RESUMEN
Metabolomics is the study of low molecular weight biochemical molecules (typically <1500 Da) in a defined biological organism or system. In case of food systems, the term "food metabolomics" is often used. Food metabolomics has been widely explored and applied in various fields including food analysis, food intake, food traceability, and food safety. Food safety applications focusing on the identification of pathogen-specific biomarkers have been promising. This chapter describes a nontargeted metabolite profiling workflow using gas chromatography coupled with mass spectrometry (GC-MS) for characterizing three globally important foodborne pathogens, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica, from selective enrichment liquid culture media. The workflow involves a detailed description of food spiking experiments followed by procedures for the extraction of polar metabolites from media, the analysis of the extracts using GC-MS, and finally chemometric data analysis using univariate and multivariate statistical tools to identify potential pathogen-specific biomarkers.
Asunto(s)
Biomarcadores , Microbiología de Alimentos , Cromatografía de Gases y Espectrometría de Masas , Listeria monocytogenes , Metabolómica , Metabolómica/métodos , Cromatografía de Gases y Espectrometría de Masas/métodos , Biomarcadores/análisis , Microbiología de Alimentos/métodos , Listeria monocytogenes/metabolismo , Listeria monocytogenes/aislamiento & purificación , Salmonella enterica/metabolismo , Escherichia coli O157/metabolismo , Escherichia coli O157/aislamiento & purificación , Enfermedades Transmitidas por los Alimentos/microbiología , MetabolomaRESUMEN
Long-term and excessive use of tetracycline hydrochloride (TC) can lead to its accumulation in the environment, which can cause water contamination, bacterial resistance, and food safety problems. 2,6-Pyridine dicarboxylic acid (DPA) is a major biomarker of Bacillus anthracis spores, and its rapid and sensitive detection is of great significance for disease prevention and counter-terrorism. A bifunctional ratiometric fluorescent nanoprobe has been fabricated to detect DPA and TC. 3,5-dicarboxyphenylboronic acid (BOP) was intercalated into layered europium hydroxide (LEuH) by the ion-exchange method and exfoliated into nanosheets as a fluorescent nanoprobe (PNP). DPA and TC could significantly enhance the red fluorescence of Eu3+ through the antenna effect under different excitation wavelengths, while the fluorescence of BOP can be used as a reference based on the constant emission intensity, realizing ratiometric detection. A low limit of detection (LOD) for the target (DPA: 9.7 nM, TC: 21.9 nM) can be achieved. In addition, visual detection of DPA and TC was realized using color recognition software based on the obvious color changes. This is the first ratiometric fluorescent nanoprobe based on layered rare-earth hydroxide (LRH) for the detection of DPA and TC simultaneously, which opens new ideas in the design of multifunctional probes.
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Bacillus anthracis , Biomarcadores , Colorantes Fluorescentes , Espectrometría de Fluorescencia , Esporas Bacterianas , Tetraciclina , Colorantes Fluorescentes/química , Espectrometría de Fluorescencia/métodos , Bacillus anthracis/aislamiento & purificación , Biomarcadores/análisis , Tetraciclina/análisis , Límite de Detección , Ácidos Picolínicos/análisis , Carbunco/diagnósticoRESUMEN
BACKGROUND: Diabetic retinopathy (DR), a chronic and progressive microvascular complication of diabetes mellitus, substantially threatens vision and is a leading cause of blindness among working-age individuals worldwide. Traditional diagnostic methods, such as ophthalmoscopy and fluorescein angiography are nonquantitative, invasive, and time consuming. Analysis of protein biomarkers in tear fluid offers noninvasive insights into ocular and systemic health, aiding in early DR detection. This study introduces a surface acoustic wave (SAW) microchip that rapidly enhances fluorescence in bead-based immunoassays for the sensitive and noninvasive DR detection from human tear samples. RESULTS: The device facilitated particle mixing for immunoassay formation and particle concentration in the droplet, resulting in an enhanced immunofluorescence signal. This detachable SAW microchip allows the disposal of the cover glass after every use, thereby improving the reusability of the interdigital transducer and minimizing potential cross-contamination. A preliminary clinical test was conducted on a cohort of 10 volunteers, including DR patients and healthy individuals. The results demonstrated strong agreement with ELISA studies, validating the high accuracy rate of the SAW microchip. SIGNIFICANCE: This comprehensive study offers significant insights into the potential application of a novel SAW microchip for the early detection of DR in individuals with diabetes. By utilizing protein biomarkers found in tear fluid, the device facilitates noninvasive, rapid, and sensitive detection, potentially revolutionizing DR diagnostics and improving patient outcomes through timely intervention and management of this vision-threatening condition.
