scholarly journals The Alterations in Methylene Blue/Light-Treated Frozen Plasma Proteins Revealed by Proteomics

2021 ◽  
pp. 1-8
Author(s):  
Tiange Wu ◽  
Xiaoning Wang ◽  
Kai Ren ◽  
Xiaochen Huang ◽  
Jiankai Liu
Keyword(s):  
Mass Spectrometry ◽  
Methylene Blue ◽  
Western Blot ◽  
Blue Light ◽  
Differentially Expressed ◽  
Enzyme Linked Immunosorbent Assay ◽  
Differentially Expressed Protein ◽  
Significant Difference ◽  
Modified Proteins ◽  
Frozen Plasma

Introduction: The aim of this study was to investigate the modified proteins in methylene blue/light-treated frozen plasma (MB-FP) compared with fresh frozen plasma (FFP) in order to gain a better application of MB/light-treated plasma in clinic transfusion. Methods: MB-FP and FFP were collected from Changchun central blood station, and a trichloroacetic acid/acetone precipitation method was used to remove albumin for the enrichment of lower abundance proteins. The plasma protein in MB-FP and FFP were separated using two-dimensional gel electrophoresis (2-DE) and the differentially expressed protein spots were analyzed using mass spectrometry. Finally, the differentially expressed proteins were tested using Western blot and enzyme-linked immunosorbent assay (ELISA). Results: Approximately 14 differentially expressed protein spots were detected in the MB-FP, and FFP was chosen as the control. After 2-DE comparison analysis and mass spectrometry, 8 significantly differentially expressed protein spots were identified, corresponding to 6 different proteins, including complement C1r subcomponent (C1R), inter-alpha-trypsin inhibitor heavy chain H4 (ITI-H4), keratin, type II cytoskeletal 1 (KRT1), hemopexin (HPX), fibrinogen gamma chain (FGG), and transthyretin (TTR). Western blot showed no significant difference in the expression level of KRT1 between MB-FP and FFP (p > 0.05). Both Western blot and ELISA indicated that the level of HPX was significantly higher in FFP than in MB-FP (p < 0.05). Conclusion: This comparative proteomics study revealed that some significantly modified proteins occur in MB-FP, such as C1R, ITI-H4, KRT1, HPX, FGG, and TTR. Our findings provide more theoretical data for using MB-FP in transfusion medicine. However, the relevance of the data for the transfusion of methylene blue/light-treated plasma remains unclear. The exact modification of these proteins and the effects of these modified proteins on their functions and their effects in clinical plasma infusion need to be further studied.

scholarly journals Serum proteins differentially expressed in early- and late-onset preeclampsia assessed using iTRAQ proteomics and bioinformatics analyses

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9753
Author(s):  
Chengcheng Tu ◽  
Feng Tao ◽  
Ying Qin ◽  
Mingzhu Wu ◽  
Ji Cheng ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Lipid Metabolism ◽  
Pregnant Women ◽  
Serum Proteins ◽  
Late Onset ◽  
Differentially Expressed ◽  
Coagulation Cascade ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Bioinformatics Analyses

Background Preeclampsia remains a serious disorder that puts at risk the lives of perinatal mothers and infants worldwide. This study assessed potential pathogenic mechanisms underlying preeclampsia by investigating differentially expressed proteins (DEPs) in the serum of patients with early-onset preeclampsia (EOPE) and late-onset preeclampsia (LOPE) compared with healthy pregnant women. Methods Blood samples were collected from four women with EOPE, four women with LOPE, and eight women with normal pregnancies, with four women providing control samples for each preeclampsia group. Serum proteins were identified by isobaric tags for relative and absolute quantitation combined with liquid chromatography–tandem mass spectrometry. Serum proteins with differences in their levels compared with control groups of at least 1.2 fold-changes and that were also statistically significantly different between the groups at P < 0.05 were further analyzed. Bioinformatics analyses, including gene ontology and Kyoto Encyclopedia of Genes and Genomes signaling pathway analyses, were used to determine the key proteins and signaling pathways associated with the development of PE and to determine those DEPs that differed between women with EOPE and those with LOPE. Key protein identified by mass spectrometry was verified by enzyme linked immunosorbent assay (ELISA). Results Compared with serum samples from healthy pregnant women, those from women with EOPE displayed 70 proteins that were differentially expressed with significance. Among them, 51 proteins were significantly upregulated and 19 proteins were significantly downregulated. In serum samples from women with LOPE, 24 DEPs were identified , with 10 proteins significantly upregulated and 14 proteins significantly downregulated compared with healthy pregnant women. Bioinformatics analyses indicated that DEPs in both the EOPE and LOPE groups were associated with abnormalities in the activation of the coagulation cascade and complement system as well as with lipid metabolism. In addition, 19 DEPs in the EOPE group were closely related to placental development or invasion of tumor cells. Downregulationof pregnancy-specific beta-1-glycoprotein 9 (PSG9) in the LOPE group was confirmed by ELISA. Conclusion The pathogenesis of EOPE and LOPE appeared to be associated with coagulation cascade activation, lipid metabolism, and complement activation. However, the pathogenesis of EOPE also involved processes associated with greater placental injury. This study provided several new proteins in the serum which may be valuable for clinical diagnosis of EOPE and LOPE, and offered potential mechanisms underpinning the development of these disorders.

