Quantitative screening of protein biomarkers of early glycation, advanced glycation, oxidation and nitrosation in cellular and extracellular proteins by tandem mass spectrometry multiple reaction monitoring

2003 ◽  
Vol 31 (6) ◽  
pp. 1417-1422 ◽  
Author(s):  
N. Ahmed ◽  
P.J. Thornalley
Keyword(s):  
Blood Plasma ◽  
Nitrosative Stress ◽  
Cellular Protein ◽  
Extracellular Proteins ◽  
Protein Glycation ◽  
Advanced Glycation ◽  
Protein Biomarkers ◽  
Renal Clearances ◽  
Quantitative Screening ◽  
In Blood Plasma

Glycation of proteins forms fructosamines and AGEs (advanced glycation end products). Oxidative and nitrosative stress leads to the formation of oxidative and nitrosative modifications. The modified amino acid residues formed in these processes are biomarkers of protein damage: some are risk markers and some may be risk factors for disease development. We developed a method for the concurrent quantitative measurement of 16 biomarkers indicative of protein glycation, oxidation and nitrosation damage using LC-MS/MS (LC with tandem MS detection). Underivatized analytes were detected free in physiological fluids and in enzymatic hydrolysates of cellular and extracellular proteins. Hydroimidazolones were the most important glycation biomarkers, and methionine sulphoxide was the most important oxidative biomarker quantitatively; 3-nitrotyrosine was the biomarker of nitrosation. Quantitative screening showed high levels of AGEs in cellular protein and moderate levels in protein of blood plasma. Levels of 3-nitrotyrosine were typically 100-fold lower than this. The major glycation adducts in blood plasma had high renal clearances in normal healthy human subjects, whereas methionine sulphoxide and 3-nitrotyrosine had low renal clearances due to further metabolism. Physiological AGEs in blood plasma were eliminated from the circulation in the kidney and not in the liver. LC-MS/MS peptide mapping was also used to locate the protein biomarkers. These studies reveal that advanced glycation is a significant modification of cellular and extracellular protein. The enzymatic defences against glycation, antioxidants and proteasomal protein degradation inside cells are probable factors regulating biomarker levels of cellular protein.

scholarly journals Quantitative screening of advanced glycation endproducts in cellular and extracellular proteins by tandem mass spectrometry

2003 ◽  
Vol 375 (3) ◽  
pp. 581-592 ◽  
Author(s):  
Paul J. THORNALLEY ◽  
Sinan BATTAH ◽  
Naila AHMED ◽  
Nikolaos KARACHALIAS ◽  
Stamatina AGALOU ◽  
...  
Keyword(s):  
Blood Plasma ◽  
Advanced Glycation Endproducts ◽  
Vascular Complications ◽  
Extracellular Proteins ◽  
Tandem Mass ◽  
Advanced Glycation ◽  
Glycation Endproducts ◽  
Advanced Glycation Endproduct ◽  
Quantitative Screening ◽  
In Blood Plasma

Glycation of proteins forms fructosamines and advanced glycation endproducts. Glycation adducts may be risk markers and risk factors of disease development. We measured the concentrations of the early glycation adduct fructosyl-lysine and 12 advanced glycation endproducts by liquid chromatography with tandem mass spectrometric detection. Underivatized analytes were detected free in physiological fluids and in enzymic hydrolysates of cellular and extracellular proteins. Hydroimidazolones were the most important glycation biomarkers quantitatively; monolysyl adducts (Nε-carboxymethyl-lysine and Nε-1-carboxyethyl-lysine) were found in moderate amounts, and bis(lysyl)imidazolium cross-links and pentosidine in lowest amounts. Quantitative screening showed high levels of advanced glycation endproducts in cellular protein and moderate levels in protein of blood plasma. Glycation adduct accumulation in tissues depended on the particular adduct and tissue type. Low levels of free advanced glycation endproducts were found in blood plasma and levels were 10–100-fold higher in urine. Advanced glycation endproduct residues were increased in blood plasma and at sites of vascular complications development in experimental diabetes; renal glomeruli, retina and peripheral nerve. In clinical uraemia, the concentrations of plasma protein advanced glycation endproduct residues increased 1–7-fold and free adduct concentrations increased up to 50-fold. Comprehensive screening of glycation adducts revealed the relative and quantitative importance of α-oxoaldehyde-derived advanced glycation endproducts in physiological modification of proteins–particularly hydroimidazolones, the efficient renal clearance of free adducts, and the marked increases of glycation adducts in diabetes and uraemia–particularly free advanced glycation endproducts in uraemia. Increased levels of these advanced glycation endproducts were associated with vascular complications in diabetes and uraemia.

