ABSTRACT
NOD1 and NOD2 as two representative members of nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) family play important roles in antimicrobial immunity. However, transcription mechanism of nod1 and nod2 and their signal circle are less understood in teleost fish. In this study, with the cloning of card9 and ripk2 in Chinese perch, the interaction between NOD1, NOD2, and CARD9 and RIPK2 were revealed through coimmunoprecipitation and immunofluorescence assays. The overexpression of NOD1, NOD2, RIPK2 and CARD9 induced significantly the promoter activity of NF-κB, IFNh and IFNc. Furthermore, it was found that nod1 and nod2 were induced by poly(I:C), type I IFNs, RLR and even NOD1/NOD2 themselves through the ISRE site of their proximal promoters. It is thus indicated that nod1 and nod2 can be classified also as ISGs due to the presence of ISRE in their proximal promoter, and their expression can be mechanistically controlled through PRR pathway as well as through IFN signaling in antiviral immune response.
Subject(s)
Fish Proteins , Nod1 Signaling Adaptor Protein , Nod2 Signaling Adaptor Protein , Receptor-Interacting Protein Serine-Threonine Kinase 2 , Signal Transduction , Animals , Nod1 Signaling Adaptor Protein/genetics , Nod1 Signaling Adaptor Protein/metabolism , Receptor-Interacting Protein Serine-Threonine Kinase 2/metabolism , Receptor-Interacting Protein Serine-Threonine Kinase 2/genetics , Nod2 Signaling Adaptor Protein/genetics , Nod2 Signaling Adaptor Protein/metabolism , Fish Proteins/genetics , Fish Proteins/metabolism , Fish Proteins/immunology , Perches/genetics , Perches/immunology , Perches/metabolism , Interferons/metabolism , Interferons/genetics , Promoter Regions, Genetic , Transcription, Genetic , Immunity, Innate/genetics , Protein BindingABSTRACT
Type IV interferon (IFN) has been shown to be a cytokine with antiviral activity in fish and amphibian. But, it has not been cloned and characterized functionally in avian species. In this study, type IV IFN, IFN-υ, and its 2 possible receptors, IFN-υR1 and IL10RB, were identified from an avian species, the mallard (Anas platyrhynchos). Mallard IFN-υ has a 531 bp open reading frame (ORF), encoding 176 amino acids (aa), and has highly conserved features as reported in different species, with an N-terminal signal peptide and a predicted multi-helix structure. The IFN-υR1 and IL10RB contain 528 and 343 aa, respectively, with IFN-υR1 protein containing JAK1 and STAT binding sites, and IL10RB containing TYK2 binding site. These 2 receptor subunits also possess 3 domains, the N-terminal extracellular domain, the transmembrane domain, and the C-terminal intracellular domain. Expression analysis indicated that IFN-υ, IFN-υR1 and IL10RB were widely expressed in examined organs/tissues, with the highest level observed in pancreas, blood, and kidney, respectively. The expression of IFN-υ, IFN-υR1 and IL10RB in liver, spleen or kidney was significantly upregulated after stimulation with polyI:C. Furthermore, recombinant IFN-υ protein induced the expression of ISGs, and the receptor of IFN-υ was verified as IFN-υR1 and IL10RB using a chimeric receptor approach in HEK293 cells. Taken together, these results indicate that IFN-υ is involved in the host innate immune response in mallard.
