Integrated bioinformatics identifies key mediators in cytokine storm and tissue remodeling during Vibrio mimicus infection in yellow catfish (Pelteobagrus fulvidraco)

IntroductionThe pathogenesis of Vibrio mimicus infection in yellow catfish (Pelteobagrus fulvidraco) remains poorly understood, particularly regarding the impact of infection with the pathogen on primary target organs such as the skin and muscle.MethodsIn this study, we aim to analyze the pathological intricacies of the skin and muscle of yellow catfish after being infected with V. mimicus using a 1/10 LC50 seven-day post-infection model. Furthermore, we have utilized integrated bioinformatics to comprehensively elucidate the regulatory mechanisms and identify the key regulatory genes implicated in this phenomenon.ResultsOur histopathological examination revealed significant pathological changes in the skin and muscle, characterized by necrosis and inflammation. Moreover, tissue remodeling occurred, with perimysium degeneration and lesion invasion into the muscle along the endomysium, accompanied by a transformation of type I collagen into a mixture of type I and type III collagens in the perimysium and muscle bundles. Our eukaryotic transcriptomic and 4D label-free analyses demonstrated a predominantly immune pathway response in both the skin and muscle, with downregulation observed in several cell signaling pathways that focused on focal adhesion-dominated cell signaling pathways. The upregulated genes included interleukins (IL)-1 and -6, chemokines, and matrix metallopeptidases (mmp)-9 and -13, while several genes were significantly downregulated, including col1a and col1a1a. Further analysis revealed that these pathways were differentially regulated, with mmp-9 and mmp-13 acting as the potential core regulators of cytokine and tissue remodeling pathways. Upregulation of NF-κB1 and FOSL-1 induced by IL-17C and Nox 1/2-based NADPH oxidase may have held matrix metallopeptidase and cytokine-related genes. Also, we confirmed these relevant regulatory pathways by qPCR and ELISA in expanded samples.DiscussionOur findings unequivocally illustrate the occurrence of a cytokine storm and tissue remodeling, mediated by interleukins, chemokines, and MMPs, in the surface of yellow catfish infected with V. mimicus. Additionally, we unveil the potential bidirectional regulatory role of MMP-9 and MMP-13. These results provide novel perspectives on the intricate immune response to V. mimicus infection in yellow catfish and highlight potential targets for developing therapies

Sodium Fluoride Arrests Renal G2/M Phase Cell-Cycle Progression by Activating ATM-Chk2-P53/Cdc25C Signaling Pathway in Mice

Background/Aims: Excessive fluoride intake can induce cytotoxicity, DNA damage and cell-cycle changes in many tissues and organs, including the kidney. However, the underlying molecular mechanisms of fluoride-induced renal cell-cycle changes are not well understood at present. In this study, we used a mouse model to investigate how sodium fluoride (NaF) induces cell-cycle changes in renal cells. Methods: Two hundred forty ICR mice were randomly assigned to four equal groups for intragastric administration of NaF (0, 12, 24 and 48 mg/kg body weight/day) for 42 days. Kidneys were taken to measure changes of the cell-cycle at 21 and 42 days of the experiment, using flow cytometry, quantitative real-time polymerase chain reaction (qRT-PCR) and western blot methods. Results: NaF, at more than 12 mg/kg body weight, induced G2/M phase cell-cycle arrest in the renal cells, which was supported by the finding of significantly increased percentages of renal cells in the G2/M phase. We found also that G2/M phase cell-cycle arrest was accompanied by up-regulation of p-ATM, p-Chk2, p-p53, p-Cdc25C, p-CDK1, p21, and Gadd45a protein expression levels; up-regulation of ATM, Chk2, p53, p21, and Gadd45a mRNA expression levels; down-regulation of CyclinB1, mdm2, PCNA protein expression levels; and down-regulation of CyclinB1, CDK1, Cdc25C, mdm2, and PCNA mRNA expression levels. Conclusion: In this mouse model, NaF, at more than 12 mg/ kg, induced G2/M phase cell-cycle arrest by activating the ATM-Chk2-p53/Cdc25C signaling pathway, which inhibits the proliferation of renal cells and development of the kidney. Activation of the ATM-Chk2-p53/Cdc25C signaling pathway is the mechanism of NaF-induced renal G2/M phase cell-cycle arrest in this model

In vitro and in vivo antiviral activity of monolaurin against Seneca Valley virus

