Last data update: Dec 09, 2024. (Total: 48320 publications since 2009)
Records 1-7 (of 7 Records) |
Query Trace: Quinones-Gonzalez J[original query] |
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Recapitulation of human pathophysiology and identification of forensic biomarkers in a translational model of chlorine inhalation injury
Achanta S , Gentile MA , Albert CJ , Schulte KA , Pantazides BG , Crow BS , Quinones-Gonzalez J , Perez JW , Ford DA , Patel RP , Blake TA , Gunn MD , Jordt SE . Am J Physiol Lung Cell Mol Physiol 2024 Chlorine gas (Cl(2)) has been repeatedly used as a chemical weapon, first in World War I and most recently in Syria. Life-threatening Cl(2) exposures frequently occur in domestic and occupational environments, and in transportation accidents. Modeling the human etiology of Cl(2)-induced acute lung injury (ALI), forensic biomarkers, and targeted countermeasures development have been hampered by inadequate large animal models. The objective of this study was to develop a translational model of Cl(2)-induced ALI in swine to understand toxico-pathophysiology and is suitable for screening potential medical countermeasures, and identify biomarkers useful for forensic analysis. Specific pathogen-free Yorkshire swine (30-40 kg) of either sex were exposed to Cl(2) (≤ 240 ppm for 1 h) or filtered air under anesthesia and controlled mechanical ventilation. Exposure to Cl(2) resulted in severe hypoxia and hypoxemia, increased airway resistance and peak inspiratory pressure, and decreased dynamic lung compliance. Cl(2) exposure resulted in increased total leucocyte and neutrophil counts in bronchoalveolar lavage fluid (BALF), vascular leakage, and pulmonary edema compared to the air-exposed group. The model recapitulated all three key histopathological features of human ALI, such as neutrophilic alveolitis, deposition of hyaline membranes, and formation of microthrombi. Free and lipid-bound 2‑chlorofatty acids and chlorotyrosine-modified proteins (3-chloro-L-tyrosine and 3,5-dichloro-L-tyrosine) were detected in plasma and lung tissue after Cl(2)‑exposure. In this study, we developed a translational swine model that recapitulates key features of human Cl(2) inhalation injury and is suitable for testing medical countermeasures, and validated chlorinated fatty acids and protein adducts as biomarkers of Cl(2) inhalation. |
Recapitulation of Human Pathophysiology and Identification of Forensic Biomarkers in a Translational Swine Model of Chlorine Inhalation Injury (preprint)
Achanta S , Gentile MA , Albert CJ , Schulte KA , Pantazides BG , Crow BS , Quinones-Gonzalez J , Perez JW , Ford DA , Patel RP , Blake TA , Gunn MD , Jordt SE . bioRxiv 2022 10 Rationale: Chlorine gas (Cl<inf>2</inf>) has been repeatedly used as a chemical weapon, first in World War I and most recently in Syria. Life-threatening Cl<inf>2</inf> exposures frequently occur in domestic and occupational environments, and in transportation accidents. There is a knowledge gap in large animal models of Cl<inf>2</inf>-induced acute lung injury (ALI) required to accurately model human etiology and for the development of targeted countermeasures Objective: To develop a translational model of Cl<inf>2</inf>-induced ALI in swine to study toxicopathophysiology and identify biomarkers useful for forensic analysis. Method(s): Specific pathogen-free Yorkshire swine (30-40 kg) of either sex were exposed to Cl<inf>2</inf> gas (<= 240 ppm for 1 h) or filtered air under anesthesia and controlled mechanical ventilation. Result(s): Exposure to Cl<inf>2</inf> resulted in severe hypoxia and hypoxemia, increased airway resistance and peak inspiratory pressure, and decreased dynamic lung compliance. Chlorine exposure resulted in increased total BALF and neutrophil counts, vascular leakage, and edema compared to the control group. The model recapitulated all three key histopathological features of human ALI, such as neutrophilic alveolitis, deposition of hyaline membranes, and formation of microthrombi. Free and lipid-bound 2-chlorofatty acids and chlorotyrosine-modified proteins (3-chloro-L-tyrosine and 3,5-dichloro-L-tyrosine) were detected in plasma and lung after Cl<inf>2</inf>-exposure. Conclusion(s): The translational model developed in this study replicates key features of humans exposed to Cl<inf>2</inf> and is suitable to test medical countermeasures. Specific biomarkers of Cl<inf>2</inf> exposure have been identified in plasma and lung tissue samples. Copyright The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license. |
Development of a clinical assay to measure chlorinated tyrosine in hair and tissue samples using a mouse chlorine inhalation exposure model
