RESUMEN
COVID-19 is a pandemic with high morbidity and mortality. In an autopsy cohort of COVID-19 patients, we found extensive accumulation of the tryptophan degradation products 3-hydroxy-anthranilic acid and quinolinic acid in the lungs, heart, and brain. This was not related to the expression of the tryptophan-catabolizing indoleamine 2,3-dioxygenase (IDO)-1, but rather to that of its isoform IDO-2, which otherwise is expressed rarely. Bioavailability of tryptophan is an absolute requirement for proper cell functioning and synthesis of hormones, whereas its degradation products can cause cell death. Markers of apoptosis and severe cellular stress were associated with IDO-2 expression in large areas of lung and heart tissue, whereas affected areas in brain were more restricted. Analyses of tissue, cerebrospinal fluid, and sequential plasma samples indicate early initiation of the kynurenine/aryl-hydrocarbon receptor/IDO-2 axis as a positive feedback loop, potentially leading to severe COVID-19 pathology. © 2021 The Authors. The Journal of Pathology published by John Wiley & Sons, Ltd on behalf of The Pathological Society of Great Britain and Ireland.
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Encéfalo/enzimología , COVID-19/enzimología , Indolamina-Pirrol 2,3,-Dioxigenasa/análisis , Pulmón/enzimología , Miocardio/enzimología , Ácido 3-Hidroxiantranílico/análisis , Adulto , Anciano , Apoptosis , Autopsia , Encéfalo/patología , COVID-19/mortalidad , COVID-19/patología , COVID-19/virología , Humanos , Quinurenina/análisis , Pulmón/patología , Persona de Mediana Edad , Miocardio/patología , Estudios Prospectivos , Ácido Quinolínico/análisis , Índice de Severidad de la Enfermedad , Triptófano/análisisRESUMEN
The simultaneous quantitative estimation of tryptophan (TRP) and its metabolites represents a great challenge because of their diverse chemical properties, e.g., presence of acidic, basic, and nonpolar functional groups and their immensely different concentrations in biological matrices. A short ultra high-performance liquid chromatography (UHPLC)-tandem mass spectrometry (MS/MS) method was validated for targeted analysis of TRP and its 11 most important metabolites derived via both kynurenine (KYN) and serotonin (SERO) pathways in human serum and cerebrospinal fluid (CSF): SERO, KYN, 3-hydroxyanthranilic acid, 5-hydroxyindoleacetic acid, anthranilic acid, kynurenic acid (KYNA), 3-hydroxykynurenine (3-HK), xanthurenic acid, melatonin, picolinic acid (PICA), and quinolinic acid (QUIN). After selecting the "best" reversed-phase column and organic modifier, DryLab®4 was used to optimize the gradient time and temperature in chromatographic separation. To achieve absolute quantification, deuterium-labeled internal standards were used. Among all compounds, 3 were analyzed in derivatized (butyl ester) forms (3-HK, PICA, and QUIN) and the remaining 9 in underivatized forms. Validation was performed in accordance with the ICH and FDA guidelines to determine the intraday and interday precision, accuracy, sensitivity, and recovery. To demonstrate the applicability of the developed UHPLC-MS/MS method, the aforementioned metabolites were analyzed in serum and CSF samples from patients with multiple sclerosis (multiple sclerosis group) and those with symptomatic or noninflammatory neurological diseases (control group). The concentration of QUIN dramatically increased, whereas that of KYNA slightly decreased in the multiple sclerosis group, resulting in a significantly increased QUIN/KYNA ratio and significantly decreased PICA/QUIN ratio.
