scholarly journals An iTRAQ-Based Comparative Proteomics Analysis of the Biofilm and Planktonic States of Aeromonas veronii TH0426

2020 ◽  
Vol 21 (4) ◽  
pp. 1450 ◽  
Author(s):  
Ying Li ◽  
Bintong Yang ◽  
Jiaxin Tian ◽  
Wuwen Sun ◽  
Guiqin Wang ◽  
...  
Keyword(s):  
Virulent Strain ◽  
Economic Losses ◽  
Planktonic Cells ◽  
Absolute Quantitation ◽  
Factors Affecting ◽  
Aeromonas Veronii ◽  
Biofilm Model ◽  
Aquaculture Industry ◽  
Isobaric Tags

Aeromonas veronii is a virulent fish pathogen that causes extensive economic losses in the aquaculture industry worldwide. In this study, a virulent strain of A. veronii TH0426 was used to establish an in vitro biofilm model. The results show that the biofilm-forming abilities of A. veronii TH0426 were similar in different media, peaking under conditions of 20 °C and pH 6. Further, isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics methods were used to compare the differential expression of A. veronii between the biofilm and planktonic cells. The results show alterations in 277 proteins, with 130 being upregulated and 147 downregulated. Pathway analysis and GO (Gene Ontology) annotations indicated that these proteins are mainly involved in metabolic pathways and the biosynthesis of secondary metabolites and antibiotics. These proteins are the main factors affecting the adaptability of A. veronii to its external environment. MRM (multiple reaction 27 monitoring) and qPCR (qPCR) were used to verify the differential proteins of the selected A. veronii. This is the first report on the biofilm and planktonic cells of A. veronii, thus contributing to studying the infection and pathogenesis of A. veronii.

scholarly journals Rhabdovirus Infection Is Dependent on Serine/Threonine Kinase AP2-Associated Kinase 1

Life ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 170
Author(s):  
Jun Luo ◽  
Yue Zhang ◽  
Yang Wang ◽  
Qing Liu ◽  
Luman Chen ◽  
...  
Keyword(s):  
Absolute Quantitation ◽  
Serine Threonine Kinase ◽  
Threonine Kinase ◽  
N2a Cells ◽  
Vesicular Stomatitis ◽  
Isobaric Tags ◽  
Potential Strategy ◽  
Prevention And Therapy

Rabies virus (RABV) causes a fatal neurological disease in both humans and animals. Understanding the mechanism of RABV infection is vital for prevention and therapy of virulent rabies infection. Our previous proteomics analysis based on isobaric tags for relative and absolute quantitation to identify factors revealed that RABV infection enhanced AP-2-associated protein kinase 1 (AAK1) in N2a cells. In this study, to further confirm the role of AAK1, we showed that RABV infection increased the transcription and expression of AAK1 in N2a cells. AAK1 knockdown significantly decreased RABV infection in both N2a and BHK-21 cells. AAK1 knockout inhibited RABV infection in N2a cells. Furthermore, inhibition of AAK1 kinase activity using sunitinib decreased RABV infection. However, AAK1 overexpression did not change RABV infection in vitro. Therapeutic administration of sunitinib did not significantly improve the survival rate of mice following lethal RABV challenge. In addition, AAK1 knockdown decreased infection in N2a cells by vesicular stomatitis virus, which is another rhabdovirus. These results indicate that rhabdovirus infection is dependent on AAK1 and inhibition of AAK1 is a potential strategy for the prevention and therapy of rabies.

scholarly journals Quantitative Proteomic Analyses of a Pathogenic Strain and Its Highly Passaged Attenuated Strain ofMycoplasma hyopneumoniae

