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Zinc (Zn) is found in many neuronal pathways in the brain and has implications for neuromodulation and cerebrovascular disease. However, the association between Zn levels and stroke risk remains controversial. Therefore, we conducted a meta-analysis to explore these relationships. A systematic literature search using PubMed, EMBASE database, and Google Scholar was performed for relevant articles from inception to August 2020. Standardized mean differences (SMDs) and 95% confidence intervals (CIs) were considered the effect sizes and statistical analyses were performed using Stata 12.0. A total of 12 studies involving 1878 cases of stroke and 1754 controls were enrolled. Overall, the meta-analysis demonstrated no significant difference in Zn levels between the stroke group and control group (SMD =-0.18, 95% CI =-0.69 to 0.32, P = 0.480). Subgroup analysis showed that type of stroke had an influence on the Zn levels. A meta-analysis of nine ischemic stroke (IS) studies, which included 1645 cases and 1585 controls, revealed that the Zn levels were significantly higher in IS patients than in controls (SMD (95% CI): 0.61(0.04, 1.19), P = 0.036), but no significant association was found between Zn levels and risk of hemorrhagic stroke (P = 0.113). Egger's test indicated no significant publication bias. This meta-analysis indicates that higher Zn levels may be associated with increased risk of IS; however, these findings should be further confirmed.

A new original Russian test kit for the detection of IgG-antibodies to the causative agent of COVID-19 - coronavirus SARS-CoV-2 by the method of enzyme-linked immunosorbent assay (ELISA) on a solid-phase «ELISA-SARS-CoV-2-AT-G» has been developed. In comparative tests with similar test systems «Vitrotest® SARS-CoV-2 IgG» (Vitrotest, Ukraine) and «Anti-SARS-Cov-2 ELISA (IgG)» (EUROIMMUN AG, Germany) high diagnostic efficiency of the new test system was shown.

Objectives Our recent survey of 44 mass spectrometry laboratories across 17 countries identified variation in internal standard (IS) choice for the measurement of serum/plasma 17α-hydroxyprogesterone (17OHP) by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The choice of IS may contribute to inter-method variations. This study evaluated the effect of two common isotopically labeled IS on the quantification of 17OHP by LC-MS/MS. Methods Three collaborating LC-MS/MS laboratories from Asia, Europe and Australia, who routinely measure serum 17OHP, compared two IS, (1) IsoSciences carbon-13 labeled 17OHP-[2,3,4-13C3], and (2) IsoSciences deuterated 17OHP-[2,2,4,6,6,21,21,21-2H]. This was performed as part of their routine patient runs using their respective laboratory standard operating procedure. Results The three laboratories measured 99, 89, 95 independent samples, respectively (up to 100 nmol/L) using the 13C- and 2H-labeled IS. The slopes of the Passing-Bablok regression ranged 0.98–1.00 (all 95% confidence interval [CI] estimates included the line of identity), and intercept of Conclusions Overall, the comparison between the results of 13C- and 2H-labeled IS for 17OHP showed good agreement, and show no clinically significant bias when incorporated into the LC-MS/MS methods employed in the collaborating laboratories.

Evidence suggests that interleukin-6 (IL-6) concentrations have an important role in suicide behavior (SB) as they are usually increased in these individuals, although no conclusive outcomes have been attained. The purpose of this study was to evaluate the IL-6 levels in plasma, serum, and cerebral spinal fluid (CSF) to determine through a meta-analysis if these levels are increased in individuals with SB in comparison to a group. We calculated the standardized mean difference and 95% confidence intervals (95% CIs). In the systematic review, 21 studies were included, while in the meta-analysis, we included nine studies. The results of our meta-analysis indicated that individuals with SB had reduced levels of IL-6 in plasma (

OBJECTIVE To determine the correlation between glucose concentrations in serum, plasma, and blood measured by a point-of-care glucometer (POCG) and serum glucose concentration measured by an automated biochemical analyzer (ABA; gold standard). SAMPLE 152 canine and 111 feline blood samples. PROCEDURES For each sample, the glucose concentration in serum, plasma, and blood was measured by a POCG and compared with the ABA-measured glucose concentration by means of the Lin concordance correlation coefficient. Results were summarized by species for all samples and subsets of samples with hyperglycemia (ABA-measured glucose concentration > 112 mg/dL for dogs and > 168 mg/dL for cats) and pronounced hyperglycemia (ABA-measured glucose concentration > 250 mg/dL for both species). The effect of PCV on correlations between POCG and ABA measurements was also assessed. RESULTS Hyperglycemia and pronounced hyperglycemia were identified in 69 and 36 canine samples and 44 and 29 feline samples, respectively. The POCG-measured glucose concentrations in serum, plasma, and blood were strongly and positively correlated with the gold standard concentration. The PCV was positively associated with the correlation between the POCG-measured blood glucose concentration and the gold standard concentration but was not associated with the correlations between the POCG-measured glucose concentrations in serum and plasma and the gold standard concentration. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that POCG-measured glucose concentrations in serum, plasma, and blood were strongly correlated with the ABA-measured serum glucose concentration, even in hyperglycemic samples. Given the time and labor required to harvest serum or plasma from blood samples, we concluded that blood was the preferred sample type for use with this POCG.

