ABSTRACT
BACKGROUND: The aim of this study was to determine the effect of human urine-derived stem cells (HUSCs) for the treatment of spinal cord injury (SCI) and investigate associated the molecular network mechanism by using bioinformatics combined with experimental validation. METHODS: After the contusive SCI model was established, the HUSC-expressed specific antigen marker was implanted into the injury site immediately, and the Basso, Beattie and Bresnahan locomotor rating scale (BBB scale) was utilized to evaluate motor function so as to determine the effect of HUSCs for the neural repair after SCI. Then, the geneCards database was used to collect related gene targets for both HUSCs and SCI, and cross genes were merged with the findings of PubMed screen. Subsequently, protein-protein interaction (PPI) network, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment, as well as core network construction, were performed using Cytoscape software. Lastly, real-time quantitative polymerase chain reaction (PCR) and immunofluorescence were employed to validate the mRNA expression and localization of 10 hub genes, and two of the most important, designated as cadherin 1 (CDH1) and integrin subunit beta 1 (ITGB1), were identified successfully. RESULTS: The immunophenotypes of HUSCs were marked by CD90+ and CD44+ but not CD45, and flow cytometry confirmed their character. The expression rates of CD90, CD73, CD44 and CD105 in HUSCs were 99.49, 99.77, 99.82 and 99.51%, respectively, while the expression rates of CD43, CD45, CD11b and HLA-DR were 0.08, 0.30, 1.34 and 0.02%, respectively. After SCI, all rats appeared to have severe motor dysfunction, but the BBB score was increased in HUSC-transplanted rats compared with control rats at 28 days. By using bioinformatics, we obtained 6668 targets for SCI and 1095 targets for HUSCs and identified a total of 645 cross targets between HUSCs and SCI. Based on the PPI and Cytoscape analysis, CD44, ACTB, FN1, ITGB1, HSPA8, CDH1, ALB, HSP90AA1 and GAPDH were identified as possible therapeutic targets. Enrichment analysis revealed that the involved signal pathways included complement and coagulation cascades, lysosome, systemic lupus erythematosus, etc. Lastly, quantificational real-time (qRT)-PCR confirmed the mRNA differential expression of CDH1/ITGB1 after HUSC therapy, and glial fibrillary acidic protein (GFAP) immunofluorescence staining showed that the astrocyte proliferation at the injured site could be reduced significantly after HUSC treatment. CONCLUSIONS: We validated that HUSC implantation is effective for the treatment of SCI, and the underlying mechanisms associated with the multiple molecular network. Of these, CDH1 and ITGB1 may be considered as important candidate targets. Those findings therefore provided the crucial evidence for the potential use of HUSCs in SCI treatment in future clinic trials.
Subject(s)
Spinal Cord Injuries , Rats , Humans , Animals , Rats, Sprague-Dawley , Spinal Cord Injuries/genetics , Spinal Cord Injuries/therapy , Spinal Cord Injuries/metabolism , Stem Cells , RNA, Messenger/metabolism , Integrins/therapeutic useABSTRACT
INTRODUCTION: The world is experiencing exponential growth in communication, especially wireless communication. Wireless connectivity has recently become a part of everyone's daily life. Recent developments in low-cost, low-power, and miniature devices contribute to a significant rise in radiofrequency-electromagnetic field (RF-EM) radiation exposure in our environment, raising concern over its effect on biological systems. The inconsistent and conflicting research results make it difficult to draw definite conclusions about how RF-EM radiation affects living things. OBJECTIVES: This study identified two micro-environments based on their level of exposure to cellular RF-EM radiation, one with significantly less exposure and another with very high exposure to RF-EM radiation. Emphasis is given to studying the metabolites in the urine samples of humans naturally exposed to these two different microenvironments to understand short-term metabolic dysregulations. METHODS: Untargeted 1H NMR spectroscopy was employed for metabolomics analyses to identify dysregulated metabolites. A total of 60 subjects were recruited with 5 ml urine samples each. These subjects were divided into two groups: one highly exposed to RF-EM (n = 30) and the other consisting of low-exposure populations (n = 30). RESULTS: The study found that the twenty-nine metabolites were dysregulated. Among them, 19 were downregulated, and 10 were upregulated. In particular, Glyoxylate and dicarboxylate and the TCA cycle metabolism pathway have been perturbed. The dysregulated metabolites were validated using the ROC curve analysis. CONCLUSION: Untargeted urine metabolomics was conducted to identify dysregulated metabolites linked to RF-EM radiation exposure. Preliminary findings suggest a connection between oxidative stress and gut microbiota imbalance. However, further research is needed to validate these biomarkers and understand the effects of RF-EM radiation on human health. Further research is needed with a diverse population.