2019 ◽  
Vol 2019 ◽  
pp. 1-18 ◽  
Author(s):  
Sha Li ◽  
Liurong Fang ◽  
Wei Liu ◽  
Tao Song ◽  
Fuwei Zhao ◽  
...  
Keyword(s):  
Inositol Phosphate ◽  
Virulent Strain ◽  
Absolute Quantification ◽  
Chronic Respiratory Disease ◽  
Economic Losses ◽  
Biological Processes ◽  
Inositol Phosphate Metabolism ◽  
Nucleoside Metabolism ◽  
Isobaric Tags ◽  
Future Work

Mycoplasma hyopneumoniaeis the causative agent of porcine enzootic pneumonia, a chronic respiratory disease in swine resulting in enormous economic losses. To identify the components that contribute to virulence and unveil those biological processes potentially related to attenuation, we used isobaric tags for relative and absolute quantification technology (iTRAQ) to compare the protein profiles of the virulentM. hyopneumoniaestrain 168 and its attenuated highly passaged strain 168L. We identified 489 proteins in total, 70 of which showing significant differences in level of expression between the two strains. Remarkably, proteins participating in inositol phosphate metabolism were significantly downregulated in the virulent strain, while some proteins involved in nucleoside metabolism were upregulated. We also mined a series of novel promising virulence-associated factors in our study compared with those in previous reports, such as some moonlighting adhesins, transporters, lipoate-protein ligase, and ribonuclease and several hypothetical proteins with conserved functional domains, deserving further research. Our survey constitutes an iTRAQ-based comparative proteomic analysis of a virulentM. hyopneumoniaestrain and its attenuated strain originating from a single parent with a well-characterized genetic background and lays the groundwork for future work to mine for potential virulence factors and identify candidate vaccine proteins.

scholarly journals Isobaric Tags for Relative and Absolute Quantitation (iTRAQ)-Based  proteomic analysis of mRNA splicing relevant proteins in aging HSPCs

2019 ◽  
Author(s):  
Xiaolan Lian ◽  
Lina Zhang
Keyword(s):  
Proteomic Analysis ◽  
Aging Process ◽  
Mrna Splicing ◽  
Absolute Quantitation ◽  
Qrt Pcr ◽  
Isobaric Tags ◽  
Spliceostatin A

Abstract Background: HSPC aging was closely associated with the organism aging, senile diseases and hematopoietic related diseases. Therefore, study on HSPC aging born great significance to further elucidate the mechanisms of aging and to treat hematopoietic disease resulted from HSPC aging. Little attention had been paid to mRNA splicing as a mechanism underlying HSPC senescence. Results: We used our lab’s patented aging model of HSPCs in vitro to analyze mRNA splicing relevant proteins alterations with iTRAQ based proteomic analysis. We found that not only the notable mRNA splicing genes such as SR, hnRNP, WBP11, Sf3b1, Ptbp1 and U2AF1 but also the scarcely reported mRNA splicing relavant genes such as Rbmxl1, Dhx16, Pcbp2, Pabpc1 were significantly down-regulated. We further verified their genes expressions by qRT-PCR. In addition, we reported the effect of Spliceostatin A (SSA), which inhibits mRNA splicing in vivo and in vitro, on HSPC aging. Conclusions: It was concluded that mRNA splicing emerged as an important vulnerability of HSPC aging. This study improved our understanding of the role of mRNA splicing in the HSPC aging process.

scholarly journals Transcriptomic Analysis of High- and Low-Virulence Bovine Pasteurella multocida in vitro and in vivo

2021 ◽  
Vol 8 ◽  
Author(s):  
Fang He ◽  
Zongling Zhao ◽  
Xiaoyan Wu ◽  
Lijie Duan ◽  
Nengzhang Li ◽  
...  
Keyword(s):  
Pasteurella Multocida ◽  
Virulent Strain ◽  
Survival Rates ◽  
Opportunistic Pathogen ◽  
Economic Losses ◽  
Sequencing Analysis ◽  
Serotype A ◽  
Naturally Occurring