Evidence indicates that patients with Alzheimer’s dementia (AD) show signs of copper (Cu) dyshomeostasis. This study aimed at evaluating the potential of Cu dysregulation as an AD susceptibility factor. We performed a meta-analysis of 56 studies investigating Cu biomarkers in brain specimens (pooled total of 182 AD and 166 healthy controls, HC) and in serum/plasma (pooled total of 2929 AD and 3547 HC). We also completed a replication study of serum Cu biomarkers in 97 AD patients and 70 HC screened for rs732774 and rs1061472 ATP7B, the gene encoding for the Cu transporter ATPase7B. Our meta-analysis showed decreased Cu in AD brain specimens, increased Cu and nonbound ceruloplasmin (Non-Cp) Cu in serum/plasma samples, and unchanged ceruloplasmin. Serum/plasma Cu excess was associated with a three to fourfold increase in the risk of having AD. Our replication study confirmed meta-analysis results and showed that carriers of the ATP7B AG haplotype were significantly more frequent in the AD group. Overall, our study shows that AD patients fail to maintain a Cu metabolic balance and reveals the presence of a percentage of AD patients carrying ATP7B AG haplotype and presenting Non-Cp Cu excess, which suggest that a subset of AD subjects is prone to Cu imbalance. This AD subtype can be the target of precision medicine-based strategies tackling Cu dysregulation.

This protocol describes the procedures where our published microwave instrument-assisted acid hydrolysis (MAAH) coupled HPAEC-PAD analysis are optimized for glycan monosaccharide composition analysis of serum/plasma samples. The optimized acid hydrolysis of serum/plasma samples takes only 10 min and 10 μl of acid and 2 μl serum/plasma samples. The monosaccharide composition analysis is subsequently accomplished by HPAEC-PAD analysis. Each step of the experimental procedures has been optimized with repeated tests of monosaccharide standards and serum samples. The described workflow takes approximately 70-90 min, up to 48 serum/plasma samples can be analyzed with one HPAEC-PAD instrument per day.

Background It is important for clinical laboratories to have protocols for investigating suspected biotin interference in patient samples. VeraPrep Biotin™ is a commercial product used to rapidly deplete biotin from serum/plasma samples. The objectives of this study were to verify that VeraPrep Biotin™: (a) does not impact immunoassay analyte recovery in control samples and (b) can effectively deplete biotin from samples (both biotin-spiked and samples from donors who ingested biotin supplements). Methods De-identified residual waste serum/plasma samples were combined to create 9 pools for each immunoassay. Plasma/serum samples (n = 23) were obtained from 6 healthy donors at varying times following ingestion of biotin (20 mg, 100 mg, or 200 mg). Nine Elecsys immunoassays were evaluated using the e 602 (Roche Diagnostics Inc.). Control, biotin-spiked (n = 10, ∼400 ng/mL), and donor samples were assayed pre- and post-VeraPrep treatment. Percentage analyte recovery [(posttreatment/pretreatment) × 100] was calculated for control samples. A laboratory-developed LC–MS/MS method was used to quantify biotin. Results In control samples (n = 81), 90–110% analyte recovery was observed post-VeraPrep treatment in over 95% of samples (77/81). The pre- and post-VeraPrep treatment biotin concentration [mean ± standard deviation (SD)] for specimens spiked with up to 500 ng/mL biotin was 357 ± 47 ng/mL and 1.0 ± 0.6 ng/mL, respectively. The mean (range) biotin concentration for the donor samples pre- and post-treatment was 166 (15–1029) ng/mL and 0.2 ( Conclusions These data demonstrate that treatment with VeraPrep Biotin™ does not affect analyte recovery in biotin-negative samples and effectively depletes both spiked and endogenous biotin in serum/plasma.

A novel method has been proposed to develop a simple, rapid, sensitive and affordable chromogenic attempt for the quantification of catalase (CAT) activity in blood samples. The method is based on the oxidation of pyrocatechol (PC) to give quinone form which by oxidative coupling with aminyl radical of 4-aminoantipyrine (4-AAP) resulting from H2O2/CAT to produce a pink colored quinone-imine product with λmax = 530 nm in a 100 mmol/L of tris buffer of pH 9.8 at room temperature (30 °C). The linearity of CAT assay was between 0.316 and 10 U/mL. The accuracy ranges for CAT having concentrations of 1.25, 5 and 7.5 μmol/L were 89–105.52, 90–107%, and 91–104.58% respectively. Within-run and between-run precision studies showed CV’s of 1.98–3.02% (n = 7) and 2.97–4.40% (n = 7), respectively. The detection and quantification limits of CAT were 0.12 and 0.225 μmol/L, respectively. The Michaelis-Menten constant and maximum velocity of the reaction was K m = 1.052 mM and V max = 0.168 μmol/min, respectively. The present method provides a convenient means for investigating the usefulness of CAT measurements in biological sample assessing the potential for free radical-induced pathology.

Sulthiame is an old antiepileptic medicine with controversial history, whose effectiveness and safety in use have been stated in some current studies. However, there is still a need for further clinical examinations for confirmation of its usefulness and tolerability in monotherapy and add-on therapy for epilepsy of various etiologies. A fully validated RP HPLC-UV method for determination of sulthiame in serum/plasma samples using desethylatrazine as the internal standard was developed. The biological fluid was prepared for analysis by a simple precipitation method with acetonitrile. The following validation parameters of the method were determined: selectivity/specificity, linearity range (0.2-50.0 μl/ml, R2 > 0.9999), limits of detection (0.19 μl/ml) and quantification (0.58 μl/ml), precision (intra-day CV 1.06% and inter-day CV 1.25%), extraction recovery (~100%), accuracy (bias, -4.61-0.80%), carryover and ruggedness. Moreover, the stability of the medicine in plasma samples under different storage conditions was also tested. The usability of the method for clinical examinations was checked by analysis of serum samples originating from 19 patients treated with sulthiame. The proposed method is appropriate for determination of sulthiame in serum/plasma samples for drug monitoring purposes, as well as for pharmacokinetic studies.

