Identification of candidate proteomic markers in the serum of medication overuse headache patients: An exploratory study

Cephalalgia ◽  
2020 ◽  
Vol 40 (10) ◽  
pp. 1070-1078
Author(s):  
Lanfranco Pellesi ◽  
Simona Guerzoni ◽  
Carlo Baraldi ◽  
Maria Michela Cainazzo ◽  
Luigi Alberto Pini ◽  
...  
Keyword(s):  
Serum Proteins ◽  
Medication Overuse ◽  
Medication Overuse Headache ◽  
Retinol Binding Protein ◽  
Pathophysiological Mechanism ◽  
Serum Samples ◽  
Two Dimensional Gel Electrophoresis ◽  
Sds Page ◽  
Liquid Chromatography Tandem Mass ◽  
Chronic Pain Conditions

Purpose of the study The pathophysiological mechanism of medication overuse headache is uncertain; no distinctive markers have been described right now. The aim of this study was to conduct proteomic analyses on serum samples from patients with medication overuse headache and healthy individuals. Specifically, mono- (SDS-PAGE) and two-dimensional gel electrophoresis (2-DE) followed by liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to evaluate changes in serum proteins. Main findings By SDS-PAGE, four over-expressed bands were revealed in patients, compared to controls. 2-DE combined with LC-MS/MS analysis allowed confirmation of some proteins preliminarily detected by SDS-PAGE: Hemopexin, alpha-1-acid glycoprotein 1, apolipoprotein A4 and haptoglobin. Moreover, other differential proteins were isolated, mostly increased in MOH patients: Alpha-1-antitrypsin, immunoglobulin heavy constant alpha 1, retinol binding protein and transthyretin. Only one protein, immunoglobulin kappa constant, was decreased in the patients’ group. Conclusions The investigation of the serum proteome can offer a better understanding about biological mechanisms underlying medication overuse headache. Specifically, medication overuse headache shares some serum biochemical markers with chronic pain conditions. Further studies might uncover the relevance of these proteins in medication overuse headache.

Diagnostic and epidemiological implications of regional differences in serum concentrations of proteins observed in six Asian cities

2004 ◽  
Vol 42 (7) ◽  
Author(s):  
Kiyoshi Ichihara ◽  
Yoshihisa Itoh ◽  
Won-Ki Min ◽  
Sook Fan Yap ◽  
Christopher W. K. Lam ◽  
...  
Keyword(s):  
Plasma Proteins ◽  
Regional Differences ◽  
Serum Proteins ◽  
Great Influence ◽  
Density Lipoprotein ◽  
Reference Intervals ◽  
Retinol Binding Protein ◽  
Serum Samples ◽  
Acute Phase Reactants ◽  
Reactive Protein

AbstractThe IFCC Committee on Plasma Proteins has been investigating regional differences for commonly assayed plasma proteins to determine whether universal reference intervals can be applied. As a part of this study, we launched an Asian project analyzing the concentrations of 13 serum proteins whose values are standardized to CRM470, and five newer analytes: retinol-binding protein (RBP), cystatin C (CysC), lightchain-κ (L-κ), and light-chain-λ (L-λ). In Tokyo, Seoul, Kuala Lumpur, Hong Kong, Taipei and Shanghai, serum samples were collected from 146 to 415 apparently healthy individuals with nearly equal gender ratios. All assays were performed in Tokyo on a Behring Nephelometer II (BN II). Seven chemical analytes (aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyltransferase (γGT), creatinine, total cholesterol (TC), triglycerides (TG) and high-density lipoprotein cholesterol (HDL-C)) were also measured. These results were used for excluding individuals with possible latent clinical disorders.Positive acute phase reactants were consistently lower, and negative ones were higher, in Tokyo than those in other cities. The most conspicuous difference was observed in C-reactive protein (CRP). There were no regional differences in transferrin, albumin, or CysC. Creatinine was much lower in Tokyo despite comparable CysC levels. ALT and γGT were higher in Shanghai, Taipei and Seoul; γGT and TG were higher in Shanghai; and HDL-C was higher in Tokyo.Gender-related differences in reference intervals were observed for immunoglobulin (Ig)M, haptoglobin, RBP, transferrin, αEnvironmental factors and lifestyle seem to have a great influence on many commonly measured analytes.

