scholarly journals Quantitative Proteomics Analysis by Isobaric Tags for Relative and Absolute Quantitation Identified Lumican as a Potential Marker for Acute Aortic Dissection

2011 ◽  
Vol 2011 ◽  
pp. 1-10 ◽  
Author(s):  
Guorong Gu ◽  
Weizhong Cheng ◽  
Chenling Yao ◽  
Jun Yin ◽  
Chaoyang Tong ◽  
...  
Keyword(s):  
Aortic Dissection ◽  
Quantitative Proteomics ◽  
Acute Aortic Dissection ◽  
Serum Marker ◽  
Serum Biomarkers ◽  
Serum Samples ◽  
Absolute Quantitation ◽  
Itraq Analysis ◽  
Potential Marker ◽  
Isobaric Tags

Acute aortic dissection (AAD) is a serious vascular disease. Currently the diagnosis relies on clinical and radiological means whereas serum biomarkers are lacking. The purpose of this study was to identify potential serum biomarkers for AAD using isobaric tags for relative and absolute quantitation (iTRAQ) approach. A total of 120 serum samples were collected from three groups: AAD patients (n=60), patients with acute myocardial infarction (AMI,n=30), and healthy volunteers (n=30), whereas the first 10 samples from each group were used for iTRAQ analysis. Using iTRAQ approach, a total of 174 proteins were identified as significantly different between AAD patients and healthy subjects. Among them, forty-six proteins increased more than twofold, full-scale analysis using serum sample for the entire 120 subjects demonstrated that Lumican level was significantly increased relative to control and AMI samples. Further, Lumican level correlated with time from onset to admission in AAD but not AMI samples. Using iTRAQ approach, our study showed that Lumican may be a potential AAD-related serum marker that may assist the diagnosis of AAD.

scholarly journals Acute Aortic Dissection Biomarkers Identified Using Isobaric Tags for Relative and Absolute Quantitation

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Ziya Xiao ◽  
Yuan Xue ◽  
Chenling Yao ◽  
Guorong Gu ◽  
Yaping Zhang ◽  
...  
Keyword(s):  
Aortic Dissection ◽  
Sensitivity And Specificity ◽  
Acute Aortic Dissection ◽  
Roc Curves ◽  
Diagnostic Value ◽  
Serum Samples ◽  
Absolute Quantitation ◽  
Isobaric Tags ◽  
Diagnostic Sensitivity And Specificity ◽  
D Dimer

The purpose of this study was to evaluate the utility of potential serum biomarkers for acute aortic dissection (AAD) that were identified by isobaric Tags for Relative and Absolute Quantitation (iTRAQ) approaches. Serum samples from 20 AAD patients and 20 healthy volunteers were analyzed using iTRAQ technology. Protein validation was performed using samples from 120 patients with chest pain. A total of 355 proteins were identified with the iTRAQ approach; 164 proteins reached the strict quantitative standard, and 125 proteins were increased or decreased more than 1.2-fold (64 and 61 proteins were up- and downregulated, resp.). Lumican, C-reactive protein (CRP), thrombospondin-1 (TSP-1), and D-dimer were selected as candidate biomarkers for the validation tests. Receiver operating characteristic (ROC) curves show that Lumican and D-dimer have diagnostic value (area under the curves [AUCs] 0.895 and 0.891,P<0.05). For Lumican, the diagnostic sensitivity and specificity were 73.33% and 98.33%, while the corresponding values for D-dimer were 93.33% and 68.33%. For Lumican and D-dimer AAD combined diagnosis, the sensitivity and specificity were 88.33% and 95%, respectively. In conclusion, Lumican has good specificity and D-dimer has good sensitivity for the diagnosis of AAD, while the combined detection of D-dimer and Lumican has better diagnostic value.

scholarly journals Identification of Protein Expression Changes in Hepatocellular Carcinoma through iTRAQ

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Yuanyuan Zhang ◽  
Xia Ying ◽  
Qian Zhao ◽  
Jinlu Ma ◽  
Dan Zhang ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Serum Levels ◽  
Serum Biomarkers ◽  
The Cancer Genome Atlas ◽  
Paraoxonase 1 ◽  
Serum Samples ◽  
Absolute Quantitation ◽  
Isobaric Tags ◽  
Tcga Dataset ◽  
Normal Controls