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Esclerosis Múltiple Recurrente-Remitente/diagnóstico , Espectrometría de Masas en Tándem/métodos , Triptófano/análisis , Adulto , Biomarcadores/análisis , Biomarcadores/metabolismo , Calibración , Estudios de Casos y Controles , Cromatografía Líquida de Alta Presión/instrumentación , Cromatografía Líquida de Alta Presión/métodos , Cromatografía Líquida de Alta Presión/normas , Femenino , Humanos , Ácido Quinurénico/análisis , Ácido Quinurénico/metabolismo , Persona de Mediana Edad , Esclerosis Múltiple Recurrente-Remitente/sangre , Esclerosis Múltiple Recurrente-Remitente/líquido cefalorraquídeo , Ácidos Picolínicos/análisis , Ácidos Picolínicos/metabolismo , Ácido Quinolínico/análisis , Ácido Quinolínico/metabolismo , Estándares de Referencia , Espectrometría de Masas en Tándem/instrumentación , Espectrometría de Masas en Tándem/normas , Triptófano/metabolismo , Adulto JovenRESUMEN
BACKGROUND: Behavioral associated disturbance involves excitotoxic quinolinate in alcohol withdrawal syndrome in man due to increase availability of tryptophan. In present study we investigated alcoholism related clinical features in relation to tryptophan and 5-HT levels in rat's model. METHODS: Locally bred male Wistar rats, weighing 200-250 g were housed separately into 6 animals/ group with 12 h light: dark cycle at room temp 22±3 °C. They were given diet ad libitum, for three days then alcohol 8% (v/v) was added into the liquid diet. Matched control rats of each group were given maltose-dextrin as a substitute of alcohol. Alcohol withdrawal syndrome was assessed after 7 hours by replacing the alcohol-containing liquid diet with tap water. RESULTS: Alcohol withdrawal group showed significant increase (p<0.001) in holo, apo, and total tryptophan 2, 3 dioxygenase enzyme activities, no significant change in brain tryptophan and 5HIAA however significant decrease (p<0.001) in brain 5HT was observed when compared with chow controls. Both alcohols administered and withdrawal groups showed significant rise in serum corticosterone by p<0.05 and p<0.001 respectively. Liver quinolinic acid concentrations were increased significantly (p<0.01) with robust increase in alcohol withdrawn rats. CONCLUSIONS: We conclude that the excitotoxin tryptophan metabolite quinolinic acid of peripheral origin plays significant role in the behavioral manifestation of the alcohol withdrawal syndrome. Tryptophan metabolites should be targeted to develop new strategies in the progress of pharmacological interventions related to alcoholism.
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Alcoholismo/metabolismo , Hígado/química , Ácido Quinolínico/análisis , Síndrome de Abstinencia a Sustancias/metabolismo , Animales , Hígado/metabolismo , Masculino , Ratas , Ratas Wistar , TriptófanoRESUMEN
Highly fluorescent nitrogen doped carbon quantum dots (NCQDs) were synthesized using microwave assisted green method. It was characterized by Transmission Electron Microscopy (TEM), FTIR, UV-Visible absorption and Photoluminiscence (PL) techniques. The NCQDs were immobilized with an enzyme named quinolinate phoshphoribosyl transferase (QPRTase). The NCQDs immobilized by QPRTase was used as a fluorescent bioprobe for the selective detection of endogenous neurotoxin quinolinic acid (QA) whose elevated level in serum is marker of many neurological disorders such as Alzheimer's, Huntington's and HIV associated dementia (HAD) as well as deficiency of vitamin B6. Steady state PL studies were carried out to measure the PL response of the fabricated fluorescent bioprobe as a function of QA concentrations in human serum samples. This probe was found applicable in linear range [3.22-51µM] with the limit of detection ~ 6.51µM. It has desirable sensitivity ~ (0.02340±0.0001) µM-1, excellent stability for ~ 7 weeks and good reproducibility. The similar response of this fluorescent bioprobe for QA detection in triple distilled water and human serum shows that it is unaffected by variation in media. Hence, this fluorescent bioprobe can be employed for QA detection in serum sample for the early detection of many diseases.
Asunto(s)
Técnicas Biosensibles/métodos , Carbono/química , Colorantes Fluorescentes/química , Neurotoxinas/sangre , Nitrógeno/química , Puntos Cuánticos/química , Ácido Quinolínico/sangre , Técnicas Biosensibles/instrumentación , Enzimas Inmovilizadas/química , Humanos , Límite de Detección , Neurotoxinas/análisis , Pentosiltransferasa/química , Ácido Quinolínico/análisis , Reproducibilidad de los ResultadosRESUMEN
Quinolinic acid (QA) is a metabolite of tryptophan degradation obtained through kynurenine pathway, produced naturally in the mammalian brain as well as in the human cerebrospinal fluid. The presence of QA ~10-40µM is a clear indicator of many neurological disorders as well as deficiency of vitamin B6 in human being. In the present work; rapid, sensitive and cost-effective bio-electrodes were prepared to detect the trace amount of endogenous neurotoxin (QA). Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) studies were carried out to measure the electrochemical response of the fabricated bio-electrodes as a function of QA concentrations. These devices were found to exhibit desirable sensitivity of ~7.86mAµM-1cm-2 in wide concentration range (6.5µM-65mM). The lower detection limit of this device is as low as 6.5µM and it has excellent storage stability of ~30 days. The capability of the proposed electrochemical bio-sensor was also checked to detect QA in the real samples (human serum). These results reveal that the use of this electrochemical bio-sensor may provide a potential platform for the detection of QA in the real samples for the prior detection of many diseases.