scholarly journals The Application of a Three-Step Proteome Analysis for Identification of New Biomarkers of Pancreatic Cancer

2011 ◽  
Vol 2011 ◽  
pp. 1-13 ◽  
Author(s):  
Mayinuer Abulaizi ◽  
Takeshi Tomonaga ◽  
Mamoru Satoh ◽  
Kazuyuki Sogawa ◽  
Kazuyuki Matsushita ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
High Performance ◽  
Binding Protein ◽  
Proteome Analysis ◽  
Serum Levels ◽  
Retinol Binding Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Two Dimensional Gel Electrophoresis ◽  
Retinol Binding Protein 4

We searched for novel tumor markers of pancreatic cancer by three-step serum proteome analysis. Twelve serum abundant proteins were depleted using immunoaffinity columns followed by fractionation by reverse-phase high-performance liquid chromatography. Proteins in each fraction were separated by two-dimensional gel electrophoresis. Then the gel was stained by Coomassie Brilliant Blue. Protein spots in which the expression levels were significantly different between cancer and normal control were identified by LC-MS/MS. One hundred and two spots were upregulated, and 84 spots were downregulated in serum samples obtained from patients with pancreatic cancers, and 58 proteins were identified by mass spectrometry. These candidate proteins were validated using western blot analysis and enzyme-linked immunosorbent assay (ELISA). As a result of these validation process, we could confirm that the serum levels of apolipoprotein A-IV, vitamin D-binding protein, plasma retinol-binding protein 4, and tetranectin were significantly decreased in patients with pancreatic cancer.

Serum Protein Profiling Distinguishes BCR-ABL+ CML from Normal and Neutrophilia Cases.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4871-4871
Author(s):  
Azim M. Mohamedali ◽  
Satyaji Sahu ◽  
Nicholas Shaun B. Thomas ◽  
Ghulam J. Mufti
Keyword(s):  
Serum Proteins ◽  
Peptide Mass Fingerprinting ◽  
Protein Profiling ◽  
Sample Collection ◽  
Serum Samples ◽  
Metal Affinity ◽  
Peptide Mass ◽  
Sds Page ◽  
And Control ◽  
Normal Controls

Abstract We sought to identify additional biomarkers for chronic myelogenous leukaemia (CML) that could be an aid to early diagnosis and also yield novel antigens for immunotherapy. To this end, we screened patient serum samples at presentation against hematologically normal controls as well as patients with neutrophilia using Surface Enhanced Laser Desorption/Ionization technology (SELDI; Ciphergen ProteinChip series 4000). A total of 84 retrospective and prospective serum samples were analysed: presentation −28, reactive neutrophilia (>15x109 neutrophils/L) − 24 and hematopoietic normal controls − 33. Patients were initially screened by routine cytogenetics and in some cases with qPCR for the BCR-ABL breakpoint. The sera samples were evaluated on 4 different array surfaces and the Immobilised Metal Affinity (IMAC) array was chosen as it bound serum proteins that distinguished CML from normal controls. As little as 1 μl serum was sufficient for each analysis. Biomarker artefacts due to variations in sample collection procedures were ruled out by analysing sera (n=4) from each group at the time of collection and 3 and 6 hours post collection. There were no significant differences in any of the biomarkers at any of the time points. The spectrum of proteins obtained from each of the 84 serum samples was averaged from duplicate runs per experiment. Using the Ciphergen Express program, a panel of 5 proteins were significantly differentially expressed in CML versus the reactive neutrophilia and normal hematopoietic controls (p<0.001). These proteins were identified by a combination of purification techniques using Q HyperD F columns, desalting using reverse phase C-18 beads and isolating the biomarker by 1D-SDS PAGE. The biomarkers were identified by peptide mass fingerprinting and confirmed by Tandem MS sequencing. These were Albumin fragment − 2.8Kd (p< 3.5 x 10−5, ROC=0.78), Fibrinogen fragments − 5.3Kd (p< 6.25 x 10−10, ROC=0.07) and 5.9Kd (p< 9.6 x 10−8, ROC=0.14), Complement 3a precursor fragment − 8.9Kd (p< 0.0015, ROC= 0.70), Platelet basic protein precursor − 10.2Kd (p< 1.5 x 10−4, ROC=0.73) and Lysozyme − 14.6Kd (p=0, ROC=0.92). Biomarkers 3, 4 and 5 were also verified by antibody capture experiments using NP-20 arrays. In a blinded test set of sera, CML, normal and neutrophilia samples were correctly classified 27/28 (96%), 32/32 (100%), 20/24 (83%) respectively using a combination of the 5.3Kd, 10.2 Kd and the 14.6 Kd markers (Biomaker Pattern software). The algorithm correctly classified 21 new samples as CML (7/8) and control (10/13). The 1/8 CML was misclassified for technical reasons. Therefore, a small number of serum biomarkers in as little as 1 μl serum can be used to distinguish between patients with CML and neutrophilia or hematopoietic normal controls. Similar analyses may be applicable to other more heterogeneous hematological malignancies.