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Retinopatía Diabética , Lágrimas , Humanos , Lágrimas/química , Retinopatía Diabética/diagnóstico , Inmunoensayo/métodos , Sonido , Técnicas Biosensibles/instrumentación , Biomarcadores/análisis , Propiedades de SuperficieRESUMEN
Oil spills from pipeline accidents can have long-lasting health effects on residents of polluted regions. Assessing the potential health risk of these accidents is crucial for effective environmental health management. This study analyzed the concentration of 2-OHNAP in urine and hair as biomarkers of PAHs exposure among the people living in a region with frequent oil pipeline incident in Iran. Fifty pairs of hair and urine samples were collected from residents along with demographic information and dietary habits via a questionnaire. The concentration of 2-OHNAP was analyzed using high performance liquid chromatography coupled with fluorescence detector (HPLC-FLD). 2-OHNAP was detected in 100% of urine and 88% of hair samples. The mean concentration of 2-OHNAP in urine was 16.65 ± 21.98 µg/g creatinine and in hair was 8.16±7.62 ng/g dry weight (dw). However, there was no significant correlations between the levels of 2-OHNAP in urine and hair. The mean values of HQ and CR were below 1 and 10-6, respectively. Moreover, some simulated health risk indices were near the threshold levels, and the carcinogenic risk above 70% of the simulated CRs was above 10-6 as well. Therefore, the health risk attributed to the exposure to the parent compound of 2-OHNAP in the study area is currently acceptable, but it is not negligible and may be worsened in the future. This study provides a valuable scientific information for regional decision makers and stakeholders about human health programs and identification of environmental health priorities.
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Cabello , Humanos , Irán/epidemiología , Cabello/química , Medición de Riesgo , Masculino , Femenino , Adulto , Contaminación por Petróleo/análisis , Exposición a Riesgos Ambientales/análisis , Persona de Mediana Edad , Biomarcadores/orina , Biomarcadores/análisis , Naftalenos/análisis , Naftalenos/orina , Monitoreo del Ambiente , Cromatografía Líquida de Alta PresiónRESUMEN
INTRODUCTION: Volatile organic compounds (VOCs) can arise from underlying metabolism and are detectable in exhaled breath, therefore offer a promising route to non-invasive diagnostics. Robust, precise, and repeatable breath measurement platforms able to identify VOCs in breath distinguishable from background contaminants are needed for the confident discovery of breath-based biomarkers. OBJECTIVES: To build a reliable breath collection and analysis method that can produce a comprehensive list of known VOCs in the breath of a heterogeneous human population. METHODS: The analysis cohort consisted of 90 pairs of breath and background samples collected from a heterogenous population. Owlstone Medical's Breath Biopsy® OMNI® platform, consisting of sample collection, TD-GC-MS analysis and feature extraction was utilized. VOCs were determined to be "on-breath" if they met at least one of three pre-defined metrics compared to paired background samples. On-breath VOCs were identified via comparison against purified chemical standards, using retention indexing and high-resolution accurate mass spectral matching. RESULTS: 1471 VOCs were present in > 80% of samples (breath and background), and 585 were on-breath by at least one metric. Of these, 148 have been identified covering a broad range of chemical classes. CONCLUSIONS: A robust breath collection and relative-quantitative analysis method has been developed, producing a list of 148 on-breath VOCs, identified using purified chemical standards in a heterogenous population. Providing confirmed VOC identities that are genuinely breath-borne will facilitate future biomarker discovery and subsequent biomarker validation in clinical studies. Additionally, this list of VOCs can be used to facilitate cross-study data comparisons for improved standardization.
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Pruebas Respiratorias , Cromatografía de Gases y Espectrometría de Masas , Compuestos Orgánicos Volátiles , Compuestos Orgánicos Volátiles/análisis , Pruebas Respiratorias/métodos , Humanos , Cromatografía de Gases y Espectrometría de Masas/métodos , Masculino , Femenino , Persona de Mediana Edad , Adulto , Biomarcadores/análisis , Anciano , Adulto Joven , EspiraciónRESUMEN
Clinical expression of coronavirus disease 2019 (COVID-19) infectionis widely variable including fatal cases and patients with mild symptoms and a rapid resolution. We studied saliva from 63 hospitalized COVID-19 patients and from 30 healthy controls by integrating large-scale proteomics, peptidomics and targeted metabolomics to assess the biochemical alterations following the infection and to obtain a set of putative biomarkers useful for noninvasive diagnosis. We used an untargeted approach by using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for proteomics and peptidomics analysis and targeted LC-multiple reaction monitoring/MS for the analysis of amino acids. The levels of 77 proteins were significantly different in COVID-19 patients. Among these, seven proteins were found only in saliva from patients with COVID-19, four were up-regulated and three were down-regulated at least five-folds in saliva from COVID-19 patients in comparison to controls. The analysis of proteins revealed a complex balance between pro-inflammatory and anti-inflammatory proteins and a reduced amount of several proteins with immune activity that possibly favours the spreading of the virus. Such reduction could be related to the enhanced activity of endopeptidases induced by the infection that in turn caused an altered balance of free peptides. In fact, on a total of 28 peptides, 22 (80%) were differently expressed in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and control subjects. The multivariate analysis of such peptides permits to obtain a diagnostic algorithm that discriminate the two populations with a high diagnostic efficiency. Among amino acids, only threonine resulted significantly different between COVID-19 patients and controls, while alanine levels were significantly different between COVID-19 patients with different severity. In conclusion, the present study defined a set of molecules to be detected with a quick and easy method based on mass spectrometry tandem useful to reveal biochemical alterations involved in the pathogenesis of such a complex disease. Data are available via ProteomeXchange with identifier PXD045612.
