scholarly journals Proteomic Approach for Identification of IgA Nephropathy-Related Biomarkers in Urine

2017 ◽  
pp. 621-632 ◽  
Author(s):  
P. PRIKRYL ◽  
L. VOJTOVA ◽  
D. MAIXNEROVA ◽  
M. VOKURKA ◽  
M. NEPRASOVA ◽  
...  
Keyword(s):  
Iga Nephropathy ◽  
Kidney Diseases ◽  
Surrogate Marker ◽  
Pilot Project ◽  
Protein Quantification ◽  
Absolute Quantitation ◽  
Proteomic Approach ◽  
Isobaric Tag ◽  
Alpha 1 Antitrypsin ◽  
Potential Biomarkers

Proteinuria is often used as a surrogate marker in monitoring and predicting outcome in patients with chronic kidney diseases, but it is non-specific. IgAN belongs to the most common primary glomerulonephritis worldwide with serious prognosis. The main aim of this work was to assess differences in urine proteins in patients with IgA nephropathy and to identify abnormal proteins as potential biomarkers of IgA nephropathy or the renal disease. In our pilot project, we selected 20 patients and compared them with 20 healthy volunteers. Protein quantification was performed using iTRAQ (isobaric tag for relative and absolute quantitation) labeling method. The peptides were separated by the isoelectric focusing method (IEF) and nano-LC with C18 column and identified by mass spectrometry using MALDI-TOF/TOF MS. Proteins´ lists obtained from IEF-LC-MS-MS/MS analysis were combined and contained 201 proteins. It was found out that 113 proteins were common in both experiments. 30 urinary proteins were significantly up- or down-regulated in patients with IgA nephropathy. We characterized potential biomarkers such as alpha-1-antitrypsin, apolipoprotein A-I, CD44 antigen or kininogen. Potential biomarkers of IgAN should be validated in further studies.

scholarly journals Integrated Proteomics and Metabolomic Analyses of Plasma Injury Biomarkers in a Serious Brain Trauma Model in Rats

2019 ◽  
Vol 20 (4) ◽  
pp. 922 ◽  
Author(s):  
Tao Song ◽  
Ying Zhu ◽  
Peng Zhang ◽  
Minzhu Zhao ◽  
Dezhang Zhao ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Diffuse Axonal Injury ◽  
Rat Plasma ◽  
Integrated Analysis ◽  
Differential Analysis ◽  
Absolute Quantitation ◽  
Proteomic Data ◽  
Isobaric Tag ◽  
Trauma Model ◽  
Potential Biomarkers

Diffuse axonal injury (DAI) is a prevalent and serious brain injury with significant morbidity and disability. However, the underlying pathogenesis of DAI remains largely unclear, and there are still no objective laboratory-based tests available for clinicians to make an early diagnosis of DAI. An integrated analysis of metabolomic data and proteomic data may be useful to identify all of the molecular mechanisms of DAI and novel potential biomarkers. Therefore, we established a rat model of DAI, and applied an integrated UPLC-Q-TOF/MS-based metabolomics and isobaric tag for relative and absolute quantitation (iTRAQ)-based proteomic analysis to obtain unbiased profiling data. Differential analysis identified 34 metabolites and 43 proteins in rat plasma of the injury group. Two metabolites (acetone and 4-Hydroxybenzaldehyde) and two proteins (Alpha-1-antiproteinase and Alpha-1-acid glycoprotein) were identified as potential biomarkers for DAI, and all may play important roles in the pathogenesis of DAI. Our study demonstrated the feasibility of integrated metabolomics and proteomics method to uncover the underlying molecular mechanisms of DAI, and may help provide clinicians with some novel diagnostic biomarkers and therapeutic targets.

SP200SERUM 2-CYSTEINE PEROXIREDOXINS: POTENTIAL BIOMARKERS OF IGA NEPHROPATHY AND LUPUS NEPHRITIS

2019 ◽  
Vol 34 (Supplement_1) ◽  
Author(s):  
Natalia Krata ◽  
Barbara Moszczuk ◽  
Bartosz Foroncewicz ◽  
Radosław Zagożdżon ◽  
Niemczyk Mariusz ◽  
...  
Keyword(s):  
Lupus Nephritis ◽  
Iga Nephropathy ◽  
Potential Biomarkers

Abstract 399: The Circadian Rhythm of Plasma Angiotensinogen in Healthy Volunteers