Subject(s)
Avian Proteins , Ducks , Interleukin-10 Receptor beta Subunit , Animals , Ducks/genetics , Interleukin-10 Receptor beta Subunit/genetics , Interleukin-10 Receptor beta Subunit/chemistry , Interleukin-10 Receptor beta Subunit/metabolism , Avian Proteins/genetics , Avian Proteins/chemistry , Avian Proteins/metabolism , Amino Acid Sequence , Phylogeny , Receptors, Interferon/genetics , Receptors, Interferon/metabolism , Receptors, Interferon/chemistry , Sequence Alignment/veterinary , Immunity, Innate , Interferons/genetics , Interferons/metabolism , Gene Expression Profiling/veterinaryABSTRACT
Type II interferons (IFNs) are a key class of molecules regulating innate and adaptive immunity in vertebrates. In the present study, two members of the type II IFNs, IFN-γ and IFNγ-rel, were identified in the blunt snout bream (Megalobrama amblycephala). The open reading frame (ORF) of IFN-γ and IFNγ-rel was found to have 564 bp and 492 bp, encoding 187 and 163 amino acids, with the first 26 and 24 amino acids being the signal peptide, respectively. IFN-γ and IFNγ-rel genes showed a high degree of similarity to their zebrafish homologues, being 76.9 % and 58.9 %, respectively. In the phylogenetic tree, IFN-γ and IFNγ-rel were clustered with homologous genes in cyprinids. In blunt snout bream, IFN-γ and IFNγ-rel were constitutively expressed in trunk kidney, head kidney, spleen, liver, heart, muscle, gill, intestine and brain and were significantly up-regulated by poly (I:C) induction in head kidney, spleen, liver, gill and intestine. Using recombinant proteins of IFN-γ and IFNγ-rel, the surface plasmon resonance (SPR) results showed that IFN-γ was bound to CRFB6, CRFB13 and CRFB17, but mainly to CRFB6 and CRFB13, whereas IFN-γrel bound mainly to CRFB17 and had no affinity with CRFB6. These results contribute to a better understanding on type II IFNs and their receptor usage in teleost fish.
Subject(s)
Cyprinidae , Zebrafish , Animals , Zebrafish/metabolism , Phylogeny , Interferon-gamma/genetics , Interferon-gamma/metabolism , Amino Acid Sequence , Fish Proteins/chemistry , Recombinant Proteins/genetics , Amino Acids/geneticsABSTRACT
Bedding materials are important for suckling buffalo calves. Treated dung has been used as a bedding material for dairy cows but the lack of an appropriate safety assessment limits its application. In this study, we evaluated the feasibility of treated dung (TD) as a bedding material for suckling calves by comparing TD with rice husk (RH) and rice straw (RS) bedding materials. The TD was prepared through high-temperature composting by Bacillus subtilis. Thirty-three newborn suckling buffalo calves (Bubalus bubalis, 40.06 ± 5.79 kg) were randomly divided into 3 bedding material groups (TD, RH, and RS) and bedded with 1 of the 3 bedding materials for 60 d. We compared cost, moisture content, bacterial counts, and microbial composition of the 3 bedding materials, and investigated growth performance, health status, behavior, rumen fermentation, and blood parameters of bedded calves. The results showed that TD contained the fewest gram-negative bacteria and coliforms on d 1 and 30 and the lowest relative abundance of Staphylococcus throughout the experiment. The RH and TD bedding materials had the lowest cost. Calves in the TD and RS groups showed a higher dry matter intake, and final body weight and average daily gain in the TD and RS groups tended to be higher than in the RH group. Calves in the TD and RS groups had a lower disease incidence (diarrhea and fever), fewer antibiotic treatments, and lower fecal score than calves in the RH group. Higher contents of IgG, IgA, and IgM were observed in calves of the TD and RS groups than in calves of the RH group on d 10, indicating higher immune ability in TD and RS groups. Furthermore, TD bedding increased the butyric acid content in the calf's rumen, whereas RS bedding increased the acetate content, which might be attributed to the longer time and higher frequency of eating bedding material in the RS group. Considering all of the above indicators, we concluded that TD is the optimal bedding material for calves based on economics, bacterial count, microbial diversity, growth performance, and health status. Our findings provide a valuable reference for bedding material choice and calf farming.