Pantazides BG , Crow BS , Quiñones-González J , Perez JW , Harvilchuck JA , Wallery JJ , Hu TC , Thomas JD , Johnson RC , Blake TA . Anal Bioanal Chem 2021 413 (6) 1765-1776 Chlorine is a toxic industrial chemical with a history of use as a chemical weapon. Chlorine is also produced, stored, and transported in bulk making it a high-priority pulmonary threat in the USA. Due to the high reactivity of chlorine, few biomarkers exist to identify exposure in clinical and environmental samples. Our laboratory evaluates acute chlorine exposure in clinical samples by measuring 3-chlorotyrosine (Cl-Tyr) and 3,5-dichlorotyrosine (Cl(2)-Tyr) using liquid chromatography tandem mass spectrometry (LC-MS/MS). Individuals can have elevated biomarker levels due to their environment and chronic health conditions, but levels are significantly lower in individuals exposed to chlorine. Historically these biomarkers have been evaluated in serum, plasma, blood, and bronchoalveolar lavage (BAL) fluid. We report the expansion into hair and lung tissue samples using our newly developed tissue homogenization protocol which fits seamlessly with our current chlorinated tyrosine quantitative assay. Furthermore, we have updated the chlorinated tyrosine assay to improve throughput and ruggedness and reduce sample volume requirements. The improved assay was used to measure chlorinated tyrosine levels in 198 mice exposed to either chlorine gas or air. From this animal study, we compared Cl-Tyr and Cl(2)-Tyr levels among three matrices (i.e., lung, hair, and blood) and found that hair had the most abundant chlorine exposure biomarkers. Furthermore, we captured the first timeline of each analyte in the lung, hair, and blood samples. In mice exposed to chlorine gas, both Cl-Tyr and Cl(2)-Tyr were present in blood and lung samples up to 24 h and up to 30 days in hair samples. |
A quantitative method to detect human exposure to sulfur and nitrogen mustards via protein adducts
Pantazides BG , Quinones-Gonzalez J , Rivera Nazario DM , Crow BS , Perez JW , Blake TA , Johnson RC . J Chromatogr B Analyt Technol Biomed Life Sci 2019 1121 9-17 Sulfur and nitrogen mustards are internationally banned vesicants listed as Schedule 1 chemical agents in the Chemical Weapons Convention. These compounds are highly reactive electrophiles that form stable adducts to a variety of available amino acid residues on proteins upon exposure. We present a quantitative exposure assay that simultaneously measures agent specific protein adducts to cysteine for sulfur mustard (HD) and three nitrogen mustards (HN1, HN2, and HN3). Proteinase K was added to a serum or plasma sample to digest protein adducts and form the target analyte, the blister agent bound to the tripeptide cysteine-proline-phenylalanine (CPF). The mustard adducted-tripeptide was purified by solid phase extraction and analyzed using isotope dilution LC-MS/MS. Product ion structures were identified using high-resolution product ion scan data for HD-CPF, HN1-CPF, HN2-CPF, and HN3-CPF. Thorough matrix comparison, analyte recovery, ruggedness, and stability studies were incorporated during method validation to produce a robust method. The method demonstrated long term-stability, precision (RSD<15%), and intra- and inter-day accuracies>85% across the reportable range of 3.00-200ng/mL for each analyte. Compared to previously published assays, this method quantitates both sulfur and nitrogen mustard exposure biomarkers, requires only 10muL of sample volume, and can use either a liquid sample or dried sample spot. |
Simultaneous measurement of 3-chlorotyrosine and 3,5-dichlorotyrosine in whole blood, serum and plasma by isotope dilution HPLC-MS-MS
Crow BS , Quinones-Gonzalez J , Pantazides BG , Perez JW , Winkeljohn WR , Garton JW , Thomas JD , Blake TA , Johnson RC . J Anal Toxicol 2016 40 (4) 264-71 Chlorine is a public health concern and potential threat due to its high reactivity, ease and scale of production, widespread industrial use, bulk transportation, massive stockpiles and history as a chemical weapon. This work describes a new, sensitive and rapid stable isotope dilution method for the retrospective detection and quantitation of two chlorine adducts. The biomarkers 3-chlorotyrosine (Cl-Tyr) and 3,5-dichlorotyrosine (Cl2-Tyr) were isolated from the pronase digest of chlorine exposed whole blood, serum or plasma by solid-phase extraction (SPE), separated by reversed-phase HPLC and detected by tandem mass spectrometry (MS-MS). The calibration range is 2.50-1,000 ng/mL (R2 ≥ 0.998) with a lowest reportable limit (LRL) of 2.50 ng/mL for both analytes, an