scholarly journals Comparison of two multiplexed technologies for profiling >1,000 serum proteins that may associate with tumor burden

F1000Research ◽  
2021 ◽  
Vol 10 ◽  
pp. 509
Author(s):  
Annie Ren ◽  
Ioannis Prassas ◽  
Vijithan Sugumar ◽  
Antoninus Soosaipillai ◽  
Marcus Bernardini ◽  
...  
Keyword(s):  
Serum Concentration ◽  
Protein Concentration ◽  
Serum Proteins ◽  
Tumor Burden ◽  
Cancer Antigen 125 ◽  
Concentration Changes ◽  
Related Proteins ◽  
Serological Tumor ◽  
Proximity Extension Assay ◽  
Single Enzyme

Background: In this pilot study, we perform a preliminary comparison of two targeted multiplex proteomics technologies for discerning serum protein concentration changes that may correlate to tumor burden in ovarian cancer (OC) patients. Methods: Using the proximity extension assay (PEA) and Quantibody® Kiloplex Array (QKA), we measured >1,000 proteins in the pre-surgical and post-surgical serum from nine OC patients (N=18 samples). We expect that proteins that have decreased significantly in the post-surgical serum concentration may correlate to tumor burden in each patient. Duplicate sera from two healthy individuals were used as controls (N=4 samples). We employed in-house ELISAs to measure five proteins with large serum concentration changes in pre- and post-surgical sera, from four of the original nine patients and the two original controls. Results: Both platforms showed a weak correlation with clinical cancer antigen 125 (CA125) data. The two multiplexed platforms showed a significant correlation with each other for >400 overlapping proteins. PEA uncovered 15 proteins, while QKA revealed 11 proteins, with more than a two-fold post-surgical decrease in at least six of the nine patients. Validation using single enzyme-linked immunosorbent assays (ELISAs) showed at least a two-fold post-surgical decrease in serum concentration of the same patients, as indicated by the two multiplex assays. Conclusion: Both methods identified proteins that had significantly decreased in post-surgical serum concentration, as well as recognizing proteins that had been implicated in OC patients. Our findings from a limited sample size suggest that novel targeted proteomics platforms are promising tools for identifying candidate serological tumor-related proteins.  However further studies are essential for the improvement of accuracy and avoidance of false results.

Serum Concentration and Skin Expression of S100A7 (Psoriasin) in Patients Suffering from Hidradenitis Suppurativa

Dermatology ◽  
2020 ◽  
pp. 1-7
Author(s):  
Aleksandra Batycka-Baran ◽  
Wojciech Baran ◽  
Danuta Nowicka-Suszko ◽  
Maria Koziol-Gałczyńska ◽  
Andrzej Bieniek ◽  
...  
Keyword(s):  
Serum Concentration ◽  
Hidradenitis Suppurativa ◽  
Protein Concentration ◽  
Normal Skin ◽  
Innate Immune ◽  
Antimicrobial Protein ◽  
Healthy Controls ◽  
Serum Concentrations ◽  
Reactive Protein

<b><i>Background:</i></b> Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease. An important role of innate immune dysregulation in the pathogenesis of HS has been highlighted. S100A7 (psoriasin) is an innate, antimicrobial protein that exerts proinflammatory and chemotactic action. <b><i>Objectives:</i></b> The objective of the study was to investigate serum concentrations of S100A7 in individuals with HS as compared to healthy controls. Further, we evaluated the expression of S100A7 in lesional HS skin as compared to perilesional (clinically uninvolved) HS skin and normal skin. <b><i>Methods:</i></b> Serum concentrations of S100A7 were evaluated with a commercially available ELISA kit. The expression of S100A7 in the skin was assessed using qRT-PCR and immunofluorescence staining. <b><i>Results:</i></b> We found increased expression of S100A7 in lesional HS skin as compared to perilesional HS skin (<i>p</i> = 0.0017). The expression of S100A7 in lesional HS skin was positively associated with serum C-reactive protein concentration and the severity of disease according to Hurley staging. The serum concentration of S100A7 in individuals with HS was decreased as compared to healthy controls and patients with psoriasis. <b><i>Conclusions:</i></b> Upregulated in lesional HS skin, S100A7 may enhance the inflammatory process and contribute to the HS pathogenesis.