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Biomarcadores , COVID-19 , Metabolómica , Proteómica , SARS-CoV-2 , Saliva , Espectrometría de Masas en Tándem , Humanos , COVID-19/virología , COVID-19/metabolismo , Saliva/química , Saliva/virología , Espectrometría de Masas en Tándem/métodos , Masculino , Femenino , Proteómica/métodos , Persona de Mediana Edad , Metabolómica/métodos , Biomarcadores/análisis , Biomarcadores/metabolismo , Adulto , Anciano , Cromatografía Liquida/métodos , Estudios de Casos y Controles , Proteoma/análisis , Proteoma/metabolismoRESUMEN
Non-alcoholic fatty liver disease (NAFLD) and myocardial infarction (MI) are two major health burdens with significant prevalence and mortality. This study aimed to explore the co-expressed genes to understand the relationship between NAFLD and MI and identify potential crucial biomarkers of NAFLD-related MI using bioinformatics and machine learning. Functional enrichment analysis was conducted, a co-protein-protein interaction (PPI) network diagram was constructed, and support vector machine-recursive feature elimination (SVM-RFE) and least absolute shrinkage and selection operator (LASSO) techniques were employed to identify one differentially expressed gene (DEG), Thrombospondin 1 (THBS1). THBS1 demonstrated strong performance in distinguishing NAFLD patients (AUC = 0.981) and MI patients (AUC = 0.900). Immuno-infiltration analysis revealed significantly lower CD8+ T cells and higher neutrophil levels in patients with NAFLD and MI. CD8+ T cells and neutrophils were effective in distinguishing NAFLD/MI from healthy controls. Correlation analysis showed that THBS1 was positively correlated with CCR (chemokine receptor), MHC class (major histocompatibility complex class), neutrophils, parainflammation, and Tfh (follicular helper T cells), and negatively correlated with CD8+ T cells, cytolytic activity, and TIL (tumor-infiltrating lymphocytes) in NAFLD and MI patients. THBS1 emerged as a novel biomarker for diagnosing NAFLD/MI in comparison to healthy controls. The results indicate that CD8+ T cells and neutrophils could serve as inflammatory immune features for differentiating patients with NAFLD/MI from healthy individuals.
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Enfermedad del Hígado Graso no Alcohólico , Trombospondina 1 , Humanos , Enfermedad del Hígado Graso no Alcohólico/inmunología , Enfermedad del Hígado Graso no Alcohólico/genética , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Trombospondina 1/genética , Trombospondina 1/metabolismo , Infarto del Miocardio/inmunología , Infarto del Miocardio/metabolismo , Infarto del Miocardio/genética , Máquina de Vectores de Soporte , Biomarcadores/metabolismo , Biomarcadores/análisisRESUMEN
INTRODUCTION: The WHO estimates a gap of about 30% between the incident (10.6 million) and notified (7.5 million) cases of tuberculosis (TB). Combined with the growing recognition in prevalence surveys of the high proportion of cases identified who are asymptomatic or paucisymptomatic, these data underscore how current symptom screening approaches and use of diagnostic tests with suboptimal performance on sputum miss large numbers of cases. Thus, the development of sputum-free biomarker-based tests for diagnosis is becoming necessary, which the WHO has already identified as a priority for new TB diagnostics.The objective of this study is to evaluate a combination of exhaled breath condensate (EBC) samples and mycobacterial lipoarabinomannan (LAM) as point-of-care (POC) assays to identify TB patients. METHODS AND ANALYSIS: This prospective diagnostic accuracy study is conducted at the TB Screening and Treatment Centre of International Center for Diarrhoeal Disease Research, Bangladesh, on a cohort of adults and adolescents >11 years of age. A total of 614 individuals with presumptive pulmonary TB based on TB signs, symptoms and radiography are being recruited from 28 August 2023. Spot sputum is collected for standard reference testing (L-J culture, GeneXpert MTB/Rif, acid-fast Bacilli microscopy) to fine-tune categorisation of TB disease status for each participant, defined as (1) definite TB (at least one positive standard reference test); (2) probable TB (not microbiologically confirmed but under TB treatment); (3) possible TB (no TB treatment but signs, symptoms and radiography suggestive of TB); (4) other respiratory disease (microbiologically not confirmed and no radiography presenting abnormalities compatible with TB); and (5) unknown (no microbiological evidence with normal/no TB abnormalities with radiography). Urine and EBC specimens will be subjected to LAM POC testing and biobanked for further investigation. Statistical analyses will include an assessment of diagnostic accuracy by constructing receiver operating curves and calculating sensitivity and specificity, as well as post-test probabilities. ETHICS AND DISSEMINATION: The study protocol was approved by the Research Review Committee as well as the Ethical Review Committee of icddr,b and recorded under a protocol reference number, PR-2301. Results will be submitted to open-access peer-reviewed journals, presented at academic meetings, and shared with national and international policymaking bodies.