scholarly journals Cardanol-Based Salophen-Modified Electrode for Analysing Nitric Oxide Bioavailability under Nitrosative Stress Conditions

2022 ◽  
Author(s):  
Aristides Reis ◽  
André Santos ◽  
Amison Souza ◽  
Luiz Arrais Junior ◽  
Heberty Facundo ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Blood Plasma ◽  
Nitrosative Stress ◽  
Signal To Noise Ratio ◽  
Transient Behavior ◽  
High Reactivity ◽  
Great Sensitivity ◽  
Modified Carbon Paste Electrode ◽  
Carbon Paste ◽  
In Blood Plasma

Abstract High levels of nitric oxide (NO) can signal nitrosative stress, but its analysis is challenging considering the high reactivity, short half-life and transient behavior of this target molecule in biological milieu. In this work, a cardanol-based salophen-modified carbon paste electrode (CDN-salophen/MCPE) was developed and successfully applied to assess NO bioavailability in blood plasma of mice under induced stress. The results revealed that the modifier improved the device performance in terms of signal-to-noise ratio, charge-transport and fouling resistance. NO reactivity on CDN-salophen/MCPE was higher in 0.1 mol L‒1 H2SO4, and the resulting redox process involves adsorption steps that control the reaction kinetics. Monitoring molecule oxidation by square-wave voltammetry (100 s−1 frequency, 30 mV amplitude, 2 mV scan increment, after electrode preconditioning at 0.9 V for 15 s for analyte accumulation), it was possible to identify and quantify NO with great sensitivity (detection and quantification limit < 0.1 µmol L‒1) and low data variance (RSD ≤ 9.4% for repeatability and reproducibility tests), through a simple, fast and reliable electroanalytical protocol. The robustness acquired with CDN-salophen/MCPE allowed to detect changes in NO content in blood plasma during nitrosative stress, proving its efficiency for research on this subject.

Removal of advanced glycation end products in clinical renal failure by peritoneal dialysis and haemodialysis

2003 ◽  
Vol 31 (6) ◽  
pp. 1394-1396 ◽  
Author(s):  
S. Agalou ◽  
N. Ahmed ◽  
A. Dawnay ◽  
P.J. Thornalley
Keyword(s):  
Renal Failure ◽  
Peritoneal Dialysis ◽  
Blood Plasma ◽  
Advanced Glycation End Products ◽  
Peritoneal Cavity ◽  
Dwell Time ◽  
Advanced Glycation ◽  
Glycation End Products ◽  
End Products ◽  
In Blood Plasma

AGEs (advanced glycation end products) accumulate markedly in the plasma of human subjects with renal failure. We investigated the efficiency of removal of AGEs from the circulation by PD (peritoneal dialysis) and HD (haemodialysis) therapy. Free AGEs were measured by LC-MS/MS in blood plasma before dialysis, in dialysis fluid effusate after a 2–12 h dwell time in the peritoneal cavity of PD subjects, and in the HD dialysate before and after HD therapy. In clinical uraemia, the concentrations of free AGEs in blood plasma were increased up to 50-fold. For example, levels of MG-H1 (methylglyoxal-derived hydroimidazolone) were: normal controls, 110±46 nM; PD subjects, 1876±676 (P<0.01); HD subjects, 5496±1138 nM (P<0.001). In PD subjects, the AGE concentration in the effusate increased with increasing dwell time, reaching a maximum at a concentration higher than that in plasma for some AGEs at 4–12 h. This may reflect AGE formation in the peritoneal cavity. In HD, AGE concentrations in HD fluid were decreased markedly from the start to the end of a dialysis session, except that levels of the methylglyoxal-derived AGEs N∊-(1-carboxyethyl)lysine and MG-H1, and of pentosidine, remained 5-fold higher than control levels. Inadequate clearance of free AGEs may be linked to the increased risk of cardiovascular disease in patients with renal failure.