Subject(s)
Metabolome , Metabolomics , Radio Waves , Humans , Male , Adult , Metabolomics/methods , Female , Radio Waves/adverse effects , Metabolome/radiation effects , Middle Aged , Electromagnetic Fields/adverse effects , Young AdultABSTRACT
Human urine-derived stem cells (hUSCs) process self-renewal and multilineage differentiation ability. Due to their non-invasive and easily available clinical source, hUSCs represent a promising alternative source of mesenchymal stem cells (MSCs) for application potential in cytotherapy. However, technical limitations, such as stemness property maintenance, have hindered hUSCs' clinical application. Certain some small molecules have been recognized with advantage in maintaining the stemness of stem cells. In this study, we identified stemness-regulated key targets of hUSCs based on the StemCellNet database, CMAP database and literature mining. Furthermore, we identified a small molecule compound, boldine, which may have the potential to promote the stemness of hUSCs. It promotes cell proliferation, multilineage differentiation and maintains stemness of hUSCs by cell viability assay, single-cell clone formation, osteogenic differentiation and stemness marker expression (OCT-4 and C-MYC). We identified that boldine may be a potential GSK-3ß inhibitor by molecular docking and confirmed that it can upregulate the level of ß-catenin and promote translocation of ß-catenin into nucleus of hUSCs using Western blotting and immunofluorescence analysis. Our study indicates boldine activates the Wnt/ß-catenin signaling pathway in hUSCs and provides an effective strategy for MSCs research and application of small molecules in maintaining the stemness of hUSCs.
Subject(s)
Aporphines , Wnt Signaling Pathway , beta Catenin , Humans , Glycogen Synthase Kinase 3 beta , Molecular Docking Simulation , Osteogenesis , Stem Cells , Cell Differentiation , Cell ProliferationABSTRACT
Polycyclic aromatic hydrocarbon (PAH) exposure is suspected to be linked to oxidative damage. Herein, ten PAH human exposure biomarkers [hydroxylated PAH metabolites (OH-PAHs)] and five oxidative stress biomarkers (OSBs) were detected in urine samples collected from participants living in a rural area (n = 181) in Northwestern China. The median molar concentration of ΣOH-PAHs in urine was 47.0 pmol mL-1. The 2-hydroxynaphthalene (2-OHNap; median: 2.21 ng mL-1) was the dominant OH-PAH. The risk assessment of PAH exposure found that hazard index (HI) values were <1, indicating that the PAH exposure of rural people in Jingyuan would not generate significant cumulative risks. Smokers (median: 0.033) obtained higher HI values than nonsmokers (median: 0.015, p < 0.01), suggesting that smokers face a higher health risk from PAH exposure than nonsmokers. Pearson correlation and multivariate linear regression analysis revealed that ΣOH-PAH concentrations were significant factors in increasing the oxidative damage to deoxyribonucleic acid (DNA) (8-hydroxy-2'-deoxyguanosine, 8-OHdG), ribonucleic acid (RNA) (8-oxo-7,8-dihydroguanine, 8-oxoGua), and protein (o, o'-dityrosine, diY) (p < 0.05). Among all PAH metabolites, only 1-hydroxypyrene (1-OHPyr) could positively affect the expression of all five OSBs (p < 0.05), suggesting that urinary 1-OHPyr might be a reliable biomarker for PAH exposure and a useful indicator for assessing the impacts of PAH exposure on oxidative stress. This study is focused on the relation between PAH exposure and oxidative damage and lays a foundation for the study of the health effect mechanism of PAHs.