Several studies suggest that variations in the concentration of plasma glycoproteins can influence cellular changes in a large number of diseases. In recent years, proton nuclear magnetic resonance (1H-NMR) has played a major role as an analytical tool for serum and plasma samples. In recent years, there is an increasing interest in the characterization of glycoproteins through 1H-NMR in order to search for reliable and robust biomarkers of disease. The objective of this review was to examine the existing studies in the literature related to the study of glycoproteins from an analytical and clinical point of view. There are currently several techniques to characterize circulating glycoproteins in serum or plasma, but in this review, we focus on 1H-NMR due to its great robustness and recent interest in its translation to the clinical setting. In fact, there is already a marker in H-NMR representing the acetyl groups of the glycoproteins, GlycA, which has been increasingly studied in clinical studies. A broad search of the literature was performed showing a general consensus that GlycA is a robust marker of systemic inflammation. The results also suggested that GlycA better captures systemic inflammation even more than C-reactive protein (CRP), a widely used classical inflammatory marker. The applications reviewed here demonstrated that GlycA was potentially a key biomarker in a wide range of diseases such as cancer, metabolic diseases, cardiovascular risk, and chronic inflammatory diseases among others. The profiling of glycoproteins through 1H-NMR launches an encouraging new paradigm for its future incorporation in clinical diagnosis.

Thyroid hormones (THs), which mainly consist of 3, 3', 5-triiodo-l-thyronine (T3) and L-thyroxine (T4), play a critical role in regulating biological processes such as growth and metabolism in various animal species. Thus, accurate measurement of T3 and T4, especially physiologically active free (protein-unbound) forms, in serum/plasma is needed for the evaluation of TH homeostasis. However, such high-precision determination of free THs is lacking for non-human species. The present study aimed to develop a highly sensitive and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of six free THs in serum/plasma, which is applicable to not only humans but also non-human species. Two different physical separation steps, ultrafiltration (UF) and equilibrium dialysis (ED), were examined to obtain the free TH fraction. Several experimental conditions were carefully optimized and validated for UF or ED using the commercially available bovine serum. As a result, UF at 1100 × g and 37 °C for 30 min with a 30 kDa ultrafiltration device (Centrifree YM-30, Millipore) yielded excellent precision (CV:10%). The optimized ED step also yielded high precision (CV:10%) and the measurement values were approximately equal to those of UF, but at least 16 h were required to reach equilibrium. Thus, UF combined with LC-MS/MS was finally chosen, in terms of the time needed for the measurement. Acceptable accuracy (recovery: 70%-110%) and intra- and inter-day precision (CV:10% and12%, respectively) were obtained, when triplicate analyses in three different days were conducted using the bovine serum. The developed analytical method was successfully applied to the determination of free THs in serum/plasma samples of humans, cats, and dogs. Furthermore, comparison with free T4 concentrations measured by a common immunoassay method evidently indicated that the ultrafiltration-LC-MS/MS method developed in this study can increase the specificity and accuracy of TH measurement.

Background: Mass spectrometry (MS)-based 17-hydroxyprogesterone (17OHP) methods show considerable variation in results in external quality assurance (EQA) programs. An understanding of the current status of MS-based serum/plasma 17OHP quantification is important to facilitate harmonization. Methods: A 50-item e-survey related to (1) laboratory characteristics, (2) pre-analytical considerations and (3) analysis of 17OHP was developed and circulated to clinical MS laboratories via professional associations in Asia Pacific, Europe and North America. Results: Forty-four laboratories from 17 countries completed the survey. Sample preparation varied between laboratories with protein precipitation and liquid-liquid extraction being the most common processes. Analyte separation was most commonly achieved by liquid chromatography (LC) using a C18 column and mobile phases of water, methanol and formic acid. The ions selected for quantification were 331>97 m/z or 331>109 m/z. Alternative transition ions were used as qualifiers. Twenty-seven of 44 respondents reported preparing their calibrators in-house and variations in material purity and matrix were evident. Nine of 44 laboratories did not participate in an EQA program, and half did not know if their method separated out isobars. The reference intervals, and also their partitioning, reported by the laboratories were highly discrepant, in some cases, by multiple folds. Conclusions: Although MS-based methods are similar in many facets, they are highly disparate. Five recommendations have been developed as an outcome of this survey to support the continued improvement of analysis of serum/plasma 17OHP by MS.

The use of lithium in medicine began in the mid-19th century, when the solubility of uric acid salts in the solution of lithium carbonate was demonstrated in vitro, which meant that lithium could be used in the treatment of gout. The use of lithium in psychiatry starts in 1949, when the effect of lithium in the treatment of mania was demonstrated. The mechanism of action of lithium is not yet completely understood. In parallel with the use of lithium in the treatment of psychiatric disorders, different methods for the determination of lithium in human samples have been developed. The first aim of the study was to determine the amount of lithium in serum samples using electrochemical methods, flame photometry and spectrophotometry and use the results to compare the results obtained using these methods. Because of the possibility of determining lithium in various media, the second aim of the study was to evaluate the clinical value of determining the concentration of lithium in erythrocytes in relation to the concentration of lithium in plasma. The third aim was to investigate the extent to which a therapeutic dose of lithium correlates with the measured concentrations of lithium in serum, plasma and erythrocytes. It was concluded that statistically there was no significant difference between the three test laboratory methods (P = 0.507). Investigating the correlation between the concentration of lithium in various media measured by different methods and the daily therapeutic dose of lithium, it was concluded that statistically a significant correlation was found only in serum lithium concentrations measured with electrochemical method (P = 0.009 ; r = 0.47). There was statistically a significant moderate correlation between the concentration of lithium in plasma and erythrocytes (P = 0.002 ; r = - 0.54), and the lithium concentration erythrocytes are higher than lithium concentrations in plasma (P = 0, 043). The range of the ratio of the concentration of lithium in erythrocytes and plasma is wide (13.25 to 111.15), and is not in correlation with the therapeutic daily dose and therefore is not a better indicator in the control of the treatment.