Rapid Inactivation of HIV-1 in Single Donor Preparations of Human Fresh Frozen Plasma by Methylene Blue/Light Treatment

Biologicals ◽  
1994 ◽  
Vol 22 (3) ◽  
pp. 227-231 ◽  
Author(s):  
Bernd Lambrecht ◽  
Stephen G. Norley ◽  
Reinhard Kurth ◽  
Harald Mohr
Keyword(s):  
Methylene Blue ◽  
Blue Light ◽  
Fresh Frozen Plasma ◽  
Light Treatment ◽  
Single Donor ◽  
Rapid Inactivation ◽  
Fresh Frozen ◽  
Frozen Plasma ◽  
Hiv 1

Multi-omics Analysis at Serum Proteomic and Metabolomic Levels Reveals Mechanisms of Moxibustion for a Mouse Model of Ankylosing Spondylitis

2020 ◽  
Author(s):  
Xiao Xu ◽  
Ting Liu ◽  
Rong-Yun Wang ◽  
Ya-Nan Shi ◽  
Jing-ming Xu ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Ankylosing Spondylitis ◽  
Histopathological Examination ◽  
Protein Profiling ◽  
Differentially Expressed ◽  
Enzyme Linked Immunosorbent Assay ◽  
Acupuncture Points ◽  
Interleukin 17 ◽  
Proteomic Approach ◽  
Tof Ms

Abstract Background: Moxibustion is widely used in ameliorating symptoms of ankylosing spondylitis (AS). The aim of this study was to identify the potential molecular profiles of moxibustion on relieving AS mice through incorporating ultra-high-performance liquid chromatography quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF/MS) based metabolomics approach and data independent acquisition-mass spectrometry (DIA-MS) based proteomic. Methods: In this research, mice with proteoglycan-induced spondylitis (PGISp) were intervened with moxibustion for four weeks at specific acupuncture points or at non-acupuncture points. Arthritis severity, histopathological examinations, cytokines and osteoblast (OB) related genes were assessed. Protein profiling and metabolite profiling of serum of mice were examined by UHPLC-Q-TOF/MS based metabolomics technology and DIA-MS based proteomic approach. A multi-omic analysis was implemented for integrating the results of the single proteomic and metabolomic. The levels of some novel proteins in significantly enriched integrated pathways were validated by the enzyme-linked immunosorbent assay (ELISA) method. Results: Our findings clearly indicate that acupuncture points’ moxibustion can significantly decrease the arthritis index (AI) score, improve the histopathological examination and reduce the serum levels of C-reactive protein (CRP), Interferon-γ (IFN-γ), Interleukin-17 (IL-17), Alkaline phosphatase (ALP) and Osteocalcin (OCN) for AS mice. A total of 21 differentially expressed metabolites and 21 differentially expressed proteins (DEPs) were identified as potential moxibustion-intervened biomarkers for AS mice. These molecules were mainly involved in six integrative canonical pathways: mineral absorption, lipid metabolism, purine metabolism, glycolysis/gluconeogenesis, glycine, serine and threonine metabolism and phenylalanine metabolism pathways. Furthermore, The ELISA analysis of four key proteins was consistent with the results of the proteomic analysis. Conclusions: Combing UHPLC-Q-TOF/MS based metabolomics approach and DIA-MS based proteomic, as a promising tool, can advance our understanding of potential mechanisms of action of moxibustion for AS from a holistic perspective.

scholarly journals Helicobacter pylori-Specific Immune Responses of Children: Implications for Future Vaccination Strategy

2002 ◽  
Vol 9 (5) ◽  
pp. 1126-1128 ◽  
Author(s):  
Günter Bode ◽  
Isolde Piechotowski ◽  
Dietrich Rothenbacher ◽  
Hermann Brenner
Keyword(s):  
Helicobacter Pylori ◽  
Confidence Interval ◽  
Immune Responses ◽  
Western Blot ◽  
Vaccination Strategy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibody ◽  
Significant Difference ◽  
Specific Igg ◽  
H Pylori