Subject(s)
Biomarkers , Oxidative Stress , Polycyclic Aromatic Hydrocarbons , Rural Population , Polycyclic Aromatic Hydrocarbons/urine , Humans , China , Risk Assessment , Biomarkers/urine , Male , Female , Environmental Exposure , Middle Aged , AdultABSTRACT
Human serum albumin (HSA) serves as a crucial indicator for therapeutic monitoring and biomedical diagnosis. In this study, a near infrared (NIR) fluorescent probe, termed BTPA, characterized a donor-π-acceptor (D-π-A) structure based on bridged triphenylamine (TPA) was developed. BTPA exhibited outstanding sensitivity and selectivity towards HSA among various analysts, with a remarkable 50-fold fluorescence enhancement with a significant Stokes shift (â¼190 nm) and a wide linear detection range of 0-20 µM of HSA. Especially, BTPA displayed selectivity for discrimination of HSA from BSA. Job's Plot analysis suggested a 1:1 stoichiometry for the formation of the BTPA-HSA complex. Displacement assays and molecular docking demonstrated that BTPA binds to subdomain IB of HSA which could effectively avoid interference from most drugs. Besides, BTPA have good biocompatibility and could detect of exogenous HSA with a relatively low fluorescence background. For practical applications, BTPA was tested for detecting HSA levels in human urine without any pretreatment, showing detection capability in the range of 0-10 µM with a fast response (<30 s), a limit of detection (LOD) of 0.12 µM and good recoveries (81.7-92.9 %), highlighting the high performance of bridged triphenylamine-based probe BTPA.
ABSTRACT
Benzothiazole (BTH), benzotriazole (BTR), and their respective derivatives (BTHs and BTRs) are emerging environmental pollutants with widespread human exposure and oncogenic potential. Studies have demonstrated adverse effects of exposure to certain BTHs and BTRs on the respiratory system. However, no study has examined the associations between exposure to BTHs and BTRs and lung cancer risk. We aimed to examine the associations between urinary concentrations of BTHs and BTRs and the risk of lung cancer in the general population from Quzhou, China. We conducted a nested case-control study in an ongoing prospective Quzhou Environmental Exposure and Human Health (QEEHH) cohort, involving 20, 694 participants who provided urine samples during April 2019-July 2020. With monthly follow-up until November 2022, 212 lung cancer cases were recruited and 1:1 matched with healthy controls based on age and sex. We estimated odds ratios (ORs) and 95% confidence intervals (CIs) of lung cancer risk associated with urinary BTHs and BTRs concentrations using conditional logistic regression models after controlling for potential covariates. We also examined effect modification by several covariates, including sex, socioeconomic status, smoking status, alcohol consumption, and dietary habit. Creatinine-corrected urinary BTH and 2-hydroxy-benzothiazole (2-OH-BTH) levels were significantly associated with the risk of lung cancer, after adjusting for a variety of covariates. Participants in the highest quartile of BTH had a 95% higher risk of lung cancer, compared with those in the lowest quartile (adjusted OR = 1.95, 95% CI: 1.08-3.49; p for trend = 0.01). Participants with higher levels of urinary 2-OH-BTH had an 83% higher risk of lung cancer than those with lower levels (adjusted OR = 1.83, 95% CI: 1.16-2.88; p for trend = 0.01). Exposure to elevated levels of BTH and 2-OH-BTH may be associated with an increased risk of lung cancer. These associations were not modified by socio-demographic characteristics.