State of hormonal regulation in experimental animals exposed to noise and vibration

2020 ◽  
pp. 680-681
Author(s):  
A. V. Lizarev ◽  
V. A. Pankov
Keyword(s):  
Blood Plasma ◽  
Hormonal Regulation ◽  
Adrenocorticotropic Hormone ◽  
Vibration Exposure ◽  
Noise And Vibration ◽  
Experimental Animals ◽  
In Blood Plasma

When exposed to noise and vibration in experimental animals there was a decrease in the content of threeiodinethyronine, thyroxin and adrenocorticotropic hormone in blood plasma after 15 and 30 days of experience. An increase in loads led to an increase in the level of threeiodinethyronine and thyroxin under vibration exposure and was normalized with noise. The content of adrenocorticotropic hormone leveled in both cases.

EFFECT OF DIETARY LEVELS OF SODIUM AND POTASSIUM ON GROWTH AND ON CONCENTRATIONS IN BLOOD PLASMA AND TISSUES OF WHITE RAT

1950 ◽  
Vol 162 (1) ◽  
pp. 182-188 ◽  
Author(s):  
J. H. Meyer ◽  
R. R. Grunert ◽  
Marie T. Zepplin ◽  
R. H. Grummer ◽  
G. Bohstedt ◽  
...  
Keyword(s):  
Blood Plasma ◽  
Sodium And Potassium ◽  
In Blood Plasma

Two SPRi biosensors for the determination of cathepsin S in blood plasma

Talanta ◽  
2021 ◽  
Vol 225 ◽  
pp. 121900
Author(s):  
Lukasz Oldak ◽  
Anna Sankiewicz ◽  
Beata Żelazowska-Rutkowska ◽  
Bogdan Cylwik ◽  
Zenon Lukaszewski ◽  
...  
Keyword(s):  
Blood Plasma ◽  
Cathepsin S ◽  
In Blood Plasma

scholarly journals DETERMINATION OF TRYPSIN INHIBITOR IN BLOOD PLASMA

1953 ◽  
Vol 204 (1) ◽  
pp. 147-152 ◽  
Author(s):  
Shirley F. McCann ◽  
M. Laskowski
Keyword(s):  
Blood Plasma ◽  
Trypsin Inhibitor ◽  
In Blood Plasma

scholarly journals The comparison of pro- and antioxidative parameters in plasma and placental tissues during early phase of placental development in cows

2021 ◽  
Author(s):  
Jacek Wawrzykowski ◽  
Monika Jamioł ◽  
Wioleta Mojsym ◽  
Marta Kankofer
Keyword(s):  
Blood Plasma ◽  
Antioxidative Enzymes ◽  
Placental Development ◽  
Blood Samples ◽  
Bovine Placenta ◽  
Total Antioxidant ◽  
Tissue Homogenates ◽  
Further Development ◽  
General Profile ◽  
In Blood Plasma

AbstractPhysiological balance between pro- and antioxidative processes is crucial for placentation and further development of fetus and placenta. Parameters of pro- and antioxidative profile may serve as markers of proper course of pregnancy. The aim of study was to assess whether the balance between pro- and antioxidative parameters during placentation phase in bovine placenta is maintained. Placental and blood samples were collected from healthy, HF, pregnant (2nd-3rd month) cows (n = 8) in slaughterhouse and in farm, respectively. Formylokinurenine and bityrosine content were measured spectrofluorimetrically in blood plasma and tissue homogenates while metabolites of lipid peroxidation, total antioxidant capacity, SH groups and activity of antioxidative enzymes (glutathione peroxidase and superoxide dismutase) were determined in examined tissues by spectrophotometry. Western blotting was used to confirm the presence of enzymatic proteins in placenta. Results: Local profile in tissues was more pronounced than general profile in blood plasma. Activities of antioxidative enzymes were significantly (p < 0.05) higher in 2nd compared to 3rd month of pregnancy in maternal part of placenta while prooxidant parameters showed opposite relationship. Obtained results showed significant differences when compared to data from non-pregnant animals or time of parturition. Further studies are necessary for elucidation of placentation phase in cows.

The convenient UPLS method for the determination of Ceftiofur in blood plasma

2020 ◽  
Author(s):  
Artem V. Baklykov ◽  
Konstantin A. Chistiakov ◽  
Dmitry S. Kopchuk ◽  
Grigory V. Zyryanov ◽  
Gennady L. Rusinov ◽  
...  
Keyword(s):  
Blood Plasma ◽  
In Blood Plasma
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