scholarly journals Peptide Charge Derivatization as a Tool for Early Detection of Preeclampsia by Mass Spectrometry—A Comparison with the ELISA Test

Molecules ◽  
2021 ◽  
Vol 26 (23) ◽  
pp. 7102
Author(s):  
Paulina Grocholska ◽  
Andrzej Konieczny ◽  
Zuzanna Kaźmierczak ◽  
Krystyna Dąbrowska ◽  
Karolina Panek-Laszczyńska ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Early Detection ◽  
Pregnant Women ◽  
Early Stage ◽  
Multiple Reaction Monitoring ◽  
Medical Diagnostics ◽  
Elisa Test ◽  
Urine Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Peptide Charge

Early detection of any preeclampsia biomarkers may lower the risk of mortality, both for a mother and a child. Our study focuses on techniques for preeclampsia biomarker identification by comparing the results of a method using liquid chromatography mass spectrometry in multiple reaction monitoring mode (LC-MS/MS) with those by the enzyme-linked immunosorbent assay (ELISA) test, as well as by comparing the obtained results with clinical data. In the proposed LC-MS/MS method a tryptic digest peptide charge derivatization strategy was used as a tool for sensitive detection of podocin, i.e., a previously discovered preeclampsia biomarker present in urine samples from pregnant women. Urine samples from pregnant women with diagnosed preeclampsia were collected at different stages of pregnancy and from healthy subjects, and then were analyzed by ELISA test and the proposed method with LC-MS/MS. Charge derivatization of the ε amino group of C-terminal lysine residues in tryptic digests by 2,4,6-triphenylpyrylium salt was performed to increase the ionization efficiency in the LC-MS/MS mode. Podocin was identified at the early stage of pregnancy, while its detection using an ELISA test was not possible. The protocol for urine sample preparation was optimized. Our results show that the proposed method by LC-MS/MS in combination with peptide charge derivatization, provides an ultrasensitive tool for diagnosis of preeclampsia, and provides earlier detection than a clinical diagnosis or ELISA test. The proposed solution may revolutionize medical diagnostics.

scholarly journals Circulating Heme Oxygenase-1: Not a Predictor of Preeclampsia but Highly Expressed in Pregnant Women Who Subsequently Develop Severe Preeclampsia

2018 ◽  
Vol 2018 ◽  
pp. 1-5 ◽  
Author(s):  
Valéria C. Sandrim ◽  
Mayara Caldeira-Dias ◽  
Heloisa Bettiol ◽  
Marco Antonio Barbieri ◽  
Viviane Cunha Cardoso ◽  
...  
Keyword(s):  
Pregnant Women ◽  
Heme Oxygenase ◽  
Clinical Symptoms ◽  
Severe Preeclampsia ◽  
Urine Samples ◽  
University Hospital ◽  
Enzyme Linked Immunosorbent Assay ◽  
Heme Oxygenase 1 ◽  
Study Cohort ◽  
Potential Biomarker

Preeclampsia is the major cause of maternal and fetal deaths worldwide. Circulating biomarker concentrations to predict preeclampsia must be determined. Therefore, the objective was to evaluate heme oxygenase-1 (HO-1) concentration in both plasma and urine samples from pregnant women before the development of preeclampsia and to identify a potential biomarker for preeclampsia development. We performed a case-control study nested in a prospective study cohort at University Hospital of the Ribeirao Preto Medical School, University of São Paulo (HCFMRP-USP), Ribeirao Preto, Brazil. Of 1400 pregnant women evaluated at 20–25 weeks of gestation, 460 delivered in hospitals outside our institution. Of 940 pregnant women who completed the protocol, 30 developed preeclampsia (cases, 14 cases of severe preeclampsia and 16 cases of mild preeclampsia). Healthy pregnant women (controls, n=90) were randomly selected from the remaining 910 participants. HO-1 concentration was evaluated in plasma/urine samples by using a commercial enzyme-linked immunosorbent assay kit. We found similar HO-1 levels in the plasma and urine for case and control groups. In the subgrouped preeclampsia, lower plasma HO-1 levels were found in mild compared with severe preeclampsia. We conclude that plasma HO-1 levels were not altered at 20–25 weeks of gestation before the manifestation of preeclampsia symptoms. Pregnant women who subsequently develop severe preeclampsia show higher expression of HO-1. This may be indicative of important underlying pathophysiologic mechanisms that differentiate between mild and severe preeclampsia and may possibly be related to a higher prooxidative status even before the development of clinical symptoms.

