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1.
Nat Commun ; 11(1): 3169, 2020 06 23.
Artigo em Inglês | MEDLINE | ID: mdl-32576825

RESUMO

Understanding tumor metabolism holds the promise of new insights into cancer biology, diagnosis and treatment. To assess human cancer metabolism, here we report a method to collect intra-operative samples of blood from an artery directly upstream and a vein directly downstream of a brain tumor, as well as samples from dorsal pedal veins of the same patients. After performing targeted metabolomic analysis, we characterize the metabolites consumed and produced by gliomas in vivo by comparing the arterial supply and venous drainage. N-acetylornithine, D-glucose, putrescine, and L-acetylcarnitine are consumed in relatively large amounts by gliomas. Conversely, L-glutamine, agmatine, and uridine 5-monophosphate are produced in relatively large amounts by gliomas. Further we verify that D-2-hydroxyglutarate (D-2HG) is high in venous plasma from patients with isocitrate dehydrogenases1 (IDH1) mutations. Through these paired comparisons, we can exclude the interpatient variation that is present in plasma samples usually taken from the cubital vein.


Assuntos
Biomarcadores Tumorais/sangue , Vasos Sanguíneos/metabolismo , Neoplasias Encefálicas/sangue , Neoplasias Encefálicas/metabolismo , Glioma/sangue , Glioma/metabolismo , Metabolômica , Acetilcarnitina/sangue , Adulto , Idoso , Agmatina/sangue , Sangue , Análise Química do Sangue , Glicemia , Neoplasias Encefálicas/diagnóstico por imagem , Neoplasias Encefálicas/genética , Feminino , Glioma/diagnóstico por imagem , Glioma/genética , Glucose , Glutamina/sangue , Glutaratos/sangue , Humanos , Isocitrato Desidrogenase/sangue , Isocitrato Desidrogenase/genética , Isocitrato Desidrogenase/metabolismo , Masculino , Pessoa de Meia-Idade , Ornitina/análogos & derivados , Ornitina/sangue , Putrescina/sangue , Uridina Monofosfato/sangue , Adulto Jovem
2.
EBioMedicine ; 49: 318-330, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31676389

RESUMO

BACKGROUND: Type 2 diabetes patients and individuals at risk of developing diabetes are characterized by metabolic inflexibility and disturbed glucose homeostasis. Low carnitine availability may contribute to metabolic inflexibility and impaired glucose tolerance. Here, we investigated whether carnitine supplementation improves metabolic flexibility and insulin sensitivity in impaired glucose tolerant (IGT) volunteers. METHODS: Eleven IGT- volunteers followed a 36-day placebo- and L-carnitine treatment (2 g/day) in a randomised, placebo-controlled, double blind crossover design. A hyperinsulinemic-euglycemic clamp (40 mU/m2/min), combined with indirect calorimetry (ventilated hood) was performed to determine insulin sensitivity and metabolic flexibility. Furthermore, metabolic flexibility was assessed in response to a high-energy meal. Skeletal muscle acetylcarnitine concentrations were measured in vivo using long echo time proton magnetic resonance spectroscopy (1H-MRS, TE=500 ms) in the resting state (7:00AM and 5:00PM) and after a 30-min cycling exercise. Twelve normal glucose tolerant (NGT) volunteers were included without any intervention as control group. RESULTS: Metabolic flexibility of IGT-subjects completely restored towards NGT control values upon carnitine supplementation, measured during a hyperinsulinemic-euglycemic clamp and meal test. In muscle, carnitine supplementation enhanced the increase in resting acetylcarnitine concentrations over the day (delta 7:00 AM versus 5:00 PM) in IGT-subjects. Furthermore, carnitine supplementation increased post-exercise acetylcarnitine concentrations and reduced long-chain acylcarnitine species in IGT-subjects, suggesting the stimulation of a more complete fat oxidation in muscle. Whole-body insulin sensitivity was not affected. CONCLUSION: Carnitine supplementation improves acetylcarnitine formation and rescues metabolic flexibility in IGT-subjects. Future research should investigate the potential of carnitine in prevention/treatment of type 2 diabetes.