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Pruebas Respiratorias , Lipopolisacáridos , Tuberculosis Pulmonar , Humanos , Lipopolisacáridos/análisis , Tuberculosis Pulmonar/diagnóstico , Pruebas Respiratorias/métodos , Estudios Prospectivos , Biomarcadores/análisis , Bangladesh , Adulto , Pruebas en el Punto de Atención , Sensibilidad y Especificidad , Sistemas de Atención de Punto , Masculino , Femenino , Adolescente , Mycobacterium tuberculosis/aislamiento & purificación , Esputo/microbiologíaRESUMEN
Alzheimer's disease (AD) is the most common neurodegenerative disorder, characterized pathologically by extracellular deposition of ß-amyloid (Aß) into senile plaques and intracellular accumulation of hyperphosphorylated tau (pTau) as neurofibrillary tangles. Clinically, AD patients show memory deterioration with varying cognitive dysfunctions. The exact molecular mechanisms underlying AD are still not fully understood, and there are no efficient drugs to stop or reverse the disease progression. In this review, we first provide an update on how the risk factors, including APOE variants, infections and inflammation, contribute to AD; how Aß and tau become abnormally accumulated and how this accumulation plays a role in AD neurodegeneration. Then we summarize the commonly used experimental models, diagnostic and prediction strategies, and advances in periphery biomarkers from high-risk populations for AD. Finally, we introduce current status of development of disease-modifying drugs, including the newly officially approved Aß vaccines, as well as novel and promising strategies to target the abnormal pTau. Together, this paper was aimed to update AD research progress from fundamental mechanisms to the clinical diagnosis and therapies.
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Enfermedad de Alzheimer , Enfermedad de Alzheimer/terapia , Enfermedad de Alzheimer/diagnóstico , Enfermedad de Alzheimer/metabolismo , Humanos , Proteínas tau/metabolismo , Péptidos beta-Amiloides/metabolismo , Animales , Biomarcadores/metabolismo , Biomarcadores/análisisRESUMEN
BACKGROUND: Sepsis is a heterogeneous syndrome. This study aimed to identify new sepsis sub-phenotypes using plasma cortisol trajectory. METHODS: This retrospective study included patients with sepsis admitted to the intensive care unit of Zhongshan Hospital Fudan University between March 2020 and July 2022. A group-based cortisol trajectory model was used to classify septic patients into different sub-phenotypes. The clinical characteristics, biomarkers, and outcomes were compared between sub-phenotypes. RESULTS: A total of 258 patients with sepsis were included, of whom 186 were male. Patients were divided into two trajectory groups: the lower-cortisol group (n = 217) exhibited consistently low and slowly declining cortisol levels, while the higher-cortisol group (n = 41) showed relatively higher levels in comparison. The 28-day mortality (65.9% vs.16.1%, P < 0.001) and 90-day mortality (65.9% vs. 19.8%, P < 0.001) of the higher-cortisol group were significantly higher than the lower-cortisol group. Multivariable Cox regression analysis showed that the trajectory sub-phenotype (HR = 5.292; 95% CI 2.218-12.626; P < 0.001), APACHE II (HR = 1.109; 95% CI 1.030-1.193; P = 0.006), SOFA (HR = 1.161; 95% CI 1.045-1.291; P = 0.006), and IL-1ß (HR = 1.001; 95% CI 1.000-1.002; P = 0.007) were independent risk factors for 28-day mortality. Besides, the trajectory sub-phenotype (HR = 4.571; 95% CI 1.980-10.551; P < 0.001), APACHE II (HR = 1.108; 95% CI 1.043-1.177; P = 0.001), SOFA (HR = 1.270; 95% CI 1.130-1.428; P < 0.001), and IL-1ß (HR = 1.001; 95% CI 1.000-1.001; P = 0.015) were also independent risk factors for 90-day mortality. CONCLUSION: This study identified two novel cortisol trajectory sub-phenotypes in patients with sepsis. The trajectories were associated with mortality, providing new insights into sepsis classification.
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Biomarcadores , Hidrocortisona , Fenotipo , Sepsis , Humanos , Masculino , Sepsis/sangre , Sepsis/clasificación , Sepsis/mortalidad , Sepsis/fisiopatología , Femenino , Hidrocortisona/sangre , Hidrocortisona/análisis , Estudios Retrospectivos , Persona de Mediana Edad , Anciano , Biomarcadores/sangre , Biomarcadores/análisis , Unidades de Cuidados Intensivos/organización & administración , Unidades de Cuidados Intensivos/estadística & datos numéricos , APACHE , Modelos de Riesgos Proporcionales , AdultoRESUMEN
The oriented design of reticular materials as emitters can significantly enhance the sensitivity of electrochemiluminescence (ECL) sensing analysis for disease markers. However, due to the structural fragility of hydrogen bonds, relational research on hydrogen-bonded organic frameworks (HOFs) has not been thoroughly conducted. Additionally, the modulation of luminescence behavior through HOFs has been rarely reported. In view of this, hydrogen-bonded biohybrid organic frameworks (HBOFs) were synthesized and recruited for ECL immunoassay applications. HBOFs was easily prepared using 6,6',6â³,6â´-(pyrene-1,3,6,8-tetrayl)tetrakis(2-naphthoic acid) as linkers via bovine serum albumin (BSA) activated hydrogen-bonded cross-linking. The material exhibited good fluorescence emission characteristics. And the highly ordered topological structure and molecular motion limitation mediated by BSA overcome aggregation-caused quenching and generate strong aggregation induced emission, expressing hydrogen-bond interaction enhanced ECL (HIE-ECL) activity with the participation of tri-n-propylamine. Furthermore, a sandwich immunosensor was constructed employing cobalt-based metal-phenolic network (CMPN) coated ferrocene nanoparticles (FNPs) as quenchers (CMPN@FNPs). Signal closure can be achieved by annihilating the excited state through electron transfer from both CMPN and FNPs. Using a universal disease marker, carcinoembryonic antigen, as the analysis model, the signal-off sensor obtained a detection limit of 0.47 pg/mL within the detection range of 1 pg/mL - 50 ng/mL. The synthesis and application of highly stable HBOFs triggered by proteins provide a reference for the development of new reticular ECL signal labels, and electron transfer model provides flexible solutions for more sensitive sensing analysis.
