The impact of fertilization on the chicken egg yolk plasma and granule proteome 24 hours post-lay at room temperature: capitalizing on high-pH/low-pH reverse phase chromatography in conjunction with tandem mass tag (TMT) technology

Abstract Redox signalling plays an important role in endothelial cell (EC) physiology and pathophysiology. Proteins sense redox signals via cysteine thiol groups. A common oxidative post-translational modification (oxPTM) on cysteine thiols is S-glutathionylation which is reversed to a free thiol state by glutaredoxin (Glrx). OxPTMs alter protein function, location and stability. Identifying which proteins undergo modification will help determine the role of redox signalling in EC function. A proteome-wide screen of human cardiac microvascular endothelial cells identified redox-sensitive proteins involved in vascular signalling mechanisms. Methods Human microvascular endothelial cells (HCMVEC, Lonza) were exposed to VEGF (50ng/ml, 24h) or hypoxia (0.5% O2, 24h) with adenoviral (ad) Glrx (Vectorlabs) overexpression or adLacZ (control). A tandem mass tag mass spectrometry system (TMT) coupled with a thiol-switch technique was used to quantify changes in redox sensitive thiol modifications. Protein lysates were treated with MMTS to alkylate unmodified thiols. Iodo-TMT six-plex probes were tagged to redox-sensitive sites after reversal of oxPTMs by DTT. Samples were pooled and processed by nLC-MS/MS. The abundance of each peptide in different conditions was compared with either adGlrx or adLacZ (control) expression to provide a ratio of changes in redox modifications. Results Iodo-TMT analysis revealed 113 unique thiol modifications identified on 78 different proteins using a ±1.5-fold threshold in a given treatment. Additionally, 44 modifications in 33 proteins were present in at least 2 different conditions, namely Glrx under VEGF and hypoxic conditions. A STRING interaction network identified clusters of 10 proteins involved in organonitrogen synthesis and 6 proteins in angiogenesis. Jagged-1 involved in the regulation of angiogenic sprouting through the Notch pathway was established as a target of redox signalling. Identified redox sensitive cysteines were found in extracellular EFG1 and the calcium binding EGF12 domains. Seven different In Silico programs (including MutationTaster, PolyPhen-2 and PANTHER) predicting the impact of substitution mutations indicated a functional affect for these redox sensitive sites, demonstrating the importance of these residues. Conclusion A non-biased proteomics approach identified novel thiol modifications on proteins involved in microvascular function. Future work will demonstrate the impact of these redox-sensitive thiol modifications on microvascular function to provide a better understanding of redox signalling in protein function and disease. FUNDunding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): MRC-DTP,European Union's Horizon 2020 research and innovation programme.

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Quantitative Analysis of Pyrazines and Their Perceptual Interactions in Soy Sauce Aroma Type Baijiu

Foods ◽

10.3390/foods10020441 ◽

2021 ◽

Vol 10 (2) ◽

pp. 441

Author(s):

Yan Yan ◽

Shuang Chen ◽

Yao Nie ◽

Yan Xu

Keyword(s):

Mass Spectrometry ◽

Quantitative Analysis ◽

Ultra Performance Liquid Chromatography ◽

Alcohol Solution ◽

Soy Sauce ◽

Tandem Mass ◽

Odor Thresholds ◽

Quantitative Results ◽

Odor Activity Value ◽

The Impact

Pyrazines are important compounds in soy sauce aroma type Baijiu (SSAB). In this work, a total of 16 pyrazines were analyzed using ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC–MS/MS) in SSAB. The quantitative results showed that 2,3,5,6-tetramethylpyrazine, 2,6-dimethylpyrazine and 2,3,5-trimethylpyrazine were the three most concentrated pyrazines. The highest odor activity value (OAV) was determined for 2-ethyl-3,5-dimethylpyrazine. Quantitative analysis combined with descriptive sensory analysis revealed that sub-threshold pyrazines (2,3-dimethylpyrazine, 2,3-diethylpyrazine, 2,3-diethyl-5-methylpyrazine and 2-acetyl-3-methylpyrazine) are significantly correlated with the roasted aroma in SSAB. Our study focused on the impact of sub-threshold pyrazines on the perception of roasted aroma in SSAB. The effect of the sub-threshold pyrazines was detected by the addition of various pyrazines in SSAB samples, despite their sub-threshold concentrations. Furthermore, the presence of sub-threshold pyrazines in dilute alcohol solution resulted in a significant reduction in the odor thresholds of supra-threshold pyrazines. Sensory investigation indicated that pyrazines have a synergistic effect on the perception of roasted aroma. The results highlighted the contribution of some pyrazines to the roasted aroma in SSAB despite their sub-threshold concentrations.