Currently, there is limited knowledge on the distribution of poly- and perfluoroalkyl substances (PFASs) in different blood matrices, particularly for novel PFASs such as polyfluoroalkyl phosphate esters (PAPs) and perfluoroalkyl phosphonates (PFPAs). To explore this, serum, plasma, and whole blood from 61 adults in Oslo, Norway were collected. The largest number of PFASs were detected in whole blood. For PAPs and PFPAs, the highest frequencies of detection and concentrations were observed in plasma. PAPs contributed to 8% of total PFASs in plasma (median, 0.81 ng mL–1). Perfluorohexylphosphonate (PFHxPA) was the dominant PFPA, regardless of blood matrix. The relative composition profiles of PFASs in blood matrices differed. For some specific PFASs such as perfluorooctanesulfonamide (PFOSA) and perfluorohexanoate (PFHxA), the highest concentrations were observed in whole blood. The PFAS concentration ratios varied between blood matrices, depending on the compounds. However, similar ratios were observed fo...

Monitoring recency of infection helps to identify current transmission in vulnerable populations for effective disease control. We have established an in-house avidity based hepatitis C virus (HCV) recency assay based on the Monolisa Anti-HCV PLUS Version 3 ELISA kit for use of dried serum/plasma spots (DS/PS) in order to distinguish recent and long-term infections. A first panel of DS/PS (n = 218; genotype 1 n = 170 and non-genotype 1 n = 48) consisting of primary and at least one follow up sample was used to analyze the temporal changes of the Avidity Index (AI) over time. Sub-panels of longitudinal DS/PS (n = 66) and acute cases (26 weeks; n = 34) were taken to calculate the Mean Duration of Recent Infection (MDRI) and the False Long-term Rate (FLTR), respectively. A second panel of DS/PS104 weeks (n = 132) and a third panel of DS/PS prepared from resolved infections (≥180 days since last positive; n = 32) were used to calculate the False Recent Rate (FRR). For all genotypes, the optimal AI cut-off was determined to be 40% resulting in an MDRI of 364 days (95% CI: 223-485). FLTR was 5.9% (95% CI: 0.7-19.7), 8.3% (95% CI: 1-27), and 0% (-) and FRR was 13.6% (95% CI: 8.3-20.7), 11.7% (95% CI: 6.6-19), and 30.6% (95% CI: 9.1-61.4) for all genotypes, genotype 1, and non-genotype 1 infections, respectively. For resolved infections, the FRR was 53.1% (95% CI: 35.8-70.4). Thus, this assay performs particularly well for genotype 1 reaching a high rate of correct discriminations between infections acquired less than a year before diagnosis and those acquired earlier by applying an AI cut-off of 40%. Due to a rapid decline in avidity post resolution of an HCV infection this assay is not recommended to be used in HCV RNA negative patients.

This study aimed to review and critically appraise the current methodological issues undermining the suitability of the measurement of serum/plasma glutathione, both in the total and reduced form, as a measure of systemic oxidative stress in chronic obstructive pulmonary disease (COPD). Fourteen relevant articles published between 2001 and 2020, in 2003 subjects, 1111 COPD patients, and 892 controls, were reviewed. Nine studies, in 902 COPD patients and 660 controls, measured glutathione (GSH) in the reduced form (rGSH), while the remaining five, in 209 COPD patients and 232 controls, measured total GSH (tGSH). In the control group, tGSH ranged between 5.7 and 7.5 µmol/L, whilst in COPD patients, it ranged between 4.5 and 7.4 µmol/L. The mean tGSH was 6.6 ± 0.9 µmol/L in controls and 5.9 ± 1.4 µmol/L in patients. The concentrations of rGSH in the control group showed a wide range, between 0.47 and 415 µmol/L, and a mean value of 71.9 ± 143.1 µmol/L. Similarly, the concentrations of rGSH in COPD patients ranged between 0.49 and 279 µmol/L, with a mean value of 49.9 ± 95.9 µmol/L. Pooled tGSH concentrations were not significantly different between patients and controls (standard mean difference (SMD) = −1.92, 95% CI −1582 to 0.0219; p = 0.057). Depending on whether the mean concentrations of rGSH in controls were within the accepted normal range of 0.5–5.0 µmol/L, pooled rGSH concentrations showed either a significant (SMD = −3.8, 95% CI −2.266 to −0.709; p < 0.0001) or nonsignificant (SMD = −0.712, 95% CI −0.627 to 0.293; p = 0.48) difference. These results illustrate the existing and largely unaddressed methodological issues in the interpretation of the serum/plasma concentrations of tGSH and rGSH in COPD.

Blood microRNAs (miRNAs) are ideal biomarkers, and blood derivatives are often collected in the scope of miRNA research projects. However, knowledge of temporal variations of miRNAs in healthy individuals is lacking. In this study, miRNA variability was measured over a 1-year period in different blood derivatives, collected every 2-3 months from two healthy donors. There is a continuum of intraindividual temporal variability, with particularly stable (coefficient of variation [CV]20%-30%) and particularly unstable (CV100%-130%) miRNAs in serum, plasma, and specific white blood cell subpopulations. The temporal intraindividual variability of miRNAs should be taken into consideration in experimental design of biospecimen collections and validation of diagnostic biomarkers.