Electroimmunoassay of alpha-2-opsonic protein during reticuloendothelial blockade

1977 ◽  
Vol 232 (3) ◽  
pp. R80-R87 ◽  
Author(s):  
F. Blumenstock ◽  
P. Weber ◽  
T. M. Saba ◽  
R. Laffin
Keyword(s):  
Serum Concentration ◽  
Phagocytic Activity ◽  
Lipid Emulsion ◽  
Protein Concentration ◽  
Normal Values ◽  
Serum Levels ◽  
Physiological Control ◽  
Opsonic Activity ◽  
Physiological Regulation ◽  
Monospecific Antiserum

Physiological regulation of reticuloendothelial (RE) phagocytic activity by a plasma opsonic factor has been documented. In the recent study, serum levels of this alpha-2-opsonic protein in rats during colloid-induced RE blockade were measured utilizing an electroimmunoassay (Rocket immunoelectrophoresis) with monospecific antiserum to the purified alpha-2-glycoprotein. RE blockade was produced by the intravenous injection of the gelatinized "RE-test-lipid emulsion" at a dose of 50 mg/100 g body wt. The opsonic activity of serum at various intervals during colloid-induced RE blockade as measured by tissue slice bioassay manifested a high correlation (r = 0.98) with the serum opsonic protein concentration as measured by the electroimmunoassay. During RE blockade (30 min), there was a rapid depletion of the opsonic alpha-2-glycoprotein to 20% of the initial preinjection levels. Serum concentration of this glycoprotein remained low for at least 2-3 h after which time its concentration progressively increased with approximation of normal values by 6 h postblockade. Opsonic protein concentration at 24 h postinjection were significantly (P less than 0.05) elevated above controls. Thus, colloid-induced RE blockade is associated with the removal of this glycoprotein from the serum and recovery from RE blockade is accompanied by a restoration of opsonin levels. The electroimmunoassay can provide a sensitive technique to monitor this humoral factor known to exert a physiological control on the RE system.

Filtration coefficient in cat hindlimb using protein concentration changes

1989 ◽  
Vol 256 (1) ◽  
pp. H186-H194 ◽  
Author(s):  
P. D. Watson ◽  
M. B. Wolf
Keyword(s):  
Protein Concentration ◽  
Venous Pressure ◽  
Filtration Coefficient ◽  
Volume Changes ◽  
Vascular Volume ◽  
Perfusate Flow ◽  
Wide Range ◽  
Concentration Changes ◽  
The Difference ◽  
Pentobarbital Sodium Anesthesia

The maximum value of capillary filtration coefficient (CFC) in maximally vasodilated cat skeletal muscle is disputed. It was hypothesized that the wide range of reported values was caused by the inability of gravimetric and volumetric measurements of tissue volume to separate transcapillary filtration from vascular volume changes. Consequently, we developed a method of measuring filtration rates from changes in venous protein concentration using Evan's blue-labeled albumin in the isolated hindlimb (pentobarbital sodium anesthesia). The filtration coefficient (PFFC) calculated from these filtration rates after a step in venous pressure should not be influenced by vascular volume changes. When the perfusate flow rate through the hindlimb was greater than 15 ml.min-1.100 g muscle-1, PFFC was 0.0085 +/- 0.0015 (SD, n = 8) ml.min-1.mmHg-1.100 g muscle-1. PFFC was observed to be unvarying from 1 to 12 min after the venous pressure elevation, in contrast to CFC values, which fall during the same period. It is argued that the difference between CFC and PFFC values is caused by vascular volume changes.

PAROTID SECRETION OF PROTEIN IN MAN

1958 ◽  
Vol 36 (10) ◽  
pp. 1001-1008 ◽  
Author(s):  
Marion H. Ferguson ◽  
H. P. Krahn ◽  
J. A. Hildes
Keyword(s):  
Total Protein ◽  
Protein Concentration ◽  
Serum Proteins ◽  
Amylase Activity ◽  
Early Morning ◽  
Total Protein Concentration ◽  
Zone Electrophoresis ◽  
Residual Radioactivity ◽  
Unstimulated Saliva ◽  
Protein Components

In unstimulated saliva, total protein concentration averaged 186 mg per 100 ml and amylase activity 146 units per 100 ml. The protein concentration was lower in the early morning than at midday. After dilute acetic acid stimulation, both total protein concentration and amylase activity were increased but the concentrations were not affected by rates of secretion above 0.1 ml per minute. Unlike protein, the potassium concentration fell with stimulation.Using zone electrophoresis on filter paper, as many as nine protein components were found, none of which corresponded to the serum proteins. The amylase activity was restricted to a component of low mobility which moved to the anode. There were two or three bands containing glycoproteins; all moved towards the cathode. There were qualitative and quantitative differences between stimulated and unstimulated secretions. Saliva collected 2 or 24 hours after a tracer dose of I131 showed less than 1% residual radioactivity after dialysis or treatment with an anion exchange resin, indicating that little if any of the salivary iodide is organically bound.

scholarly journals Immune checkpoint-related serum proteins and genetic variants predict outcomes of localized prostate cancer, a cohort study

2020 ◽  
Author(s):  
Qinchuan Wang ◽  
Yuanqing Ye ◽  
Hao Yu ◽  
Shu-Hong Lin ◽  
Huakang Tu ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Mrna Expression ◽  
Immune Checkpoint ◽  
Serum Proteins ◽  
Meta Analysis ◽  
Genetic Variations ◽  
Localized Prostate Cancer ◽  
Nucleotide Polymorphisms ◽  
Clinical Predictors ◽  
Related Proteins