Subject(s)
Composting , Female , Animals , Cattle , Buffaloes , Temperature , Body Weight , Bedding and Linens , Health Status , Diet/veterinary , Animal Feed/analysis , Weaning , RumenABSTRACT
The class II cytokine receptor family members are receptors of class 2 helical cytokines in mammals, and are named cytokine receptor family B (CRFB) in fish. In zebrafish, sixteen members, including CRFB1, CRFB2 and CRFB4-17 were reported. With the availability of genome sequence, a total of nineteen CRFBs was identified in the blunt snout bream (Megalobrama amblycephala), including CRFB1, CRFB2, CRFB4-17 with the presence of three CRFB9 isoforms, and two CRFB14 isoforms. These CRFB molecules contain well conserved features, such as fibronectin type III (FNIII) domain, transmembrane and intracellular domains as other class II cytokine receptors, and are phylogenetically grouped into thirteen clades with their homologues from other species of fish. The CRFB genes were constitutively expressed in organs/tissues examined in the fish. The finding of more CRFB members in the bream may provide clues to understand possible receptor-ligand interaction and their diversity from an evolutionary point of view.
Subject(s)
Cyprinidae , Zebrafish , Animals , Cyprinidae/genetics , Fish Proteins/genetics , Protein Isoforms , Receptors, Cytokine , Zebrafish/geneticsABSTRACT
AIM: To establish and assess a computed tomography (CT)-based radiomics nomogram for identifying malignant and benign Bosniak IIF masses. MATERIALS AND METHODS: In total, 150 patients with Bosniak IIF masses were separated into a training set (n=106) and a test set (n=44) in a ratio of 7:3. A radiomics signature was calculated based on extracted features from the three phases of CT images. A clinical model was constructed based on clinical characteristics and CT features, and a nomogram incorporating the radiomics signature and independent clinical variables was established. The calibration ability, discrimination accuracy, and clinical value of the nomogram model were assessed. RESULTS: Twelve features derived from CT images were applied to establish the radiomics signature. The performance levels of three machine-learning models were improved by adding the synthetic minority oversampling technique algorithm. The optimised machine learning model was a combination of the minimum redundancy maximum relevance-least absolute shrinkage and selection operator feature screening method + logistic regression classifier + synthetic minority oversampling technique algorithm, which demonstrated excellent identification ability on the test set (area under the curve [AUC], 0.970; 95% confidence interval [CI], 0.940-1.000). The nomogram model displayed outstanding discrimination ability on the test set (AUC, 0.972; 95% CI, 0.942-1.000). CONCLUSIONS: The CT-based radiomics nomogram was useful for discriminating between malignant and benign Bosniak IIF masses, which improved the precision of preoperative diagnosis.
Subject(s)
Algorithms , Nomograms , Humans , Area Under Curve , Calibration , Tomography, X-Ray ComputedABSTRACT
In mammals, type II interferon (IFN; i.e. IFN-γ) signalling transduces through its specific receptors IFN-γR1 and IFN-γR2. In an osteoglossiform fish, the arapaima Arapaima gigas, three type II IFNs, IFN-γ-like, IFN-γ and IFN-γrel, and their four possible receptor subunits IFN-γR1-1, IFN-γR1-2, IFN-γR2-1 and IFN-γR2-2 were identified in this study. The three type II IFN genes are composed of four exons and three introns, and they all contain IFN-γ signature motif and signal peptide, with the presence of potential nuclear localization signal (NLS) in IFN-γ-like and IFN-γ. The IFN-γR1-1, IFN-γR1-2, IFN-γR2-1 and IFN-γR2-2 are composed of seven exons and six introns, with predicted IFN-γR1-1 and IFN-γR1-2 proteins containing JAK1 and STAT1 binding sites, and IFN-γR2-1 and IFN-γR2-2 containing JAK2 binding sites. Gene synteny analysis showed that the type II IFN and their receptor loci are duplicated in arapaima. All these genes were expressed constitutively in all organs/tissues examined, and responded to the stimulation of polyI:C. The prokaryotic recombinant IFN-γ-like, IFN-γ and IFN-γrel proteins can significantly induce the upregulation of immune-related genes in trunk kidney leucocytes. The ligand-receptor relationship analyses revealed that recombinant IFN-γ-like, IFN-γ, and IFN-γrel transduce downstream signalling through IFN-γR1-1/IFN-γR2-1, IFN-γR1-2/IFN-γR2-2, and IFN-γR1-1, respectively, in xenogeneic cells with the overexpression of original or chimeric receptors. In addition, tyrosine (Y) 366 and Y377 in the intracellular region may be essential for the function of IFN-γR1-2 and IFN-γR1-1, respectively. The finding of type II IFN system in A. gigas thus provides different knowledge in understanding the diversity and evolution of type II IFN ligand-receptor relationships in vertebrates.