Background. Hepatocellular carcinoma (HCC) is a malignant tumor associated with a poor prognosis. Serum biomarkers of HCC have the potential to improve the diagnosis, provide a means to monitor the tumors, and predict their malignancy. Proteins that are expressed differentially between HCC patients and normal controls have the potential to be biomarkers. Method. Serum samples from 10 confirmed HCC patients and 10 controls were collected. The differentially expressed proteins in the serum were identified using an isobaric tags for relative and absolute quantitation- (iTRAQ-) based method. Potential serum biomarkers were validated by ELISA in another 20 HCC patients and 20 controls. Their expression data in HCC were extracted from The Cancer Genome Atlas (TCGA) dataset. Results. A total of 260 proteins were measured in the serum of HCC patients and compared to those in sex- and age-matched normal controls. Forty-one proteins displayed significant changes, with 26 being downregulated and 15 being upregulated. Upregulated proteins included alpha-1-antitrypsin (A1AT) and peroxiredoxin 2 (PRDX2), and downregulated proteins included paraoxonase 1 (PON1) and C-reactive protein (CRP). We then used ELISA to measure serum levels of A1AT, PRDX2, PON1, and CRP in another 20 patients with HCC and found that only PON1 levels were consistent with the iTRAQ result. In TCGA dataset, PON1 expression was downregulated in HCC tissues (P<0.001) and low expression of PON1 was associated with poor survival in HCC patients (P<0.001). Conclusions. PON1 could act as a biomarker for HCC to assist in the diagnosis of HCC.

scholarly journals Comparative Proteomic Investigation of Plasma Reveals Novel Potential Biomarker Groups for Acute Aortic Dissection

2020 ◽  
Vol 2020 ◽  
pp. 1-13
Author(s):  
Na Cheng ◽  
Hao Wang ◽  
Weizong Zhang ◽  
Heng Wang ◽  
Xiang Jin ◽  
...  
Keyword(s):  
Aortic Dissection ◽  
Acute Aortic Dissection ◽  
Density Lipoprotein ◽  
Interaction Network ◽  
Enrichment Analysis ◽  
Absolute Quantitation ◽  
Molecular Change ◽  
Amyloid A ◽  
Potential Biomarker ◽  
Isobaric Tags

Acute aortic dissection (AAD) is a catastrophic cardiovascular disease with high disability and mortality due to multiple fatal complications. However, the molecular changes of the serum proteome after AAD are not very clear. Here, we performed isobaric tags for relative and absolute quantitation- (iTRAQ-) based comparative proteomic analysis to investigate the proteome profile changes after AAD by collecting plasma samples from 20 AAD patients and 20 controls. Out of the 345 identified proteins, 266 were considered as high-quality quantified proteins (95%confident peptides≥2), of which 25 proteins were accumulated and 12 were reduced in AAD samples. Gene ontology enrichment analysis showed that the 25 AAD-accumulated proteins were enriched in high-density lipoprotein particles for the cellular component category and protein homodimerization acidity for the molecular function category. Protein-protein interaction network analysis showed that serum amyloid A proteins (SAAs), complement component proteins, and carboxypeptidase N catalytic chain proteins (CPNs) possessed the key nodes of the network. The expression levels of six selected AAD-accumulated proteins, B2-GP1, CPN1, F9, LBP, SAA1, and SAA2, were validated by ELISA. Moreover, ROC analysis showed that the AUCs of B2-GP1 and CPN1 were 0.808 and 0.702, respectively. Our data provide insights into molecular change profiles in proteome levels after AAD and indicate that B2-GP1 and CPN1 are potential biomarkers for AAD.

scholarly journals Identification of Vinculin as a Potential Diagnostic Biomarker for Acute Aortic Dissection Using Label-Free Proteomics

2020 ◽  
Vol 2020 ◽  
pp. 1-8
Author(s):  
He-Qing Wang ◽  
Hang Yang ◽  
Qian Tang ◽  
Yi-Chen Gong ◽  
Yuan-Hao Fu ◽  
...  
Keyword(s):  
Aortic Dissection ◽  
Acute Aortic Dissection ◽  
Early Stage ◽  
Serum Biomarkers ◽  
Label Free ◽  
Serum Samples ◽  
Proteomics Analysis ◽  
Proteomics Approach ◽  
Healthy Control ◽  
High Level