scholarly journals Exploration of candidate serum biomarkers potentially related to the chronic pain condition in Medication–overuse headache

BMC Neurology ◽  
2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Lanfranco Pellesi ◽  
Elisa Bellei ◽  
Simona Guerzoni ◽  
Maria Michela Cainazzo ◽  
Carlo Baraldi ◽  
...  
Keyword(s):  
Neuropathic Pain ◽  
Correlation Coefficient ◽  
Trigeminal Nerve ◽  
Chronic Headache ◽  
Serum Proteins ◽  
Medication Overuse ◽  
Medication Overuse Headache ◽  
Serum Levels ◽  
Serum Concentrations ◽  
Healthy Individuals

Abstract Background Medication Overuse Headache (MOH) is a prevalent and disabling disorder resulting from the overuse of analgesic drugs, triptans or other acute headache medications. In previous proteomic studies, several proteins have been found at high concentrations in the urine of MOH patients and in the serum of rats with neuropathic pain. The aim of this study was to compare the serum levels of lipocalin-type Prostaglandin D2 synthase (L-PGDS), Vitamin D-binding protein (VDBP), apolipoprotein E (APOE) and apolipoprotein A1 (APOA1) in MOH patients and healthy individuals, further exploring their relationship with cutaneous pain thresholds (CPTs) in the territories innervated by the trigeminal nerve. Methods Sixty-nine MOH patients and 42 age- and sex-matched healthy volunteers were enrolled in the study. Von Frey-like filaments were applied to the skin territories innervated by the trigeminal nerve, to determine the CPTs. L-PGDS, VDBP, APOE and APOA1 were quantified in the serum by Enzyme-linked Immunosorbent Assay (ELISA). Clinical and laboratory data were collected. Comparisons between MOH patients and healthy individuals were performed using independent t test or χ2 test. To correlate serum proteins with CPTs, Pearson correlation coefficient or Spearman’s rank correlation coefficient were used. Results CPTs were lower among MOH patients. L-PGDS, VDBP and APOE had significantly different serum concentrations between groups (p < 0.01), but no correlation was found with CPTs. APOA1 serum concentrations did not differ between patients and healthy individuals. Conclusions L-PGDS, VDBP and APOE had abnormal serum levels in MOH patients, confirming their alteration in some conditions of chronic headache and neuropathic pain. However, they had no relationship with CPTs. The in-depth study of serum proteins represents a promising approach for a better understanding of MOH, as well as the detection of candidate biomarkers for chronic headache or the risks associated with overuse medications.

Identification of Candidate Biomarkers for HCV Leading to Hepatocellular Carcinoma Differential Stages From Serum Samples

2020 ◽  
Vol 16 (3) ◽  
pp. 262-272
Author(s):  
Amber Afroz ◽  
Saba Saleem ◽  
Kalsoom Sughra ◽  
Sabaz Ali Khan ◽  
Nadia Zeeshan
Keyword(s):  
Hepatocellular Carcinoma ◽  
Hepatitis C ◽  
Serum Proteins ◽  
Sodium Dodecyl ◽  
Fold Increase ◽  
Clinical Staging ◽  
Serum Samples ◽  
Liver Malignancy ◽  
Sds Page ◽  
Staging Systems