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Técnicas Biosensibles , Técnicas Electroquímicas , Enlace de Hidrógeno , Mediciones Luminiscentes , Albúmina Sérica Bovina , Técnicas Biosensibles/métodos , Inmunoensayo/métodos , Técnicas Electroquímicas/métodos , Mediciones Luminiscentes/métodos , Humanos , Albúmina Sérica Bovina/química , Animales , Estructuras Metalorgánicas/química , Límite de Detección , Bovinos , Metalocenos/química , Compuestos Ferrosos/química , Anticuerpos Inmovilizados/química , Biomarcadores/análisis , Cobalto/químicaRESUMEN
OBJECTIVE: The aim of our study was to investigate whether a 1-month-long milk-free diet results in a reduction in faecal calprotectin (FC) and faecal-zonulin-related proteins (FZRP) in children with milk-protein-induced allergic proctocolitis (MPIAP). MATERIALS AND METHODS: This is a single-centre, prospective, observational cohort study involving 86 infants with MPIAP, aged 1-3 months, and 30 healthy controls of the same age. The FC and FZRP were marked using the ELISA method (IDK® Calprotectin or Zonulin ELISA Kit, Immunodiagnostik AG, Bensheim, Germany). The diagnosis of MPIAP was confirmed with an open milk challenge test. RESULTS: FFC and FZRP proved useful in evaluating MPIAP treatment with a milk-free diet, and the resolution of allergic symptoms and a significant (p = 0.0000) decrease in the concentrations of both biomarkers were observed after 4 weeks on the diet. The FC and FZRP concentrations were still higher than in the control group. A high variability of FC concentrations was found in all the study groups. An important limitation is the phenomenon of FZRP not being produced in all individuals, affecting one in five infants. CONCLUSIONS: FC and FZRP can be used to monitor the resolution of colitis in infants with MPIAP treated with a milk-free diet, indicating a slower resolution of allergic inflammation than of allergic symptoms. The diagnosis of MPIAP on the basis of FC concentrations is subject to considerable error, due to the high individual variability of this indicator. FZRP is a better parameter, but this needs further research, as these are the first determinations in infants with MPIAP.
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Biomarcadores , Heces , Haptoglobinas , Complejo de Antígeno L1 de Leucocito , Hipersensibilidad a la Leche , Proteínas de la Leche , Proctocolitis , Precursores de Proteínas , Humanos , Complejo de Antígeno L1 de Leucocito/análisis , Complejo de Antígeno L1 de Leucocito/metabolismo , Heces/química , Lactante , Femenino , Estudios Prospectivos , Masculino , Biomarcadores/análisis , Proctocolitis/diagnóstico , Haptoglobinas/análisis , Haptoglobinas/metabolismo , Hipersensibilidad a la Leche/diagnóstico , Hipersensibilidad a la Leche/dietoterapia , Precursores de Proteínas/análisis , Precursores de Proteínas/metabolismo , Proteínas de la Leche/análisis , Toxina del Cólera/análisis , Estudios de SeguimientoRESUMEN
BACKGROUND: Magnetic ionic liquids (MILs) have been explored in dispersive liquid-liquid microextraction (DLLME). Their usage allows to substitute centrifugation and/or filtration steps by a quick magnetic separation. Besides, effervescence-assisted DLLME is one of the most known options to improve the dispersion of the extractant in the sample, while allowing to avoid the consumption of external energy during dispersion. Despite these interesting features, only one study incorporates MILs containing the tetrachloroferrate anion in effervescence tablets. These MILs are highly viscous and liquid at room temperature, thus compromising the stability of the tablets when used as extraction microdevices in effervescence-assisted DLLME, and only allowing their use in the conventional MIL-DLLME mode. RESULTS: A new class of effervescence tablets containing a Ni(II)-based MIL, that is solid at room temperature, is here proposed. This type of tablets permits their use, for first time, in the in situ DLLME mode, occurring through the transformation of a water-soluble MIL into a water-insoluble MIL microdroplet. This way, the tablet formulation included: the MIL, the metathesis reagent lithium bis[(trifluoromethyl)sulfonyl]imide, NaH2PO4 and K2CO3 as effervescence precursors salts, and Na2SO4 as salting-out and desiccating agent. The method is combined with high-performance liquid-chromatography and both fluorescence and ultraviolet detection, for the determination of monohydroxylated polycyclic aromatic hydrocarbons (OH-PAHs) and benzophenones (BPs), as biomarkers in urine. The method simply involved the addition of the effervescence tablet to the sample, thus taken place simultaneously the effervescence process and the metathesis reaction, without requiring any external energy consumption. The method presented limits of detection down to 10 ng L-1 for OH-PAHs and to 0.60 µg L-1 for BPs, inter-day relative standard deviations lower than 17 %, and average relative recoveries of 94 % in urine. The determined OH-PAHs contents in urine were between 0.40 and 16 µg L-1, and between 17.8 and 334 µg L-1 for BPs. SIGNIFICANCE: We have developed the first MIL-based effervescence tablets that are completely solid, thus improving the stability and robustness of these microdevices with respect to previously reported tablets involving MILs, while permitting to perform into the in situ DLLME mode (thus gaining in extraction efficiency). This approach including the MIL-based effervescence tablets constitutes an alternative on-site platform for the analysis of urine, as satisfactory precision, accuracy, and sensitivity are achieved despite not involving any external energy input within the analytical sample preparation setup. This method also constitutes the first application of MIL-based effervescence tablets for bioanalysis.