Hypertension ◽  
2012 ◽  
Vol 60 (suppl_1) ◽  
Author(s):  
Kumiko Kaifu ◽  
Hiroyuki Kobori ◽  
Yoko Nishijima ◽  
Akira Nishiyama ◽  
Masakazu Kohno
Keyword(s):  
Blood Pressure ◽  
Circadian Rhythm ◽  
Healthy Volunteers ◽  
Pulse Rate ◽  
Capillary Tube ◽  
Kidney Diseases ◽  
Surrogate Marker ◽  
Recumbent Position ◽  
Time Points ◽  
Significant Difference

Background: We have previously reported that urinary angiotensinogen (AGT) excretion did not have a circadian rhythm and could be a novel biomarker for the activity of the renin-angiotensin system (RAS) in kidney. However, there have been few reports investigating the circadian rhythm of plasma AGT in human body. Thus, this study was performed to examine the circadian rhythm in plasma AGT in human. METHODS: Evaluating RAS in clinical practice is generally performed in a recumbent position after a 30-minute stabilization period. However, to determine the necessity of recumbent position, we first compared plasma AGT concentrations measured right after waking up and after a 5-minute sitting rest. Next, we examined the circadian rhythm of plasma AGT in 43 healthy volunteers who had shown no abnormalities in the medical examinations in 2011. Plasma AGT was measured at three time points (9 a.m., 1 p.m., and 4 p.m.) in the above volunteers. Blood was collected by a micro hematocrit capillary tube with heparin, frozen for storage after centrifugation, and thawed for the measurement of plasma AGT using an ELISA kit. Results: There was no significant difference between the plasma AGT values of the two measuring methods (P = 0.1202, n = 5). Based on the result, we performed blood sampling after a 5-minute sitting rest in the volunteers consisting of 17 men and 26 women. Average blood pressure was 116.3/75.1 mmHg at 9 a.m., 116.3/71.9 mmHg at 1 p.m., and 115.5/70.1 mmHg at 4 p.m.; average pulse rate was 78.7/min at 9 a.m., 77.1/min at 1 p.m., and 73.3/min at 4 p.m. Blood pressure and pulse rate did not change throughout the day. Average plasma AGT was 20.4 ± 6.0 ng/ml at 9 a.m., 20.7 ± 5.0 ng/ml at 1 p.m., and 19.8 ± 6.4 ng/ml at 4 p.m. Plasma AGT did not show a circadian rhythm (P = 0.3803). Conclusion: We found in this study that plasma AGT did not have a circadian rhythm. We also found that plasma AGT was not affected by daily life actions. Thus, future patients may not be required to rest nor wait for certain time points before measuring plasma AGT. We also have to unveil the normal AGT levels and the influence on the levels by diseases. As we think that plasma AGT and ratio of urinary AGT to plasma AGT can be a new surrogate marker of hypertension and kidney diseases, we further need to go into this research area.

scholarly journals A cellular and proteomic approach to assess proteins extracted from cryopreserved human amnion in the cultivation of corneal stromal keratocytes for stromal cell therapy

2019 ◽  
Vol 6 (1) ◽  
Author(s):  
Beau J. Fenner ◽  
Nur Zahirah B. M. Yusoff ◽  
Matthias Fuest ◽  
Lei Zhou ◽  
Francisco Bandeira ◽  
...  
Keyword(s):  
Epithelial Mesenchymal Transition ◽  
Protein Extraction ◽  
Culture Media ◽  
Cell Metabolism ◽  
Absolute Quantitation ◽  
Human Amnion ◽  
Mesenchymal Transition ◽  
Proteomic Approach ◽  
Isobaric Tagging ◽  
Human Keratocytes

Abstract Background Human corneal stromal keratocytes propagated in culture media supplemented with human amnion extract (AME) can correct early corneal haze in an animal model. Clinical application of cultivated keratocytes is limited by infectious disease screening before amnion products can be used in humans. It remains unclear if AME from cryopreserved versus fresh human amnion can support human keratocyte propagation, and which components of the extract promote keratocyte growth. Methods Three placentas were collected for the preparation of fresh and cryopreserved amnion tissues followed by homogenization and protein extraction. AME protein profiles were studied using isobaric tagging for relative and absolute quantitation (iTRAQ) proteomics. Enriched gene ontology (GO) terms and functional classes were identified. Primary human keratocytes from 4 donor corneas were cultured in media supplemented with fresh AME (F-AME) or cryopreserved AME (C-AME). Cell viability, proliferation and keratocyte marker expression were examined by confocal immunofluorescence and flow cytometry. Results AME proteomics revealed 1385 proteins with similar expression levels (between 0.5- and 2-fold) between F- and C-AME, while 286 proteins were reduced (less than 0.5-fold) in C-AME. Enriched GO term and biological pathway analysis showed that those proteins with comparable expression between F-AME and C-AME were involved in cell metabolism, epithelial-mesenchymal transition, focal adhesion, cell-extracellular matrix interaction, cell stress regulation and complement cascades. Human corneal stromal keratocytes cultured with F-AME or C-AME showed similar morphology and viability, while cell proliferation was mildly suppressed with C-AME (P > 0.05). Expression of aldehyde dehydrogenase 3A1 (ALDH3A1) and CD34 was similar in both cultures. Conclusion AME from cryopreserved amnion had limited influence on keratocyte culture. It is feasible to use protein extract from cryopreserved amnion to propagate human keratocytes for potential translational applications.