Subject(s)
Benzothiazoles , Lung Neoplasms , Triazoles , Humans , Case-Control Studies , Lung Neoplasms/urine , Lung Neoplasms/chemically induced , Lung Neoplasms/epidemiology , Triazoles/urine , Male , Middle Aged , Female , Benzothiazoles/urine , Aged , China/epidemiology , Environmental Exposure , Adult , Environmental Pollutants/urine , Prospective StudiesABSTRACT
Health exposure to benzotriazole ultraviolet stabilizers (BUVSs) may pose diverse toxic impacts on health. Presently, the occurrence of BUVSs in human urine remains inadequately understood. This study analyzed 13 kinds of BUVSs in human urine (n = 182) from the general Chinese adult participants. Totally, nine BUVSs were measurable in these human urine samples. Among the detected BUVSs, 2-(2H-benzotriazol-2-yl)-p-cresol (UV-P) was the most predominant BUVS in the human urine, with the mean concentration of 1.6 µg/g creatinine (Subject(s)
Triazoles
, Humans
, Male
, Female
, Triazoles/urine
, Adult
, Middle Aged
, Young Adult
, Aged
, Ultraviolet Rays
, China
, Environmental Pollutants/urine
, Biological Monitoring/methods
ABSTRACT
Immunoglobulin nephropathy (IgAN) stands as the most prevalent primary glomerular nephropathy globally, typically diagnosed through an invasive renal biopsy. Emerging research suggests the significant involvement of chiral amino acids in kidney disease progression. This study introduces a nonderivative LC-tandem mass spectrometry approach, offering efficient separation outcomes within 15 min for identifying chiral amino acids in human urine samples. Subsequently, using this method, the analysis of l- and d-amino acids in the urine of both patients with IgAN and healthy individuals was conducted. Fourteen d-amino acids and 20 l-amino acids were identified in the urine samples obtained from 17 patients with IgAN and 21 healthy individuals. The results indicated notable variances in the concentrations of both l- and d-amino acids between the IgAN and healthy control groups. In contrast to the healthy group, the IgAN group exhibited higher mean urine concentrations of most l-amino acids and lower concentrations of d-amino acids. Furthermore, correlations between amino acids and clinical markers were investigated. These results propose a novel method for monitoring trace amino acids in urine samples and introduce a new concept for potential markers of IgAN.
Subject(s)
Amino Acids , Glomerulonephritis, IGA , Tandem Mass Spectrometry , Humans , Tandem Mass Spectrometry/methods , Amino Acids/urine , Glomerulonephritis, IGA/urine , Chromatography, Liquid/methods , Male , Adult , Female , Middle Aged , Reproducibility of Results , Biomarkers/urine , Stereoisomerism , Linear Models , Case-Control Studies , Young AdultABSTRACT
Amlodipine (AM) is a long active calcium channel blocker used to relax blood vessels by preventing calcium ion transport into the vascular walls and its supporting molecules acetaminophen (AP) and ascorbic acid (AA) are recommended for hypertension control and prevention. Considering their therapeutic importance and potential side effects due to over dosage, we have fabricated a sensor for individual and simultaneous determination of AA, AP, and AM in pharmaceuticals and human urine using novel Zn-doped Ca2CuO3 nanoparticles modified glassy carbon electrode (GCE). Optimally doped Ca2CuO3 (2.5 wt% Zn at Cu site) enhanced the detection of target molecules over much wider concentration ranges of 50 to 3130 µM for AA, 0.25 to 417 µM for AP, and 0.8 to 354 µM for AM with the corresponding lowest detection limits of 14 µM, 0.05 µM, and 0.07 µM, respectively. Furthermore, the Zn-Ca2CuO3/GCE exhibited excellent selectivity and high sensitivity even in the presence of several potential interfering agents. The usefulness of the developed electrode was tested using an amlodipine besylate tablet and urine samples of seven hypertension patients under medication. The results confirmed the presence of a significant amount of AP and AM in six patients' urine samples indicating that the personalized medication is essential and the quantum of medication need to be fixed by knowing the excess medicines excreted through urine. Thus, the Zn-Ca2CuO3/GCE with a high recovery percentage and good sensitivity shall be useful in the pharmaceutical and biomedical sectors.
Subject(s)
Acetaminophen , Amlodipine , Ascorbic Acid , Copper , Electrodes , Hypertension , Zinc , Amlodipine/urine , Amlodipine/analysis , Humans , Ascorbic Acid/urine , Copper/chemistry , Acetaminophen/urine , Zinc/chemistry , Zinc/urine , Hypertension/drug therapy , Hypertension/urine , Electrochemical Techniques/methods , Electrochemical Techniques/instrumentation , Limit of Detection , Metal Nanoparticles/chemistry , Nanoparticles/chemistry , Carbon/chemistryABSTRACT
Electrocatalytic oxidation of urea (UOR) is a potential energy-saving hydrogen production technology that can replace oxygen evolution reaction (OER). Therefore, CoSeP/CoP interface catalyst is synthesized on nickel foam using hydrothermal, solvothermal, and in situ template methods. The strong interaction of tailored CoSeP/CoP interface promotes the hydrogen production performance of electrolytic urea. During the hydrogen evolution reaction (HER), the overpotential can reach 33.7 mV at 10 mA cm-2 . The cell voltage can reach 1.36 V at 10 mA cm-2 in the overall urea electrolytic process. Notably, the overall urine electrolysis performance of the catalyst in the human urine medium can reach 1.40 V at 10 mA cm-2 and can exhibit durable cycle stability at 100 mA cm-2 . Density functional theory (DFT) proves that the CoSeP/CoP interface catalyst can better adsorb and stabilize reaction intermediates CO* and NH* on its surface through a strong synergistic effect, thus enhancing the catalytic activity.