Assuntos
Acetilcarnitina/metabolismo , Carnitina/farmacologia , Suplementos Nutricionais , Voluntários Saudáveis , Músculo Esquelético/metabolismo , Acetilcarnitina/sangue , Composição Corporal/efeitos dos fármacos , Carnitina/sangue , Feminino , Teste de Tolerância a Glucose , Glicogênio/metabolismo , Humanos , Hiperinsulinismo/sangue , Resistência à Insulina , Cinética , Masculino , Metaboloma , Pessoa de Meia-Idade , Consumo de Oxigênio/efeitos dos fármacos
3.
J Mass Spectrom ; 54(11): 878-884, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31652368

RESUMO

Overcoming the detrimental effects of sweet spots during crystallization is an important step to improve the quantitative abilities of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. In this study, we introduce MALDI targets, which exhibit a channel design to reduce sweet spot phenomena and improve reproducibility. The size of the channels was 3.0 mm in length, 0.35 mm in depth, and 0.40 mm in width, adjusted to the width of the implemented laser beam. For sample deposition, the matrix/sample mixture was homogenously deposited into the channels using capillary action. To demonstrate the proof-of-principle, the novel plates were used for the quantification of acetyl-L-carnitine in human blood plasma using a combined standard addition and isotope dilution method. The results showed that the reproducibility of acetyl-L-carnitine detection was highly improved over a conventional MALDI-MS assay, with RSD values of less than 5.9% in comparison with 15.6% using the regular MALDI method. The limits of quantification using the new plates were lowered approximately two-fold in comparison with a standard rastering approach on a smooth stainless-steel plate. Matrix effects were also assessed and shown to be negligible. The new assay was subsequently applied to the quantification of acetyl-L-carnitine in human plasma samples.


Assuntos
Acetilcarnitina/sangue , Cristalização , Humanos , Limite de Detecção , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Aço Inoxidável/química , Propriedades de Superfície
4.
Nutrients ; 11(4)2019 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-31010169

RESUMO

The prospero homeobox 1 (PROX1) gene may show pleiotropic effects on metabolism. We evaluated postprandial metabolic alterations dependently on the rs340874 genotypes, and 28 non-diabetic men were divided into two groups: high-risk (HR)-genotype (CC-genotype carriers, n = 12, 35.3 ± 9.5 years old) and low-risk (LR)-genotype (allele T carriers, n = 16, 36.3 ± 7.0 years old). Subjects participated in two meal-challenge-tests with high-carbohydrate (HC, carbohydrates 89%) and normo-carbohydrate (NC, carbohydrates 45%) meal intake. Fasting and 30, 60, 120, and 180 min after meal intake plasma samples were fingerprinted by liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). In HR-genotype men, the area under the curve (AUC) of acetylcarnitine levels was higher after the HC-meal [+92%, variable importance in the projection (VIP) = 2.88] and the NC-meal (+55%, VIP = 2.00) intake. After the NC-meal, the HR-risk genotype carriers presented lower AUCs of oxidized fatty acids (-81-66%, VIP = 1.43-3.16) and higher linoleic acid (+80%, VIP = 2.29), while after the HC-meal, they presented lower AUCs of ornithine (-45%, VIP = 1.83), sphingosine (-48%, VIP = 2.78), linoleamide (-45%, VIP = 1.51), and several lysophospholipids (-40-56%, VIP = 1.72-2.16). Moreover, lower AUC (-59%, VIP = 2.43) of taurocholate after the HC-meal and higher (+70%, VIP = 1.42) glycodeoxycholate levels after the NC-meal were observed. Our results revealed differences in postprandial metabolites from inflammatory and oxidative stress pathways, bile acids signaling, and lipid metabolism in PROX1 HR-genotype men. Further investigations of diet-genes interactions by which PROX1 may promote T2DM development are needed.


Assuntos
Ácidos e Sais Biliares/sangue , Diabetes Mellitus Tipo 2/genética , Carboidratos da Dieta/metabolismo , Proteínas de Homeodomínio/genética , Inflamação/sangue , Metabolismo dos Lipídeos/genética , Polimorfismo de Nucleotídeo Único , Proteínas Supressoras de Tumor/genética , Acetilcarnitina/sangue , Adulto , Alelos , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/etiologia , Dieta , Interação Gene-Ambiente , Genótipo , Humanos , Ácidos Linoleicos/sangue , Masculino , Refeições , Metaboloma , Ornitina/sangue , Estresse Oxidativo , Período Pós-Prandial , Transdução de Sinais , Esfingosina/sangue
5.
J Eur Acad Dermatol Venereol ; 33(7): 1378-1385, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30803039