This paper on the biological tests carried out on serum/plasma samples from donors of human body material (HBM) is the result of a project of the working Group of Superior Health Council of Belgium formed with experts in the field of HBM and infectious serology. Indeed, uncertainty about the interpretation of biological test results currently leads to the sometimes unjustified cancelling of planned donations or the rejection of harvested HBM, whilst more sophisticated diagnostic algorithms would still allow the use of organs or HBM that would otherwise have been rejected. NAT tests will not be discussed in this publication. In the first part some general aspects as the need for a formal agreement between the Tissue Establishment l and the laboratory responsible for the biological testing, but also some specifications regarding testing material, the choice of additional biological tests, and some general aspects concerning interpretation and reporting are discussed. In a second part, detailed information and recommendations concerning the interpretation are presented for each of the mandatory tests (human immunodeficiency virus, hepatitis B virus, hepatitis C virus and syphilis) is presented. A number of not mandatory, but regularly used optional serological tests (e.g. for the detection of antibodies to Toxoplasma gondii, Epstein–Barr virus, human T cell leukemia virus and cytomegalovirus) are also extensively discussed. Although the project was meant to provide clarification and recommendations concerning the Belgian legislation, the majority of recommendations are also applicable to testing of donors of tissues and cells in other (European) countries., SCOPUS: ar.j, SCOPUS: er.j, info:eu-repo/semantics/published

INTRODUCTION: Glioblastoma (GBM) is a malignant primary brain tumour with dismal prognosis. Treatment monitoring remains a challenge in clinical routine, since brain imaging cannot reliably differentiate between true progression and treatment-associated changes. In this project, we evaluate different methods of extracellular vesicles (EV) purification, in order to specifically isolate GBM-EVs from human serum/plasma and introduce EVs, as well as cell-free RNA as possible biomarkers for treatment monitoring in GBM patients. METHODS: EVs from primary GBM cells and the Gaussia luciferase expressing Gli36-GLuc cells were isolated via size-exclusion chromatography (SEC) and ultracentrifugation. EV-surface markers were evaluated by flow cytometry. Gli36-GLuc EVs containing GLuc mRNA were spiked in healthy plasma. Thereafter, plasma EVs were isolated via ultracentrifugation, SEC and immunoprecipitation. Subsequently, RNA was isolated from vesicles and evaluated for GLuc levels via qRT-PCR. Total cell-free RNA from serum of GBM patients was tested for different mRNAs and micro-RNAs at different disease stages. RESULTS: EVs from GBM cells expressed high levels of CD29 and CD44, when compared to EVs from healthy donor plasma. Gli36-GLuc EVs spiked in healthy plasma were more effectively isolated with CD44-based immunoprecipitation than with ultracentrifugation or SEC, as shown by higher GLuc RNA levels in the corresponding vesicles. When compared to total cell-free RNA extracted from this plasma, RNA from EVs exhibited a higher GLuc yield. In cell-free RNA from GBM patients, MGMT levels alone were not capable of detecting progressive disease. CONCLUSIONS: 1. CD44 could serve as a novel, promising target for GBM-EV and be utilised for immunoprecipitation-based EV capturing. 2. Using the appropriate EV purification method possibly affects their potential as biomarkers for GBM. 3. MGMT levels alone in cell-free RNA of GBM patients did not correlate with disease status contrary to previous reports.

BACKGROUND AND OBJECTIVE The evidence regarding the association between serum/plasma vitamin D (VitD) concentrations and systemic lupus erythematosus (SLE) is inconsistent. The study was based on relevant results from literatures that were identified and evaluated. The aim of this meta-analysis is to determine circulating VitD in SLE patients and explore influencing factors. METHODS Studies examining VitD levels in SLE patients were identified through targeted searches in the PubMed and EMBASE databases (up to December 2017). Data extracted from eligible studies was synthesized to calculate the standardized mean difference (SMD), odds ratio (OR), and 95% confidence interval (CI). A fixed or a random effects model was applied to calculate the pooled SMDs and ORs depending on heterogeneity across studies. RESULTS A total of 24 studies, including 6017 patients and 18,417 controls were included. The pooled analysis suggested that VitD levels were significantly lower in SLE patients compared with those in controls [SMD= -0.09, 95%CI= -0.12 to -0.06, P < 0.001]. When the studies were stratified by ethnicity, VitD concentrations were also significantly lower in Asian, Caucasian and African patients. When the studies were stratified by age, gender, VitD level was lower in patients than that in controls. Subgroup analyses stratified by measurement type (expect for radioimmunoassay) also demonstrated consistent results. Moreover, VitD insufficiency was more prevalent in SLE patients than healthy controls [OR=6.57, 95%CI=4.64-9.29]. CONCLUSION Compared with healthy controls, SLE patients had lower concentration of VitD. Additionally, the prevalence of VitD insufficiency is more common in SLE patients.