ABSTRACT We analyzed the specific anti-Helicobacter pylori immunoglobulin G (IgG) antibody profile for a sample of 824 asymptomatic schoolchildren in southern Germany (mean age, 10.7 ± 0.65 years) with an H. pylori-specific IgG enzyme-linked immunosorbent assay and Western blot analysis. The prevalence of infection was 19.8% (95% confidence interval, 17.1 to 22.7%). The immunoresponses were characterized predominantly by antibodies against low-molecular-mass antigens of 14 and 29 kDa, with a significant difference between children of German and Turkish nationalities (P = 0.0012 and P < 0.0001, respectively).

scholarly journals VDAC1 and RhoA in Plasma Exosomes Influence the Severity of Adolescent Idiopathic Scoliosis

2020 ◽  
Author(s):  
Huizhen Li ◽  
Nan Shen ◽  
Lin Mao ◽  
Meijia Chen ◽  
Xuan Zhou ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Adolescent Idiopathic Scoliosis ◽  
Tandem Mass Spectrometry ◽  
Idiopathic Scoliosis ◽  
Differentially Expressed ◽  
Tandem Mass ◽  
Healthy Controls ◽  
Differentially Expressed Proteins ◽  
Differentially Expressed Protein

Abstract Background: Adolescent idiopathic scoliosis (AIS) is the most common spine deformity, but biomarkers for its condition are lacking. Rhodopsin A (RhoA) and voltage-dependent anion-selective channel 1 (VDAC1) in plasma exosomes were defined as differentially expressed proteins between AIS patients and healthy controls. The purpose of this study was to assess exosomes as biomarkers for the occurrence and progression of AIS. Methods：We recruited 10 AIS patients and 8 healthy controls to detect expressed proteins from plasma by liquid chromatography coupled to tandem mass spectrometry. Plasma samples were analyzed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Pathway analysis identified that the VDAC1 and RhoA proteins were alterations expressed in the AIS patients, with the most different alteration was found in extracellular exosomes. Ultracentrifugation was carried out to isolate exosomes from plasma. Verification of the most differentially expressed protein was accessed by Western blot analysis and bioinformatics analysis was performed to predict the pathway of it.Results: 42 of significantly differentially expressed proteins were found in all subjects, and 17 proteins had significant difference. The differentially expressed proteins were enriched in plasma exosomes, and some proteins, such as FN1, were upregulated and others, such as VDAC1, RhoA and AHNAK, were downregulated in the AIS patients. Furthermore, ultracentrifugation was carried out to isolate exosomes from plasma, and RhoA and VDAC1 proteins in plasma exosomes were verified to downregulate by western blot. KEGG signaling pathways were used to predict potential pathways involved in the RhoA and VDAC1 proteins in the AIS patients. We found that the RhoA protein influences AIS probably through the chemokine signaling pathway, platelet activation and cAMP signaling pathway, and the VDAC1 protein is a key factor that participates in the necroptosis pathway, acting on the development of AIS.Conclusions: Consequently, this study mapped a profile of plasma protein, found the differentially expressed protein in AIS, which indicating that plasma exosomes, as a novel biomarker with high specificity, could be associated with the severity of AIS.

Immunoreactivity of Tissue Plasminogen Activator and of Its Inhibitor Complexes

1989 ◽  
Vol 61 (03) ◽  
pp. 409-414 ◽  
Author(s):  
M Rånby ◽  
G Nguyen ◽  
P Y Scarabin ◽  
M Samama
Keyword(s):  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Degradation Products ◽  
Gel Filtration ◽  
Free Form ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plasma Samples ◽  
Significant Difference ◽  
Tissue Plasminogen ◽  
Pai 1

SummaryAn enzyme linked immunosorbent assay (ELISA) based on goat polyclonal antibodies against human tissue plasminogen activator (tPA) was evaluated. The relative immunoreactivity of tPA in free form and tPA in complex with inhibitors was estimated by ELISA and found to be 100, 74, 94, 92 and 8l% for free tPA and tPA in complex with PAI-1, PAI-2, α2-antiplasmin and C1-inhibitor, respectively. Addition of tPA to PAI-1 rich plasma resulted in rapid and total loss of tPA activity without detectable loss of ELISA response, indicating an immunoreactivity of tPA in tPA/PAI-1 complex of about l00%. Three different treatments of citrated plasma samples (acidification/reneutralization, addition of 5 mM EDTA or of 0.5 M lysine) prior to determination by ELISA all resulted in increased tPA levels. The fact that the increase was equally large in all three cases along with good analytical recovery of tPA added to plasffi, supported the notion that all tPA antigen present in plasma samples is measured by the ELISA. Analysis by ELISA of fractions obtained by gel filtration of plasma from a patient undergoing tPA treatment identified tPA/inhibitor complexes and free tPA but no low molecular weight degradation products of tPA. Determinations of tPA antigen were made at seven French clinical laboratories on coded and randomized plasma samples with known tPA antigen content. For undiluted samples there was no significant difference between the tPA levels found and those known to be present. The between-assay coefficient of variation was 7 to 10%. In conclusion, the ELISA appeared suited for determination of total tPA antigen in human plasma samples.