RESUMO

BACKGROUND: Global metabolomics analysis can provide substantial information on energy metabolism, physiology, possible diagnostic biomarkers and intervention strategies for pathogens. OBJECTIVE: To gain a better understanding of the mechanisms of syphilis and analysis of serum metabolite profiles in syphilis patients. METHODS: We conducted an untargeted metabolomics analysis of serum from 20 syphilis patients and 20 healthy controls. RESULTS: A total of 2890 molecular features were extracted from each sample, and the peak intensity of each feature was obtained. Distinct differential metabolites were identified by principal component analysis, partial least squares-discriminant analysis and hierarchical clustering analysis. Furthermore, five metabolites were identified as significantly different by Student's t-test, including trimethylamine N-oxide, l-arginine, lysoPC(18:0), betaine and acetylcarnitine. KEGG analysis showed that these differential metabolites were in various pathways, including Chagas disease, fatty acid biosynthesis, primary bile acid biosynthesis, Salmonella infection, ABC transporters, glycerophospholipid metabolism and choline metabolism. Among them, trimethylamine N-oxide was 3.922 times in patients with syphilis than healthy controls. CONCLUSION: Trimethylamine N-oxide may be used as an indicator to distinguish between syphilis patients and healthy controls. The changes in these metabolites suggest that Treponema pallidum affects the normal metabolic activity of host cells, providing some clues for elucidating the pathogenesis of T. pallidum.


Assuntos
Acetilcarnitina/sangue , Arginina/sangue , Betaína/sangue , Metilaminas/sangue , Sífilis/sangue , Biomarcadores/sangue , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Redes e Vias Metabólicas , Metabolômica , Pessoa de Meia-Idade , Análise de Componente Principal , Sífilis/microbiologia
6.
Crit Care Med ; 47(2): 210-218, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30379669

RESUMO

OBJECTIVES: Recent metabolomic studies of sepsis showed that increased circulatory acylcarnitines were associated with worse survival. However, it is unknown whether plasma carnitine and acylcarnitines can reflect the severity of sepsis, and the role of specific acylcarnitines in prognostic assessment need further confirmation. This study aimed to clarify these questions. DESIGN: Prospective multicenter cohort studies with derivation and validation cohort design. SETTING: ICUs at two medical centers and three regional hospitals in Taiwan. PATIENTS: Patients with sepsis and acute organ dysfunction were enrolled. Recruitment of the derivation (n = 90) and validation cohorts (n = 120) occurred from October 2010 through March 2012 and January 2013 through November 2014, respectively. INTERVENTIONS: Plasma samples were collected immediately after admission, and the levels of carnitine and acylcarnitines were measured by ultra-high performance liquid chromatography-mass spectrometry. MEASUREMENTS AND MAIN RESULTS: In the derivation cohort, increased plasma levels of short- and medium-chain acylcarnitines were significantly associated with hepatobiliary dysfunction, renal dysfunction, thrombocytopenia, and hyperlactatemia. However, acetylcarnitine is the only acylcarnitine significantly correlating with various plasma cytokine concentrations and also associated with blood culture positivity and 28-day mortality risk. The association between plasma acetylcarnitine and multiple organ dysfunction severity, blood culture positivity, and 28-day mortality, was confirmed in the validation cohort. Patients with high plasma acetylcarnitine (≥ 6,000 ng/mL) had significantly increased 28-day mortality compared with those with plasma acetylcarnitine less than 6,000 ng/mL (52.6% vs 13.9%; hazard ratio, 5.293; 95% CI, 2.340-11.975; p < 0.001 by Cox proportional hazard model). CONCLUSIONS: We confirm that plasma acetylcarnitine can reflect the severity of organ dysfunction, inflammation, and infection in sepsis and can serve as a prognostic biomarker for mortality prediction.