accuracy of ≥93% and an LOD of 0.443 ng/mL for Cl-Tyr and 0.396 ng/mL for Cl2-Tyr. Inter- and intra-day precision of quality control samples had coefficients of variation of ≤10% and ≤7.0%, respectively. Blood and serum samples from 200 healthy individuals and 175 individuals with chronic inflammatory disease were analyzed using this method to assess background levels of chlorinated tyrosine adducts. Results from patients with no known inflammatory disease history (healthy) showed baseline levels of <LRL-4.26 ng/mL Cl-Tyr and <LRL Cl2-Tyr. Patients with inflammatory disease had baseline levels of <LRL-15.4 ng/mL Cl-Tyr and <LRL-5.22 ng/mL Cl2-Tyr. Blood exposed to 2.02 ppm chlorine gas for 15 min produced 941 ng/mL Cl-Tyr and 223 ng/mL Cl2-Tyr. This high-throughput method has been developed and analytically validated for the diagnosis of human exposure to chlorine. |
Simplified method for quantifying sulfur mustard adducts to blood proteins by ultrahigh pressure liquid chromatography-isotope dilution tandem mass spectrometry
Pantazides BG , Crow BS , Garton JW , Quinones-Gonzalez JA , Blake TA , Thomas JD , Johnson RC . Chem Res Toxicol 2015 28 (2) 256-61 Sulfur mustard binds to reactive cysteine residues, forming a stable sulfur-hydroxyethylthioethyl [S-HETE] adduct that can be used as a long-term biomarker of sulfur mustard exposure in humans. The digestion of sulfur mustard-exposed blood samples with proteinase K following total protein precipitation with acetone produces the tripeptide biomarker [S-HETE]-Cys-Pro-Phe. The adducted tripeptide is purified by solid phase extraction, separated by ultrahigh pressure liquid chromatography, and detected by isotope dilution tandem mass spectrometry. This approach was thoroughly validated and characterized in our laboratory. The average interday relative standard deviation was ≤9.49%, and the range of accuracy was between 96.1 and 109% over a concentration range of 3.00 to 250. ng/mL with a calculated limit of detection of 1.74 ng/mL. A full 96-well plate can be processed and analyzed in 8 h, which is 5 times faster than our previous 96-well plate method and only requires 50 muL of serum, plasma, or whole blood. Extensive ruggedness and stability studies and matrix comparisons were conducted to create a robust, easily transferrable method. As a result, a simple and high-throughput method has been developed and validated for the quantitation of sulfur mustard blood protein adducts in low volume blood specimens which should be readily adaptable for quantifying human exposures to other alkylating agents. |
Simultaneous measurement of tabun, sarin, soman, cyclosarin, VR, VX, and VM adducts to tyrosine in blood products by isotope dilution UHPLC-MS/MS
Crow BS , Pantazides BG , Quinones-Gonzalez J , Garton JW , Carter MD , Perez JW , Watson CM , Tomcik DJ , Crenshaw MD , Brewer BN , Riches JR , Stubbs SJ , Read RW , Evans RA , Thomas JD , Blake TA , Johnson RC . Anal Chem 2014 86 (20) 10397-405 This work describes a new specific, sensitive, and rapid stable isotope dilution method for the simultaneous detection of the organophosphorus nerve agents (OPNAs) tabun (GA), sarin (GB), soman (GD), cyclosarin (GF), VR, VX, and VM adducts to tyrosine (Tyr). Serum, plasma, and lysed whole blood samples (50 muL) were prepared by protein precipitation followed by digestion with Pronase. Specific Tyr adducts were isolated from the digest by a single solid phase extraction (SPE) step, and the analytes were separated by reversed-phase ultra high performance liquid chromatography (UHPLC) gradient elution in less than 2 min. Detection was performed on a triple quadrupole tandem mass spectrometer using time-triggered selected reaction monitoring (SRM) in positive electrospray ionization (ESI) mode. The calibration range was characterized from 0.100-50.0 ng/mL for GB- and VR-Tyr and 0.250-50.0 ng/mL for GA-, GD-, GF-, and VX/VM-Tyr (R2 ≥ 0.995). Inter- and intra-assay precision had coefficients of variation of ≤17 and ≤10%, respectively, and the measured concentration accuracies of spiked samples were within 15% of the targeted value for multiple spiking levels. The limit of detection was calculated to be 0.097, 0.027, 0.018, 0.074, 0.023, and 0.083 ng/mL for GA-, GB-, GD-, GF-, VR-, and VX/VM-Tyr, respectively. A convenience set of 96 serum samples with no known nerve agent exposure was screened and revealed no baseline values or potential interferences. This method provides a simple and highly specific diagnostic tool that may extend the time postevent that a confirmation of nerve agent exposure can be made with confidence. |
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