This study aimed to explore associations between semen quality and trace element level in serum and seminal plasma in Yorkshire boars. Semen quality of 112 Yorkshire boars was assessed for 13 weeks to calculate semen utilization rate, which was then divided into three categories: low utilization rate group (LG,60% utilization rate), medium utilization rate group (MG, 60-80%), and high utilization rate group (HG,80%). After grouping, serum and seminal plasma samples of selected boars were collected to determine concentrations of 10 elements including Ca, Mg, Cu, Fe, Zn, Mn, Se, Cr, Pb and Cd using inductively coupled plasma mass spectrometry. Results showed the increase of semen utilization rate was accompanied by the increase of sperm motility and the decrease of abnormal sperm rate among three groups (P 0.01). Serum Fe concentration in LG boars was lower than that in HG boars (P 0.05). Regression analysis revealed serum Fe concentration was positively correlated with sperm motility (r = 0.251; P 0.05), while negatively correlated with abnormal sperm rate (r = -0.207; P 0.05). However, MG and HG boars had lower serum Se concentration than LG boars (P 0.05), and serum Se concentration contribution to sperm motility varied in a quadratic manner (Sperm motility = -0.0004 Se

Objectives Fibroblast growth factor-21 (FGF-21) plays an important role in glucose and lipid metabolism. This study aims to systemically review the evidence regarding the relationship between the FGF-21 levels and type 2 diabetes mellitus (T2DM), as well as the related influential factors. Methods Research related to plasma/serum FGF-21 levels in patients with T2DM and healthy controls were searched in PubMed, EMBASE and The Cochrane Library databases (up to 31 March 2017). Pooled standard mean difference (SMD) with 95% CI was calculated by fixed-effect or random-effect model analysis. Heterogeneity test was performed by the Q-statistic and quantified using I2, and publication bias was evaluated using a funnel plot and Egger’s linear regression test. Results In total, 317 articles were obtained after searching databases, and 11 studies with 866 patients with T2DM and 629 controls were finally included. Meta-analysis revealed that, compared with the control group, the T2DM group had a significantly higher plasma/serum FGF-21 level (p < 0.001), with the SMD of 1.34% and 95% CI (0.70 to 1.98). Meta-regression analysis and subgroup analyses suggested that body mass index (BMI), triglycerides (TG) and total cholesterol (TC) were likely related to the observed FGF-21 differences between two groups. Conclusions Overall, our study suggests that patients with T2DM have significantly higher plasma/serum FGF-21 levels, and the FGF-21 levels were influenced by BMI, TC and TG.

The analysis of aggregates of therapeutic proteins is crucial in order to ensure efficacy and patient safety. Typically, the analysis is performed in the finished formulation to ensure that aggregates are not present. An important question is, however, what happens to therapeutic proteins, with regard to oligomerization and aggregation, after they have been administrated (i.e., in the blood). In this paper, the separation of whole blood, plasma, and serum is shown using asymmetric flow field-flow fractionation (AF4) with a minimum of sample pre-treatment. Furthermore, the analysis and size characterization of a fluorescent antibody in blood plasma using AF4 are demonstrated. The results show the suitability and strength of AF4 for blood analysis and open new important routes for the analysis and characterization of therapeutic proteins in the blood. Electronic supplementary material The online version of this article (10.1007/s00216-018-1127-2) contains supplementary material, which is available to authorized users.

Extracellular RNAs are initiating increased interest due to their potentials in serving as novel biomarkers, mediators of intercellular communication, and therapeutic applications. As a newly emerging field, one of the main obstacles is the lack of standardized protocols for RNA isolations. Here we describe protocols for commercially available kits that have been modified to yield consistent results for isolation of extracellular RNA from both whole serum/plasma and extracellular vesicle-enriched serum/plasma samples.

Proteomics is one of the modern tools for in-depth study of the protein and peptide composition of complex protein mixtures and also has major applications in the field of animal science. Blood-derived fluids such as serum and plasma are rather unique biological samples as their protein content is contributed by the summation of all cellular proteome sets in the organism. Thus, they are a valuable source of information, reflecting the physiopathological status of the individuals. Being easy to obtain, they were and still are often the samples of choice to study physiology, investigate or diagnose diseases, as well as monitor and compare the influence of potentially harmful substances in different species.

// Teresa Moran 1 , Eudald Felip 1 , Joaquim Bosch-Barrera 2 , Itziar de Aguirre 3 , Jose Luis Ramirez 3 , Carles Mesia 4 , Enric Carcereny 1 , Diana Roa 2 , Elia Sais 2 , Yolanda Garcia 5 , Remei Blanco 6 , Silvia Sanchez 7 , Claudia Rosa Villacorta 7 , Cristina Queralt 3 , Jose Maria Velarde 8 and Rafael Rosell 3 1 Medical Oncology Department, Catalan Institute of Oncology - Badalona, Hospital Universitari Germans Trias i Pujol, Badalona, Universitat Autonoma de Barcelona (UAB), Barcelona, Spain 2 Medical Oncology Department, Catalan Institute of Oncology-Girona, Hospital Universitari Doctor Trueta Girona, Girona, Spain 3 Molecular Biology Laboratory of Cancer Dr. Rosell, Can Ruti Campus: Institute Germans Trias i Pujol (IGTP), Catalan Institute of Oncology, Badalona, Spain 4 Medical Oncology Department, Catalan Institute of Oncology - Hospital Duran i Reynalds, l’Hospitalet de Llobregat, Barcelona, Spain 5 Medical Oncology Department, Hospital Parc Tauli, Sabadell, Barcelona, Spain 6 Medical Oncology Department, Consorci Sanitari de Terrassa, Terrassa, Barcelona, Spain 7 Research Nurse Team, Catalan Institute of Oncology - Badalona, Hospital Universitari Germans Trias i Pujol, Badalona, Spain 8 Statistics Department, Fundacio Germans Trias i Pujol, Badalona, Spain Correspondence to: Teresa Moran, email: mmoran@iconcologia.net Keywords: EGFR-T790M mutation; serum/plasma; osimertinib; acquired resistance; EGFR tyrosine kinase inhibitors Received: March 19, 2018 Accepted: May 02, 2018 Published: June 05, 2018 ABSTRACT Background: Osimertinib is efficacious in lung cancer patients with epidermal growth factor receptor ( EGFR ) mutations and acquired resistance (AR) to EGFR tyrosine kinase inhibitors due to EGFR -T790M mutation (T790M). We sought to describe T790M changes in serum/plasma during osimertinib therapy and correlate these changes with treatment outcomes. Material and methods: Serum/plasma from EGFR -mutant lung cancer patients with T790M-AR was collected before and during osimertinib treatment. Changes in T790M were evaluated using a peptide-nucleic acid-PCR assay, and correlated with clinical and radiographic response. Results: Thirteen patients were included. Median time on osimertinib treatment was 10.6 months with a median progression-free survival of 13.6 months. Best response to osimertinib was partial response (PR), stable disease (SD) or progression (PD) in 46.1%, 30.8% and 23.1% of patients, respectively. Most of the patients were paucisymptomatic at baseline. Symptom improvement was reported in 66.6% of responder patients; while symptoms remained stable in 75% of patients with SD, and 66% of patients with PD had clinical deterioration. Three patterns of T790M changes during osimertinib treatment were identified. T790 remained detectable with PD or a short-lasting SD in 15.4% of the patients. T790M disappeared in 69.2% of patients with PR or SD. T790M disappeared, despite clinical and/or radiographic progression in 15.4% of the patients. Conclusion: Changes of T790M in serum/plasma in EGFR -mutant lung cancer patients with T790M-AR might be a useful marker of symptomatic and radiographic outcome to osimertinib. Longer follow-up is needed to establish if subsequent emergence of T790M could be a marker of resistance.