Assuntos
Acetilcarnitina/sangue , Insuficiência de Múltiplos Órgãos/sangue , Sepse/sangue , Idoso , Biomarcadores/sangue , Carnitina/sangue , Feminino , Humanos , Masculino , Insuficiência de Múltiplos Órgãos/etiologia , Insuficiência de Múltiplos Órgãos/mortalidade , Modelos de Riscos Proporcionais , Estudos Prospectivos , Sepse/complicações , Sepse/mortalidade , Taiwan/epidemiologia
7.
J Pharm Biomed Anal ; 165: 101-111, 2019 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-30522064

RESUMO

Although major adverse effects associated with nonsteroidal anti-inflammatory drugs (NSAIDs) are gastric injury, assessment of NSAIDs-induced gastrointestinal adverse effects is mostly dependent on endoscopy due to the lack of plasma biomarkers. Several amino acids associated with collagenase activity and gastric mucosal mass have been suggested as plasma biomarker candidates for gastric injury. Therefore, this study aimed to develop a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the plasma biomarker candidates, i.e., acetylcarnitine, proline, hydroxyproline, citrulline, and arginine and evaluate their potential as a biomarker for NSAIDs-induced gastric injury. The method utilized simple protein precipitation with methanol and D4-citrulline as an internal standard (IS). The assay resulted in the lower limit of quantification (LLOQ) of 0.1 µg/mL for acetylcarnitine and 1 µg/mL for proline, hydroxyproline, citrulline, and arginine in the surrogate blank plasma. The intra- and inter-day accuracy ranged 82.5-111.2% for acetylcarnitine, 95.4-103.3% for proline, 98.9-106.4% for hydroxyproline, 99.5-103.5% for citrulline, and 87.4-105.3% for arginine. The precision was within 6.17%, 3.63%, 6.20%, 6.31%, and 6.17% for acetylcarnitine, proline, hydroxyproline, citrulline, and arginine, respectively. The developed assay was successfully applied to monitor the changes of the plasma levels of the five amino acids in rats and Beagle dogs following repeated oral administrations of aceclofenac. In rats, plasma concentrations of proline, hydroxyproline, and citrulline were significantly reduced after 4 days of aceclofenac administration compared to the control group. In dogs, plasma concentrations of proline and citrulline were significantly decreased after 7 days of aceclofenac administration compared to those obtained after the first aceclofenac administration. These data indicate that plasma levels of proline, hydroxyproline, and citrulline may be used as quantitative biomarkers of NSAIDs-induced gastric damage. The present assay could also be utilized to monitor the changes of these amino acids as potential indicators for various physiological and pathophysiological conditions.


Assuntos
Anti-Inflamatórios não Esteroides/toxicidade , Cromatografia Líquida/métodos , Diclofenaco/análogos & derivados , Espectrometria de Massas em Tandem/métodos , Acetilcarnitina/sangue , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Arginina/sangue , Biomarcadores/sangue , Citrulina/sangue , Diclofenaco/administração & dosagem , Diclofenaco/toxicidade , Cães , Gastroenteropatias/sangue , Gastroenteropatias/induzido quimicamente , Gastroenteropatias/diagnóstico , Hidroxiprolina/sangue , Limite de Detecção , Masculino , Prolina/sangue , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes
8.
Proc Natl Acad Sci U S A ; 115(34): 8627-8632, 2018 08 21.
Artigo em Inglês | MEDLINE | ID: mdl-30061399

RESUMO

The lack of biomarkers to identify target populations greatly limits the promise of precision medicine for major depressive disorder (MDD), a primary cause of ill health and disability. The endogenously produced molecule acetyl-l-carnitine (LAC) is critical for hippocampal function and several behavioral domains. In rodents with depressive-like traits, LAC levels are markedly decreased and signal abnormal hippocampal glutamatergic function and dendritic plasticity. LAC supplementation induces rapid and lasting antidepressant-like effects via epigenetic mechanisms of histone acetylation. This mechanistic model led us to evaluate LAC levels in humans. We found that LAC levels, and not those of free carnitine, were decreased in patients with MDD compared with age- and sex-matched healthy controls in two independent study centers. Secondary exploratory analyses showed that the degree of LAC deficiency reflected both the severity and age of onset of MDD. Moreover, these analyses showed that the decrease in LAC was larger in patients with a history of treatment-resistant depression (TRD), among whom childhood trauma and, specifically, a history of emotional neglect and being female, predicted the decreased LAC. These findings suggest that LAC may serve as a candidate biomarker to help diagnose a clinical endophenotype of MDD characterized by decreased LAC, greater severity, and earlier onset as well as a history of childhood trauma in patients with TRD. Together with studies in rodents, these translational findings support further exploration of LAC as a therapeutic target that may help to define individualized treatments in biologically based depression subtype consistent with the spirit of precision medicine.