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Biomarcadores , Líquidos Iónicos , Microextracción en Fase Líquida , Comprimidos , Líquidos Iónicos/química , Microextracción en Fase Líquida/métodos , Comprimidos/química , Biomarcadores/orina , Biomarcadores/análisis , Humanos , Límite de Detección , Fenómenos MagnéticosRESUMEN
OBJECTIVE: This study aims to analyse the association between the baseline microbial load of selected periodontopathogenic bacteria collected from gingival crevicular fluid (GCF) and the primary outcome of steps I and II therapy. MATERIALS AND METHODS: 222 patients with stage III periodontitis were included into this retrospective analysis that received steps 1 and 2 periodontal therapy without adjunctive systemic antibiotics. Baseline GCF samples were quantitatively analysed using ELISA-based kits for levels of periodontopathogens (Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa), Prevotella intermedia (Pi), Fusobacterium nucleatum (Fn), Treponema denticola (Td), and Tannerella forsythia (Tf)) and associated with the primary therapy outcome using a "treat-to-target" therapy endpoint (TE) defined as ≤ 4 sites with PD ≥ 5 mm six months after therapy. RESULTS: 38.2% of the patients achieved TE. Patients failing to achieve TE revealed significantly increased levels of Pg, Fn, and Tf at baseline (Pg: p = 0.010, Fn: p = 0.008 Tf: p = 0.004). Multivariate binary logistic regression adjusted for sex, mean probing depth, diabetes, and current smoking status showed an independent relationship between Tf and the TE (aOR 2.570, p = 0.023). CONCLUSION: Increased microbial load is associated with decreased responsiveness to therapy. The findings suggest that specifically baseline Tf levels are associated with poorer treatment outcomes and might improve the accuracy of periodontal diagnosis. CLINICAL RELEVANCE: The findings of this study support the concept of a critical biomass that is sufficient to induce and maintain an immune response within the periodontal pocket, which ultimately leads to irreversible tissue destruction. However, calculating this level in advance may serve as an early indicator for intervention. KEY FINDING: Baseline Tannerella forsythia levels are associated with poorer treatment outcome.
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Biomarcadores , Líquido del Surco Gingival , Humanos , Femenino , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Líquido del Surco Gingival/microbiología , Líquido del Surco Gingival/química , Resultado del Tratamiento , Biomarcadores/análisis , Ensayo de Inmunoadsorción Enzimática , Carga Bacteriana , Adulto , Treponema denticola/aislamiento & purificación , Porphyromonas gingivalis/aislamiento & purificación , Fusobacterium nucleatum/aislamiento & purificación , Tannerella forsythia/aislamiento & purificación , Periodontitis/microbiología , Periodontitis/terapia , Aggregatibacter actinomycetemcomitans/aislamiento & purificación , Prevotella intermedia/aislamiento & purificaciónRESUMEN
Passive smoking from environmental tobacco smoke not only increases the risk of lung cancer and cardiovascular disease but may also be a stressor triggering neuropsychiatric and other disorders. To prevent these diseases, understanding the relationship between passive smoking and stress is vital. In this study, we developed a simple and sensitive method to simultaneously measure nicotine (Nic) and cotinine (Cot) as tobacco smoke exposure biomarkers, and cortisol (CRT), serotonin (5-HT), melatonin (MEL), dopamine (DA), and oxytocin (OXT) as stress-related biomarkers. These were extracted and concentrated from saliva by in-tube solid-phase microextraction (IT-SPME) using a Supel-Q PLOT capillary as the extraction device, then separated and detected within 6 min by liquid chromatography-tandem mass spectrometry (LC-MS/MS) using a Kinetex Biphenyl column (Phenomenex Inc., Torrance, CA, USA). Limits of detection (S/N = 3) for Nic, Cot, CRT, 5-HT, MEL, DA, and OXT were 0.22, 0.12, 0.78, 0.39, 0.45, 1.4, and 3.7 pg mL-1, respectively, with linearity of calibration curves in the range of 0.01-25 ng mL-1 using stable isotope-labeled internal standards. Intra- and inter-day reproducibilities were under 7.9% and 14.6% (n = 5) relative standard deviations, and compound recoveries in spiked saliva samples ranged from 82.1 to 106.6%. In thirty nonsmokers, Nic contents positively correlated with CRT contents (R2 = 0.5264, n = 30), while no significant correlation was found with other biomarkers. The standard deviation of intervals between normal beats as the standard measure of heart rate variability analysis negatively correlated with CRT contents (R2 = 0.5041, n = 30). After passive smoke exposure, Nic levels transiently increased, Cot and CRT levels rose over time, and 5-HT, DA, and OXT levels decreased. These results indicate tobacco smoke exposure acts as a stressor in nonsmokers.