IntroductionSurveillance of the Seneca Valley virus (SVV) shows a disproportionately higher incidence on Chinese pig farms. Currently, there are no vaccines or drugs to treat SVV infection effectively and effective treatment options are urgently needed.MethodsIn this study, we evaluated the antiviral activity of the following medium-chain fatty acids (MCFAs) or triglycerides (MCTs) against SVV: caprylic acid, caprylic monoglyceride, capric monoglyceride, and monolaurin.ResultsIn vitro experiments showed that monolaurin inhibited viral replication by up to 80%, while in vivo studies showed that monolaurin reduced clinical manifestations, viral load, and organ damage in SVV-infected piglets. Monolaurin significantly reduced the release of inflammatory cytokines and promoted the release of interferon-γ, which enhanced the viral clearance activity of this type of MCFA.DiscussionTherefore, monolaurin is a potentially effective candidate for the treatment of SVV infection in pigs

Characteristics of pathology and transcriptome profiling reveal features of immune response of acutely infected and asymptomatic infected of carp edema virus in Koi

Koi sleepy disease (KSD) is a high mortality and infection viral disease caused by carp edema virus (CEV), which was a serious threat to aquaculture of common carp and export trade of Koi worldwide. Asymptomatic infection is an important cause of the difficulty in preventing KSD and its worldwide spread, because asymptomatic infection can be activated under appropriate condition. However, the understanding of the molecular correlates of these infections is still unknown. The purpose of this study was to compare the pathology change, enzyme activity, immunoglobulin activity, host and viral gene expression differences in acutely infected and cohabiting asymptomatic Koi infected with CEV. Healthy Koi were used as a control. The gross pathology, histopathology and ultrastructural pathology showed the difference and characteristics damage to the tissues of Koi under different infection conditions. Periodic Acid-Schiff stain (PAS), enzyme activity and immunoglobulin activity revealed changes in the immune response of gill tissue between acutely infected, asymptomatic infected and healthy Koi. A total of 111 and 2484 upregulated genes and 257 and 4940 downregulated genes were founded in healthy Koi vs asymptomatic infected Koi and healthy Koi vs acutely infected Koi, respectively. Additionally, 878 upregulated genes and 1089 downregulated genes were identified in asymptomatic vs. acutely infected Koi. Immune gene categories and their corresponding genes in different comparison groups were revealed. A total of 3, 59 and 28 immune-related genes were identified in the group of healthy Koi vs asymptomatic infected Koi, healthy Koi vs acutely infected Koi and asymptomatic infected Koi vs acutely infected Koi, respectively. Nineteen immune-related genes have the same expression manner both in healthy Koi vs acutely infected Koi and asymptomatic Koi vs acutely infected Koi, while 9 immune-related genes were differentially expressed only in asymptomatic Koi vs acutely infected Koi, which may play a role in viral reactivation. In addition, 8 differentially expressed genes (DEGs) were validated by quantitative reverse transcription PCR (RT-qPCR), and the results were consistent with the RNA-Seq results. In conclusion, the data obtained in this study provide new evidence for further elucidating CEV-host interactions and the CEV infection mechanism and will facilitate the implementation of integrated strategies for controlling CEV infection and spread

Investigation of the anti‐pseudorabies virus activity of interferon lambda 3 in cultured porcine kidney epithelial cells

Abstract Background It has been reported Interferon‐λ (IFN‐λ) has stronger antiviral effect than other interferons. IFN‐λ can induce antiviral interferon stimulated genes (ISGs) in epithelia to protect against virus. Pseudorabies virus (PRV) infection in pigs resulting in fatal encephalitis in newborn piglets, respiratory disorders in finishing pigs, reproductive disorders in sows and other symptoms. Objectives Since the effect of IFN‐λ on inhibiting PRV proliferation is still unknown. Inthis study, we investigate the relative contribution of porcine IFN‐λ3 toward controlling the infection of PRV in vitro. Our findings may provide a new insight for the prevention and treatment of PRV. Methods Therefore, the antiviral assay, western blot, qRT‐PCR and ELISA assay were used to investigating the contribution of IFN‐λ against PRV in PK‐15 cells. Results Here, we demonstrate that the replication of PRV in PK‐15 cells was inhibited after pre‐treatment with IFN‐λ3, and such inhibition was dose dependent. Overexpression of IFN‐λ3 receptor (IFNLR) also restricted virus titre in PK‐15 cells. In addition, IFN‐λ3 also increased the mRNA and protein expression of interferon‐stimulated genes 15 (ISG15), 2′‐5′‐oligoadenylate synthase 1 (OAS1), IFN‐inducible transmembrane 3 (IFITM3) and myxoma resistance protein 1 (Mx1) in PRV‐infected PK‐15 cells. Other than modulation ISGs, IFN‐λ specifically activated IFN‐γ mRNA expression not IFN‐α or IFN‐β. Conclusions IFN‐λ3 had antiviral activity against PRV and the upregulation of ISGs and IFN‐γ mRNA expression may be the mechanism of IFN‐λ3's antiviral activities. Thus, IFN‐λ3 has a decisive function that greatly limits PRV replication in PK‐15 cells. Our study explores the antiviral activity of IFN‐λ3 on PRV for the first time