Acute aortic dissection (AAD) is an emergent vascular disease. Currently, its diagnosis depends on clinical and radiological investigations but lacking of serum biomarkers. In this study, we aimed to identify potential serum biomarkers for AAD using label-free proteomics approach. A total of 90 serum samples were collected from three groups: patients with acute aortic dissection (AAD, n=30), patients with acute myocardial infarction (AMI, n=30), and healthy controls (n=30), and the first four samples from each group were selected for label-free proteomics analysis. Using label-free approach, a total of 22 differentially expressed proteins were identified in the serum samples of the AAD group, of which 15 were upregulated and 7 were downregulated as compared to the AMI and healthy control groups. The most prominent increased protein was vinculin, which was selected to validate in total samples. The level of vinculin was significantly elevated in AAD patients (15.8 ng/ml, IQR: 9.3-19.9 ng/ml) than that in AMI patients (8.6 ng/ml, IQR:5.3-11.4 ng/ml) and healthy volunteers (5.3 ng/ml, IQR:2.8-7.6 ng/ml), P<0.0001. Furthermore, the concentration of vinculin both increased in type A and B dissection. At the early stage of AAD, vinculin maintained a high level to 48 hours compared with that of AMI. Our study demonstrated that vinculin may play a role in the early diagnosis of AAD.

scholarly journals Partial Proteome Map of Campylobacter Jejuni Strain Nctc11168 by Gel-Free Proteomics Analysis

2014 ◽  
Vol 2 (4) ◽  
pp. 464-477
Author(s):  
Zilun Shi ◽  
Chris Dawson ◽  
Stephen L.W. On ◽  
Malik Altaf Hussain
Keyword(s):  
Campylobacter Jejuni ◽  
Foodborne Pathogen ◽  
Brain Heart Infusion ◽  
Cell Protein ◽  
Absolute Quantitation ◽  
Whole Cell ◽  
Itraq Analysis ◽  
Proteomic Approach ◽  
Whole Cell Protein ◽  
Isobaric Tags

A proteome map of the foodborne pathogen Campylobacter jejuni NCTC11168 was analyzed using a state-of-the-art gel-free proteomic approach for the first time. A whole cell protein extract was prepared from the C. jejuni strain NCTC11168 grown in brain heart infusion (BHI) broth at 42°C under microaerobic conditions. A gel-free technique using isobaric tags for relative and absolute quantitation (iTRAQ) was employed to create a protein expression profile of the strain. Liquid chromatography-mass spectrometry (LC-MS/MS) was used to identify the proteins. Protein functionalities were searched to classify them. A total of 235 proteins were identified in the whole cell protein fraction of C. jejuni NCTC11168 cells using iTRAQ analysis. Functional grouping of the identified proteins showed that forty percent of these proteins were associated with energy metabolism, protein synthesis and genetic information processing. iTRAQ was faster, easier and proved more sensitive than two-dimensional gel-based proteomics approaches previously applied to C. jejuni, making it an attractive tool for further studies of cellular physiological response. DOI: http://dx.doi.org/10.3126/ijasbt.v2i4.11253  Int J Appl Sci Biotechnol, Vol. 2(4): 464-477 

scholarly journals Identification of Proteins Implicated in the Increased Heart Rate in ShenSongYangXin-Treated Bradycardia Rabbits by iTRAQ-Based Quantitative Proteomics

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Zhouying Liu ◽  
Jian Huang ◽  
Youping Huo ◽  
Jing Gong ◽  
Yinhui Zhang ◽  
...  
Keyword(s):  
Heart Rate ◽  
Quantitative Proteomics ◽  
Sinoatrial Node ◽  
Absolute Quantitation ◽  
Treatment Groups ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Isobaric Tags ◽  
Altered Proteins ◽  
Chinese Medicine Treatment