ABSTRACT
Bladder outlet obstruction (BOO) is a common disease that always make the bladder develops from inflammation to fibrosis. This study was to investigate the effect of exosomes from human urine-derived stem cells (hUSCs) on bladder fibrosis after BOO and the underlying mechanism. The BOO mouse model was established by inserting a transurethral catheter, ligation of periurethral wire, and removal of the catheter. Mouse primary bladder smooth muscle cells (BSMCs) were isolated and treated with TGFß1 to mimic the bladder fibrosis model in vitro. Exosomes from hUSCs (hUSC-Exos) were injected into the bladder of BOO mice and added into the culture of TGFß1-induced BSMCs. The associated factors in mouse bladder tissues and BSMCs were detected. It was confirmed that the treatment of hUSC-Exos alleviated mouse bladder fibrosis and down-regulated fibrotic markers (a-SMA and collagen III) in bladder tissues and TGFß1-induced BSMCs. Overexpression of NRF1 in hUSC-Exos further improved the effects of hUSC-Exos on bladder fibrosis both in vivo and in vitro. TGFßR1 was a target of NRF1 and miR-301b-3p, and miR-301b-3p was a target of NRF1. It was next characterized that hUSC-Exos carried NRF1 to up-regulate miR-301B-3p, thereby reducing TGFßR1level. Our results illustrated that hUSC-Exos carried NRF1 to alleviate bladder fibrosis through regulating miR-301b-3p/TGFßR1 pathway.
Subject(s)
Exosomes , MicroRNAs , Urinary Bladder Neck Obstruction , Humans , Mice , Animals , Urinary Bladder/metabolism , Exosomes/genetics , Exosomes/metabolism , Urinary Bladder Neck Obstruction/genetics , Urinary Bladder Neck Obstruction/metabolism , Urinary Bladder Neck Obstruction/pathology , Stem Cells/metabolism , MicroRNAs/genetics , MicroRNAs/metabolism , FibrosisABSTRACT
Bile acids (BAs) are a complex class of metabolites that have been described as specific biomarkers of gut microbiota activity. The development of analytical methods allowing the quantification of an ample spectrum of BAs in different biological matrices is needed to enable a wider implementation of BAs as complementary measures in studies investigating the functional role of the gut microbiota. This work presents results from the validation of a targeted ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the determination of 28 BAs and six sulfated BAs, covering primary, secondary, and conjugated BAs. The analysis of 73 urine and 20 feces samples was used to test the applicability of the method. Concentrations of BAs in human urine and murine feces were reported, ranging from 0.5 to 50 nmol/g creatinine and from 0.012 to 332 nmol/g, respectively. Seventy-nine percent of BAs present in human urine samples corresponded to secondary conjugated BAs, while 69% of BAs present in murine feces corresponded to primary conjugated BAs. Glycocholic acid sulfate (GCA-S) was the most abundant BA in human urine samples, while taurolithocholic acid was the lowest concentrated compound detected. In murine feces, the most abundant BAs were α-murocholic, deoxycholic, dehydrocholic, and ß-murocholic acids, while GCA-S was the lowest concentrated BA. The presented method is a non-invasive approach for the simultaneous assessment of BAs and sulfated BAs in urine and feces samples, and the results will serve as a knowledge base for future translational studies focusing on the role of the microbiota in health.
