosen for further studies due to its putative role in immune response. The interaction between LvNotch and LvCSN1 was confirmed by Far-Western blot and GST pull-down analyses. In vivo knockdown of LvNotch resulted in an increase in LvCSN1 expression in hemocytes, which suggest that the COP9 signalosome complex might be negatively regulated by LvNotch. In addition, in vivo silencing of LvNotch upregulated the expression of LvDorsal, LvTNFSF and LvCrustin2 (NF-κB pathway related-genes), while their expression decreased after LvCSN1 depletion. Collectively, the current results indicate that LvNotch negatively regulates the NF-κB pathway by modulating LvCSN1 in shrimp. SIGNIFICANCE: Although the Notch signaling pathway has been implicated in the regulation of immune response in vertebrates and invertebrates, the functions and immune-related interacting networks of Notch in shrimp immune response remain unknown. In this study, twenty-one proteins including COP9 signalosome complex subunit 1 (CSN1) were identified as potential interacting partners of LvNotch. Further analysis revealed that LvNotch negatively regulates the NF-κB pathway by binding to CSN1 and modulating its expression. These findings for the first time suggest that the Notch signaling pathway has cross-talk with the NF-κB pathway in shrimp as part of the immune response.


Assuntos
NF-kappa B , Penaeidae , Animais , Proteínas de Artrópodes , Complexo do Signalossomo COP9 , Cromatografia Líquida , Espectrometria de Massas em Tandem
19.
Front Immunol ; 11: 574721, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33224140

RESUMO

Aquaculture production of crustaceans (mainly shrimp and crabs) has expanded globally, but disease outbreaks and pathogenic infections have hampered production in the last two decades. As invertebrates, crustaceans lack an adaptive immune system and mainly defend and protect themselves using their innate immune system. The immune system derives energy and metabolites from nutrients, with amino acids constituting one such source. A growing number of studies have shown that amino acids and their metabolites are involved in the activation, synthesis, proliferation, and differentiation of immune cells, as well as in the activation of immune related signaling pathways, reduction of inflammatory response and regulation of oxidative stress. Key enzymes in amino acid metabolism have also been implicated in the regulation of the immune system. Here, we reviewed the role played by amino acids and their metabolites in immune-modulation in crustaceans. Information is inferred from mammals and fish where none exists for crustaceans. Research themes are identified and the relevant research gaps highlighted for further studies.


Assuntos
Aminoácidos/imunologia , Crustáceos/imunologia , Sistema Imunitário/imunologia , Imunidade Inata , Aminoácidos/metabolismo , Animais , Crustáceos/metabolismo , Sistema Imunitário/metabolismo , Transdução de Sinais , Especificidade da Espécie
20.
Int J Ophthalmol ; 13(10): 1621-1628, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33078114

RESUMO

AIM: To provide a detailed description of the natural history of persistent subretinal fluid (SRF) after successful repair of rhegmatogenous retinal detachment (RRD) and its association with visual outcome. METHODS: This was a prospective long-term follow-up for eyes undergoing scleral buckling (SB) surgery for macula-off RRD. Examinations were carried out preoperatively and postoperatively at 1, 3, 6, 9 and 12mo, until persistent SRF had completely resolved. One month postoperatively, optical coherence tomography (OCT) was used to classify SRF into three patterns: bleb-like loculated (BL), shallow-diffused (SD), and multiple blebs (MB). Serial OCT imaging was used to evaluate morphological changes in SRF until its complete disappearance. Patients were divided into two groups depending on the presence or absence of persistent SRF. RESULTS: A total of 59 patients (59 eyes) were included. There were no statistical differences between two groups at baseline, except for the proportion of patients with high myopia and a younger age. One month after surgery, OCT detected persistent SRF in 49 eyes (83.1%). The 3 morphological patterns of SRF were observed in 27 eyes (55.1%) with BL, 13 eyes (26.5%) with SD, and 9 eyes (18.4%) with MB. The mean time for complete absorption differed significantly across the three SRF patterns (F=8.097, P=0.001), which was 8.8±6.1, 20.1±12.1, and 16.7±10.2mo in BL, SD, and MB, respectively. In 9 of the 13 eyes with SD, the pattern transformed into MB type. In cases involving MB, the size and number of blebs decreased gradually until they had been completely absorbed. Eyes with persistent SRF were more likely to demonstrate disruption of the ellipsoid zone (49.0% vs 10%, P=0.034). The final best-corrected visual acuity of two groups was 0.37±0.11 (with SRF) vs 0.34±0.12 (without SRF) logMAR (P=0.499), respectively. CONCLUSION: High preoperative myopia and younger age are associated with persistent SRF. BL is the most commonly observed pattern with the shortest duration and gradually disappeared. Most cases involving SD SRF transform into MB type during resolution. The size and number of the MBs decrease gradually until they were completely absorbed. The absence of persistent SRF may contribute to slow visual recovery in the short-term but does not influence the final visual outcome.

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