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Determination of multi pesticide residues in leaf and needle samples using a modified QuEChERS approach and gas chromatography-tandem mass spectrometry

Analytical Methods ◽

10.1039/d0ay02329a ◽

2021 ◽

Author(s):

Arnelle Löbbert ◽

Sonja Schanzer ◽

Henrik Krehenwinkel ◽

Franz Bracher ◽

Christoph Müller

Keyword(s):

Mass Spectrometry ◽

Gas Chromatography ◽

Tandem Mass Spectrometry ◽

Forest Ecosystems ◽

Tandem Mass ◽

Modified Quechers ◽

Chromatography Tandem Mass Spectrometry ◽

The Impact ◽

Impact Of Pesticides

A novel, validated QuEChERS-based GC-MS/MS method was developed, which will allow the assessment of the impact of pesticides on forest ecosystems.

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Assessment of the impact of hydrolysis on bound sulfonamide residue determination in honey using stable isotope dilution ultrahigh performance liquid chromatography tandem mass spectrometry

Food Chemistry ◽

10.1016/j.foodchem.2021.130094 ◽

2021 ◽

pp. 130094

Author(s):

Yan Zhang ◽

Xiu Qin Li ◽

Zhen Guo ◽

Xia Zhou ◽

Shuang Qing Li ◽

...

Keyword(s):

Mass Spectrometry ◽

Liquid Chromatography ◽

Stable Isotope ◽

Tandem Mass ◽

Stable Isotope Dilution ◽

Residue Determination ◽

Chromatography Tandem Mass Spectrometry ◽

Liquid Chromatography Tandem Mass ◽

Ultrahigh Performance Liquid Chromatography ◽

The Impact

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COMPARISON OF ANTIVENOM POTENTIAL OF CHICKEN EGG YOLK ANTIBODIES GENERATED AGAINST BENTONITE AND ADJUVENT COATED VENOMES OF COMMON POISONOUS SNAKE IN INDIA

Bangladesh Journal of Veterinary Medicine ◽

10.3329/bjvm.v7i1.5070 ◽

1970 ◽

Vol 7 (1) ◽

pp. 259-267 ◽

Cited By ~ 1

Author(s):

S. Meenatchisundaram ◽

R. Selvakumaran ◽

G. Parameswari ◽

A. Michael

Keyword(s):

Room Temperature ◽

Deae Cellulose ◽

Egg Yolk ◽

Precipitation Method ◽

Enzyme Linked Immunosorbent Assay ◽

Snake Venoms ◽

Chicken Egg ◽

Ammonium Sulphate Precipitation ◽

Egg Yolk Antibodies ◽

Chicken Egg Yolk

Antivenom antibodies were generated in white leghorn chicken against bentonite and adjuvant coated venoms of Common Indian Poisonous Snakes (Cobra, Krait, Russell's viper and Saw Scaled viper).The antivenom from immunized chicken egg yolk were purified by polyethylene glycol (PEG) and ammonium sulphate precipitation method and further purified by DEAE cellulose ion exchange column chromatography and concentrated by polyvinyl pyrolidone powder at room temperature. The titer of antibodies was estimated using Enzyme Linked Immunosorbent Assay (ELISA).The chickens immunized with Freund's complete adjuvant showed slightly higher titre when compared to bentonite. Inhibition of lethal, edema, haemorrhagic, procoagulant and phospholipase A2 and fibrinolytic activities of snake venoms were determined. The chicken egg yolk antivenom was effective in neutralization of these toxic and enzymatic activities of venom. The median effective dose (ED50) of chicken egg yolk antibodies raised against adjuvant coated venoms showed effective neutralizing venom toxicity when compared to the antibodies raised using bentonite coated venoms.

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The proteins cleaved by endogenous tryptic proteases in normal EDTA plasma by C18 collection of peptides for liquid chromatography micro electrospray ionization and tandem mass spectrometry

10.32920/14640405.v1 ◽

2021 ◽

Author(s):

Jaimie Dufresne ◽

Angelique Florentinus-Mefailoski ◽

Juliet Ajambo ◽

Ammara Ferwa ◽

Peter Bowden ◽

...