The present study tries to identify proteins implicated in bradycardia rabbits in hearts after ShenSongYangXin (SSYX, a traditional Chinese medicine) treatment. Eighteen adult rabbits were randomly assigned to three groups: sham, model, and SSYX treatment groups. Heart rate was recorded in rabbits and proteins were isolated from ventricular muscle. We used isobaric tags for elative and absolute quantitation (iTRAQ) coupled with two-dimensional liquid chromatography-tandem mass spectrometry to identify altered proteins after SSYX treatment. The heart rate decreased after six weeks due to the injury of the sinoatrial node in the model group. This effect was partially reversed by 4-week SSYX treatment. A total of a2988 proteins were quantified by performing the iTRAQ-based experiments. Of these, 86 proteins were differentially expressed according to our criteria (42 upregulated and 44 downregulated). The identification of key proteins implicated in the treatment of bradycardia could serve as a foundation to better understand and further explore the molecular mechanism of SSYX treatment.

scholarly journals Differentiating Two Closely Related Alexandrium Species Using Comparative Quantitative Proteomics

Toxins ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 7
Author(s):  
Bryan John J. Subong ◽  
Arturo O. Lluisma ◽  
Rhodora V. Azanza ◽  
Lilibeth A. Salvador-Reyes
Keyword(s):  
Quantitative Proteomics ◽  
Polyketide Synthase ◽  
Harmful Algal Bloom ◽  
Feedback Mechanism ◽  
Type I ◽  
Absolute Quantitation ◽  
Alexandrium Minutum ◽  
2D Dige ◽  
Negative Feedback Mechanism ◽  
Isobaric Tags

Alexandrium minutum and Alexandrium tamutum are two closely related harmful algal bloom (HAB)-causing species with different toxicity. Using isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics and two-dimensional differential gel electrophoresis (2D-DIGE), a comprehensive characterization of the proteomes of A. minutum and A. tamutum was performed to identify the cellular and molecular underpinnings for the dissimilarity between these two species. A total of 1436 proteins and 420 protein spots were identified using iTRAQ-based proteomics and 2D-DIGE, respectively. Both methods revealed little difference (10–12%) between the proteomes of A. minutum and A. tamutum, highlighting that these organisms follow similar cellular and biological processes at the exponential stage. Toxin biosynthetic enzymes were present in both organisms. However, the gonyautoxin-producing A. minutum showed higher levels of osmotic growth proteins, Zn-dependent alcohol dehydrogenase and type-I polyketide synthase compared to the non-toxic A. tamutum. Further, A. tamutum had increased S-adenosylmethionine transferase that may potentially have a negative feedback mechanism to toxin biosynthesis. The complementary proteomics approach provided insights into the biochemistry of these two closely related HAB-causing organisms. The identified proteins are potential biomarkers for organismal toxicity and could be explored for environmental monitoring.

scholarly journals Quantitative Proteomics Analysis of Differentially Expressed Proteins in Serum of Former Uranium Miners by Isobaric Tags for the Relative and Absolute Quantitation

Dose-Response ◽  
2021 ◽  
Vol 19 (4) ◽  
pp. 155932582110561
Author(s):  
Xuhong Dang ◽  
Haipeng Lin ◽  
Yayi Yuan ◽  
Biao Yang ◽  
Juancong Dong ◽  
...  
Keyword(s):  
Radiation Damage ◽  
Process Analysis ◽  
Density Lipoprotein ◽  
Differentially Expressed ◽  
Enzyme Linked Immunosorbent Assay ◽  
Differentially Expressed Proteins ◽  
Serum Samples ◽  
Absolute Quantitation ◽  
Uranium Miners ◽  
Isobaric Tags

The carcinogenicity of radon has been convincingly documented through epidemiological studies of underground miners. However, there is a lack of early warning indicators for radon radiation damage. In this study, mixed serum samples of 3 groups were collected from 27 underground uranium miners and seven aboveground miners according to the radiation exposure dose. The differentially expressed proteins in the serum were identified using the isobaric tags for the relative and absolute quantitation (iTRAQ)-based method. Some differentially expressed proteins were validated by enzyme-linked immunosorbent assay (ELISA) in 84 underground and 32 aboveground miners. A total of 25 co-differentially expressed proteins in 2 underground miner groups were screened, of which 9 were downregulated and 13 were upregulated. Biological process analysis of these proteins using Metascape showed that 5 GO terms were enriched, such as negative regulation of very-low-density lipoprotein particle clearance, endocytosis, and regulated exocytosis. The results of the ELISA for the expression levels of GCN1, CIP2A, and IGHV1-24 in the serum of 116 miners’ serum showed that the levels of GCN1 and CIP2A were consistent with the iTRAQ results. In conclusion, APOC1, APOC2, APOC3, ORM1, ORM2, ANTXR1, GCN1, and CIP2A may be potential early markers of radon radiation damage.