scholarly journals Comprehensive Detection of Disorders of Purine and Pyrimidine Metabolism by HPLC with Electrospray Ionization Tandem Mass Spectrometry

2006 ◽  
Vol 52 (6) ◽  
pp. 1127-1137 ◽  
Author(s):  
Susen Hartmann ◽  
Jürgen G Okun ◽  
Christiane Schmidt ◽  
Claus-Dieter Langhans ◽  
Sven F Garbade ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Electrospray Ionization ◽  
Multiple Reaction Monitoring ◽  
Dihydropyrimidine Dehydrogenase ◽  
Urine Samples ◽  
Tandem Mass ◽  
Pyrimidine Metabolism ◽  
Phosphoribosyl Transferase ◽  
Ionization Tandem Mass Spectrometry

Abstract Background: Clinical presentation and disease severity in disorders of purine and pyrimidine metabolism vary considerably. We present a method that allows comprehensive, sensitive, and specific diagnosis of the entire spectrum of abnormalities in purine and pyrimidine metabolism in 1 analytical run. Methods: We used reversed-phase HPLC electrospray ionization tandem mass spectrometry to investigate 24 metabolites of purine and pyrimidine metabolism in urine samples from healthy persons and from patients with confirmed diagnoses of inherited metabolic disorders. Urine samples were filtered and diluted to a creatinine concentration of 0.5 mmol/L. Stable-isotope–labeled internal standards were used for quantification. The metabolites were analyzed by multiple-reaction monitoring in positive and negative ionization modes. Results: Total time of analysis was 20 min. Recovery (n = 8) of a compound after addition of a known concentration was 85%–133%. The mean intraday variation (n = 10) was 12%. The interday variation (n = 7) was ≤17%. Age-related reference intervals were established for each compound. Analysis of patient urine samples revealed major differences in tandem mass spectrometry profiles compared with those of control samples. Twelve deficiencies were reliably detected: hypoxanthine guanine phosphoribosyl transferase, xanthine dehydrogenase, purine nucleoside phosphorylase, adenylosuccinate lyase, uridine monophosphate synthase, adenosine deaminase, adenine phosphoribosyl transferase, molybdenum cofactor, thymidine phosphorylase, dihydropyrimidine dehydrogenase, dihydropyrimidinase, and β-ureidopropionase. Conclusion: This method enables reliable detection of 13 defects in purine and pyrimidine metabolism in a single analytical run.

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.

scholarly journals Microscopic Examination of Urine Samples as the Early Detection of Asymptomatic Urinary Tract Infection in Pregnant Women: A Cross-Sectional Study

2019 ◽  
pp. 208-212
Author(s):  
Yeva Rosana ◽  
Dwiana Ocviyanti ◽  
Rahmah Amran
Keyword(s):  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Tract Infection ◽  
Early Diagnosis ◽  
Pregnant Women ◽  
Microscopic Examination ◽  
Urine Culture ◽  
Early Stage ◽  
Urine Samples ◽  
P Value