Subject(s)
Interferon-gamma , Mammals , Animals , Interferon-gamma/genetics , LigandsABSTRACT
As an important proinflammation and immunomodulatory cytokine, IL-18 has been reported in several species of fish, but its receptor subunits, IL-18Rα and IL-18Rß, and its decoy receptor, IL-18BP, have not been functionally characterized in fish. In the present study, IL-18Rα, IL-18Rß and IL-18BP were cloned from rainbow trout Oncorhynchus mykiss, and they possess common conserved domains with their mammalian orthologues. In tested organs/tissues, IL-18Rα and IL-18Rß exhibit basal expression levels, and IL-18BP has a pattern of constitutive expression. When transfected with diï¬erent combinations of chimeric receptors in HEK293T cells, recombinant IL-18 (rIL-18) can induce the activation of NF-κB only when pcDNA3.1-IL-18Rα/IL-1R1 and pcDNA3.1-IL-18Rß/IL-1RAP were both expressed. On the other hand, recombinant receptors, including rIL-18BP, rIL-18Rα-ECD-Fc and rIL-18Rß-ECD-Fc can down-regulate significantly the activity of NF-κB, suggesting the participation of IL-18Rα, IL-18Rß and IL-18BP in rainbow trout IL-18 signal transduction. Co-IP assays indicated that IL-18Rß may form a complex with MyD88, IRAK4, IRAK1, TRAF6 and TAB2 in HEK293T cells, indicating that IL-18Rß, in IL-18 signalling pathway, is associated with these signalling molecules. In conclusion, IL-18Rα, IL-18Rß and IL-18BP in rainbow trout are conserved in function and signalling pathway with their mammalian orthologues.
Subject(s)
Oncorhynchus mykiss , Humans , Animals , Receptors, Interleukin-18/metabolism , Oncorhynchus mykiss/metabolism , Carrier Proteins , Interleukin-18/genetics , Interleukin-18/metabolism , NF-kappa B/metabolism , HEK293 Cells , MammalsSubject(s)
Anastomotic Leak , Gastrectomy , Anastomosis, Surgical , Anastomotic Leak/surgery , Drainage , Humans , SuctionABSTRACT
N-myc-interactor (Nmi) belongs to interferon (IFN) stimulated genes (ISGs) and is involved in the regulation of physiological processes including viral infection, inflammatory response, apoptosis and tumorigenesis in mammals. However, the function of Nmi in teleost fish remains to be explored. In this study, an Nmi homologue was characterized from mandarin fish Siniperca chuatsi. The mandarin fish Nmi shares two conserved functional Nmi/IFP35 homology domains (NIDs) with mammalian Nmi protein in its C-terminal domain and a coiled coil region (CC) in its N-terminal domain, with its genomic DNA sequence consisting of nine exons and eight introns. Subcellular localization analysis shows that mandarin fish Nmi is a cytoplasmic protein and that its localization is dependent on the CC and NID1 regions. High and constitutive mRNA level of Nmi was observed in all examined tissues, with the highest level being observed in blood. In addition, the Nmi gene was significantly induced in various organs/tissues following the infection of infectious spleen and kidney necrosis virus (ISKNV), and its mRNA and protein level was also significantly induced in vitro after the treatment of IFNh, IFNc, as well as IFN-γ. The dual luciferase activity analysis indicated that the Nmi promoter was activated by the three type I IFNs through interferon-stimulated response element (ISRE) sites, and it can be also transcriptionally activated by IFN-γ via IRF1 which can activate the expression of Nmi through ISRE. Taken together, it is demonstrated in this study that the transcription of Nmi in mandarin fish can be regulated by type I and type II IFNs, thus confirming that Nmi in fish is also an ISG, and is involved in antiviral and IFN-induced innate immunity.