Pasteurella multocida is a gram-negative opportunistic pathogen that causes various diseases in poultry, livestock, and humans, resulting in huge economic losses. Pasteurella multocida serotype A CQ6 (PmCQ6) is a naturally occurring attenuated strain, while P. multocida serotype A strain CQ2 (PmCQ2) is a highly virulent strain isolated from calves. Compared with PmCQ2, it was found that bacterial loads and tissue lesions of lung tissue significantly decreased and survival rates significantly improved in mice infected with PmCQ6 by intranasal infection. However, comparative genome analysis showed that the similarity between the two strains is more than 99%. To further explore the virulence difference mechanism of PmCQ2 and PmCQ6, transcriptome sequencing analysis of the two strains was performed. The RNA sequencing analysis of PmCQ2 and PmCQ6 showed a large number of virulence-related differentially expressed genes (DEGs) in vivo and in vitro. Among them, 38 virulence-related DGEs were significantly up-regulated due to PmCQ6 infection, while the number of PmCQ2 infection was 46, much more than PmCQ6. In addition, 18 virulence-related DEGs (capsule, iron utilization, lipopolysaccharide, and outer membrane protein-related genes) were up-regulated in PmCQ2 infection compared to PmCQ6 infection, exhibiting a higher intensive expression level in vivo. Our findings indicate that these virulence-related DEGs (especially capsule) might be responsible for the virulence of PmCQ2 and PmCQ6, providing prospective candidates for further studies on pathogenesis.

scholarly journals Development, Biological Characterization, and Immunological Evaluation of Virosome Vaccine against Newcastle Disease in Pakistan

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Muhammad Hidayat Rasool ◽  
Asif Mehmood ◽  
Muhammad Saqalein ◽  
Muhammad Atif Nisar ◽  
Ahmad Almatroudi ◽  
...  
Keyword(s):  
Newcastle Disease ◽  
Fluorescent Protein ◽  
Virulent Strain ◽  
Negative Control ◽  
Economic Losses ◽  
Experimental Conditions ◽  
In Vivo Evaluation ◽  
Inactivated Vaccines

Newcastle disease (ND) is a highly fatal, infectious, viral disease, and despite immunization with live and inactivated vaccines, the disease is still endemic, causing heavy morbidity and mortality leading to huge economic losses to the poultry industry in Pakistan. Therefore, the present study was aimed for the first time in the country at using novel virosomal technology to develop the ND vaccine using an indigenous highly virulent strain of the virus. ND virosome was prepared using Triton X-100, and SM2 Bio-Beads were used to remove the detergent and reconstitute the viral membrane into virosome. Confirmation was done by transmission electron microscopy and protein analysis by SDS-PAGE. In vitro cell adhesion property was observed by incorporating green fluorescent protein (GFP), producing plasmid into virosome and in vitro cell culture assay. Sterility, safety, and stability of the vaccine were tested before in vivo evaluation of immunogenicity and challenge protection study in commercial broiler. The virosome vaccine was administered (30 μg/bird) at days 7 and 14 through the intranasal route in comparison with commercially available live and inactivated ND vaccines. Results revealed significantly high ( p < 0.05 ) and clinically protective hemagglutination inhibition (HI) antibody titers at 7, 14, 21, and 28 days postimmunization with the virosome vaccine in comparison to the negative control. The GMTs were comparable to live and inactivated vaccines with nonsignificant ( p > 0.05 ) differences throughout the experiment. Antibody levels increased in all vaccinated groups gradually from the 7th day and were maximum at 28th-day postvaccination. In the virosome-administered group, GMT was 83.18 and 77.62 at 21st and 28th-days postvaccination, respectively. Challenge revealed 100%, 90%, and 80% protection in virosome, live, and inactivated vaccinated groups, respectively. Under given experimental conditions, we can conclude that ND virosome vaccine prepared from the indigenous virus was found to be safe and immunogenic.