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Técnicas Biosensibles/instrumentación , Técnicas Electroquímicas/instrumentación , Ácido Quinolínico/sangre , Electrodos , Enzimas Inmovilizadas/química , Diseño de Equipo , Grafito/química , Humanos , Límite de Detección , Neurotoxinas/análisis , Neurotoxinas/sangre , Oxidación-Reducción , Pentosiltransferasa/química , Ácido Quinolínico/análisisRESUMEN
BACKGROUND: The aim of this exploratory study is to gain for the first time a more comprehensive picture of the impact of changes of quinolinic acid concentrations on depressive symptomatology during and after IFN-α therapy. METHODS: The quinolinic acid concentrations of 35 HCV patients are examined in a prospective survey over the entire period of IFN-α treatment as well as three months later at six different times (baseline, one, three, six and nine months after the beginning of IFN-α treatment, and after the end of treatment). RESULTS: During IFN-α treatment Hamilton Depression Rating Scale scores rise significantly. At the same time there is greater activity of indoleamine 2,3-dioxygenase, with a resulting increase in plasma kynurenine concentrations. Compared to baseline values quinolinic acid concentrations increase significantly during therapy, reflecting an increased neurotoxic challenge. In addition, patients with higher scores in the Hamilton Depression Rating Scale at six and nine months after starting therapy show significantly higher levels of quinolinic acid concentration. CONCLUSIONS: The increase of quinolinic acid during IFN-α therapy might contribute to depressive symptomatology through the neurotoxic challenge caused by quinolinic acid. Subsequently, our exploratory study results support the inflammatory hypothesis of depression. The awareness of relevant risk factors of IFN-α treatment-induced depression is essential to develop preventative treatment strategies.
Asunto(s)
Antivirales/efectos adversos , Depresión/inducido químicamente , Hepatitis C Crónica/tratamiento farmacológico , Interferón-alfa/efectos adversos , Ácido Quinolínico/análisis , Adulto , Antivirales/uso terapéutico , Depresión/metabolismo , Femenino , Hepatitis C Crónica/metabolismo , Hepatitis C Crónica/psicología , Humanos , Interferón-alfa/uso terapéutico , Quinurenina/sangre , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Escalas de Valoración PsiquiátricaRESUMEN
Postmortem and positron emission tomography studies have indicated the pathophysiological involvement of microglial cells in schizophrenia. We hypothesized that the microglial production of quinolinic acid (QUIN), an endogenous N-methyl-d-aspartate receptor (NMDAR) agonist, may be linked to the previously described glutamatergic deficits in the hippocampus of schizophrenia patients. We performed a semi-quantitative assessment of QUIN-immunoreactive microglial cells in schizophrenia patients and matched controls in the CA1, CA2/3, and dentate gyrus (DG) area of the posterior hippocampal formation. Complementary immunostaining of the commonly used microglial surface marker HLA-DR was performed in adjacent histological sections. Fewer QUIN-immunoreactive microglial cells were observed in the CA1 hippocampal subregion of schizophrenia patients compared to controls (left p=0.028, right p=0.018). No significant diagnosis-dependent changes were observed in the CA2/3 and DG regions. These results were controlled for potential confounds by age, duration of disease, autolysis time, psychotropic medication, and hippocampal volume. No diagnosis-related differences were observed for the overall density of microglial cells (HLA-DR expression). Our findings suggest that reduced microglial QUIN content in the hippocampal CA1 region is associated with schizophrenia. We hypothesize that this association may contribute to impaired glutamatergic neurotransmission in the hippocampus of schizophrenia patients.