Subject(s)
Fish Diseases , Interferon Type I , Iridoviridae , Perciformes , Animals , Antiviral Agents , Fish Proteins/metabolism , Fishes , Interferon Type I/genetics , Interferon Type I/metabolism , Mammals/genetics , RNA, MessengerABSTRACT
Fish retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) are critical RNA sensors in cytoplasm and are involved in antiviral innate immunity. However, some species of fish lack RIG-I gene, and the function of RLR members in RIG-I-absent fish is poorly understood. In the present study, MDA5, LGP2 and MAVS genes were identified in commercially important snakehead Channa argus. But, RIG-I gene was not found in this fish, and a systematic analysis of RLRs in available genome database of fish indicated the absence of RIG-I in the Acanthomorphata, Clupeiformes and Polypteriformes, suggesting that loss events of RIG-I gene may have occurred independently three times in the evolutionary history of fish. The MDA5, LGP2 and MAVS in snakehead have conserved protein domains and genomic location based on sequence, phylogenetic and syntenic analyses. These genes are constitutively expressed in healthy fish and can be induced by polyinosinic and polycytidylic acid (poly(I:C)) stimulation in vitro. It is further revealed that the snakehead MDA5 and LGP2 have binding capacity with dsRNA, such as poly(I:C), and MDA5 can interact with MAVS, implying the antiviral function of MDA5 in the RIG-I-absent fish.
Subject(s)
RNA Helicases , Signal Transduction , Animals , DEAD Box Protein 58/genetics , DEAD Box Protein 58/metabolism , Immunity, Innate , Interferon-Induced Helicase, IFIH1/genetics , Interferon-Induced Helicase, IFIH1/metabolism , Phylogeny , RNA Helicases/genetics , RNA, Double-StrandedABSTRACT
OBJECTIVES: This study aimed to evaluate associations between mental distress and COVID-19-related changes in behavioral outcomes and potential modifiers (age, gender, educational attainment) of such associations. STUDY DESIGN: This was a cross-sectional study. METHODS: An online survey using anonymous network sampling was conducted in China from April to May 2020 using a 74-item questionnaire. A national sample of 10,545 adults in 31 provinces provided data on sociodemographic characteristics, COVID-19-related mental distress, and changes in behavioral outcomes. Structural equation models were used for data analyses. RESULTS: After adjusting for covariates, greater mental distress was associated with increased smoking (odds ratio [OR] = 1.42, 95% confidence interval [CI]: 1.20-1.68 and OR = 1.54, 95% CI: 1.31-1.82 per one standard deviation [SD] increase in mental distress) and alcohol consumption (OR = 1.67, 95% CI: 1.45-1.92 and OR = 1.47, 95% CI: 1.24-1.75 per one SD increase in mental distress) among current smokers and drinkers and with both increased and decreased physical activity (ORs ranged from 1.32 to 1.56). Underweight adults were more likely to lose body weight (≥1 kg; OR = 1.63, 95% CI: 1.30-2.04), whereas overweight adults were more likely to gain weight (OR = 1.61, 95% CI: 1.46-1.78) by the same amount. Association between mental distress and change in physical activity was stronger in adults aged ≥40 years (ORs ranged from 1.43 to 2.05) and those with high education (ORs ranged from 1.43 to 1.77). Mental distress was associated with increased smoking in males (OR = 1.60, 95% CI: 1.37-1.87) but not females (OR = 1.11, 95% CI: 0.82-1.51). CONCLUSIONS: Greater mental distress was associated with some positive and negative changes in behavioral outcomes during the pandemic. These findings inform the design of tailored public health interventions aimed to mitigate long-term negative consequences of mental distress on outcomes.