Identification of HnRNP M as a novel biomarker for colorectal carcinoma by quantitative proteomics

2014 ◽  
Vol 306 (5) ◽  
pp. G394-G403 ◽  
Author(s):  
Shuijiao Chen ◽  
Jie Zhang ◽  
Lunxi Duan ◽  
Yu Zhang ◽  
Cui Li ◽  
...  
Keyword(s):  
Colorectal Carcinoma ◽  
Cell Cycle Progression ◽  
Regional Lymph Node ◽  
Cancer Recurrence ◽  
Epithelial Tissue ◽  
Absolute Quantitation ◽  
Itraq Analysis ◽  
Isobaric Tags ◽  
Carcinogenic Process

Colorectal carcinoma (CRC) is one of the most common cancers in the world, and identification of new CRC biomarkers will be helpful for the diagnosis and treatment of CRC. For isobaric tags for relative and absolute quantitation (iTRAQ) analysis, fresh CRC and adjacent, colonic adenoma, ulcerative colitis, Crohn's disease, and noncancerous colonic epithelial tissue were obtained from patients at the 2nd Xiangya Hospital of Central South University, China. The function of heterogeneous nuclear ribonucleoprotein M (HnRNP M) during the proliferation, invasion, and metastasis of CRC cells in vitro was evaluated. One hundred and twenty-six differentially expressed proteins were identified by iTRAQ analysis. The expression of HnRNP M exhibited progressive changes during the carcinogenic process and was validated by Western blot. The upregulation of HnRNP M correlated with cancer recurrence and regional lymph node metastasis. Furthermore, biological role exploration suggests that HnRNP M positively regulates cell cycle progression, promotes cell growth and invasion in vitro, and increases the colony-forming ability of LS174T cells. The present data demonstrate that the upregulation of HnRNP M is involved in human colorectal epithelial carcinogenesis and may serve as a carcinoma biomarker for CRC.

scholarly journals Singapore Grouper Iridovirus Disturbed Glycerophospholipids Homeostasis: Cytosolic Phospholipase A2 Was Essential for Virus Replication

2021 ◽  
Vol 22 (22) ◽  
pp. 12597
Author(s):  
Na Ni ◽  
Jiaying Zheng ◽  
Wenji Wang ◽  
Linyong Zhi ◽  
Qiwei Qin ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Ectopic Expression ◽  
Cytosolic Phospholipase A2 ◽  
Economic Losses ◽  
Key Enzymes ◽  
Cytosolic Phospholipase ◽  
Aquaculture Industry ◽  
Infected Cells ◽  
First Time

Singapore grouper iridovirus (SGIV), belonging to genus Ranavirus, family Iridoviridae, causes great economic losses in the aquaculture industry. Previous studies demonstrated the lipid composition of intracellular unenveloped viruses, but the changes in host-cell glyceophospholipids components and the roles of key enzymes during SGIV infection still remain largely unknown. Here, the whole cell lipidomic profiling during SGIV infection was analyzed using UPLC-Q-TOF-MS/MS. The lipidomic data showed that glycerophospholipids (GPs), including phosphatidylcholine (PC), phosphatidylserine (PS), glycerophosphoinositols (PI) and fatty acids (FAs) were significantly elevated in SGIV-infected cells, indicating that SGIV infection disturbed GPs homeostasis, and then affected the metabolism of FAs, especially arachidonic acid (AA). The roles of key enzymes, such as cytosolic phospholipase A2 (cPLA2), 5-Lipoxygenase (5-LOX), and cyclooxygenase (COX) in SGIV infection were further investigated using the corresponding specific inhibitors. The inhibition of cPLA2 by AACOCF3 decreased SGIV replication, suggesting that cPLA2 might play important roles in the process of SGIV infection. Consistent with this result, the ectopic expression of EccPLA2α or knockdown significantly enhanced or suppressed viral replication in vitro, respectively. In addition, the inhibition of both 5-LOX and COX significantly suppressed SGIV replication, indicating that AA metabolism was essential for SGIV infection. Taken together, our results demonstrated for the first time that SGIV infection in vitro disturbed GPs homeostasis and cPLA2 exerted crucial roles in SGIV replication.

