scholarly journals Understanding the Toxin Effects of β-Zearalenol and HT-2 on Bovine Granulosa Cells Using iTRAQ-Based Proteomics

Animals ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 130
Author(s):  
Lian Li ◽  
Min Yang ◽  
Chengmin Li ◽  
Fangxiao Yang ◽  
Genlin Wang
Keyword(s):  
Granulosa Cells ◽  
Absolute Quantification ◽  
P Value ◽  
Toxic Response ◽  
Human Food Chain ◽  
Response Profile ◽  
Isobaric Tags ◽  
Shock Proteins ◽  
Cellular Components ◽  
Marker Molecule

Zearalenone (ZEA) and T-2 are the most common mycotoxins in grains and can enter the animal and human food-chain and cause many health disorders. To elucidate the toxic response profile, we stimulated bovine granulosa cells (GCs) with β-zearalenol or HT-2. Using isobaric tags for relative and absolute quantification (iTRAQ)-based proteomic, 178 and 291 differentially expressed proteins (DEPs, fold change ≥ 1.3 and p-value < 0.05) in β-zearalenol and HT-2 groups were identified, respectively. Among these DEPs, there were 66 common DEPs between β-zearalenol and HT-2 groups. These 66 DEPs were associated with 23 biological processes terms, 14 molecular functions terms, and 19 cellular components terms. Most heat shock proteins (HSPs) were involved in the toxic response. Reactive oxygen species accumulation, the endoplasmic reticulum (ER)-stress related marker molecule (GRP78), and apoptosis were activated. β-zearalenol and HT-2 inhibited oestradiol (E2) production. These results emphasized the important function of HSPs, clarified oxidative stress, and demonstrated the caspase-3 signaling cascade involved in mycotoxin-treated toxic response, along with decreased E2 production. This study offers new insights into the toxicity of β-zearalenol and HT-2 on ovarian granulosa cells.

scholarly journals Study on Differential Protein Expression in Natural Selenium-Enriched and Non-Selenium-Enriched Rice Based on iTRAQ Quantitative Proteomics

Biomolecules ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 130 ◽  
Author(s):  
Rui Zeng ◽  
Muhammad Farooq ◽  
Li Wang ◽  
Yang Su ◽  
Tengda Zheng ◽  
...  
Keyword(s):  
Amino Acid ◽  
Differential Expression ◽  
Starch Synthesis ◽  
Oxygen Metabolism ◽  
Absolute Quantification ◽  
Stress Effect ◽  
Cysteine Synthase ◽  
Liquid Chromatography Tandem Mass ◽  
Isobaric Tags ◽  
Shock Proteins

This work was designated to scrutinize the protein differential expression in natural selenium-enriched and non-selenium-enriched rice using the Isobaric-tags for relative and absolute quantification (iTRAQ) proteomics approach. The extracted proteins were subjected to enzyme digestion, desalting, and identified by iTRAQ coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology. High pH C18 separation analysis was performed, and the data were then analyzed by Protein PilotTM (V4.5) search engine. Protein differential expression was searched out by comparing relatively quantified proteins. The analysis was conducted using gene ontology (GO), cluster of orthologous groups of proteins (COG) and Kyoto encyclopedia of genes and genomes (KEGG) metabolic pathways. A total of 3235 proteins were detected and 3161 proteins were quantified, of which 401 were differential proteins. 208 down-regulated and 193 up-regulated proteins were unveiled. 77 targeted significant differentially expressed proteins were screened out for further analysis, and were classified into 10 categories: oxidoreductases, transferases, isomerases, heat shock proteins, lyases, hydrolases, ligases, synthetases, tubulin, and actin. The results indicated that the anti-stress, anti-oxidation, active oxygen metabolism, carbohydrate and amino acid metabolism of natural selenium-enriched rice was higher than that of non-selenium rice. The activation of the starch synthesis pathway was found to be bounteous in non-selenium-enriched rice. Cysteine synthase (CYS) and methyltransferase (metE) might be the two key proteins that cause amino acid differences. OsAPx02, CatC, riPHGPX, HSP70 and HSP90 might be the key enzymes regulating antioxidant and anti-stress effect differences in two types of rice. This study provides basic information about deviations in protein mechanism and secondary metabolites in selenium-enriched and non-selenium-enriched rice.

iTRAQ-Based Quantitative Proteomic Analysis of Pseudostellaria heterophylla from Geo-Authentic Habitat and Cultivated Bases