Keyword(s):

Mass Spectrometry ◽

Liquid Chromatography ◽

Tandem Mass Spectrometry ◽

Electrospray Ionization ◽

Room Temperature ◽

Cellular Proteins ◽

Tandem Mass ◽

Temperature Plasma ◽

Tryptic Peptides ◽

Gene Symbols

The tryptic peptides from ice cold versus room temperature plasma were identified by C18 liquid chromatography and micro electrospray ionization tandem mass spectrometry (LC–ESI–MS/MS). Samples collected on ice showed low levels of endogenous tryptic peptides compared to the same samples incubated at room temperature. Plasma on ice contained peptides from albumin, complement, and apolipoproteins and others that were observed by the X!TANDEM and SEQUEST algorithms. In contrast to ice cold samples, after incubation at room temperature, greater numbers of tryptic peptides from well characterized plasma proteins, and from cellular proteins were observed. A total of 583,927 precursor ions and MS/MS spectra were correlated to 94,669 best fit peptides that reduced to 22,287 correlations to the best accession within a gene symbol and to 7174 correlations to at least 510 gene symbols with ≥ 5 independent MS/MS correlations (peptide counts) that showed FDR q-values ranging from E−9 (i.e. FDR = 0.000000001) to E−227. A set of 528 gene symbols identified by X!TANDEM and SEQUEST including C4B showed ≥ fivefold variation between ice cold versus room temperature incubation. STRING analysis of the protein gene symbols observed from endogenous peptides in normal plasma revealed an extensive protein-interaction network of cellular factors associated with cell signalling and regulation, the formation of membrane bound organelles, cellular exosomes and exocytosis network proteins. Taken together the results indicated that a pool of cellular proteins, or protein complexes, in plasma are apparently not stable and degrade soon after incubation at room temperature.

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National Academy of Clinical Biochemistry Laboratory Medicine Practice Guidelines: Follow-Up Testing for Metabolic Disease Identified by Expanded Newborn Screening Using Tandem Mass Spectrometry; Executive Summary

Clinical Chemistry ◽

10.1373/clinchem.2009.131300 ◽

2009 ◽

Vol 55 (9) ◽

pp. 1615-1626 ◽

Cited By ~ 54

Author(s):

Dennis J Dietzen ◽

Piero Rinaldo ◽

Ronald J Whitley ◽

William J Rhead ◽

W Harry Hannon ◽

...

Keyword(s):

Mass Spectrometry ◽

Amino Acids ◽

Tandem Mass Spectrometry ◽

Organic Acids ◽

Newborn Screening ◽

Tandem Mass ◽

Confirmatory Testing ◽

The Us ◽

The Impact

Abstract Background: Almost all newborns in the US are screened at birth for multiple inborn errors of metabolism using tandem mass spectrometry. Screening tests are designed to be sufficiently sensitive so that cases are not missed. The NACB recognized a need for standard guidelines for laboratory confirmation of a positive newborn screen such that all babies would benefit from equal and optimal follow-up by confirmatory testing. Methods: A committee was formed to review available data pertaining to confirmatory testing. The committee evaluated previously published guidelines, published methodological and clinical studies, clinical case reports, and expert opinion to support optimal confirmatory testing. Grading was based on guidelines adopted from criteria derived from the US Preventive Services Task Force and on the strength of recommendations and the quality of the evidence. Three primary methods of analyte measurement were evaluated for confirmatory testing including measurement of amino acids, organic acids, and carnitine esters. The committee graded the evidence for diagnostic utility of each test for the screened conditions. Results: Ample data and experience were available to make strong recommendations for the practice of analyzing amino acids, organic acids, and acylcarnitines. Likewise, strong recommendations were made for the follow-up test menu for many disorders, particularly those with highest prevalence. Fewer data exist to determine the impact of newborn screening on patient outcomes in all but a few disorders. The guidelines also provide an assessment of developing technology that will fuel a refinement of current practice and ultimate expansion of the diseases detectable by tandem mass spectrometry. Conclusions: Guidelines are provided for optimal follow-up testing for positive newborn screens using tandem mass spectrometry. The committee regards these tests as reliable and currently optimal for follow-up testing. .

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