Search for Melanoma Markers in Plasma and Serum Samples

2005 ◽  
Vol 11 (3) ◽  
pp. 353-360 ◽  
Author(s):  
Roberta Seraglia ◽  
Susanna Vogliardi ◽  
Graziella Allegri ◽  
Stefano Comai ◽  
Mario Lise ◽  
...  
Keyword(s):  
High Frequency ◽  
Healthy Subjects ◽  
Ionic Species ◽  
Low Molecular Weight ◽  
Ionization Mass ◽  
Plasma Samples ◽  
Serum Samples ◽  
Blood Samples ◽  
Melanoma Patients ◽  
Diagnostic Ions

Fourteen blood samples from patients with melanomas and 11 blood samples from healthy subjects were analyzed by matrix-assisted laser desorption/ionization mass spectrometry. The study focussed on species of low molecular weight, in the 800–5000 Da range, present in plasma and sera. While for healthy subjects plasma samples lead to the production of a higher number of ionic species, for melanoma patients a high number of diagnostic ions, present with high frequency and with quite high relative abundance, are present, in particular, in serum samples and, to a lesser extent, also in plasma. Since plasma samples are obtained more easily in comparison to sera, it is possible to suggest that plasma can also be used for these studies.

Stability of antioxidant vitamins in whole human blood during overnight storage at 4°C and frozen storage up to 6 months

2018 ◽  
Vol 88 (3-4) ◽  
pp. 151-157 ◽  
Author(s):  
Scott W. Leonard ◽  
Gerd Bobe ◽  
Maret G. Traber
Keyword(s):  
Ascorbic Acid ◽  
Whole Blood ◽  
Healthy Subjects ◽  
Frozen Storage ◽  
Blood Collection ◽  
Plasma Samples ◽  
Optimal Conditions ◽  
Plasma Ascorbic Acid ◽  
Blood Samples ◽  
Sample Handling

Abstract. To determine optimal conditions for blood collection during clinical trials, where sample handling logistics might preclude prompt separation of erythrocytes from plasma, healthy subjects (n=8, 6 M/2F) were recruited and non-fasting blood samples were collected into tubes containing different anticoagulants (ethylenediaminetetra-acetic acid (EDTA), Li-heparin or Na-heparin). We hypothesized that heparin, but not EDTA, would effectively protect plasma tocopherols, ascorbic acid, and vitamin E catabolites (α- and γ-CEHC) from oxidative damage. To test this hypothesis, one set of tubes was processed immediately and plasma samples were stored at −80°C, while the other set was stored at 4°C and processed the following morning (~30 hours) and analyzed, or the samples were analyzed after 6 months of storage. Plasma ascorbic acid, as measured using HPLC with electrochemical detection (LC-ECD) decreased by 75% with overnight storage using EDTA as an anticoagulant, but was unchanged when heparin was used. Neither time prior to processing, nor anticoagulant, had any significant effects upon plasma α- or γ-tocopherols or α- or γ-CEHC concentrations. α- and γ-tocopherol concentrations remained unchanged after 6 months of storage at −80°C, when measured using either LC-ECD or LC/mass spectrometry. Thus, refrigeration of whole blood at 4°C overnight does not change plasma α- or γ-tocopherol concentrations or their catabolites. Ascorbic acid is unstable in whole blood when EDTA is used as an anticoagulant, but when whole blood is collected with heparin, it can be stored overnight and subsequently processed.

scholarly journals Influences of Blood Sample Processing on Low–Molecular-Weight Proteome Identified by Surface-Enhanced Laser Desorption/Ionization Mass Spectrometry

2005 ◽  
Vol 51 (9) ◽  
pp. 1637-1649 ◽  
Author(s):  
Rosamonde E Banks ◽  
Anthea J Stanley ◽  
David A Cairns ◽  
Jennifer H Barrett ◽  
Paul Clarke ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Laser Desorption ◽  
Ionization Mass ◽  
Plasma Samples ◽  
Laser Desorption Ionization ◽  
Serum Samples ◽  
Affinity Capture ◽  
Desorption Ionization ◽  
Elapsed Time ◽  
Surface Enhanced