Abstract Objectives:To investigate the role of microscopic examination of urine sample in supporting early diagnosis of asymptomatic urinary tract infection (UTI) in pregnant women. To compare correspondence between microscopic examination and urine culture result as a gold standard diagnostic modality to support the diagnosis of UTIs and as an evidence-based to start empirical therapy. Methods: Microscopic analysis was conducted in 74 centrifuged and non-centrifuged urine samples from 317 pregnant who came to six healthcare centres in Jakarta, which showed a positive result of nitrite examination.  The results of the microscopic examination of bacteriuria and leukocyturia were compared with a urine culture. Results: Sensitivity of centrifuged bacteriuria was the highest among the other microscopic parameters, which was 74% with the p-value of 0.009. Combination of bacteriuria and leukocyturia≥3/HPF dan≥5/HPF have increased the specificity with the value of 91.5% and 93.6% in non-centrifuged urine. Conclusion: This result showed that the best method of microscopic examination for early diagnosis of asymptomatic urinary tract infection in pregnant women is the detection of bacteriuria in centrifuged urine. Combination of bacteriuriaandleukocyturiatest, as well as leukocyturia≥3/HPF and≥5/HPF,  can be used to rule out the diagnosis of UTI at an early stage. Keywords: asymptomatic urinary tract infection, bacteriuria, leukocyturia,microscopic examination, urine culture.   Abstrak Tujuan: Mengetahui peran pemeriksaan mikroskopis sampel urin dalam mendukung diagnosis dini infeksi saluran kemih asimptomatik (ISK) pada perempuan hamil. Membandingkan kesesuaian antara pemeriksaan mikroskopis dan hasil kultur urin sebagai modalitas diagnostik standar emas untuk mendukung diagnosis ISK dan sebagai dasar bukti untuk memulai terapi empiris. Metode: Penelitian uji mikroskopik dilakukan pada 74 sampel urin disentrifugasi dan tidak sentrifugasi, dari 317 sampel urin perempuan hamil yang berobat ke-enam puskesmas di Jakarta dengan uji nitritpositif. Hasil uji mikroskopik bakteri uria dan leukosit uria dibandingkan dengan hasil kultururin. Hasil: Sensitivitas bakteriuria yang disentrifugasi menunjukan hasil yang paling baik dibandingkan dengan parameter uji mikroskopik lain, yaitu 74% dengan nilai p yang bermakna sebesar 0,009.  Kombinasi bakteri uria dan leukosituria ≥3/LPB dan ≥5/LPB dapat meningkatkan spesifisitas uji dengan nilai 91,5% dan 93,6% pada urin yang  tidak disentrifugasi. Kesimpulan: Hasil menunjukkan bahwa bakteri uria pada urin yang disentrifugasi, merupakan metode yang paling baik untuk membantu diagnosis dini ISK tidak bergejala pada perempuan hamil. Uji kombinasi bakteri uria dan leukosituria, serta uji leukosituria ≥3/LPB dan ≥5/LPB dapat dimanfaatkan untuk membantu secara dini menyingkirkan orang yang tidak mengalami  ISK. Kata kunci: bakteriuria, infeksi saluran kemih tidak bergejala, kultur urin,leukosituria, uji mikroskopik

scholarly journals Microcystins Presence in Mussels(M. galloprovincialis)and Water of Two Productive Mediterranean’s Lagoons (Sardinia, Italy)

2017 ◽  
Vol 2017 ◽  
pp. 1-7 ◽  
Author(s):  
Elena Baralla ◽  
Maria Vittoria Varoni ◽  
Tiziana Sedda ◽  
Valeria Pasciu ◽  
Antonello Floris ◽  
...  
Keyword(s):  
Public Health ◽  
Mass Spectrometry ◽  
Extraction Method ◽  
Mytilus Galloprovincialis ◽  
Solid Phase ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Phase Extraction ◽  
Mass Spectrometry Detection ◽  
The World

Microcystins (MCs) are hepatotoxins harmful for animal and human health. The most toxic type between them is MC-LR whose presence has been investigated in different reservoirs all around the world. In this work microcystins were monitored in spring and summer in water and mussels(Mytilus galloprovincialis)of two Sardinia lagoons: Cabras and Calich lagoons. A Solid Phase Extraction method was developed to clean and concentrate samples before the Enzyme Linked Immunosorbent Assay (ELISA) and the following Mass Spectrometry detection. MCs presence was detected using the screening ELISA test in both lagoons. MCs peak was revealed in July for water and mussels belonging to Cabras lagoon (0.75±0.07 ng/L in water and0.12±0.04 ng/g ww in mussels). In water of Calich lagoon there was a constant trend in the concentration of MCs during the considered months, while there was a MCs peak in July (0.6±0.5 ng/g ww) in mussels. The following LC-MS/MS analysis did not reveal MC-LR presence in all analyzed samples. These results can be useful to enrich knowledge on public health and consumer’s safeguard.

scholarly journals The Seroprevalence of Dengue Virus Infection and Its Association With Iron (Fe) Level in Pregnant Women in Guangzhou, China

2021 ◽  
Vol 8 ◽  
Author(s):  
Jiong Wang ◽  
Jiufeng Sun ◽  
Limei Sun ◽  
Yufeng Ye ◽  
Hanwei Chen ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Pregnant Women ◽  
Inductively Coupled Plasma ◽  
Denv Infection ◽  
Viral Disease ◽  
Urine Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Air Conditioner ◽  
Serum Samples ◽  
Igg Antibody