Subject(s)
Bile Acids and Salts , Tandem Mass Spectrometry , Humans , Mice , Animals , Bile Acids and Salts/analysis , Tandem Mass Spectrometry/methods , Sulfates/analysis , Chromatography, High Pressure Liquid/methods , Feces/chemistryABSTRACT
The benefits of combining drift time ion mobility (DTIMS) with liquid chromatography-high-resolution mass spectrometry (HRMS) have been reported for metabolomics but the use of differential time mobility spectrometry (DMS) is less obvious due to the need for rapid scanning of the DMS cell. Drift DTIMS provides additional precursor ion selectivity and collisional cross-section information but the separation resolution between analytes remains cell- and component-dependent. With DMS, the addition of 2-propanol modifier can improve the selectivity but on cost of analyte MS response. In the present work, we investigate the liquid chromatography-mass spectrometry (LC-MS) analysis of a mix of 50 analytes, representative for urine and plasma metabolites, using scanning DMS with the single modifiers cyclohexane (Ch), toluene (Tol), acetonitrile (ACN), ethanol (EtOH), and 2-propanol (IPA), and a binary modifier mixture (cyclohexane/2-propanol) with emphasis on selectivity and signal sensitivity. 1.5% IPA in the N2 stream was found to suppress the signal of 50% of the analytes which could be partially recovered with the use of IPA to 0.05% as a Ch/IPA mixture. The potential to use the separation voltage/compensation voltage/modifier (SV/CoV/Mod) feature as an additional analyte identifier for qualitative analysis is also presented and applied to a data-independent LCxDMS-SWATH-MS workflow for the analysis of endogenous metabolites and drugs of abuse in human urine samples from traffic control.
Subject(s)
2-Propanol , Metabolomics , Humans , Mass Spectrometry/methods , Chromatography, Liquid/methods , Spectrum AnalysisABSTRACT
Nonylphenol (NP), nonylphenol monoethoxylate (NP1EO), and nonylphenol diethoxylate (NP2EO) are widely used in various daily products and have been cataloged as endocrine-disrupting chemicals. Due to their high lipophilicity and low biodegradability, these compounds remain in the environment and enter the human body through the food chain. Growing concerns regarding the potential negative effects of NP, NP1EO, and NP2EO on human health have raised the need for biomonitoring to investigate human exposure to these compounds. In this study, a simultaneous analysis method using solid-phase extraction (SPE) combined with gas chromatography-mass spectrometry (GC-MS) was established by controlling the background contamination of NP, NP1EO, and NP2EO, which are ubiquitous in laboratory environments. The proposed method showed proper linearity of over 0.999 and a recovery greater than 85.8% for all analytes. Accuracy and precision were verified in ranges of 92.97-116.30% and 0.65-9.29%, respectively. The detection limits for NP, NP1EO, and NP2EO were 0.0363 µg L-1, 0.0401 µg L-1, and 0.0364 µg L-1, respectively, which were suitable for determining the trace analytes in human urine. Therefore, this simple and integrated analytical method was applied to measure the free and total forms of the target analytes in 25 human urine samples collected in Korea. Overall, free NP, NP1EO, and NP2EO were detected with average contents of 3.94 ± 4.14 µg L-1, 4.63 ± 2.62 µg L-1, and 0.293 ± 0.638 µg L-1, respectively, and with total NP, NP1EO, and NP2EO contents of 6.14 ± 8.24 µg L-1, 5.99 ± 2.91 µg L-1, and 0.806 ± 1.10 µg L-1, respectively. These data showed that these compounds are prevalent in human urine, and indicate the need for further studies.
ABSTRACT
Analyzing the composition of (human) urine plays a major role in the fields of biology and medicine. Organic molecules (such as urea, creatine) and ions (such as chloride, sulfate) are the major compounds present in urine, the quantification of which allows for the diagnosis of a subject's health condition. Various analytical methods have been reported for studying urine components and validated on the basis of known and referenced compounds. The present work introduces a new method able to simultaneously determine both major organic molecules and ions contained in urine, by combining ion chromatography using a conductimetric detector with mass spectroscopy. The analysis of organic and ionized compounds (anionic and cationic) was achieved in double injections. For quantification, the standard addition method was used. Human urine samples were pre-treated (diluted and filtered) for IC-CD/MS analysis. The analytes were separated in 35 min. Calibration ranges (0-20 mg.L-1) and correlation coefficients (> 99.3%) as well as detection (LODs < 0.75 mg.L-1) and quantification (LOQs < 2.59 mg.L-1) limits were obtained for the main organic molecules (lactic, hippuric, citric, uric, oxalic acids, urea, creatine, and creatinine) and ions (chloride, sulfate, phosphate, sodium, ammonium, potassium, calcium, and magnesium) contained in urine. The intra- and inter-day accuracies of the analytes consistently ranged from 0.1 to 5.0%, and the precision was within 4.0%. For all analytes, no significant matrix effects were observed, and recoveries ranged from 94.9 to 102.6%. Finally, quantitative results of analytes were obtained from 10 different human urine samples.