2019 ◽  
Vol 16 (3) ◽  
pp. 231-245
Author(s):  
Yujiao Hua ◽  
Chengcheng Wang ◽  
Shengnan Wang ◽  
Zixiu Liu ◽  
Xunhong Liu ◽  
...  
Keyword(s):  
Binding Proteins ◽  
Large Subunit ◽  
Absolute Quantification ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Realtime Pcr ◽  
Pseudostellaria Heterophylla ◽  
Response To Stress ◽  
Isobaric Tags ◽  
Shock Proteins

Background: Pseudostellaria heterophylla is an important tonic traditional Chinese medicine. However, the molecular changes in the herb from geo-authentic habitat and cultivated bases remain to be explored. Objective: The purpose of this research was to study differences in P. heterophylla from geo-authentic habitat and cultivated bases. Methods: High-throughput technologies of transcriptomic and proteomic were used to identify proteins. Isobaric Tags for Relative and Absolute Quantification (iTRAQ) MS/MS has been utilized to evaluate changes in P. heterophylla from geo-authentic habitat and cultivated bases. Results: In this study, a total of 3775 proteins were detected, and 140 differentially expressed proteins were found in P. heterophylla from geo-authentic habitat and cultivated bases. 44 significantly differential expressed proteins were identified based on functional analysis classified into nine categories. Five differentially expressed proteins were confirmed at the gene expression level by Quantitative realtime PCR. Catabolic metabolism, carbohydrate metabolism, and response to stress of oxidoreductases and transferases in P. heterophylla from geo-authentic habitat were stronger than in those from cultivated bases, but protein folding and response to stress of heat shock proteins, isomerases, rubisco large subunit-binding proteins, chaperone proteins, and luminal-binding proteins in herbs from cultivated bases were more active. ADG1 and TKTA could be the critical proteins to regulate sucrose; MFP2 and CYS may be the crucial proteins that control the metabolism of fatty acids and amino acids. Conclusion: These results will provide the basic information for exploring the differences in secondary metabolites in P. heterophylla from geo-authentic habitat and cultivated bases and the protein mechanism of its quality formation.

scholarly journals Study on inhibitory effect of cinnamaldehyde on Salmonella based on iTRAQ technology

2020 ◽  
Author(s):  
Lizi Yin ◽  
Han Chen ◽  
Xuewen He ◽  
Zhongqiong Yin ◽  
Hualin Fu ◽  
...  
Keyword(s):  
Salmonella Typhimurium ◽  
Absolute Quantification ◽  
Inhibitory Effect ◽  
Kegg Pathways ◽  
Therapeutic Drugs ◽  
Liquid Chromatography Tandem Mass ◽  
New Ideas ◽  
Isobaric Tags ◽  
Novel Drug ◽  
Cellular Components

Abstract BackgroundSalmonella is one of the most concerned pathogenic bacterium worldwide. In our previous experiments, cinnamaldehyde showed prominent antibacterial effects on Salmonella Typhimurium (S. Typhimurium), indicating that it could be developed as a novel therapeutic drugs for Salmonella.ResultsIn this assay, the mechanism of cinnamaldehyde inhibiting S. Typhimurium was investigated. We studied the antibacterial mechanism of cinnamaldehyde using isobaric tags for relative and absolute quantification (iTRAQ) with two-dimensional liquid chromatography/tandem mass spectrometry (2D-LC-MS/MS), combining with bioinformatics. There were 2,212 proteins detected by iTRAQ, of which 73 proteins were up-regulated, and 82 of proteins were down-regulated. The differently expressed proteins were connected with 10 cellular components, 9 molecular functions and 13 biological processes as well as 57 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. There were some differently expressed proteins involved into biosynthesis and metabolism of amino, redox reaction, energy metabolism, toxins and drug-resistance. Cinnamaldehyde may have the potential to become a novel drug for Salmonella infection.ConclusionsBased on the iTRAQ technology, this paper explores the mechanism of cinnamaldehyde on Salmonella typhimurium at the protein level, digs the target of action, lays a theoretical foundation for further exploration, and provides new ideas for the clinical use and development of cinnamaldehyde.Therefore, cinnamaldehyde may have the potential to be used combined with antibiotics.

scholarly journals The Relationship Between Adversity Quotient and Career Adaptability of Internship Nursing Students