Dengue fever is regarded as the most prevalent mosquito-borne viral disease in humans. However, information of dengue virus (DENV) infection in pregnant women and the influence factors remain unclear. In this study, we extracted information of 2,076 pregnant women from the Prenatal Environment and Offspring Health (PEOH) birth cohort conducted since 2016 in Guangzhou, China. Peripheral blood and clean midstream urine samples of participants were collected during their hospitalization for childbirth. Indirect enzyme-linked immunosorbent assay (ELISA) was used to detect immunoglobulin G (IgG) antibodies of DENV in serum samples, and inductively coupled plasma mass spectrometry (ICP-MS) was applied to determine the Fe concentrations in the urine samples, which were then adjusted for by urine creatinine and transformed by natural logarithm (ln-Fe). The seroprevalence of DENV IgG antibody in all included participants was 2.22% (46/2,076). We observed higher seroprevalence of IgG antibody in women aged ≥35 years (2.9%), education ≤ 12 years (2.5%), yearly income per capita &lt;100,000 yuan (2.4%), no use of air-conditioner (2.4%), no use of mosquito coils (2.3%), and no exercise during pregnancy (4.1%). A U-shaped relationship was found between ln-Fe concentration and the risk of positive IgG antibody. Compared with women with ln-Fe concentration of 2.0–2.9 μg/g creatinine, slightly higher risks of positive IgG antibody were found among women with ≤2.0 (RR = 4.16, 95% CI: 0.78, 19.91), 3.0–3.9 (RR = 1.93, 95% CI: 0.65, 7.08), 4.0–4.9 (RR = 2.19, 95% CI: 0.65, 8.51), and ≥5.0 μg/g creatinine of ln-Fe (RR = 2.42, 95% CI: 0.46, 11.33). Our findings suggested that the seroprevalence of dengue IgG antibody in pregnant women was comparable to the general population in Guangzhou, China. The risk of DENV infection may be associated with maternal demographic characteristics and behaviors. Both maternal low and high Fe concentrations may be positively associated with the risk of DENV infection.

Time course of the antibody response in humans compared with rats experimentally infected with hepatic alveolar echinococcosis

2012 ◽  
Vol 88 (1) ◽  
pp. 24-31 ◽  
Author(s):  
K. Yamano ◽  
M. Miyoshi ◽  
A. Goto ◽  
S. Kawase
Keyword(s):  
Early Detection ◽  
Antibody Response ◽  
Time Course ◽  
Alveolar Echinococcosis ◽  
Early Stage ◽  
Enzyme Linked Immunosorbent Assay ◽  
Intermediate Species ◽  
Accidental Ingestion ◽  
Hepatic Alveolar Echinococcosis

AbstractHuman alveolar echinococcosis (AE) is caused by the accidental ingestion ofEchinococcus multiloculariseggs. Early detection is essential as surgical resection is the only treatment for a complete cure. However, details are unclear about changes in the antibody response during the initial stages of infection, yet such information is useful for early serodiagnosis. Therefore, a long-term investigation was performed into the time course of the antibody response before ‘positive’ detection. Patient sera were used for enzyme-linked immunosorbent assay (ELISA) and Western blotting (WB) analysis using crude antigens extracted fromE. multilocularisprotoscoleces. Rats were experimentally infected with AE and similarly analysed by ELISA and WB. Among the markers for diagnoses, the 18 kDa band (main marker) appeared last in the preoperative stages and disappeared first after operation in a WB test. Although the 18 kDa antigen can be useful as a marker for AE diagnosis, it cannot contribute to the detection of some patients before the 18 kDa band appearance. To avoid misdiagnosis, different diagnostic antigens such as the 26–28 and 7–8 kDa bands should also be considered. These bands tend to appear earlier than the 18 kDa band and thus offer the potential for early detection of AE. We first observed changes in the antibody response in a relatively early stage after infection in human AE cases. Notably, changes in the antibody response of two intermediate species were similar. These findings provide valuable information for the early detection of human AE cases in the future.

scholarly journals The Value of A Seven-Autoantibody Panel Combined With The Mayo Model In The Differential Diagnosis of Pulmonary Nodules 