Asunto(s)
Región CA1 Hipocampal/química , Agonistas de Aminoácidos Excitadores/análisis , Microglía/química , Ácido Quinolínico/análisis , Receptores de N-Metil-D-Aspartato/agonistas , Esquizofrenia/metabolismo , Adulto , Región CA1 Hipocampal/inmunología , Región CA1 Hipocampal/patología , Recuento de Células , Femenino , Ácido Glutámico/fisiología , Antígenos HLA-DR/análisis , Hipocampo/química , Hipocampo/inmunología , Hipocampo/patología , Humanos , Masculino , Microglía/inmunología , Persona de Mediana Edad , Neuroinmunomodulación/fisiología , Especificidad de Órganos , Esquizofrenia/inmunología , Esquizofrenia/patología , Transmisión SinápticaRESUMEN
BACKGROUND: Patients with allergic asthma have exacerbations which are frequently caused by rhinovirus infection. The antiviral tryptophan-catabolising enzyme indoleamine 2,3-dioxygenase (IDO) is induced by interferon-γ and suppressed by Th2 mediators interleukin (IL)-4 and IL-13. We hypothesised that local IDO activity after viral airway infection is lower in patients with allergic asthma than in healthy controls. OBJECTIVE: To determine whether IDO activity differs between patients with allergic asthma and healthy individuals before and after rhinovirus infection. METHODS: Healthy individuals and patients with allergic asthma were experimentally infected with low-dose (10 TCID50) rhinovirus 16. Blood, bronchoalveolar lavage fluid and exhaled breath condensate (for mass spectrometry by UPLC-MS/MS) were obtained before and after rhinovirus challenge. RESULTS: IDO activity was not induced by rhinovirus infection in either group, despite increases in cold scores. However, baseline pulmonary IDO activity was lower in patients with allergic asthma than in healthy individuals. In contrast, systemic tryptophan and its catabolites were markedly higher in patients with allergic asthma. Moreover, systemic quinolinic acid and tryptophan were associated with eosinophil cationic protein (r=0.43 and r=0.78, respectively) and eosinophils (r=0.38 and r=0.58, respectively) in bronchoalveolar lavage fluid and peak asthma symptom scores after rhinovirus challenge (r=0.53 and r=0.64, respectively). CONCLUSIONS: Rhinovirus infection by itself induces no IDO activity, but the reduced pulmonary IDO activity in patients with allergic asthma at baseline may underlie a reduced control of viral infections. Notably, the enhanced systemic catabolism of tryptophan in patients with allergic asthma was strongly related to the outcome of rhinovirus challenge in asthma and may serve as a prognostic factor.
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Asma/complicaciones , Asma/enzimología , Indolamina-Pirrol 2,3,-Dioxigenasa/metabolismo , Infecciones por Picornaviridae/complicaciones , Rhinovirus , Triptófano/sangre , Adulto , Asma/fisiopatología , Biomarcadores/análisis , Biomarcadores/sangre , Pruebas Respiratorias , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Estudios de Casos y Controles , Citocinas/análisis , Progresión de la Enfermedad , Proteína Catiónica del Eosinófilo/análisis , Eosinófilos , Femenino , Humanos , Quinurenina/análisis , Quinurenina/sangre , Masculino , Óxido Nítrico/análisis , Peroxidasa/análisis , Infecciones por Picornaviridae/virología , Estudios Prospectivos , Ácido Quinolínico/análisis , Ácido Quinolínico/sangre , Triptófano/análisis , Adulto Joven , ortoaminobenzoatos/análisis , ortoaminobenzoatos/sangreRESUMEN
This article describes the development of a reliable CZE-ESI-MS method to simultaneously separate and quantitate three specific metabolites (3-hydroxyanthranilic acid (3-HAA), quinolinic acid (QA), and picolinic acid (PA)) of the kynurenine pathway (KP) of tryptophan catabolism. Using a covalently bonded sulfonated capillary, the parameters such as pH, type of background electrolyte, type of organic solvent, nebulizer pressure as well as both negative and positive ESI-MS modes were optimized to achieve the best Rs and S/N of three KP metabolites. The developed CZE-ESI-MS assay provided high resolution of PA/QA, high specificity, a total analysis time of 10 min with satisfactory intraday and interday repeatability of migration time and peak areas. Under optimized CZE-ESI-MS conditions, the calibration curves over a concentration range of 19-300 µM for 3-HAA and QA, and 75-300 µM for PA were simultaneously generated. The method was successfully applied for the first time to profile the concentrations of initial substrate, 3-HAA, and its eventual products, PA and QA, formed in the complex multienzyme system. As the ratio of two enzymes, 3-hydroxyanthranilate 3,4-dioxygenase (HAO) and α-amino-ß-carboxymuconate-ε-semialdehyde decarboxylase (ACMSD) decreases, the concentration of QA approaches essentially zero indicating that all ACMS formed by the action of HAO is consumed by ACMSD rather than its spontaneous decay to QA.