2018 ◽  
Vol 1 (1) ◽  
pp. 24
Author(s):  
Annida Nur Shalihah ◽  
Kurniawan Yudianto ◽  
Nur Oktavia Hidayati
Keyword(s):  
Nursing Students ◽  
Nursing Student ◽  
P Value ◽  
Career Adaptability ◽  
Contributing Factors ◽  
Response Profile ◽  
Spearman Correlation Test ◽  
High Level ◽  
The Relationship ◽  
Very High

A competitive job market is a challenge that needs to be faced by every university graduate. It is expected that every graduate has capability to keep face with expansion in job sectors. To be able to adapt to the job situation, a specific intelligence to face in facing every challenge and obstacle namely adversity quotient, is required. This study aimed to identify the correlation between adversity quotient (AQ) and career adaptability of internship nursing students. This study was descriptive correlational. Samples were recruited using a total sampling tecnique (n=142). Data were collected using a questionnaire consisting of a demographic survey, an Adversity Response Profile (ARP) Quick TakeTM and a Career Adapt-Abilities Scale (CAAS). Data were analyzed using descriptive statistics and Rank Spearman correlation test. The result of this study indicated that no students were in quitter category, only 0,7% of them were in quitter to camper transition, 76,8% of them were campers, 21,8% of the students were in camper to climber transition, 0,7% of all respondents were climbers. No students had a very low career adaptability, only 1,4% of them had a low career adaptability, while the proportions of students with a high and very high career adaptability were 75,4% and 23,2 % respectively. No significant correlation was found between AQ and career adaptability (p value >0,05; Spearman’s ρ +0,122). It can be concluded that most of internship nursing students at Faculty of Nursing Universitas Padjadjaran were campers and had a high level of career adaptability. There was also no significant correlation between AQ and career adaptability. Further research needs to explore about contributing factors related to career adaptability in internship nursing students.Keywords: Adversity quotient, career adaptability, internship nursing student. Hubungan Adversity Quotient dan Adaptabilitas Karier pada Mahasiswa Profesi NersAbstrakDalam menghadapi pasar kerja yang semakin ketat, setiap lulusan universitas diharapkan memiliki kemampuan untuk menghadapi tantangan dan beradaptasi dengan perkembangan pada dunia kerja. Untuk mengatasi hal tersebut diperlukan adanya kecerdasan dalam menghadapi setiap tantangan dan hambatan yang disebut adversity quotient (AQ). Penelitian ini bertujuan untuk mengetahui hubungan AQ dan adaptabilitas karier pada mahasiswa profesi ners. Penelitian ini menggunakan metode deskriptif korelatif. Responden penelitian berjumlah 181orang mahasiswa program profesi ners angkatan 32 dan 33 di Fakultas Keperawatan Universitas Padjadjaran yang di ambil dengan teknik total sampling dengan response rate 78,45%. Instrumen yang digunakan pada penelitian ini yaitu Adversity Response Profile (ARP) Quick TakeTM dan Career Adapt-Abilities Scale (CAAS). Teknik analisis data yang digunakan dalam penelitian ini menggunakan skoring ARP QuickTakeTM, skoring CAAS, serta uji korelasi Rank Spearman. Hasil penelitian menunjukkan bahwa tidak seorangpun responden yang berada pada kategori quitter, sebanyak 0,7% dari responden berada pada kategori transisi quitter ke camper, pada kategori camper sebanyak 76,8%, pada kategori transisi camper ke climber sebanyak 21,8%, dan pada kategori climber sebanyak 0,7%. Tidak ada mahasiswa dengan adaptabilitas karier sangat rendah, hanya 1,4% mahasiswa dengan adaptabilitas karier rendah, sedangkan mahasiswa dengan adaptabilitas karier tinggi dan sangat tinggi masing-masing proporsinya sebanyak 75,4% dan 23,2%. Tidak terdapat hubungan bermakna antara AQ dan adaptabilitas karier (p value >0,05; Spearman’s ρ +0,122). Simpulan dari penelitian ini yaitu sebagian besar mahasiswa profesi ners Fakultas Keperawatan Universitas Padjadjaran memiliki AQ dengan kategori camper dan memiliki adaptabilitas karier tinggi. Meskipun demikian, tidak terdapat hubungan bermakna antara AQ dan adaptabilitas karier. Bagi peneliti selanjutnya diharapkan untuk melakukan penelitian terkait faktor-faktor yang mempengaruhi adaptabilitas karier pada mahasiswa profesi ners.Kata kunci: Adversity quotient, adaptabilitas karier, mahasiswa profesi ners.

scholarly journals iTRAQ-Based Proteomics Analysis of Autophagy-Mediated Responses against MeJA in Laticifers of Euphorbia kansui L.