scholarly journals Protein Profiling of Microdissected Pancreas Carcinoma and Identification of HSP27 as a Potential Serum Marker

2007 ◽  
Vol 53 (4) ◽  
pp. 629-635 ◽  
Author(s):  
Christian Melle ◽  
Günther Ernst ◽  
Niko Escher ◽  
Daniel Hartmann ◽  
Bettina Schimmel ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Serum Marker ◽  
Pancreatic Tissue ◽  
Protein Profiling ◽  
Serum Samples ◽  
Pancreas Carcinoma ◽  
Tissue Samples ◽  
Potential Marker ◽  
Preoperative Serum ◽  
Surface Enhanced

Abstract Background: Patients with pancreatic adenocarcinomas have a poor prognosis because of late clinical manifestation and the tumor’s aggressive nature. We used proteomic techniques to search for markers of pancreatic carcinoma. Methods: We performed protein profiling of microdissected cryostat sections of 9 pancreatic adenocarcinomas and 10 healthy pancreatic tissue samples using ProteinChip technology (surface-enhanced laser desorption/ionization). We identified proteins by use of 2-dimensional gel electrophoresis, peptide fingerprint mapping, and immunodepletion and used immunohistochemistry for in situ localization of the proteins found. We used ELISA to quantify these proteins in preoperative serum samples from 35 patients with pancreatic cancer and 37 healthy individuals. Results: From among the differentially expressed signals that were detected by ProteinChip technology, we identified 2 proteins, DJ-1 and heat shock protein 27 (HSP27). We then detected HSP27 in sera of patients by use of ELISA, indicating a sensitivity of 100% and a specificity of 84% for the recognition of pancreatic cancer. Conclusions: The detection of DJ-1 and HSP27 in pure defined tissue and the retrieval of HSP27 in serum by antibody-based methods identifies a potential marker for pancreatic cancer.

Proteome Analysis of Testicular Tissue in Varicocele Rats Using iTRAQ Labeling Technology

2020 ◽  
Author(s):  
lu xian feng ◽  
zhao wei ◽  
liu jian rong ◽  
wang dan feng ◽  
yuan cai xia ◽  
...  
Keyword(s):  
Protein Transport ◽  
Proteome Analysis ◽  
Testicular Tissue ◽  
P Value ◽  
Absolute Quantitation ◽  
Tissue Samples ◽  
Itraq Analysis ◽  
Potential Biomarker ◽  
Liquid Chromatography Tandem Mass ◽  
Isobaric Tags

Abstract Purpose Varicocele (VC) is considered as the main cause of male infertility, clear and definite molecular markers of varicocele disease are helpful for early prevention and timely treatment. This study was in order to examine the effect of varicocele on protein expression in testicular tissue.Methods The testicular tissue samples of normal rats and varicocele rats were used for proteomic analysis and functional bioinformatics analysis. The proteins with a fold change of > 1.2 or < 1.2, and with a P-value < 0.05 were used to identify up- and downregulation proteins between the varicocele and control rats, and two target proteins were selected and verified with western blot.Results It was found that seminiferous epithelium was disordered and spermatogenic cells were injured seriously in varicocele group. A total of 65 differentially expressed proteins were identified compared with normal group by liquid chromatography tandem mass spectrometry (LC-MS/MS) and isobaric tags for relative and absolute quantitation (iTRAQ) analysis, including 31 up-regulated proteins and 34 down-regulated proteins, respectively. Functions of those proteins were mainly related to the following processes: signal transduction, protein cycle, transmembrane transport, protein transport, vesicular mediated transport, cell division, and membrane tissue. Two down regulated proteins of ATPase and Cu2+-transporting alpha (ATP7A) and Calcium and integrin-binding protein 1 (CIB1) in varicocele rats were selected and confirmed.Conclusion Protein spectrum of testicular tissue in varicocele rats was different from that in normal rats. Low expressions of ATP7A and CIB1 may be affect testicular spermatogenic function and can be used as a potential biomarker for testicular tissue to maintain normal spermatogenic function.

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