Abstract Background The clinical predictors and biological mechanisms for localized prostate cancer (PCa) outcomes remain mostly unknown. We aim to evaluate the role of serum immune-checkpoint-related (ICK) proteins and genetic variations in predicting outcomes of localized PCa. Methods We profiled the serum levels of 14 ICK-related proteins (BTLA, GITR, HVEM, IDO, LAG-3, PD-1, PD-L1, PD-L2, Tim-3, CD28, CD80, 4-1BB, CD27, and CTLA-4) in 190 patients with localized PCa. The genotypes of 97 single nucleotide polymorphisms (SNPs) from 19 ICK-related genes were analyzed in an extended population (N = 1762). Meta-data from ArrayExpress and TCGA was employed to validate and to probe functional data. Patients were enrolled and tumor aggressiveness, biochemical recurrence (BCR), and progression information were obtained. Statistical analyses were performed analyzing associations between serum biomarkers, genotypes, mRNA and outcomes. Results We showed that serum (s)BTLA and sTIM3 levels were associated with PCa aggressiveness (P < 0.05). sCD28, sCD80, sCTLA4, sGITR, sHVEM and sIDO correlated with both BCR and progression risks (all P < 0.05). We further identified ICK variants were significantly associated with aggressiveness, BCR and progression. Among them, 4 SNPs located in CD80 (rs7628626, rs12695388, rs491407, rs6804441) were not only associated with BCR and progression risk, but also correlated with sCD80 level (P < 0.01). rs491407 was further validated in an independent cohort. The CD80 mRNA expression was associated with BCR (HR, 1.85, 95% CI 1.06–3.22, P = 0.03) in meta-analysis of validation cohorts. Conclusion We highlight the prognostic value of serum ICK-related proteins for predicting aggressiveness, BCR and progression of PCa. The genetic variations and mRNA expression in CD80 could be predictors and potential targets of localized PCa.

scholarly journals Paraoxonase 1 Phenotype and Protein N-Homocysteinylation in Patients with Rheumatoid Arthritis: Implications for Cardiovascular Disease

Antioxidants ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 899
Author(s):  
Jolanta Parada-Turska ◽  
Grażyna Wójcicka ◽  
Jerzy Beltowski
Keyword(s):  
Rheumatoid Arthritis ◽  
Protein Concentration ◽  
Serum Proteins ◽  
Density Lipoprotein ◽  
Cardiovascular Complications ◽  
Paraoxonase 1 ◽  
Pon1 Activity ◽  
Phenyl Acetate ◽  
Control Subjects ◽  
Increased Risk

Paraoxonase 1 (PON1) is the high density lipoprotein-associated esterase which inhibits the development of atherosclerosis by metabolizing lipid peroxidation products as well as hydrolyzing proatherogenic metabolite of homocysteine (Hcy), Hcy thiolactone, which otherwise reacts with lysine groups of proteins, thus forming N-Hcy-protein in a process referred to as protein N-homocysteinylation. Rheumatoid arthritis (RA) is the chronic inflammatory autoimmune disease associated with increased risk of cardiovascular complications, but the underlying mechanisms are incompletely understood. We examined PON1 status and N-homocysteinylation of serum proteins in patients with RA. Blood was collected from 74 RA patients and 70 control subjects. PON1 activity was measured toward synthetic (paraoxon, phenyl acetate) and natural (Hcy thiolactone) substrates. PON1 protein concentration was measured by ELISA. Total Hcy as well as N-Hcy-protein were measured in serum as well. PON1 activity toward Hcy thiolactone was lower in RA patients than in control subjects which was accompanied by increased concentration of N-Hcy-protein despite normal total Hcy concentration. PON1 protein concentration was unchanged in the RA group, but the specific enzyme activity was reduced. When RA patients were categorized according to the DAS28-ESR score, PON1 concentration and enzymatic activity were lower whereas N-Hcy-protein was higher in those with high disease activity. PON1 activity and Hcy thiolactone were correlated with DAS28-ESR score and myeloperoxidase concentration. In conclusion, RA is associated with deficiency of PON1 activity and increased protein N-homocyseinylation which may contribute to accelerated development of cardiovascular diseases.

scholarly journals Cardiovascular Disease-Related Serum Proteins in Workers Occupationally Exposed to Polycyclic Aromatic Hydrocarbons