2020 ◽  
Author(s):  
Zhougui Ling ◽  
Jifei Chen ◽  
Zhongwei Wen ◽  
Xiaomou Wei ◽  
Rui Su ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Early Detection ◽  
Sensitivity And Specificity ◽  
Detection Rate ◽  
Early Stage ◽  
Pulmonary Nodules ◽  
Enzyme Linked Immunosorbent Assay ◽  
Clinical Value ◽  
Chi Square ◽  
Early Stage Lung Cancer

Abstract BACKGROUND: Identifying malignant pulmonary nodules and detecting early-stage lung cancer (LC) could reduce mortality. This study investigated the clinical value of a seven-autoantibody (7-AAB) panel in combination with the Mayo model for the early detection of LC and distinguishing benign from malignant pulmonary nodules (MPNs).METHODS: The concentrations of the elements of a 7-AAB panel were quantitated by enzyme-linked immunosorbent assay (ELISA) in 806 participants. The probability of MPNs was calculated using the Mayo predictive model. The 7-AAB panel and the Mayo model performances were analyzed by receiver operating characteristic (ROC) analyses, and the difference between groups was evaluated by Chi-square tests (χ2).RESULTS: The combined area under the ROC curve (AUC) for all 7 AABs was higher than that of a single one. The 7-AAB panel sensitivities were 67.5% in the stage I-II LC patients and 60.3% in stage III-IV patients, with a specificity of 89.6% for the healthy controls and 83.1% for benign lung disease patients. The detection rate of the 7-AAB panel in the early-stage LC patients was higher than that of traditional tumor markers. The AUC of the 7-AAB panel combined with the Mayo model was higher than that of the 7-AAB panel alone or the Mayo model alone in distinguishing MPN from benign nodules. For early-stage MPN, the sensitivity and specificity of the combination were 93.5% and 58.0%, respectively. For advanced-stage MPN, the sensitivity and specificity of the combination were 91.4% and 72.8%, respectively. The combination of the 7-AAB panel with the Mayo model significantly improved the detection rate of MPN, but the positive predictive value (PPV) and the specificity were not improved when compared with either the 7-AAB panel alone or the Mayo model alone.CONCLUSION: Our study confirmed the clinical value of the 7-AAB panel for the early detection of lung cancer, and in combination with the Mayo model, could be used to distinguish benign from malignant pulmonary nodules.

scholarly journals Urine Metabolite Profiling of Human Colorectal Cancer by Capillary Electrophoresis Mass Spectrometry Based on MRB

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Jin-Lian Chen ◽  
Jing Fan ◽  
Liu-Shui Yan ◽  
Hui-Qin Guo ◽  
Jing-Jing Xiong ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Mass Spectrometry ◽  
Capillary Electrophoresis ◽  
Cancer Patients ◽  
Advanced Colorectal Cancer ◽  
Early Stage ◽  
Urine Samples ◽  
Urine Metabolites ◽  
Colorectal Cancer Patients ◽  
Capillary Electrophoresis Mass Spectrometry

Aim. The study was to investigate the metabolic profile of urine metabolites and to elucidate their clinical significance in patients with colorectal cancer.Methods. Colorectal cancers from early stage and advanced stage were used in this study. Urine samples of colorectal cancer patients and healthy adults were collected and subjected to capillary electrophoresis mass spectrometry based on moving reaction boundary analysis. The metabolic data were analyzed by SPSS 17.0 to find urinary biomarkers for colorectal cancer.Results. The results indicated that the urine metabolic profiling of colorectal cancer patients had significant changes compared with the normal controls, and there were also differences between early stage and advanced colorectal cancer patients. Compared with the control group, the levels of isoleucine, valine, arginine, lactate acid and leucine increased(P<0.05), but those of histidine, methionine, serine, aspartic acid, citric acid, succinate, and malic acid decreased in urine samples from colorectal cancer(P<0.05). Furthermore, the levels of isoleucine and valine were lower in urine of patients with advanced colorectal cancer than those in early stage colorectal cancer(P<0.05).Conclusion. The technique of capillary electrophoresis mass spectrometry based on MRB could reveal the significant metabolic alterations during progression of colorectal cancer, and the method is feasible and may be useful for the early diagnosis of colorectal cancer.

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