Subject(s)
COVID-19 , Pandemics , Adult , China/epidemiology , Cross-Sectional Studies , Humans , Male , SARS-CoV-2ABSTRACT
In fish, type I IFNs are classified into three groups, i.e. Group I, Group II and Group III, which are further divided into seven subgroups according to the number of conservative cysteines, phylogenetic relationship, and probably their receptor complexes. In the present study, four type I IFNs and four cytokine receptor family B members (CRFBs) were identified in the Asian arowana, Scleropages formosus, an ancient species in the Osteoglossomorpha with commercial and conservation values. According to multiple sequence alignment and phylogenetic relationship, the four type I IFNs are named as IFNa1, IFNa2, IFNb and IFNc, with the former two belonging to Group I, and the latter two to Group II. The four receptors are named as CRFB1, CRFB2, CRFB5a and CRFB5b. The IFNs and their possible receptor genes are widely expressed in examined organs/tissues, and are induced following the stimulation of polyinosinic polycytidylic acid (polyI:C) in vivo. It was found that IFNa1, IFNa2, IFNb and IFNc use preferentially the receptor complexes, CRFB1 and CRFB5b, CRFB1 and CRFB5b, CRFB2 and CRFB5a, and CRFB2 and CRFB5b, respectively, indicating the evolutionary diversification in the interaction of type I IFNs and their receptors in this ancient fish species, S. formosus.
Subject(s)
Fish Proteins/immunology , Fishes/immunology , Interferon Type I/immunology , Receptors, Interferon/immunology , Amino Acid Sequence , Animals , Fishes/genetics , Gene Expression , Interferon Type I/genetics , Phylogeny , Receptors, Interferon/geneticsABSTRACT
As a key molecule in innate immune signalling pathway, interleukin (IL)-1 receptor-associated kinase 1 (IRAK1) mediates downstream signalling cascades in immune response. In the present study, an IRAK1 orthologue was characterized from rainbow trout (Oncorhynchus mykiss), with a 2115 bp open reading frame (ORF), encoding a protein of 704 amino acids (aa). Multiple alignments showed that IRAK1 contains highly conserved features among different species, with a conservative N-terminal death domain (DD) and a C-terminal conserved serine/threonine protein kinase (STKc) domain. Expression analysis indicated that IRAK1 was widely expressed in examined organs/tissues, with the highest level observed in muscle and lowest in stomach. In RTG-2 cell line, the induced expression of IRAK1 was observed following the stimulation by the fish bacterial pathogen Flavobacterium columnare. Luciferase activity assays revealed that IRAK1 induced significantly the activity of NF-κB in Human embryonic kidney 293T (HEK293T) cell line; but after co-transfected with rainbow trout IL-1 receptor-associated kinase 4 (IRAK4), the induction was significantly down-regulated when compared with the expression of IRAK1 alone. Co-immunoprecipitation (Co-IP) assays indicated that IRAK1 was associated with rainbow trout myeloid differentiation factor 88 (MyD88), IRAK4 and TNF receptor associated factor 6 (TRAF6) in transfected HEK293T cells, and may form a complex with MyD88, IRAK4 and TRAF6 during the signalling pathway.
Subject(s)
Fish Proteins/genetics , Flavobacteriaceae Infections/metabolism , Flavobacterium/physiology , Interleukin-1 Receptor-Associated Kinases/genetics , Muscles/metabolism , Oncorhynchus mykiss/immunology , Animals , Cloning, Molecular , Conserved Sequence , Fish Proteins/metabolism , HEK293 Cells , Humans , Interleukin-1 Receptor-Associated Kinases/metabolism , Myeloid Differentiation Factor 88/metabolism , NF-kappa B/metabolism , Signal Transduction , TranscriptomeABSTRACT
In vertebrates, a total of eleven interferon (IFN) regulatory factors (IRFs), IRF1 to IRF11 are reported, with the conserved presence of IRF1 to IRF9 in all classes of vertebrates. However, IRF10 has been reported only in fish and birds, and IRF11 seems to be a fish specific IRF member. In this study, IRF11 in mandarin fish Siniperca chuatsi was found upregulated following virus infection, and IRF11 was localized constitutively in nucleus as revealed through immunofluorescence test. The overexpression and/or luciferase reporter assays showed that IRF11 can induce transcriptionally the ISRE activity, and the expression of type I IFNs, IFNc and IFNh, as well as the IFN-stimulated gene, Mx, thus inhibiting the Siniperca chuatsi rhabdovirus (SCRV) replication as indicated in the reduced expression of virus protein genes. It is thus suggested that IRF11 in mandarin fish and probably in other teleost fish can exert its antiviral effect through the upregulation of type I IFNs and ISGs.