Severe Inherited “Homozygous” Protein C Deficiency in a Newborn Infant

1984 ◽  
Vol 52 (01) ◽  
pp. 053-056 ◽  
Author(s):  
A Estellés ◽  
I Garcia-Plaza ◽  
A Dasí ◽  
J Aznar ◽  
M Duart ◽  
...  
Keyword(s):  
Newborn Infant ◽  
Protein C ◽  
Skin Lesions ◽  
Fresh Frozen Plasma ◽  
Clinical Syndrome ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein C Deficiency ◽  
Fresh Frozen ◽  
Frozen Plasma

SummaryA relapsing clinical syndrome of skin lesions and disseminated intravascular coagulation (DIC) that showed remission with the infusion of fresh frozen plasma is described in a newborn infant with homozygous deficiency of protein C antigen.This patient presented since birth a recurrent clinical picture of DIC and ecchymotic skin lesions that resembled typical ecchymosis except for the fact that they showed immediate improvement with the administration of fresh frozen plasma. Using an enzyme linked immunosorbent assay method, the determination of protein C antigen levels in the patient, without ingestion of coumarin drugs, showed very low values (<1%).No other deficiencies in the vitamin-K-dependent factors or in anti thrombin III, antiplasmin, and plasminogen were found. Seven relatives of the infant had heterozygous deficiency in protein C antigen (values between 40-55%), without clinical history of venous thrombosis. The pedigree analysis of this family suggests an autosomal recessive pattern of inheritance for the clinical phenotype, although an autosomal dominant pattern has been postulated until now in other reported families.We conclude that our patient has a homozygous deficiency in protein C and this homozygous state may be compatible with survival beyond the neonatal period.

An Enzyme Immunoassay (ELISA) for the Quantitation of Human Factor VII

1986 ◽  
Vol 56 (03) ◽  
pp. 250-255 ◽  
Author(s):  
C Boyer ◽  
M Wolf ◽  
C Rothschild ◽  
M Migaud ◽  
J Amiral ◽  
...  
Keyword(s):  
Human Factor ◽  
Solid Phase ◽  
Factor Vii ◽  
Enzyme Linked Immunosorbent Assay ◽  
Poor Correlation ◽  
Vein Thrombosis ◽  
Coagulant Activity ◽  
Significant Difference ◽  
Large Populations ◽  
Deep Vein

SummaryA new solid phase enzyme-linked immunosorbent assay (ELISA) was developed for the quantitation of human Factor VII antigen (F VII Ag), using a monospecific rabbit anti-F VII antiserum. Anti-F VII F(ab′)2 fragments were adsorbed to polystyrene plates. The binding of serial dilutions of control or test plasma, containing F VII, was detected by incubation with peroxidase-labeled anti- FV II IgG followed by the addition of hydrogen peroxyde and O-phenylenediamine. This ELISA is specific, sensitive (detection limit: 0.05%) and accurate (coefficient of variation: 1.5-4% for within- and 1.6-9% for between-assays). F VII coagulant activity (F VII C) and F VII Ag were determined in large populations of controls and patients. In normal plasma (n = 38), F VII Ag ranged from 83 to 117% and the correlation coefficient between F VII Ag and F VII C was 0.94. In patients with severe (F VII C inf. 1%) congenital F VII deficiency (n = 5), F VII Ag was undetectable in two cases (inf. 0.05%) and markedly reduced (0.35 to 5.6%) in the three other cases. In patients with liver cirrhosis (n = 15), F VII Ag ranged from 21 to 59% and was in good correlation with F VII C (r = 0.84). In dicoumarol treated patients (n = 15), the levels of F VII Ag ranged from 51% to 79% and a poor correlation (r = 0.52) with F VIIC was observed. In “compensated” DIC (n = 5), levels of F VII Ag varied from 60 to 186%, with significantly higher F VII C levels (from 143 to 189%). In contrast, in “decompensated” DIC (n = 7), low F VII Ag and F VII C levels were observed (from 7 to 27%). In patients with deep-vein thrombosis (n = 25), high levels of F VII Ag (from 102 to 136%) and F VII C (from 110 to 150%) were demonstrated. In surgical patients, no significant difference was observed before and one day after intervention.

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