BACKGROUND: Supplementation of formula with bovine milk fat globule membranes has been shown to narrow the gap in immunological and cognitive development between breast-fed and formula-fed infants. METHOD: In a double-blinded randomized controlled trial 160 formula-fed infants received an experimental formula (EF), supplemented with bovine milk fat globule membranes, or standard formula until 6 months of age. A breast-fed reference group was recruited. Lipidomic analyses were performed on plasma and erythrocyte membranes at 6 months and on serum at 4 and 12 months of age. RESULTS: At 6 months of age, we observed a significant separation in the plasma lipidome between the two formula groups, mostly due to differences in concentrations of sphingomyelins (SM), phosphatidylcholines (PC), and ceramides, and in the erythrocyte membrane lipidome, mostly due to SMs, PEs and PCs. Already at 4 months, a separation in the serum lipidome was evident where SMs and PCs contributed. The separation was not detected at 12 months. CONCLUSIONS: The effect of MFGM supplementation on the lipidome is likely part of the mechanisms behind the positive cognitive and immunological effects of feeding the EF previously reported in the same study population.

Background Many clinical laboratories require that specimens for serum and urine osmolality determination be processed within 3 h of sampling or need to arrive at the laboratory on ice. This protocol is based on the World Health Organization report on sample storage and stability, but the recommendation lacks good supporting data. We studied the effect of storage temperature and time on osmolality measurements. Methods Blood and urine samples were obtained from 16 patients and 25 healthy volunteers. Baseline serum, plasma and urine osmolality measurements were performed within 30 min. Measurements were then made at 3, 6, 12, 24 and 36 h on samples stored at 4–8℃ and room temperature. We compared baseline values with subsequent measurements and used difference plots to illustrate changes in osmolality. Results At 4–8℃, serum and plasma osmolality were stable for up to 36 h. At room temperature, serum and plasma osmolality were very stable for up to 12 h. At 24 and 36 h, changes from baseline osmolality were statistically significant and exceeded the total allowable error of 1.5% but not the reference change value of 4.1%. Urine osmolality was extremely stable at room temperature with a mean change of less than 1 mosmol/kg at 36 h. Conclusions Serum and plasma samples can be stored at room temperature for up to 36 h before measuring osmolality. Cooling samples to 4–8℃ may be useful when delays in measurement beyond 12 h are anticipated. Urine osmolality is extremely stable for up to 36 h at room temperature.

Objective—To investigate the correlation between glucose concentrations in serum, plasma, and whole blood measured by a point-of-care glucometer (POCG) and serum glucose concentration measured by a biochemical analyzer. Design—Prospective clinical study. Samples—96 blood samples from 80 dogs and 90 blood samples from 65 cats. Procedures—Serum, plasma, and whole blood were obtained from each blood sample. The glucose concentrations in serum, plasma, and whole blood measured by a POCG were compared with the serum glucose concentration measured by a biochemical analyzer by use of the Lin concordance correlation coefficient (ρc) and Bland-Altman plots. Results—For both canine and feline samples, glucose concentrations in serum and plasma measured by the POCG were more strongly correlated with the serum glucose concentration measured by the biochemical analyzer (ρc, 0.98 for both canine serum and plasma; ρc, 0.99 for both feline serum and plasma) than was that in whole blood (ρc, 0.62 for canine samples; ρc, 0.90 for feline samples). The mean difference between the glucose concentrations determined by the biochemical analyzer and the POCG in serum, plasma, and whole blood was 0.4, 0.3, and 31 mg/dL, respectively, for canine samples and 7, 6, and 32 mg/dL, respectively, for feline samples. Conclusions and Clinical Relevance—Results indicated that use of a POCG to measure glucose concentrations in serum or plasma may increase the accuracy and reliability of diagnostic and treatment decisions associated with glucose homeostasis disorders in dogs and cats. (J Am Vet Med Assoc 2015;246:1327–1333)