Subject(s)
Chlorides , Creatine , Humans , Mass Spectrometry , Chromatography , Organic Chemicals , Sulfates , Urea , Chromatography, High Pressure Liquid/methodsABSTRACT
Humans are often exposed to phthalates and their alternatives, on account of their widespread use in PVC as plasticizers, which are associated with harmful human effects. While targeted biomonitoring provides quantitative information for exposure assessment, only a small portion of phthalate metabolites has been targeted. This results in a knowledge gap in human exposure to other unknown phthalate compounds and their metabolites. Although the non-targeted analysis (NTA) approach is capable of screening a broad spectrum of chemicals, there is a lack of harmonized workflow in NTA to generate reproducible data within and between different laboratories. The objective of this study was to compare two different NTA data acquisition modes, the data-dependent (DDA) and independent (DIA) acquisition (DDA), as well as two data analysis approaches, based on diagnostic ions and Compound Discoverer software for the prioritization of candidate precursors and identification of unknown compounds in human urine. Liquid chromatography coupled to high-resolution mass spectrometry was used for sample analysis. The combination of three-diagnostic-ion extraction and DDA data acquisition was able to improve data filtering and data analysis for prioritizing phthalate metabolites. With DIA, 25 molecular features were identified in human urine, while 32 molecular features were identified in the same urine samples using DDA data. The number of molecular features identified with level 1 confidence was 11 and 9 using DIA and DDA data, respectively. The study demonstrated that besides sample preparation, the impact of data acquisition must be taken into account when developing a NTA method and a consistent protocol for evaluating such an impact is necessary.
Subject(s)
Phthalic Acids , Humans , Chromatography, Liquid , Mass Spectrometry , Phthalic Acids/chemistry , Data AnalysisABSTRACT
Nicotine is the most prominent psychoactive/addictive chemical substance consumed worldwide among young players in team sports. Moreover, urinary nicotine discharge and nicotine-based products disposal in environmental waters has been unavoidable in recent years. Therefore, sensitive monitoring of nicotine content in environmental waters and human urine samples is essential. In this study, we developed a miniaturized novel green, low-cost, sensitive, in-syringe-based semi-automated fast drug extraction (FaDEx) protocol coupled with gas chromatography-flame ionization detection (GC-FID) for the efficient environmental and bio-monitoring of nicotine in aqueous samples. The FaDEx method consists of two steps; firstly, the target analyte was extracted using dimethyl carbonate (a green solvent) and extraction salts. After that, the extraction solvent was passed automatically through the solid-phase extraction cartridge at a constant flow rate for the cleanup process to achieve the sensitive nicotine analysis by GC-FID. Under optimized experimental conditions, the developed method showed excellent linearity over the concentration ranges between 20-2000 ng mL-1 with a correlation coefficient >0.99. The detection and quantification limits were 4 and 20 ng mL-1, respectively. The presented method was applied to monitor and assess nicotine exposure in sports-persons' urine and environmental water samples. The method accuracy and precision in terms of relative recovery and relative standard deviation (for triplicate analysis) were 85.4-110.2% and ≤8%, respectively. Finally, the impact of our procedure on the environment from a green analytical chemistry view was assessed using a novel metric system called AGREE, and obtained the greenness score of 0.87, indicating its an efficient alternative green analytical protocol for routine environmental and bio-monitoring of nicotine in environmental and biological samples.