scholarly journals In VitroDestruction of Streptococcus pneumoniae Biofilms with Bacterial and Phage Peptidoglycan Hydrolases

2011 ◽  
Vol 55 (9) ◽  
pp. 4144-4148 ◽  
Author(s):  
Mirian Domenech ◽  
Ernesto García ◽  
Miriam Moscoso
Keyword(s):  
Cell Wall ◽  
Streptococcus Pneumoniae ◽  
Cooperative Effect ◽  
Planktonic Cells ◽  
Content Type ◽  
Biofilm Model ◽  
Streptococcus Oralis ◽  
Peptidoglycan Hydrolases ◽  
Cell Wall Hydrolases

ABSTRACTHost- and phage-coded cell wall hydrolases have been used to fightStreptococcus pneumoniaegrowing as planktonic cellsin vitroas well as in animal models. Until now, however, the usefulness of these enzymes in biofilm-grown pneumococci has gone untested. The antipneumococcal activity of different cell wall hydrolases produced byS. pneumoniaeand a number of its phages was examined in anin vitrobiofilm model. The major pneumococcal autolysin LytA, anN-acetylmuramoyl-l-alanine amidase, showed the greatest efficiency in disintegratingS. pneumoniaebiofilms. The phage-encoded lysozymes Cpl-1 and Cpl-7 were also very efficient. Biofilms formed by the close pneumococcal relativesStreptococcus pseudopneumoniaeandStreptococcus oraliswere also destroyed by the phage endolysins but not by theS. pneumoniaeautolysin LytA. A cooperative effect of LytA and Cpl-1 in the disintegration ofS. pneumoniaebiofilms was recorded.

Vibrio harveyi virulence gene expression in vitro and in vivo during infection in black tiger shrimp Penaeus monodon

2020 ◽  
Vol 139 ◽  
pp. 153-160
Author(s):  
S Peeralil ◽  
TC Joseph ◽  
V Murugadas ◽  
PG Akhilnath ◽  
VN Sreejith ◽  
...  
Keyword(s):  
Gene Expression ◽  
Virulence Genes ◽  
Vibrio Harveyi ◽  
Penaeus Monodon ◽  
Challenge Test ◽  
Virulence Gene ◽  
Economic Losses ◽  
Virulence Gene Expression

Luminescent Vibrio harveyi is common in sea and estuarine waters. It produces several virulence factors and negatively affects larval penaeid shrimp in hatcheries, resulting in severe economic losses to shrimp aquaculture. Although V. harveyi is an important pathogen of shrimp, its pathogenicity mechanisms have yet to be completely elucidated. In the present study, isolates of V. harveyi were isolated and characterized from diseased Penaeus monodon postlarvae from hatcheries in Kerala, India, from September to December 2016. All 23 tested isolates were positive for lipase, phospholipase, caseinase, gelatinase and chitinase activity, and 3 of the isolates (MFB32, MFB71 and MFB68) showed potential for significant biofilm formation. Based on the presence of virulence genes, the isolates of V. harveyi were grouped into 6 genotypes, predominated by vhpA+ flaB+ ser+ vhh1- luxR+ vopD- vcrD+ vscN-. One isolate from each genotype was randomly selected for in vivo virulence experiments, and the LD50 ranged from 1.7 ± 0.5 × 103 to 4.1 ± 0.1 × 105 CFU ml-1. The expression of genes during the infection in postlarvae was high in 2 of the isolates (MFB12 and MFB32), consistent with the result of the challenge test. However, in MFB19, even though all genes tested were present, their expression level was very low and likely contributed to its lack of virulence. Because of the significant variation in gene expression, the presence of virulence genes alone cannot be used as a marker for pathogenicity of V. harveyi.

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