Abstract Background: Profiling approaches in proteomics, such as surface-enhanced laser desorption/ionization (SELDI) mass spectrometry, are used in disease marker discovery. The aim of this study was to investigate the potential influence of selected preanalytical factors on the results obtained. Methods: Plasma samples anticoagulated with EDTA, citrate, or heparin, and serum samples from healthy volunteers were profiled by SELDI on CM10, immobilized metal affinity capture (IMAC) array with copper, and H50 chip surfaces. Using linear mixed-effects models, we examined the influence of elapsed time between venipuncture and sample separation (immediate to 24 h) and the type of serum tube used (Greiner Vacuette activator, gel serum separator, or plain tubes). We analyzed purified platelets, as well as platelet-poor and platelet-rich plasma samples treated with calcium and/or thrombin to determine the platelet contribution, directly or via the clotting process, to the profiles generated. We then used cluster analysis to identify samples with similar peak profiles. Results: Different plasma types and sera could be distinguished on the basis of cluster analyses of their spectral profiles. Elapsed time between venipuncture and separation of plasma and serum from blood samples altered the profiles obtained, particularly for serum samples and particularly on IMAC chips. The type of serum collection tube also affected the profiles because of differences in clotting time. In vitro manipulation of platelets revealed that specific peaks in IMAC profiles of serum appeared to be derived directly from platelets. Several other peaks, including some of those exhibiting time-dependent changes, arose during the clotting process. Conclusion: Preanalytical variables, such as sample handling, can markedly influence results.

scholarly journals Double venipuncture is not required for adequate S-100B determination in melanoma patients

BioTechniques ◽  
2020 ◽  
Vol 69 (5) ◽  
pp. 371-378
Author(s):  
Samantha Damude ◽  
Anneke C Muller Kobold ◽  
Esther Bastiaannet ◽  
Schelto Kruijff ◽  
Harald J Hoekstra ◽  
...  
Keyword(s):  
Cancer Staging ◽  
Stage Iii ◽  
Tube System ◽  
Serum Samples ◽  
Blood Samples ◽  
Significant Difference ◽  
Melanoma Patients ◽  
The Mean ◽  
Stage Iii Melanoma

S-100B is used in melanoma follow-up. This serum biomarker is also present in adipocytes; therefore, subcutaneous adipocytes trapped in the needle before performing a venipuncture could contaminate the serum. The aim was to study the influence of adipocyte contamination on blood samples used for S-100B analysis, possibly resulting in falsely elevated S-100B values. A total of 294 serum samples were collected from 147 American Joint Committee on Cancer staging stage III melanoma patients. The mean difference between the first (dummy) and second tubes was 0.003 μg/l (p = 0.077), with a decrease in the second tube. Compared with the second tube, the S-100B level was higher in the first tube in 33.3% of the samples, equal in 36.8% of the samples and lower in 29.9% of the samples. No significant difference between the two consecutively drawn tubes was found. There seems to be no necessity of implementing a dummy tube system for accurate S-100B determination in melanoma patients.

Comparative Study on the Anticoagulant Effects of Low Molecular Weight Heparins as Measured by Whole Blood Act, Anti-Xa and a Newly Developed Prothrombinase Induced Clotting Time (PiCT) Assay.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4148-4148
Author(s):  
D. Hoppensteadt ◽  
M. Tobu ◽  
R. Wahi ◽  
O. Iqbal ◽  
J. M. Walenga ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Whole Blood ◽  
Low Molecular Weight ◽  
Plasma Samples ◽  
Clotting Time ◽  
Low Molecular Weight Heparins ◽  
Blood Samples ◽  
Good Relationship ◽  
Surgical Conditions ◽  
Better Than