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Biomarcadores , Saliva , Microextracción en Fase Sólida , Espectrometría de Masas en Tándem , Contaminación por Humo de Tabaco , Humanos , Saliva/química , Saliva/metabolismo , Biomarcadores/análisis , Contaminación por Humo de Tabaco/efectos adversos , Contaminación por Humo de Tabaco/análisis , Espectrometría de Masas en Tándem/métodos , Microextracción en Fase Sólida/métodos , Cromatografía Liquida/métodos , Masculino , Adulto , Femenino , Hidrocortisona/análisis , Hidrocortisona/metabolismo , Serotonina/análisis , Serotonina/metabolismo , Nicotina/análisis , Cotinina/análisis , Cromatografía Líquida con Espectrometría de MasasRESUMEN
BACKGROUND: The breath carbon isotope ratio (CIR) was recently identified as a noninvasive candidate biomarker of short-term added sugars (AS) intake. OBJECTIVES: This study aimed to better understand the potential of the breath CIR as a dietary biomarker. We evaluated the effects of short-term and long-term intakes of AS, animal protein (AP), and related variables on breath CIR, in the context of typical dietary intake patterns. METHODS: We conducted a 15-d controlled feeding study of 100 adults (age 18-70 y, 55% females) in Phoenix, AZ. Participants were provided individualized diets that approximated habitual food intakes and recorded the timing of food consumption. Three breath samples (fasting, midday, and evening) were collected on each of 3 nonconsecutive study days. We modeled the effects of dietary intake in each of 8 h preceding collection of the breath sample on breath CIR with a linear mixed model, which also included 15-d mean intakes, sex, age, and BMI. RESULTS: Median (IQR) intakes of AS and AP in our study were 65 (38) and 67 (33) g/d, respectively. Midday and evening breath CIRs correlated strongly with each other (0.80) and with fasting breath CIR (0.77 and 0.68, respectively). In our linear mixed models, breath CIR increased by AS consumed 1-4 h before sample collection, AP consumed 3-6 h before sample collection, and 15-d intakes of AS and AP, all with similar effect sizes. The breath CIR was also inversely associated with 15-d intakes of intrinsic sugars and plant protein; thus, associations with 15-d intakes were particularly strong when expressed proportionally as the AS ratio (added sugars/total sugars) and AP ratio (animal protein/total protein). CONCLUSIONS: The breath CIR is a promising measure of long-term intakes of AS and AP, especially as proportional intakes. Approaches to increase specificity would benefit the further development of this biomarker.
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Biomarcadores , Pruebas Respiratorias , Isótopos de Carbono , Humanos , Femenino , Adulto , Masculino , Persona de Mediana Edad , Adulto Joven , Anciano , Adolescente , Biomarcadores/metabolismo , Biomarcadores/análisis , Proteínas en la Dieta/administración & dosificación , Proteínas en la Dieta/análisis , Azúcares de la Dieta/administración & dosificación , DietaRESUMEN
Introduction: Idiopathic pulmonary fibrosis (IPF) affects the tissue surrounding the alveoli and occurs when the lung tissue becomes thick and stiff for unknown reasons. Clinical findings are fairly well settled, but the molecular mechanisms of IPF are still poorly known. Materials and Methods: To further our understanding, we collected publicly available transcriptome dataset from IPF cohorts, grouped them according to sampling method [bronchoalveolar lavage (BAL), biopsy, blood], and performed comparative meta-transcriptome study to (I) unravel key pathways (II), set out differences in discovered genes, pathways, and functional annotation with respect to the sampling method, and (III) find biomarkers for early diagnosis. Result: The resulting lists are also compared with DisGeNet reported genes, earlier work, and Kyoto encyclopedia of genes and genomes (KEGG) pathways. Several pathways are shared among BAL and biopsy samples while blood samples point to alternative pathways, indicating the noise in information obtained from these samples. Conclusions: Common to all sampling methods, interleukin-10 pathway and extracellular signaling pathways are pointed as further targets.