Subject(s)
Cell Nucleus/metabolism , Fish Proteins/metabolism , Interferon Regulatory Factors/metabolism , Interferon Type I/metabolism , Perciformes/immunology , Rhabdoviridae Infections/immunology , Rhabdoviridae/physiology , Animals , Antiviral Agents/metabolism , Fish Proteins/genetics , Gene Expression Regulation , Interferon Regulatory Factors/genetics , Interferon Type I/genetics , Protein TransportABSTRACT
Interferon (IFN)-stimulated genes (ISGs) exert multiple functions in immune system, and IFN-induced protein 35 (IFP35), which is a member of ISG, has been suggested to be involved in numerous cellular activities including the regulation of antiviral immunity in mammals. However, the role of IFP35 in fish innate immunity remains largely unknown. In the present study, we characterized the IFP35 gene in mandarin fish Siniperca chuatsi, which contains two conserved Nmi/IFP35 homology domains (NIDs) at C-terminus, but no leucine zipper motif, with its genomic DNA sequence consisting of eight exons and seven introns. High and constitutive mRNA level of IFP35 was observed in all examined tissues, with the highest level being observed in gills. Moreover, the IFP35 gene was significantly induced in vivo for 120 h following the infection of infectious spleen and kidney necrosis virus (ISKNV), and its mRNA and protein level was also significantly induced in vitro following the treatment of poly I:C, IFNh, IFNc, as well as IFN-γ. The subcellular localization results indicated that exogenous IFP35 protein was mainly located in cytoplasm, while endogenous IFP35 protein was transferred into, or aggregated around, the nucleus with the induction of poly I:C or IFNs. The dual luciferase activity analysis indicated that the IFP35 promoter was activated by type I and type II IFNs through ISRE site. It is considered that IFP35 in fish is involved in antiviral, as well as in IFN-induced innate immunity.
Subject(s)
Fish Diseases/immunology , Fish Proteins/genetics , Fishes/immunology , Interferons/metabolism , Intracellular Signaling Peptides and Proteins/genetics , Animals , Cell Nucleus/metabolism , Cloning, Molecular , Fish Diseases/virology , Fish Proteins/metabolism , Fishes/virology , Gene Expression Regulation/immunology , HEK293 Cells , HeLa Cells , Humans , Immunity, Innate , Intracellular Signaling Peptides and Proteins/metabolism , Poly I-C/immunology , Promoter Regions, Genetic/geneticsABSTRACT
AIM: To construct and validate a radiomics-based machine-learning method for preoperative prediction of distant metastasis (DM) from soft-tissue sarcoma. MATERIALS AND METHODS: Seventy-seven soft-tissue sarcomas were divided into a training set (n=54) and a validation set (n=23). The performance of three feature selection methods (ReliefF, least absolute shrinkage and selection operator [LASSO], and regularised discriminative feature selection for unsupervised learning [UDFS]) and four classifiers, random forest (RF), logistic regression (LOG), K nearest neighbour (KNN), and support vector machines (SVMs), were compared for predicting the likelihood of DM. To counter the imbalance in the frequencies of DM, each machine-learning method was trained first without subsampling, then with the synthetic minority oversampling technique (SMOTE). The performance of the radiomics model was assessed using area under the receiver-operating characteristic curve (AUC) and accuracy (ACC) values. RESULTS: The performance of the LASSO and SVM algorithm combination used with SMOTE was superior to that of the algorithm combination alone. The combination of SMOTE with feature screening by LASSO and SVM classifiers had an AUC of 0.9020 and ACC of 91.30% in the validation dataset. CONCLUSION: A machine-learning model based on radiomics was favourable for predicting the likelihood of DM from soft-tissue sarcoma. This will help decide treatment strategies.