New approaches are increasingly being used for studying and evaluating coronary heart disease (CHD), especially since the irruption of metabolomics. The classical approach is to use enzymatically-measured standard lipids and these are still the main markers for assessing risk of CHD. Since metabolomics relies on advanced analytical technologies, such as MS and NMR, using them to estimate standard lipids would be of great interest because there is no need for additional biochemical measures. The present study evaluates partial least squares and N-way partial least squares regression models to predict standard lipid concentrations by using serum and plasma sample sets from various clinical centres. Information provided by editing NMR techniques and 2D diffusion NMR was incorporated in these models using four different data structures. Firstly, the models were calibrated and validated with three of the four sample sets (n = 591) involved. Then the best estimation models were selected and applied to the left-out sample set. This evaluation of a new sample set gave correlation coefficients of predicted versus biochemical variables above 0.86 and %rRMSE lower than 18 %. These values are similar to those found by other studies although, in our case, the results are more general because we used a higher number of samples (n = 785) from different sample sets, different clinical centres and different blood matrices (serum and plasma). Finally, we compared the performance of NMR predicted lipids and enzymatically measured lipids in a clinical case study.

MicroRNAs (miRNAs) are a group of small RNAs with a fundamental role in the regulation of gene expression. These RNAs have been shown to participate in various cellular and physiological processes, including cellular development, apoptosis, proliferation, and differentiation. Aberrant expression of several miRNAs was found to be involved in a large variety of neoplasms, including hepatocellular carcinoma (HCC). Previous studies have shown the existence of a large amount of stable miRNAs in human serum/plasma, which laid the foundation for studying the role of serum/plasma miRNAs in the diagnosis and prognosis of HCC. Here, we review the recent progress in research on serum miRNAs as biomarkers for HCC in Chinese patients.

The discovery of clinically relevant biomarkers using gel-based proteomics has proven extremely challenging, principally because of the large dynamic range of protein abundances in biofluids such as blood and the fact that only a small number of proteins constitute the vast majority of total blood protein mass. Various separation, depletion, enrichment, and quantitative developments coupled with improvements in gel-based protein quantification technologies, specifically difference gel electrophoresis (DIGE), have contributed to significant improvements in the detection and identification of lower abundance proteins. One of these enrichment technologies, Proteominer, will be the focus of this chapter. The Proteominer technology a utilizes hexapeptide bead library with huge diversity to bind and enrich low-abundance proteins but at the same time suppressing the concentration of high-abundance proteins in subsequent analysis.

In-depth exploration and characterization of human serum and plasma proteomes is an attractive strategy for the identification of potential prognostic or diagnostic biomarkers. The possibility of analyzing larger numbers of samples in a high-throughput fashion has markedly increased with affinity-based microarrays, thus providing higher statistical power to these biomarker studies. Here, we describe a protocol for high-density serum and plasma reverse phase protein arrays (RPPAs). We demonstrate how a biobank of 12,392 samples was immobilized and analyzed on a single microarray slide, allowing high-quality profiling of abundant target proteins across all samples in one assay.

Immunoaffinity enrichment combined with LC-MS/MS enables identification of Post-translational modifications (PTMs) from serum/plasma samples without abundant protein depletion. Here we described the workflow in details in identifying various types of PTMs such as lysine acetylation and arginine methylation from cancer serum. The method described is compatible with all common proteomic analysis platforms and quantitative methods.

The purpose of this study, the direct separation of aminoalkanol derivatives I and II of 1,7-dimethyl-8,9-diphenyl-4-azatricyclo[5.2.1.0(2,6) ]dec-8-ene-3,5,10-trione, which was found in earlier studies as potential anticancer drugs, were performed. Capillary electrophoresis offers the possibility of fast, cheap, and reproducible separations for compounds I and II. In this paper, the simultaneous separation of I and II by capillary zone electrophoresis has been achieved within 8 min by use of 50 mM phosphate buffer of pH 2.5. Analysis of the two compounds in the serum plasma standards was conducted. Limits of detection of I and II by UV absorbance at 200 nm were achieved in the range of 156.3-156.6 ng/mL. The method was validated for linearity, accuracy, precision, limits of detection, and quantification. The calibration equation revealed a good linear relationship (r(2) = 0.998-0.999). Sufficient recovery was observed in the range of 96.3-99.5%. The method showed good reproducibility with intra- and interday precision of 0.97 and 1.76%, respectively. The quantification limits for the compounds were in the range of 477.0-479.8 ng/mL. The proposed method was applied to the analysis of real serum samples.

A formulation of tobacco extract containing 4% nicotine (TE) and similar nicotine formulation containing vehicle and 4% nicotine (NF) were evaluated using animal inhalation assays. Two 4-h inhalation exposures at 1 and 2 mg/L aerosol exposure concentrations, respectively, of the tobacco extract with 4% nicotine formulation showed that the LC50 was greater than 2 mg/L, the maximum concentration tested. All inhalation exposures were conducted using the capillary aerosol generator (CAG). Increasing aerosol TPM concentrations (0, 10, 50, 200, 1000 mg/m(3) TE and 0, 50, 200, 500, 1000 mg/m(3) NF) were generated via the CAG and used to expose groups of male and female rats for 4-h per day for 14 days. In life monitors for potential effects included clinical observations, weekly body weights and food consumption. Post mortem evaluations included gross tissue findings, hematology, clinical chemistry, serum plasma and nicotine levels, absolute and normalized organ and tissue weights, and histopathology of target organs. Treatment-related changes were observed in body weights, hematology, clinical chemistry, organ weights and histopathological findings for TE at the 200 and 1000 mg/m(3) exposure levels, and in the 500 and 1000 mg/m(3) exposure groups for NF. Under the conditions of these studies, the no-observed-adverse-effect level in the rat was approximately 50 mg/m(3) for the TE aerosol-exposed groups, and approximately 200 mg/m(3) in the NF aerosol-exposed groups.