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Ácido 3-Hidroxiantranílico/análisis , Electroforesis Capilar/instrumentación , Electroforesis Capilar/métodos , Complejos Multienzimáticos/química , Ácidos Picolínicos/análisis , Ácido Quinolínico/análisis , Espectrometría de Masa por Ionización de Electrospray/métodos , 3-Hidroxiantranilato 3,4-Dioxigenasa/metabolismo , Ácido 3-Hidroxiantranílico/metabolismo , Tampones (Química) , Carboxiliasas/metabolismo , Concentración de Iones de Hidrógeno , Complejos Multienzimáticos/metabolismo , Ácidos Picolínicos/metabolismo , Presión , Ácido Quinolínico/metabolismo , Reproducibilidad de los ResultadosAsunto(s)
Biomarcadores/análisis , Investigación Biomédica/métodos , Animales , Química Encefálica , Humanos , Oxitocina/análisis , Oxitocina/orina , Ácido Quinolínico/análisis , Serotonina/análisis , Serotonina/sangre , Serotonina/líquido cefalorraquídeo , Triptaminas/análisis , Vasopresinas/análisis , Vasopresinas/orinaRESUMEN
The kynurenine pathway has been implicated as a major component of the neuroinflammatory response to brain injury and neurodegeneration. We found that the neurotoxic kynurenine pathway intermediate quinolinic acid (QUIN) is rapidly expressed, within 24 h, by reactive microglia following traumatic injury to the rodent neocortex. Furthermore, administration of the astrocytic protein metallothionein attenuated this neuroinflammatory response by reducing microglial activation (by approximately 30%) and QUIN expression. The suppressive effect of MT was confirmed upon cultured cortical microglia, with 1 mug/ml MT almost completely blocking interferon-gamma induced activation of microglia and QUIN expression. These results demonstrate the neuroimmunomodulatory properties of MT, which may have therapeutic applications for the treatment of traumatic brain injury.
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Lesiones Encefálicas/patología , Regulación de la Expresión Génica/efectos de los fármacos , Metalotioneína/farmacología , Microglía/efectos de los fármacos , Ácido Quinolínico/metabolismo , Análisis de Varianza , Animales , Lesiones Encefálicas/tratamiento farmacológico , Lesiones Encefálicas/metabolismo , Recuento de Células/métodos , Células Cultivadas , Corteza Cerebral/citología , Medios de Cultivo Condicionados/farmacología , Relación Dosis-Respuesta a Droga , Ferritinas/metabolismo , Cromatografía de Gases y Espectrometría de Masas , Proteína Ácida Fibrilar de la Glía/metabolismo , Interferón gamma/farmacología , Microglía/química , Neocórtex/metabolismo , Neocórtex/patología , Neuronas/patología , Ácido Quinolínico/análisis , Ratas , Ratas WistarRESUMEN
The brain levels of the endogenous excitotoxin quinolinic acid (QUIN) and its bioprecursor, the free radical generator 3-hydroxykynurenine (3-HK), are elevated in early stage Huntington disease (HD). We now examined the status of these metabolites in three mouse models of HD. In R6/2 mice, 3-HK levels were significantly and selectively elevated in the striatum, cortex and cerebellum starting at 4 weeks of age. In contrast, both 3-HK and QUIN levels were increased in the striatum and cortex of the full-length HD models, beginning at 8 months (YAC128) and 15 months (Hdh(Q92) and Hdh(Q111)), respectively. No changes were seen in 13-month-old shortstop mice, which show no signs of motor or cognitive dysfunction or selective neuropathology. These results demonstrate both important parallels and intriguing differences in the progressive neurochemical changes in these HD mouse models and support the hypothesis that QUIN may play a role in the striatal and cortical neurodegeneration of HD.
Asunto(s)
Enfermedad de Huntington/metabolismo , Quinurenina/análogos & derivados , Quinurenina/metabolismo , Ácido Quinolínico/análisis , Factores de Edad , Animales , Química Encefálica , Cromatografía de Gases , Cromatografía Líquida de Alta Presión , Modelos Animales de Enfermedad , Humanos , Ácido Quinurénico/metabolismo , Quinurenina/análisis , Ratones , Ratones Transgénicos , Ácido Quinolínico/metabolismoRESUMEN
OBJECTIVE: We evaluated the utility of neopterin and quinolinic acid (QUIN) as surrogate measures of disease activity in juvenile idiopathic inflammatory myopathies (IIMs). METHODS: Plasma and first morning void urine samples were measured for neopterin and QUIN using commercial ELISA, HPLC, or gas chromatography-mass spectrometry in 45 juvenile IIM patients and 79 healthy controls. Myositis disease activity assessments were obtained. RESULTS: Plasma and urine neopterin and QUIN concentrations were increased in juvenile IIM patients compared with healthy controls (P <0.017). Urine neopterin and QUIN highly correlated with each other (r(s) = 0.73; P <0.0001). Urine neopterin and QUIN correlated moderately with myositis disease activity assessments, including physician and parent global activity assessments, muscle strength testing, functional assessments (Childhood Myositis Assessment Scale, Childhood Health Assessment Questionnaire), skin global activity, and edema on magnetic resonance imaging (r(s) = 0.42-0.62; P <0.05), but generally not with muscle-associated enzymes in serum. Urine neopterin or QUIN, in combination with either serum lactate dehydrogenase (LD) or aspartate aminotransferase (AST), significantly predicted global disease activity (R(2) =0.40-0.56; P <0.002), and both were more sensitive to change than these serum enzymes (standardized response means, -0.41 to -0.48). CONCLUSIONS: Urinary neopterin and QUIN are candidate measures of disease activity in juvenile IIM patients and add significantly to the prediction of global disease activity in combination with serum LD or AST values. Measurement of these markers in first morning void urine specimens appears to be as good as, or possibly better than, measurements of their concentrations in plasma.