Abstract Low Molecular Weight Heparins (LMWHs) are currently developed for anticoagulation in various intravenous indications such as DVT treatment and percutaneous interventions (PCI). Traditionally, these drugs are administered in anti-Xa units. The purpose of this study was to compare the relative levels of anticoagulation produced by different LMWHs in whole blood ACT (Hemochron), an amidolytic anti-Xa method and a plasma-based global anticoagulant assay, the PiCT test (Pentapharm, Basel, Switzerland). Various LMWHs such as dalteparin, enoxaparin, reviparin and a synthetic pentasaccharide were supplemented to freshly drawn native blood samples to simulate patient samples containing 1 U/ml anti-Xa levels. Three generic versions of enoxaparin were also included in this study. In addition to the ACT measurement in whole blood, citrated plasma samples were also tested for APTT, Heptest, PiCT, amidolytic anti-Xa and IIa activities, and protease generation assays. All agents produced assay-based effects on different tests, which were not proportional to the adjusted anti-Xa activity. Correlation of the ACT data resulted in a good relationship with PiCT for the commercially available branded products(r2=0.97). ACT and Heptest (r2=0.51), ACT and anti-Xa (r2<0.3), ACT and anti-IIa (r2= 0.93). The correlation of PiCT and Heptest was (r2=0.73) better than ACT and Heptest. However, the generic versions of enoxaparin did not provide similar results. PiCT and Heptest also gave the good correlation. These results clearly suggest that for the global anticoagulant actions of LMWHs, anti-Xa measurements are of limited value and should not be used to asses the anticoagulant efficacy of LMWHs in PCI and other surgical conditions. These differences may be more obvious in the generic versions of LMWHs, which are apparently produced using pharmacopeial specifications. Thus, it is important to include a clot-based method in the cross referencing and standardization of LMWHs.

scholarly journals Metabolomics Approach for Validation of Self-Reported Ibuprofen and Acetaminophen Use

Metabolites ◽  
2018 ◽  
Vol 8 (4) ◽  
pp. 55
Author(s):  
Kristine Dennis ◽  
Brian Carter ◽  
Susan Gapstur ◽  
Victoria Stevens
Keyword(s):  
Mass Spectrometry ◽  
Self Report ◽  
Untargeted Metabolomics ◽  
Plasma Samples ◽  
Serum Samples ◽  
Over The Counter ◽  
Blood Samples ◽  
Cutoff Value ◽  
Analgesic Use ◽  
Metabolomics Analysis

Over-the-counter analgesic use is common and is typically assessed through self-report; therefore, it is subject to misclassification. Detection of drug metabolites in biofluids offers a viable tool for validating self-reported analgesic use. Thus, the aim of this study was to determine the utility of a metabolomics approach for the validation of acetaminophen and ibuprofen use in blood samples. Untargeted mass spectrometry-based metabolomics analysis was conducted in serum samples from 1547 women and plasma samples from 556 men. The presence of two metabolites each for acetaminophen and ibuprofen at levels at or above a defined cutoff value was used to determine concordance with self-reported use. For acetaminophen use based on the presence of both acetaminophen and acetamidophenylglucuronide, concordance was 98.5–100% among individuals reporting use today, and 79.8–91.4% for those reporting never or rare use. Ibuprofen use based on the presence of both carboxyibuprofen and hydroxyibuprofen resulted in concordance of 51.3–52.5% for individuals reporting use today and 99.4–100% for those reporting never or rare use. Our findings suggest that an untargeted metabolomics approach in blood samples may be useful for validating self-reported acetaminophen use. However, this approach appears unlikely to be suitable for validating ibuprofen use.

Results for serum and plasma compared in 15 selected radioassays.

1978 ◽  
Vol 24 (1) ◽  
pp. 137-139 ◽  
Author(s):  
N P Kubasik ◽  
H E Sine
Keyword(s):  
Plasma Samples ◽  
Serum Samples ◽  
Blood Samples ◽  
Clinical Interpretation ◽  
Sufficient Magnitude

Abstract We evaluated the results for serum vs. plasma samples for 15 selected radioassay procedures, using 19 manufacturers' kits. Blood samples were collected with heparin, oxalate-fluoride, or ethylenediaminetetraacetate anticoagulants and compared with serum samples. Differences were demonstrated between serum and plasma which may be of sufficient magnitude to alter clinical interpretation of the results. Assays also demonstrated significant differences based on the kit manufacturer and procedure used.

scholarly journals Comparison of Trace Elements Levels in Atrial Fibrillation Patients and Healthy Group

2020 ◽  
pp. 113-119
Author(s):  
Zahra Bazargani ◽  
Manzar Banoo Shojaeifard ◽  
Gholam Abbas Valizadeh
Keyword(s):  
Atrial Fibrillation ◽  
Trace Elements ◽  
Blood Serum ◽  
Atomic Absorption ◽  
Healthy Subjects ◽  
Statistical Analyses ◽  
Serum Samples ◽  
Blood Samples ◽  
Healthy Group ◽  
Significant Difference