Subject(s)
Bone Neoplasms/secondary , Liver Neoplasms/secondary , Lung Neoplasms/secondary , Magnetic Resonance Imaging/methods , Preoperative Care/methods , Sarcoma/diagnostic imaging , Sarcoma/pathology , Algorithms , Bone Neoplasms/diagnosis , Female , Humans , Image Processing, Computer-Assisted/methods , Liver Neoplasms/diagnosis , Lung Neoplasms/diagnosis , Machine Learning , Male , Middle Aged , Predictive Value of Tests , Reproducibility of Results , Sarcoma/surgery , Support Vector MachineABSTRACT
Galectin-8 and galectin-9 belong to tandem repeat-type galectins, and in the present study, these two genes were cloned in mandarin fish Siniperca chuatsi. The open reading frame (ORF) of the mandarin fish galectin-8 and galectin-9 contains 942, and 1008 bp, encoding 313 and 335 amino acids, respectively. As a conserved feature, an N-terminal carbohydrate recognition domain (CRD), and a C-terminal CRD were observed in each of the two galectins in mandarin fish. In healthy fish, galectin-8 and -9 were constitutively expressed in all organs/tissues examined, and their expression can be induced following the stimulation of LPS and poly(I:C). It is obvious that galectin-8 had a higher increase at mRNA level following the stimulation of poly(I:C). It is further demonstrated that mandarin fish galectin-8 inhibited the growth of Flavobacterium columnare and Streptococcus agalactiae, and in addition to the two species of bacteria, galectin-9 inhibited also the growth of Edwardsiella piscicida, which provides the basis for further understanding their antibacterial role in immune response of mandarin fish.
Subject(s)
Fish Diseases/immunology , Galectins/genetics , Galectins/immunology , Immunity, Innate/genetics , Perciformes/genetics , Perciformes/immunology , Amino Acid Sequence , Animals , Edwardsiella/physiology , Enterobacteriaceae Infections/immunology , Enterobacteriaceae Infections/veterinary , Fish Proteins/chemistry , Fish Proteins/genetics , Fish Proteins/immunology , Flavobacteriaceae Infections/immunology , Flavobacteriaceae Infections/veterinary , Flavobacterium/physiology , Galectins/chemistry , Gene Expression Profiling/veterinary , Head Kidney/immunology , Leukocytes/immunology , Phylogeny , Sequence Alignment/veterinary , Streptococcal Infections/immunology , Streptococcal Infections/veterinary , Streptococcus agalactiae/physiologyABSTRACT
AIM: To prospectively evaluate multiple b-value diffusion-weighted imaging (DWI) in differentiating malignant from benign breast lesions. MATERIALS AND METHODS: The study cohort included 103 patients who underwent 3 T magnetic resonance imaging (MRI). The conventional sequences included T1-weighted dynamic contrast-enhanced, T1-weighted and T2-weighted fat-suppressed sequences, single b-value (b=0, 1000 s/mm2) DWI, and multiple b-values (12 values, from 0 to 3,000 s/mm2) DWI. Pathological diagnosis of breast lesions was based on the latest World Health Organization (WHO) guide on the pathology and immunohistochemistry of the breast. SPSS Statistics V19.0 was used for the statistics analysis. RESULTS: The following parameters were calculated: apparent diffusion coefficient (ADC), tissue diffusivity (D), perfusion fraction (f), pseudo-diffusion coefficient (D∗), distributed diffusion coefficient (DDC), and alpha (α) by the same radiologist twice (interval time of 3 months). There was good inter/intra-observer agreement for each of the parameters. The D, D∗, f, DDC, and α values were significantly different among malignant tumours, benign lesions, and normal breast tissue (p<0.05). CONCLUSION: D, f, DDC, α, and ADC values have good sensitivity and specificity, respectively. In addition, the combined use of D and f or DDC and α has good diagnostic performance. Thus, the applications of the new multi-b DWI variables or combined variables are promising.