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Líquido del Lavado Bronquioalveolar , Perfilación de la Expresión Génica , Fibrosis Pulmonar Idiopática , Transcriptoma , Humanos , Fibrosis Pulmonar Idiopática/genética , Fibrosis Pulmonar Idiopática/patología , Biopsia , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/química , Leucocitos Mononucleares/metabolismo , Biomarcadores/análisis , Transducción de Señal/genética , Lavado BroncoalveolarRESUMEN
Background: Acne vulgaris (AV), a chronic inflammatory pilosebaceous disorder, affects 80-90% of teenagers. This study aimed to discover lipid profiles and biomarkers of the rabbit ear acne model, and investigate the mechanism of isotretinoin in treating acne at the lipid level. Methods: Untargeted lipidomic analysis using ultra-high performance liquid chromatography system (UHPLC) coupled to q-extraction plus was performed to identify skin lipid metabolites in blank control (groups C), model group (group M) and isotretinoin group (group T). Multivariate statistical analysis was used to process the lipidomics data. Results: A total of 43 lipid classes comprising 6989 lipid species were identified from the mass spectrometry data. The orthogonal partial least squares discriminant analysis (OPLS-DA) model demonstrated significant separation in skin lipidomic profiles between group M and group C. With variable influence on projection (VIP) > 1.0 and P-value < 0.05, 299 significantly different lipid metabolites were identified. These lipid metabolites consisted mainly of ceramides (Cer) (53.85%), phosphatidylethanolamines (PE) (9.03%), phosphatidylcholines (PC)(5.35%), and sphingomyelin (SM)(4.01%). Combining with AUC ≥ 0.9 as the elected criteria, Cer (d18;1_24:0), zymosterol (ZyE)(33:5), Cer (t43:1), ZyE (33:6), ZyE (24:7), and ZyE (35:6) have "high" accuracy. Isotretinoin treatment normalized 25 lipid metabolites in the acne model. Conclusion: Our findings provide new insights into the role of lipid metabolism in the pathogenesis of acne and the action mechanism of isotretinoin.
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Acné Vulgar , Biomarcadores , Modelos Animales de Enfermedad , Isotretinoína , Lipidómica , Lípidos , Isotretinoína/farmacología , Acné Vulgar/tratamiento farmacológico , Acné Vulgar/metabolismo , Animales , Conejos , Biomarcadores/metabolismo , Biomarcadores/análisis , Lípidos/análisis , Cromatografía Líquida de Alta Presión , Masculino , Fármacos Dermatológicos/farmacología , Fármacos Dermatológicos/uso terapéuticoRESUMEN
Age related cataracts (ARC) represent the main reason for blindness globally. The lens epithelial cells (LECs) participate not only in the metabolism of many substances in the lens but also in maintaining lens transparency. This study used lipidomics to investigate the metabolic differences in LECs of ARC patients with different severity, aiming at identifying potential metabolic biomarkers of ARC. Patients diagnosed with ARC and underwent cataract surgery at Shanghai Tongren Hospital were selected to participate in this study, which were classified as mild ARC group and severe ARC group. During their cataract surgery, anterior lens capsules(LCs) containing LECs were obtained. The lipidomics of LECs were analyzed using the liquid chromatographymass spectrometry (LC-MS). Potential pathways of lipids were searched for using databases such as the Kyoto Encyclopedia of Genes and Genomes (KEGG) and MetaboAnalyst platform. In LEC lipids, 26 lipids have been identified as potential biomarkers between mild ARC and severe ARC, with AUC values of 0.67-0.94. The pathway analysis results revealed that the Glycerophospholipid (GPL) metabolism was significantly influenced, indicating that these metabolic markers contribute significantly to regulating this pathway. The LEC metabolic spectrum demonstrates a proficient ability to differentiate between patients with varying levels of cataracts. Herein, we have successfully identified potential metabolic biomarkers and pathways that have proven to be valuable in enhancing our understanding of ARC pathogenesis. The finding has translational value for developing new cataract treatment methods in the future.
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Catarata , Células Epiteliales , Cristalino , Lipidómica , Humanos , Catarata/metabolismo , Catarata/patología , Células Epiteliales/metabolismo , Células Epiteliales/patología , Masculino , Femenino , Anciano , Cristalino/metabolismo , Cristalino/patología , Biomarcadores/metabolismo , Biomarcadores/análisis , Persona de Mediana Edad , Metabolismo de los Lípidos , Cromatografía LiquidaRESUMEN
The purpose of this study was to analyze the relationship between the intra-articular inflammatory response and any associated systemic inflammatory response following knee injury requiring operative management. Patients undergoing primary knee arthroscopy provided synovial fluid, blood, and urine samples immediately prior to surgery. Samples were analyzed using a multiplex magnetic bead immunoassay for the concentrations of cytokines and growth factors that have been shown to be associated with post-injury inflammation. One hundred and fifty-one patients undergoing arthroscopic management of meniscus, ACL, and focal chondral lesions were included in the analysis. After correction for multiple tests, there were no statistically significant correlations between synovial fluid biomarkers and biomarkers in plasma or urine for any of the intra-articular pathologies assessed. This analysis suggests that the most accurate measurement of the post-injury inflammatory response must be sampled from the intra-articular space. In the post-traumatic knee, there is no substitute for synovial fluid biomarker analysis.