Background: Hepatocellular carcinoma (HCC) is one of the most deadly liver malignancy found and Hepatitis C virus (HCV) is a prominent risk factor for this disease. Prognosis of HCC is poor; initiate the need of markers to discover therapeutic targets in HCC. Introduction: Clinical staging systems of HCC composed of tumor characteristics along with liver function test are important in prognosis but they are not precise. Molecular profiling can lead to a better understanding of the physiopathology of HCC and can help in the development of novel therapeutic approaches. Methods: 64 HCC serum samples (shifted from HCV) were graded into stage I- IV; along with +ive (3 Hepatitis C) and -ive control (2 healthy persons). Proteins were separated by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) and differential mRNA expression from serum samples of different HCC stages was confirmed by Real Time Polymerase Chain Reaction (qPCR). Results: HCC serum proteins displayed differential expression of glutathione s-transferase (GST), glypican-3 (GPC3), vitronectin (VTN), and clusterin (CLU) by SDS-PAGE. GST was expressed in -ive control, while GPC3 was found in both -ive and +ive control. The qPCR analysis, display more than 0.07 fold decrease in GST in I-IV HCC stages. The highest increase in HCC stages was observed by GPC3; about 4 fold increase in I-IV stages. VTN show 1.7-3.4 fold; while CLU show 2-3.5 fold increase in four stages of HCC. Conclusion: GPC3, VTN and CLU in combination can be good potential markers for differentiating stages (I-IV) of HCC.

scholarly journals Exploration of candidate serum biomarkers potentially related to the chronic pain condition in Medication Overuse Headache

2019 ◽  
Author(s):  
Lanfranco Pellesi ◽  
Elisa Bellei ◽  
Simona Guerzoni ◽  
Maria Michela Cainazzo ◽  
Carlo Baraldi ◽  
...  
Keyword(s):  
Neuropathic Pain ◽  
Correlation Coefficient ◽  
Trigeminal Nerve ◽  
Chronic Headache ◽  
Serum Proteins ◽  
Medication Overuse ◽  
Medication Overuse Headache ◽  
Serum Levels ◽  
Serum Concentrations ◽  
Healthy Individuals

Abstract Background Medication Overuse Headache (MOH) is a prevalent and disabling disorder resulting from the overuse of analgesic drugs, triptans or other acute headache medications. In previous proteomic studies, several proteins have been found at high concentrations in the urine of MOH patients and in the serum of rats with neuropathic pain. The aim of this study was to compare the serum levels of lypocalin-type Prostaglandin D2 synthase (L-PGDS), Vitamin D-binding protein (VDBP), apolipoprotein E (APOE) and apolipoprotein A1 (APOA1) in MOH patients and healthy individuals, further exploring their relationship with cutaneous pain thresholds (CPTs) in the territories innervated by the trigeminal nerve. Methods 69 MOH patients and 42 age- and sex-matched healthy volunteers were enrolled in the study. Von Frey-like filaments were applied to the skin territories innervated by the trigeminal nerve, to determine the CPTs. L-PGDS, VDBP, APOE and APOA1 were quantified in the serum by Enzyme-linked Immunosorbent Assay (ELISA). Clinical and laboratory data were collected. Comparisons between MOH patients and healthy individuals were performed using independent t test or χ2 test. To correlate serum proteins with CPTs, Pearson correlation coefficient or Spearman's rank correlation coefficient were used. Results CPTs were lower among MOH patients. L-PGDS, VDBP and APOE had significantly different serum concentrations between groups (p < 0.01), but no correlation was found with CPTs. APOA1 serum concentrations did not differ between patients and healthy individuals. Conclusions L-PGDS, VDBP and APOE had abnormal serum levels in MOH patients, confirming their alteration in some conditions of chronic headache and neuropathic pain. However, they had no relationship with CPTs. The in-depth study of serum proteins represents a promising approach for a better understanding of MOH, as well as the detection of candidate biomarkers for chronic headache or the risks associated with overuse medications.

scholarly journals Computer-supported Detection of M-Components and Evaluation of Immunoglobulins after Capillary Electrophoresis