Background: The aim of this study is to measure the concentration of Zinc (Zn), copper (Cu)  and selenium (Se) in the blood serum of Atrial Fibrillation (AF) patients as compared to healthy subjects in both genders. Methods: This study, conducted on patients with AF (n = 50) and controls (n = 24) were evaluated by available method. Blood samples were taken from the patients and analysis of trace elements was performed by atomic absorption spectrophotometer. Statistical analyses were done using IBM-SPSS 22.0. Results: Zn and Se concentrations were evaluated in AF patients and healthy group. Zn and Se concentration in AF patients is much lower than in healthy group (p<0.008, p<0.000). AF patients showed a significantly higher Cu concentration than the healthy group (p<0.000). The data of the present study revealed that the concentrations of all trace elements had a significant difference in the serum of AF patients with respect to the healthy group (p<0.05). Conclusion: We found that concentrations of Zn & Se levels were significantly decrease in the in the serum samples of AF patients compared with the controls group.

Osteocalcin concentrations in plasma prepared with different anticoagulants

1991 ◽  
Vol 37 (2) ◽  
pp. 281-284 ◽  
Author(s):  
Michael J Power ◽  
Bernadette O'Dwyer ◽  
Eugene Breen ◽  
Patrick F Fottrell
Keyword(s):  
Enzyme Immunoassay ◽  
Sodium Citrate ◽  
Serum Osteocalcin ◽  
Plasma Samples ◽  
Serum Samples ◽  
Blood Samples ◽  
Freeze Thaw ◽  
Significant Difference

Abstract We investigated the effects on plasma osteocalcin concentrations of different anticoagulants used to collect the blood samples. Plasma osteocalcin concentrations measured by enzyme immunoassay and radioimmunoassay are influenced by the nature of the anticoagulants used. The most significant difference between concentrations found in plasma and serum was seen with oxalate/fluoride anticoagulant, which reduced osteocalcin concentrations to 37.3% of serum values. This is probably related to increased hemolysis with this anticoagulant compared with osteocalcin concentrations in plasma prepared with other anticoagulants. Samples prepared with sodium citrate (0.105 mol/L) or lithium heparin gave values 92.4% and 83.6% of those obtained with matched serum samples. Osteocalcin concentrations were relatively stable in plasma and serum at -20 degrees C for two freeze/thaw cycles. In blood from 100 patients there was a good correlation between osteocalcin concentrations in serum and plasma (lithium heparin) (r2 = 0.831); the slope and intercept (+/- SE) were 0.924 +/- 0.04 and 4.92 +/- 1.25 micrograms/L, respectively. However, in 10 patients, serum osteocalcin concentrations were two- to threefold higher than those in matched plasma samples.

scholarly journals PRRSV detection by qPCR in processing fluids and serum samples collected in a positive stable breeding herd following mass vaccination of sows with a modified live vaccine

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
A. Lebret ◽  
P. Berton ◽  
V. Normand ◽  
I. Messager ◽  
N. Robert ◽  
...  
Keyword(s):  
The United States ◽  
Live Vaccine ◽  
Mass Vaccination ◽  
Respiratory Syndrome Virus ◽  
Serum Samples ◽  
Blood Samples ◽  
Stability Monitoring ◽  
Breeding Herd ◽  
Modified Live Vaccine ◽  
Processing Fluid

AbstractIn the last two decades, in France, Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) stabilization protocols have been implemented using mass vaccination with a modified live vaccine (MLV), herd closure and biosecurity measures. Efficient surveillance for PRRSV is essential for generating evidence of absence of viral replication and transmission in pigs. The use of processing fluid (PF) was first described in 2018 in the United States and was demonstrated to provide a higher herd-level sensitivity compared with blood samples (BS) for PRRSV monitoring. In the meantime, data on vertical transmission of MLV viruses are rare even as it is a major concern. Therefore, veterinarians usually wait for several weeks after a sow mass vaccination before starting a stability monitoring. This clinical study was conducted in a PRRSV-stable commercial 1000-sow breed-to-wean farm. This farm suffered from a PRRS outbreak in January 2018. After implementing a stabilisation protocol, this farm was controlled as stable for more than 9 months before the beginning of the study. PF and BS at weaning were collected in four consecutive batches born after a booster sow mass MLV vaccination. We failed to detect PRRSV by qPCR on PF and BS collected in a positive-stable breeding herd after vaccination with ReproCyc® PRRS EU (Boehringer Ingelheim, Ingelheim, Germany).

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