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Miositis/diagnóstico , Neopterin/análisis , Ácido Quinolínico/análisis , Adolescente , Biomarcadores/sangre , Biomarcadores/orina , Niño , Preescolar , Femenino , Humanos , Masculino , Neopterin/sangre , Neopterin/orina , Ácido Quinolínico/sangre , Ácido Quinolínico/orina , Factores de TiempoRESUMEN
Quinolinic, picolinic, and nicotinic acids and nicotinamide are end products of the kynurenine pathway from l-tryptophan and are intermediates in the biosynthesis of nicotinamide adenine dinucleotide. These compounds are involved in complex interrelationships with inflammatory and apoptotic responses associated with neuronal cell damage and death in the central nervous system. To facilitate the study of these compounds, we have utilized gas chromatography-mass spectrometry in electron capture negative ionization mode for their concurrent trace quantification in a single sample. Deuterium-labeled quinolinic, picolinic, and nicotinic acids were used as internal standards and the compounds were converted to their hexafluoroisopropyl esters prior to chromatography. Nicotinamide was readily quantified after conversion to nicotinic acid using gas-phase hydrolysis-a process which did not affect the deuterated internal standards. The on-column limit of quantification was less than 1 fmol for each of the analytes and calibration curves were linear. A packed column liner was used in the gas chromatograph inlet to effectively eliminate sample interference effects in the analysis of trace (femtomolar) levels of quinolinic acid. The method enables rapid and specific concurrent quantification of quinolinic, picolinic, and nicotinic acids in tissue extracts and physiological and culture media.
Asunto(s)
Cromatografía de Gases y Espectrometría de Masas/métodos , Niacina/análisis , Niacinamida/análisis , Ácidos Picolínicos/análisis , Ácido Quinolínico/análisis , Animales , Femenino , Feto/química , Humanos , Quinurenina/metabolismo , Ovinos/sangreRESUMEN
OBJECTIVE: The effect of maternal tryptophan loading on fetal plasma and brain, kynurenic acid, and quinolinic acid concentrations was compared in late gestation fetal sheep with either chronically embolized or nonembolized placentas. STUDY DESIGN: The placentas of 4 ewes were embolized by daily injection of mucopolysaccharide microspheres into the umbilical artery from 120 days gestation in amounts sufficient to reduce the fetal arterial PO2 to < or = 12 mm Hg. Four fetuses with nonembolized placentas were the control group. At 135 to 138 days gestation, the ewe received an infusion of tryptophan (100 mg/kg, intravenously) or an equivalent volume of saline solution (100 mL) over 2 hours. Maternal and fetal arterial blood samples were obtained between 2 and 48 hours from the start of the infusion for the measurement of plasma tryptophan and kynurenine metabolites. Brains were then obtained from embolized and nonembolized fetuses 24 hours after a further maternal tryptophan loading experiment and from nonembolized non-tryptophan-treated fetuses for analysis of regional kynurenic acid and quinolinic acid content. RESULTS: Maternal tryptophan infusion resulted in a significant increase of kynurenine in fetal plasma, but this increase was significantly smaller in fetuses with an embolized placenta compared with a nonembolized placenta. Both kynurenic acid and quinolinic acid levels increased significantly in fetal plasma, with no differences between the groups. Kynurenic acid and quinolinic acid levels were increased in all regions of the fetal brain after maternal tryptophan loading, but these increases were greater in the fetuses with an embolized placenta, compared with a nonembolized placenta. CONCLUSION: Fetal tryptophan and kynurenine metabolism is significantly altered when placental function is chronically compromised in late gestation. The decreased production of kynurenine from tryptophan may result from the compromise of hepatic function in the fetus, whereas the increased production of kynurenic acid and quinolinic acid in the brain is likely to reflect alterations of metabolism of tryptophan and kynurenine to these neuroactive products by glial cells in the fetal brain.