2019 ◽  
Vol 20 (15) ◽  
pp. 3770
Author(s):  
Fang ◽  
Yao ◽  
Zhang ◽  
Tian ◽  
Wang ◽  
...  
Keyword(s):  
Developmental Process ◽  
Cysteine Proteinase ◽  
Defense Responses ◽  
Absolute Quantification ◽  
Pcr Analysis ◽  
Control Group ◽  
Proteomics Analysis ◽  
Meja Treatment ◽  
Euphorbia Kansui ◽  
Isobaric Tags

Autophagy is a well-defined catabolic mechanism whereby cytoplasmic materials are engulfed into a structure termed the autophagosome. Methyl jasmonate (MeJA), a plant hormone, mediates diverse developmental process and defense responses which induce a variety of metabolites. In plants, little is known about autophagy-mediated responses against MeJA. In this study, we used high-throughput comparative proteomics to identify proteins of latex in the laticifers. The isobaric tags for relative and absolute quantification (iTRAQ) MS/MS proteomics were performed, and 298 proteins among MeJA treated groups and the control group of Euphorbia kansui were identified. It is interesting to note that 29 significant differentially expressed proteins were identified and their associations with autophagy and ROS pathway were verified for several selected proteins as follows: α-L-fucosidase, β-galactosidase, cysteine proteinase, and Cu/Zn superoxide dismutase. Quantitative real-time PCR analysis of the selected genes confirmed the fact that MeJA might enhance the expression of some genes related to autophagy. The western blotting and immunofluorescence results of ATG8 and ATG18a which are two important proteins for the formation of autophagosomes also demonstrated that MeJA could promote autophagy at the protein level. Using the electron microscope, we observed an increase in autophagosomes after MeJA treatment. These results indicated that MeJA might promote autophagy in E. kansui laticifers; and it was speculated that MeJA mediated autophagy through two possible ways: the increase of ROS induces ATG8 accumulation and then aotophagosome formation, and MeJA promotes ATG18 accumulation and then autophagosome formation. Taken together, our results provide several novel insights for understanding the mechanism between autophagy and MeJA treatment. However, the specific mechanism remains to be further studied in the future.

scholarly journals Comparative Proteomic Analysis of Dipsacus asperoides Roots from Different Habitats in China

Molecules ◽  
2020 ◽  
Vol 25 (16) ◽  
pp. 3605
Author(s):  
Haijun Jin ◽  
Hua Yu ◽  
Haixia Wang ◽  
Jia Zhang
Keyword(s):  
Proteomic Analysis ◽  
Absolute Quantification ◽  
Pcr Analysis ◽  
Allene Oxide ◽  
Allene Oxide Cyclase ◽  
Protein Levels ◽  
Depth Analysis ◽  
Differential Proteins ◽  
Isobaric Tags

Dipsacus asperoides is a kind of Chinese herbal medicine with beneficial health properties. To date, the quality of D. asperoides from different habitats has shown significant differences. However, the molecular differences in D. asperoides from different habitats are still unknown. The aim of this study was to investigate the differences in protein levels of D. asperoides from different habitats. Isobaric tags for relative and absolute quantification (iTRAQ) and 2DLC/MS/MS were used to detect statistically significant changes in D. asperoides from different habitats. Through proteomic analysis, a total of 2149 proteins were identified, of which 42 important differentially expressed proteins were screened. Through in-depth analysis of differential proteins, the protein metabolism energy and carbohydrate metabolism of D. asperoides from Hubei Province were strong, but their antioxidant capacity was weak. We found that three proteins, UTP-glucose-1-phosphate uridylyltransferase, allene oxide cyclase, and isopentyl diphosphate isomerase 2, may be the key proteins involved in dipsacus saponin VI synthesis. Eight proteins were found in D. asperoides in response to environmental stress from different habitats. Quantitative real-time PCR analysis confirmed the accuracy and authenticity of the proteomic analysis. The results of this study may provide the basic information for exploring the cause of differences in secondary metabolites in different habitats of D. asperoides and the protein mechanism governing differences in quality.