Histopathological Changes and Inflammatory Response in Specific Pathogen-Free (SPF) with Porcine Circovirus Type 3 Infection

Since the first report of PCV3 virus infection in 2016, it has been linked to multisystemic inflammation, reproductive failure, cardiac pathology, and clinical indications resembling porcine dermatitis and nephropathy syndrome (PDNS). However, the pathogenesis and clinical significance of PCV3 is still unclear. In this study, a PCV3 infection model was created using SPF pigs, and histopathology and fluorescence quantitative PCR were utilized to examine PCV3’s pathogenicity. Reductions in body weight gain and fever were observed during this study. However, other clinical signs such as Dermatitis and Nephropathy Syndrome were not observed through the study. Viremia was detected in the PCV3-inoculated group from 17 days post-inoculation (p.i.) until the end of the study. Nasal shedding was detected from 21 to 35 dpi and fecal shedding was detected during 25–33 days and 39 days, respectively. Gross lesions and histological evaluation were detected in various tissues and organs, including the lung, heart, kidney, lymph nodes, spleen, liver, small intestine, and testis. The heart, lung, liver, kidney, lymph nodes, and spleen showed pathological changes. The pathological features include swelling, inflammation, cell degeneration, necrosis, and hemorrhage. The lesions are consistent with multisystemic inflammation. Tissue viral load results showed only heart, lung, liver, kidney, lymph nodes, and spleen was positive by qRT-PCR. Moreover, the pro-inflammation cytokines in serum increased a lot in the PCV3-inoculated group compared to the control group, demonstrating that the induced inflammation response may be the cause of tissue damage in PCV3-infection. This study demonstrated that PCV3 can produce mild pathological damage to multiple organs, especially multisystemic inflammatory cell infiltration and prolonged viremia, viral shedding in nasal secretions. This is the first in vivo characterization of PCV3 infection in the SPF piglets model using isolated PCV3 strain, and this is also the first time to show the gross and pathological lesion with all tissue and organs in the PCV3-inoculated group. Our findings might serve as a starting point for more investigation into PCV3’s pathogenic mechanism

Antiviral Activity of Porcine IFN-λ3 and IFN-α against Porcine Rotavirus In Vitro

Interferons (IFNs) play a major role in the host’s antiviral innate immunity. In response to viral infection, IFNs bind their receptors and initiate a signaling cascade, leading to the accurate transcriptional regulation of hundreds of IFN-stimulated genes (ISGs). Porcine rotavirus (PoRV) belongs to genus Rotavirus of the Reoviridae family; the infection is a global epidemic disease and a major threat to the pig industry. In this study, we found that IFN-λ3 inhibited the replication of PoRV in both MA104 cells and IPEC-J2 cells, and this inhibition was dose-dependent. Furthermore, the antiviral activity of IFN-λ3 was more potent in IPEC-J2 cells than in MA104 cells. Further research showed that IFN-λ3 and IFN-α might inhibit PoRV infection by activating ISGs, i.e., MxA, OASL and ISG15, in IPEC-J2 cells. However, the co-treatment of IFN-λ3 and IFN-α did not enhance the antiviral activity. Our data demonstrated that IFN-λ3 had antiviral activity against PoRV and may serve as a useful antiviral candidate against PoRV, as well as other viruses in swine