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Ácido Quinurénico/análisis , Insuficiencia Placentaria/metabolismo , Ácido Quinolínico/análisis , Triptófano/farmacología , Animales , Modelos Animales de Enfermedad , Embolia , Femenino , Sangre Fetal/química , Enfermedades Fetales/diagnóstico , Enfermedades Fetales/etiología , Retardo del Crecimiento Fetal/etiología , Retardo del Crecimiento Fetal/fisiopatología , Infusiones Intravenosas , Placenta/irrigación sanguínea , Insuficiencia Placentaria/etiología , Embarazo , Preñez , Probabilidad , Valores de Referencia , Sensibilidad y Especificidad , OvinosRESUMEN
Activated macrophages produce quinolinic acid (QUIN), a neurotoxin, in several inflammatory brain diseases including AIDS dementia complex. We hypothesized that IL1-beta, IL6, transforming growth factor (TGF-beta2 and platelet activating factor could increase macrophage QUIN production. And that the HIV-1 proteins Nef, Tat and gp41 may also increase synthesis of QUIN by macrophages. At 72 h there were significant increases in QUIN production in the cells stimulated with PAF (914 +/- 50 nM) and Nef (2781 +/- 162 nM), with somewhat less production by Tat stimulation (645 +/- 240 nM). The increases in QUIN production approximated in vitro concentrations of QUIN shown to be neurotoxic and correlated closely with indoleamine 2,3-dioxygenase induction. IL1-beta, IL6, TGF-beta2 and gp41 stimulation produced no significant increase in QUIN production. These results suggest that some of the neurotoxicity of PAF, nef and tat may be mediated by QUIN.
Asunto(s)
Productos del Gen nef/farmacología , Productos del Gen tat/farmacología , Macrófagos/metabolismo , Factor de Activación Plaquetaria/farmacología , Ácido Quinolínico/metabolismo , Complejo SIDA Demencia/etiología , Complejo SIDA Demencia/metabolismo , Inducción Enzimática/efectos de los fármacos , Proteína gp41 de Envoltorio del VIH/farmacología , Humanos , Interferón gamma/farmacología , Interleucina-1/farmacología , Interleucina-6/farmacología , Macrófagos/química , Macrófagos/efectos de los fármacos , Ácido Quinolínico/análisis , Factor de Crecimiento Transformador beta/farmacología , Factor de Crecimiento Transformador beta2 , Triptófano Oxigenasa/metabolismo , Productos del Gen nef del Virus de la Inmunodeficiencia Humana , Productos del Gen tat del Virus de la Inmunodeficiencia HumanaRESUMEN
OBJECTIVE: This study was undertaken to examine the effects of chronic embolization of the umbilical circulation during late gestation on regional concentrations of quinolinic acid and kynurenic acid (neuroactive products of tryptophan catabolism) and of the astrocyte-associated glial fibrillary acidic protein in the fetal brain. STUDY DESIGN: Pregnant ewes bearing fetuses with long-term catheter placement were treated daily with injections of either saline solution (n = 4; control group) or mucopolysaccharide microspheres (n = 5; embolized group) into the umbilical circulation through a femoral artery catheter between 120 and 140 days' gestation. The fetuses in the embolized group received sufficient microspheres each day to reduce and maintain the femoral arterial PO2 at < or =12 mm Hg. Autopsies were performed at 140 days' gestation to obtain the fetal brain for chemical analysis. RESULTS: Umbilical embolization resulted in nonacidemic hypoxia and hypoglycemia at 140 days' gestation. Quinolinic acid concentrations in the embolized group were significantly increased in the medulla, pons, midbrain, hypothalamus, and hippocampus, whereas kynurenic acid concentrations in the embolized group were reduced in the hippocampus and hypothalamus. There were significant reductions in glial fibrillary acidic protein contents in the occipitoparietal cortex, hippocampus, and pons in the embolized group. CONCLUSION: Placental compromise during late pregnancy had effects on kynurenine metabolism and astrocyte function in some regions of the fetal sheep brain. We suggest that these changes increase the vulnerability of the brain to asphyxial injury during late gestation and the perinatal period.