2001 ◽  
Vol 47 (1) ◽  
pp. 110-117 ◽  
Author(s):  
Magnus Jonsson ◽  
Joyce Carlson ◽  
Jan-Olof Jeppsson ◽  
Per Simonsson
Keyword(s):  
Capillary Electrophoresis ◽  
Decision Support ◽  
Protein Separation ◽  
Serum Proteins ◽  
Cost Effective ◽  
Clinical Samples ◽  
Serum Samples ◽  
Computerized Decision Support ◽  
Cost Effective Method ◽  
Mathematical Algorithms

Abstract Background: Electrophoresis of serum samples allows detection of monoclonal gammopathies indicative of multiple myeloma, Waldenström macroglobulinemia, monoclonal gammopathy of undetermined significance, and amyloidosis. Present methods of high-resolution agarose gel electrophoresis (HRAGE) and immunofixation electrophoresis (IFE) are manual and labor-intensive. Capillary zone electrophoresis (CZE) allows rapid automated protein separation and produces digital absorbance data, appropriate as input for a computerized decision support system. Methods: Using the Beckman Paragon CZE 2000 instrument, we analyzed 711 routine clinical samples, including 95 monoclonal components (MCs) and 9 cases of Bence Jones myeloma, in both the CZE and HRAGE systems. Mathematical algorithms developed for the detection of monoclonal immunoglobulins (MCs) in the γ- and β-regions of the electropherogram were tested on the entire material. Additional algorithms evaluating oligoclonality and polyclonal concentrations of immunoglobulins were also tested. Results: CZE electropherograms corresponded well with HRAGE. Only one IgG MC of 1 g/L, visible on HRAGE, was not visible after CZE. Algorithms detected 94 of 95 MCs (98.9%) and 100% of those visible after CZE. Of 607 samples lacking an MC on HRAGE, only 3 were identified by the algorithms (specificity, 99%). Algorithms evaluating total gammaglobulinemia and oligoclonality also identified several cases of Bence Jones myeloma. Conclusions: The use of capillary electrophoresis provides a modern, rapid, and cost-effective method of analyzing serum proteins. The additional option of computerized decision support, which provides rapid and standardized interpretations, should increase the clinical availability and usefulness of protein analyses in the future.

scholarly journals Antibody in Middle Ear Fluid of Children Originates Predominantly from Sera and Nasopharyngeal Secretions

2012 ◽  
Vol 19 (10) ◽  
pp. 1593-1596 ◽  
Author(s):  
Ravinder Kaur ◽  
Thomas Kim ◽  
Janet R. Casey ◽  
Michael E. Pichichero
Keyword(s):  
Middle Ear ◽  
Acute Otitis Media ◽  
Secretory Iga ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Two Dimensional Gel Electrophoresis ◽  
Middle Ear Fluid ◽  
Iga Antibodies ◽  
Iga Antibody ◽  
Entire Array

ABSTRACTThe human middle ear is devoid of any immunocompetent cells in normal mucosa. We sought to determine the source of antibody present in the middle ear of children. Total IgG, IgA, and secretory IgA antibodies were determined by enzyme-linked immunosorbent assay from the nasopharyngeal, middle ear, and serum samples of children with acute otitis media. The two-dimensional gel electrophoresis pattern of the entire array of IgA antibodies in the nasal wash (NW) and middle ear fluid (MEF) was compared from the MEF and NW samples using isoelectric focusing and Western blotting. The total IgG and IgA antibodies in the MEF and NW samples of 137 children were compared. The ratio of IgG to IgA in the MEF was significantly different (P< 0.008) compared to NW because IgA levels were higher and IgG levels lower in NW. The IgG/IgA ratio of MEF resembled serum consistent with transudation to the MEF. Small amounts of secretory IgA were detected in MEF but the electrophoresis patterns of the entire array of IgA antibodies in the MEF and NW were virtually identical in each child evaluated; thus, IgA in MEF derived predominantly from serum and the nasopharynx by reflux via the Eustachian tube. The IgG/IgA antibody levels in the MEF and the same composition of IgA antibody in the MEF and NW identifies the predominant source of antibody in the MEF as a transudate of serum combined with nasal secretions refluxed from the nasopharynx in children.

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