scholarly journals Quantitative proteomic analysis of G-protein signalling inStagonospora nodorumusing isobaric tags for relative and absolute quantification

PROTEOMICS ◽  
2010 ◽  
Vol 10 (1) ◽  
pp. 38-47 ◽  
Author(s):  
Tammy Casey ◽  
Peter S. Solomon ◽  
Scott Bringans ◽  
Kar-Chun Tan ◽  
Richard P. Oliver ◽  
...  
Keyword(s):  
G Protein ◽  
Proteomic Analysis ◽  
Absolute Quantification ◽  
Quantitative Proteomic Analysis ◽  
G Protein Signalling ◽  
Isobaric Tags

scholarly journals Monocyte Proteomics Reveals Involvement of Phosphorylated HSP27 in the Pathogenesis of Osteoporosis

2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Bhavna Daswani ◽  
Manoj Kumar Gupta ◽  
Shubhangi Gavali ◽  
Meena Desai ◽  
Gajanan J. Sathe ◽  
...  
Keyword(s):  
Absolute Quantification ◽  
Osteoclast Formation ◽  
Mineral Density ◽  
Monocyte Migration ◽  
Receptor Activator ◽  
Lower Chamber ◽  
Isobaric Tags ◽  
First Time ◽  
Upper Chamber ◽  
Increased Susceptibility

Peripheral monocytes, precursors of osteoclasts, have emerged as important candidates for identifying proteins relevant to osteoporosis, a condition characterized by low Bone Mineral Density (BMD) and increased susceptibility for fractures. We employed 4-plex iTRAQ (isobaric tags for relative and absolute quantification) coupled with LC-MS/MS (liquid chromatography coupled with tandem mass spectrometry) to identify differentially expressed monocyte proteins from premenopausal and postmenopausal women with low versus high BMD. Of 1801 proteins identified, 45 were differentially abundant in low versus high BMD, with heat shock protein 27 (HSP27) distinctly upregulated in low BMD condition in both premenopausal and postmenopausal categories. Validation in individual samples (n=80) using intracellular ELISA confirmed that total HSP27 (tHSP27) as well as phosphorylated HSP27 (pHSP27) was elevated in low BMD condition in both categories (P<0.05). Further, using transwell assays, pHSP27, when placed in the upper chamber, could increase monocyte migration (P<0.0001) and this was additive in combination with RANKL (receptor activator ofNFkBligand) placed in the lower chamber (P=0.05). Effect of pHSP27 in monocyte migration towards bone milieu can result in increased osteoclast formation and thus contribute to pathogenesis of osteoporosis. Overall, this study reveals for the first time a novel link between monocyte HSP27 and BMD.

scholarly journals iTRAQ-Based Comparative Proteomics Analysis of Urolithiasis Rats Induced by Ethylene Glycol

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Yanan Cao ◽  
Bin Duan ◽  
Xiaowei Gao ◽  
E. Wang ◽  
Zhitao Dong
Keyword(s):  
Ethylene Glycol ◽  
Kidney Stones ◽  
Enrichment Analysis ◽  
Absolute Quantification ◽  
Rat Models ◽  
Protein Protein Interaction ◽  
Parallel Reaction Monitoring ◽  
High Prevalence ◽  
Isobaric Tags ◽  
And Control

Nephrolithiasis is a frequent chronic urological condition with a high prevalence and recurrence rate. Proteomics studies on urolithiasis rat models are highly important in characterizing the pathophysiology of kidney stones and identifying potential approaches for preventing and treating kidney stones. The isobaric tags for relative and absolute quantification (iTRAQ) were performed to identify differentially expressed proteins (DEPs) in the kidney between urolithiasis rats and control rats. The results showed that 127 DEPs (85 upregulated and 42 downregulated) were identified in urolithiasis and control rats. The functions of DEPs were predicted by Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, and protein–protein interaction (PPI) network analysis. The expression of four upregulated proteins (Tagln, Akr1c9, Spp1, and Fbn1) and four downregulated proteins (Hbb, Epb42, Hmgcs2, and Ca1) were validated by parallel reaction monitoring (PRM). Proteomics studies of ethylene glycol-induced urolithiasis rat models using iTRAQ and PRM helped to elucidate the molecular mechanism governing nephrolithiasis and to identify candidate proteins for the treatment of kidney stones.

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