Asunto(s)
Química Encefálica , Encéfalo/embriología , Proteína Ácida Fibrilar de la Glía/análisis , Ácido Quinurénico/análisis , Insuficiencia Placentaria/metabolismo , Ácido Quinolínico/análisis , Animales , Embolia , Femenino , Arteria Femoral , Edad Gestacional , Microesferas , Placenta/irrigación sanguínea , Insuficiencia Placentaria/etiología , Embarazo , OvinosRESUMEN
Interferon-beta(1b) (IFN-beta(1b)) has limited efficacy in the treatment of relapsing-remitting multiple sclerosis (RRMS). The kynurenine pathway (KP) is chiefly activated by IFN-gamma and IFN-alpha, leading to the production of a variety of neurotoxins. We sought to determine whether IFN-beta(1b) induces the KP in human monocyte-derived macrophages, as one explanation for its limited efficacy. Serial dilutions of IFN-beta(1b) (at concentrations comparable to those found in the sera of IFN-beta(1b)-treated patients) were added to human macrophage cultures. Supernatants were collected at various time points and assayed for the KP end product, quinolinic acid (QUIN). The effect of IFN-beta(1b) on the KP enzymes indoleamine 2,3-dioxygenase (IDO), 3-hydroxyanthranilate dioxygenase (3HAO), and quinolinate phosphoribosyltransferase (QPRTase) mRNA expression was assessed by semiquantitative RT-PCR. IFN-beta(1b) (> or =10 IU/ml) led to increased mRNA expression of both IDO and QUIN production (7901 +/- 715 nM) after 72 h at 50 IU/ml IFN-beta(1b) (p < 0.0001). This study demonstrates that IFN-beta(1b), in pharmacologically relevant concentrations, induces KP metabolism in human macrophages and may be a limiting factor in its efficacy in the treatment of MS. Inhibitors of the KP may be able to augment the efficacy of IFN-beta in MS.
Asunto(s)
Interferón beta/farmacología , Quinurenina/metabolismo , Macrófagos/metabolismo , Células Cultivadas , Relación Dosis-Respuesta a Droga , Humanos , Interferón beta-1a , Interferon beta-1b , Interferón beta/biosíntesis , Interferón beta/uso terapéutico , Interferón gamma/biosíntesis , Interferón gamma/genética , Interferón gamma/farmacología , Cinética , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Modelos Químicos , Esclerosis Múltiple/tratamiento farmacológico , Ácido Quinolínico/análisis , ARN Mensajero/biosíntesisRESUMEN
Quinolinic acid (2,3-pyridine dicarboxylic acid), a biological intermediate in nicotinamide adenine dinucleotide (NAD) biosynthesis in microbes and mammals and a brain excitotoxin, is not fluorescent nor electrochemically active and its detection sensitivity by UV absorption is comparatively low. Quinolinic acid was successfully derivatized in water-based samples by monodansylcadaverine, a fluorescence tag, and analysed by high-performance liquid chromatography (HPLC). No extraction procedure was needed and quinolinic acid was activated by water-soluble carbodiimide and derivatized under mild conditions. As little as 3 pmol (500 pg) of quinolinic acid in 5 microliter of artificial cerebrospinal fluid sample volume could be derivatized and detected at a signal to noise ratio of 3:1. Thus, detection on a mass basis by HPLC after fluorescence derivatization is about 300 times as sensitive as direct determination of quinolinic acid by UV absorbance (500 pg vs 150 ng). A variety of activators, fluorescent tags and reaction solvents and conditions were tested but found to be less effective.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Colorantes Fluorescentes/química , Ácido Quinolínico/análisis , Cadaverina/análogos & derivados , Cadaverina/química , NAD/biosíntesis , Ácido Quinolínico/líquido cefalorraquídeo , Sensibilidad y Especificidad , AguaRESUMEN
Experimental tumors of the central nervous system were investigated with antibodies to quinolinate to assess the cellular distribution of this endogenous neurotoxin. In advanced F98 and RG-2 glioblastomas and E367 neuroblastomas in the striatum of rats, variable numbers of quinolinate immunoreactive cells were observed in and around the tumors, with the majority being present within tumors, rather than brain parenchyma. The stained cells were morphologically variable, including round, complex, rod-shaped, and sparsely dendritic cells. Neuroblastoma and glioma cells were unstained, as were neurons, astrocytes, oligodendrocytes, ependymal cells, endothelial cells, and cells of the choroid plexus and leptomeninges. Glial fibrillary acidic protein immunoreactivity was strongly elevated in astrocytes surrounding the tumors. Dual labeling immunohistochemistry with antibodies to quinolinate and glial fibrillary acidic protein demonstrated that astrocytes and the cells containing quinolinate immunoreactivity were morphologically disparate and preferentially distributed external and internal to the tumors, respectively, and no dual labeled cells were observed. Lectin histochemistry with Griffonia simplicifolia B4 isolectin and Lycopersicon esculentum lectin demonstrated numerous phagocytic macrophages and reactive microglia in and around the tumors whose distribution was similar to that of quinolinate immunoreactive cells, albeit much more numerous. Dual labeling studies with antibodies to quinolinate and the lectins demonstrated partial codistribution of these markers, with most double-labeled cells having the morphology of phagocytes. The present findings suggest the possibility that quinolinate may serve a functional role in a select population of inflammatory cell infiltrates during the immune response to brain neoplasms.