Cloning and Expression Analysis of CMB1 Gene in Pepper

【Objective】CMB1 gene plays an important role in the regulation of inflorescence structure and fruit ripening in plants, but the function and regulatory mechanism of CMB1 gene in pepper have been rarely reported. Therefore, the role of transcription factors related to the regulation of carotenoid synthesis and CMB1 gene in pepper was studied so as to provide a reference for pepper breeding.【Method】The coding sequences of CMB1 were cloned by the homology sequence method, and the expression patterns of CMB1 in fruits of red and yellow peppers with wrinkled skin at different development stages (green ripening stage, color change stage, mature stage) were analyzed by real-time quantitative PCR.【Result】The ORF length of CMB1 gene was 732 bp in both red pepper and yellow pepper, encoding 243 amino acid residues. Sequence alignment showed that the mutation occurred at the 11th site of CMB1 base sequence, G mutated to T, resulting in amino acid sequence mutated from G (Glycine) to V (Valine). Bioinformatics analysis revealed that the CMB1-encoded protein of wrinkled skin red pepper had a molecular mass of 20 923.21, a theoretical isoelectric point of 5.59, an instability coefficient of 57.28, and a total mean hydrophilicity (gravimetric) index of -1.001. The CMB1-encoded protein of wrinkled skin yellow pepper had a molecular mass of 27 958.47, a theoretical isoelectric point of 7.10, an instability coefficient of 53.54, and a total mean hydrophilicity (gravimetric) index of -0.809. Both CMB1 of wrinkled skin red and yellow peppers were unstable hydrophobic proteins, both contained 38 phosphorylation sites, both were devoid of transmembrane structures and signal peptides, and both had typical MADS-box structural domains and K-box structural domains. Evolutionary tree analysis revealed that pepper CMB1 was most closely related to Solanaceae and most distantly related to Leguminosae. The results of fluorescence quantitative PCR showed that the expression of CMB1 increased sequentially in the three stages of green ripening, color change, and maturity of pepper fruits, and the expression of CMB1 in the three stages was significantly higher in wrinkled skin red pepper than in wrinkled skin yellow pepper.【Conclusion】Pepper CMB1 is a member of MADS-box, and its expression in pepper fruit is similar to that of carotenoid in pepper, suggesting that CMB1 is mainly involved in regulating the synthesis of carotenoid like capsanthin

A Novel Viscosity-Temperature Model of Glass-Forming Liquids by Modifying the Eyring Viscosity Equation

Many models have been created and attempted to describe the temperature-dependent viscosity of glass-forming liquids, which is the foundational feature to lay out the mechanism of obtaining desired glass properties. Most viscosity models were generated along with several impact factors. The complex compositions of commercial glasses raise challenges to settle these parameters. Usually, this issue will lead to unsatisfactory predicted results when fitted to a real viscosity profile. In fact, the introduction of the reliable viscosity-temperature data to viscosity equations is an effective approach to obtain the accurate parameters. In this paper, the Eyring viscosity equation, which is widely adopted for molecular and polymer liquids, was applied in this case to calculate the viscosity of glass materials. On the basis of the linear variation of molar volume with temperature during glass cooling, a modified temperature-dependent Eyring viscosity equation was derived with a distinguished mathematical expression. By means of combining high-temperature viscosity data and the glass transition temperature (Tg), nonlinear regression analysis was employed to obtain the accurate parameters of the equation. In addition, we have demonstrated that the different regression methods exert a great effect on the final prediction results. The viscosity of a series of glasses across a wide temperature range was accurately predicted via the optimal regression method, which was further used to verify the reliability of the modified Eyring equation

Apoptosis and DNA damage mediated by ROS involved in male reproductive toxicity in mice induced by Nickel

Nickel (Ni) is the most important environmental pollution in the world. Ni has been confirmed to have multi-organ toxicology and carcinogenicity. Recently, Ni also can impair the male reproductive system, however, its precious mechanism still has not been clarified. The current work found that nickel chloride (NiCl2) induced histopathological lesions in testis. And, the Johnsen's score, seminiferous tubule diameter, and spermatogenic epithelium thickness were decreased in NiCl2-treated mice. The number of spermatogonium, primary spermatocyte, and round spermatid also were significantly reduced after Ni treatment. Next the potential molecular mechanism was measured. NiCl2 treatment elevated ROS production in the testis. Additionally, NiCl2 was found to induce apoptosis with features including up-regulation of Bax, cleaved-caspase-3, cleaved-caspase-8, caspase-9, and caspase-12, while down-regulation of Bcl-2 expression. In the meantime, the marker protein of DNA damage γ-H2AX was significantly increased in NiCl2-primed mice testis. To clarify effects of reactive oxygen species (ROS) in apoptosis and DNA damage induced by NiCl2, NiCl2 was used to co-treat antioxidant NAC (N-Acetyl-L-cysteine). NAC weakened ROS production induced by NiCl2, and played an inhibition role in apoptosis and DNA damage. Moreover, co-treatment using NiCl2 and NAC group also eliminated spermatogenesis disorders. In summary, research results reveal the relations of spermatogenesis disorder induced by NiCl2 with apoptosis and DNA damage mediated by ROS and apoptosis in the testis