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 Serum Proteome Alterations in Patients with Cognitive Impairment after Traumatic Brain Injury Revealed by iTRAQ-Based Quantitative Proteomics

2017 ◽  
Vol 2017 ◽  
pp. 1-13 ◽  
Author(s):  
Xin-gui Xiong ◽  
Qinghua Liang ◽  
Chunhu Zhang ◽  
Yang Wang ◽  
Wei Huang ◽  
...  
Keyword(s):  
Traumatic Brain Injury ◽  
Cognitive Impairment ◽  
Brain Injury ◽  
Bioinformatic Analysis ◽  
Serum Samples ◽  
Absolute Quantitation ◽  
Proteomic Approach ◽  
Healthy Control ◽  
Isobaric Tagging ◽  
Experimental Findings

Background. Cognitive impairment is the leading cause of traumatic brain injury- (TBI-) related disability; however, the underlying pathogenesis of this dysfunction is not completely understood. Methods. Using an isobaric tagging for relative and absolute quantitation- (iTRAQ-) based quantitative proteomic approach, serum samples from healthy control subjects, TBI patients with cognitive impairment, and TBI patients without cognitive impairment were analysed to identify differentially expressed proteins (DEPs) related to post-TBI cognitive impairment. In addition, DEPs were further analysed using bioinformatic platforms and validated using enzyme-linked immunosorbent assays (ELISA). Results. A total of 56 DEPs were identified that were specifically related to TBI-induced cognitive impairment. Bioinformatic analysis revealed that a wide variety of cellular and metabolic processes and some signaling pathways were involved in the pathophysiology of cognitive deficits following TBI. Five randomly selected DEPs were validated using ELISA in an additional 105 cases, and the results also supported the experimental findings. Conclusions. Despite limitations, our findings will facilitate further studies of the pathological mechanisms underlying TBI-induced cognitive impairment and provide new methods for the research and development of neuroprotective agents. However, further investigation on a large cohort is warranted.

scholarly journals Proteomic Analysis of Beef Tenderloin and Flank Assessed Using an Isobaric Tag for Relative and Absolute Quantitation (iTRAQ)

Animals ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 150
Author(s):  
Zhaomin Lei ◽  
Jianping Wu ◽  
Deyin Zhang ◽  
Ting Liu ◽  
Shengguo Zhao ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
Expression Patterns ◽  
Tca Cycle ◽  
Absolute Quantification ◽  
Absolute Quantitation ◽  
Oxidation Reduction ◽  
The Tca Cycle ◽  
Isobaric Tag ◽  
Isobaric Tags

Herein, we performed a proteomic analysis of tenderloin and flank steaks from Simmental cattle using the isobaric tags for a relative and absolute quantification (iTRAQ) approach. We identified 17 amino acids in both steaks, and Gly, Cys, Ile, Lys, and Pro differed most in abundance between the steak types (p < 0.05). A comparison of the expression patterns in steaks revealed 128 differentially expressed proteins (DEPs), of which 44 were up-regulated and 84 were down-regulated. Furthermore, 27 DEPs (p < 0.05) were subjected to gene ontology (GO) analysis, and many were found to be related to oxidation-reduction, metabolism, hydrogen ion transmembrane transport, transport, the tricarboxylic acid (TCA) cycle, mitochondrial electron transport, and the conversion of nicotinamide adenine dinucleotide (NADH) to ubiquinone. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis also implicated these DEPs in various signalling pathways, including oxidative phosphorylation, cardiac muscle contraction, the TCA cycle, biosynthesis, and the metabolism. These findings provide a new insight into key proteins involved in the determination of amino acid composition in beef.

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