2019 ◽  
Vol 171 (1) ◽  
pp. 235-246 ◽  
Author(s):  
Ayman Alhamdow ◽  
Christian Lindh ◽  
Maria Albin ◽  
Per Gustavsson ◽  
Håkan Tinnerberg ◽  
...  
Keyword(s):  
Cardiovascular Disease ◽  
Polycyclic Aromatic Hydrocarbons ◽  
Aromatic Hydrocarbons ◽  
Serum Proteins ◽  
Linear Regression Analysis ◽  
Differentially Expressed ◽  
Incidence And Mortality ◽  
Polycyclic Aromatic ◽  
Pah Exposure ◽  
Related Proteins

AbstractChimney sweeps have higher incidence and mortality of cardiovascular disease (CVD), likely related to their exposure to polycyclic aromatic hydrocarbons (PAH). In order to identify underlying mechanisms of PAH-related CVD, we here investigated whether PAH exposure was associated with levels of putative CVD-related proteins in serum among currently working chimney sweeps. We enrolled 116 chimney sweeps and 125 unexposed controls, all nonsmoking male workers from Sweden. We measured monohydroxylated PAH metabolites in urine by liquid chromatography coupled to tandem mass spectrometry and a panel of 85 proteins in serum using proximity extension assay. Linear regression analysis adjusted for age and body mass index showed that 25 proteins were differentially expressed between chimney sweeps and the controls (p < .05, adjusted for false discovery rate). Of the 25 proteins, follistatin (FS), prointerleukin-16 (IL-16), and heat shock protein beta-1 (HSP 27) showed positive associations with the monohydroxylated metabolites of PAH in a dose-response manner (p < .05). Pathway and gene ontology analyses demonstrated that the differentially expressed proteins were mainly involved in inflammatory response and immunological functions, such as leukocyte migration, cell movement of leukocytes, and adhesion of immune cells. In conclusion, we found a number of putative CVD-related proteins differentially expressed, between PAH-exposed and unexposed individuals, and mainly involved in inflammation and immune function. Our data warrant protective measures to reduce PAH exposure and longitudinal investigations of the protein profile in chimney sweeps and other occupational groups exposed to PAH.

Quantitative Electrophoresis of Serum Proteins on Paper

1956 ◽  
Vol 2 (5) ◽  
pp. 303-319 ◽  
Author(s):  
Moses Wurm ◽  
Frederick H Epstein
Keyword(s):  
Protein Concentration ◽  
Moving Boundary ◽  
Serum Proteins ◽  
Paper Electrophoresis ◽  
Bromphenol Blue ◽  
Good Resolution ◽  
Confidence Limits ◽  
Protein Patterns ◽  
Electrophoretic Patterns ◽  
Normal Human

Abstract 1. A procedure for paper electrophoresis has been described which gives highly reproducible protein patterns with good resolution and freedom from distortions. 2. Densitometry of protein bands on paper stained with bromphenol blue or Amidoschwarz 10B reveals that the logarithm of protein concentration is proportional to optical density and that Beer's law does not apply. Electrophoretic patterns of normal human serum evaluated in this manner give values in close agreement with those obtained by moving-boundary electrophoresis. 3. Confidence limits were determined for both methods.

scholarly journals Competition between antagonistic complement factors for a single protein on N. meningitidis rules disease susceptibility

eLife ◽  
2014 ◽  
Vol 3 ◽  
Author(s):  
Joseph JE Caesar ◽  
Hayley Lavender ◽  
Philip N Ward ◽  
Rachel M Exley ◽  
Jack Eaton ◽  
...  
Keyword(s):  
Meningococcal Disease ◽  
Serum Proteins ◽  
Association Studies ◽  
Factor H ◽  
Complement Factor H ◽  
Host Susceptibility ◽  
Genome Wide Association Studies ◽  
Complement Factor ◽  
Bacterial Surface ◽  
Related Proteins

Genome-wide association studies have found variation within the complement factor H gene family links to host susceptibility to meningococcal disease caused by infection with Neisseria meningitidis (<xref ref-type="bibr" rid="bib4">Davila et al., 2010</xref>). Mechanistic insights have been challenging since variation within this locus is complex and biological roles of the factor H-related proteins, unlike factor H, are incompletely understood. N. meningitidis subverts immune responses by hijacking a host-immune regulator, complement factor H (CFH), to the bacterial surface (<xref ref-type="bibr" rid="bib25">Schneider et al., 2006</xref>; <xref ref-type="bibr" rid="bib17">Madico et al., 2007</xref>; <xref ref-type="bibr" rid="bib27">Schneider et al., 2009</xref>). We demonstrate that complement factor-H related 3 (CFHR3) promotes immune activation by acting as an antagonist of CFH. Conserved sequences between CFH and CFHR3 mean that the bacterium cannot sufficiently distinguish between these two serum proteins to allow it to hijack the regulator alone. The level of protection from complement attack achieved by circulating N. meningitidis therefore depends on the relative levels of CFH and CFHR3 in serum. These data may explain the association between genetic variation in both CFH and CFHR3 and susceptibility to meningococcal disease.

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