Subject(s)
Nicotine , Water , Humans , Nicotine/analysis , Biological Monitoring , Limit of Detection , Solid Phase Extraction/methods , Solvents , Psychotropic Drugs/analysisABSTRACT
Given that the spontaneous precipitation of minerals caused by urea hydrolysis and abundant organic compounds, membrane fouling became a major obstacle for urine recovery by membrane distillation (MD). Herein, this study developed a combined system (TAP-MD) by integrating thermally activated peroxydisulfate (TAP) and MD process to inhibit membrane fouling and improve separation efficiency. Based on the TAP-MD system, the separation performance was improved significantly, improving nutrient recovery efficiency and quality of reclaimed water. More than 80% of water could be recovered from urine, and about 94.13% of total ammonia nitrogen (TAN), 99.02% of total nitrogen (TN), 100% of total phosphate (TP), and 100% of K+ were rejected. The mechanism for alleviating urine-induced fouling was systematically and intensively studied. With TAP pretreatment, the TAN concentration of pretreated urine was kept at a low level steadily and the pH was at neutral or weakly acidic. Hence, inorganic scaling represented by carbonate and phosphate precipitates were significantly inhibited by creating unfavorable solvent environment for crystallization with TAP pretreatment. Additionally, aromatic proteins were found as the main organic foulants. According to the secondary structure of protein, the proteins were degraded by the cleavage of peptide bonds by TAP pretreatment. Meanwhile, the hydrophilicity of protein increased, which reduced the hydrophobic interaction of protein and membrane surface and thus alleviated protein-induced membrane fouling. This study revealed the inorganic and organic foulants in urine that caused membrane fouling and demonstrated the mechanism of membrane fouling alleviation by TAP-MD system. The experimental results will be instrumental in better understanding the mechanisms of membrane fouling induced by urine and optimize MD process for resource recovery from urine.
ABSTRACT
Urinary extracellular vesicles (uEVs) are promising biomarkers for various diseases. However, many tools measuring uEVs rely on time-consuming uEV isolation methods, which could induce sample bias. This study demonstrates the detection of single uEVs without isolation using imaging flow cytometry (IFCM). Unstained urine samples contained auto-fluorescent (A-F) particles when characterized with IFCM. Centrifugation successfully removed A-F particles from the unprocessed urine. Based on the disappearance of A-F particles, a gate was defined to distinguish uEVs from A-F particles. The final readouts of IFCM were verified as single EVs based on detergent treatment and serial dilutions. When developing this protocol to measure urine samples with abnormally high protein levels, 25 mg/mL dithiothreitol (DTT) showed improved uEV recovery over 200 mg/mL DTT. This study provides an isolation-free protocol using IFCM to quantify and phenotype single uEVs, eliminating the hindrance and influence of A-F particles, protein aggregates, and coincidence events.
Subject(s)
Extracellular Vesicles , Flow Cytometry , Extracellular Vesicles/metabolism , Biomarkers/metabolismABSTRACT
Hydrothermal carbonization (HTC) is the most cost-effective, environmentally friendly, and efficient physicochemical and biochemical process for converting biomass to products with added value. The objective and novelty of this work is to produce and investigate the qualities of hydrochar fuel (as a solid fuel) from cow manure using human urine as a solvent in order to find a suitable replacement for conventional fuel (i.e., coal). HTC based studies were conducted in batch, at three different reaction temperatures (180 °C, 200 °C, and 220 °C) and two different reaction periods (2 and 4 h). For kinetic analysis and reaction mechanism of the combustion behavior of the produced hydrochar, the model free kinetic methods and the z-master plot were used. From the model free kinetics methods, it was observed that the resultant optimum average activation energy and pre-exponential factor for the produced hydrochar at 180 °C and 2 h reaction period (HTC_180_2) were â¼120 kJ/mol and â¼5.59 × 1025 sec-1, respectively. In addition, the little variation between ΔEα and ΔHα (â¼10 kJ/mol) suggests that the combustion of produced hydrochar (HTC_180_2) occurred with minimal energy use. Furthermore, the hydrochar exhibited its highest heating value at 200 °C for 4 h (HTC_200_4) which was 1.44 times higher than the raw dung (13.4 MJ/kg) due to the HTC process. The produced hydrochar demonstrated a significant improvement compared to the conventional solvent, i.e. water.