Assuntos
Acetilcarnitina/sangue , Acetilcarnitina/deficiência , Transtorno Depressivo Maior/sangue , Adulto , Fatores Etários , Idoso , Carnitina/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores Sexuais
9.
Eur J Vasc Endovasc Surg ; 55(5): 703-713, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29526653

RESUMO

OBJECTIVES: Deep vein thrombosis (DVT) is a major health problem, responsible for significant morbidity and mortality. The identification of a simple and effective diagnostic biomarker of DVT remains a challenge. Metabolomics have recently emerged as a new powerful scientific tool to characterise metabolic phenotypes of complex diseases and investigate small molecules in biofluids. The aim of the study was to identify the blood and vein wall metabolomic signature of DVT in a murine experimental model. METHODS: An established inferior vena cava ligation mouse model of DVT (n=10) was used and compared with sham surgery controls (n=10). Comprehensive untargeted metabolic profiling of serum and vein wall extracts was undertaken using liquid chromatography coupled mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR) spectroscopy. RESULTS: Multivariate and univariate statistical analysis demonstrated a differential metabolic profile when comparing DVT mice and control animals. Serum from DVT mice was characterised by differential concentrations of adenosine (decreased in DVT mice 9.6 fold), adenine (decreased 10.6 fold), and tricyclic acid cycle (TCA) intermediates, including citrate, succinate, and fumarate (1.5, 2.3, and 2.8 fold decreases, respectively). l-carnitine was found to be of greater abundance in the serum of DVT animals (67.0 fold change). A number of lipid moiety classes, including sphingomyelins, phosphatidylcholines, and triglycerides, were differentially abundant. Several metabolites were found in vein wall, including acetylcarnitine (increased in DVT mice 1.9 fold), adenosine (increased 2.2 fold), and ceramide (increased 2.7 fold). Correlation analysis illustrated the biochemical relationships between assigned metabolites, with the discriminatory molecules being highly correlated with each other, in both serum and vein wall. CONCLUSIONS: The present findings demonstrate that metabolic dysregulations in DVT centre on energy metabolism, sphingolipid, and adenosine metabolism, representing a DVT specific metabolite signature in a murine experimental model.


Assuntos
Biomarcadores , Metabolômica/métodos , Veia Cava Inferior/metabolismo , Trombose Venosa/sangue , Acetilcarnitina/sangue , Acetilcarnitina/metabolismo , Adenosina/sangue , Adenosina/metabolismo , Animais , Biomarcadores/sangue , Biomarcadores/metabolismo , Cromatografia Líquida/métodos , Modelos Animais de Doenças , Metabolismo Energético , Espectroscopia de Ressonância Magnética/métodos , Camundongos , Esfingomielinas/sangue , Esfingomielinas/metabolismo , Estatística como Assunto , Ácido Succínico/sangue , Ácido Succínico/metabolismo , Trombose Venosa/diagnóstico
10.
Clin J Am Soc Nephrol ; 13(3): 436-444, 2018 03 07.
Artigo em Inglês | MEDLINE | ID: mdl-29444900

RESUMO

BACKGROUND AND OBJECTIVES: In-center, extended duration nocturnal hemodialysis has been associated with variable clinical benefits, but the effect of extended duration hemodialysis on many established uremic solutes and other components of the metabolome is unknown. We determined the magnitude of change in metabolite profiles for patients on extended duration nocturnal hemodialysis. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: In a 52-week prospective, observational study, we followed 33 patients receiving conventional thrice weekly hemodialysis who converted to nocturnal hemodialysis (7-8 hours per session, three times per week). A separate group of 20 patients who remained on conventional hemodialysis (3-4 hours per session, three times per week) served as a control group. For both groups, we applied liquid chromatography-mass spectrometry-based metabolite profiling on stored plasma samples collected from all participants at baseline and after 1 year. We examined longitudinal changes in 164 metabolites among those who remained on conventional hemodialysis and those who converted to nocturnal hemodialysis using Wilcoxon rank sum tests adjusted for multiple comparisons (false discovery rate <0.05). RESULTS: On average, the nocturnal group had 9.6 hours more dialysis per week than the conventional group. Among 164 metabolites, none changed significantly from baseline to study end in the conventional group. Twenty-nine metabolites changed in the nocturnal group, 21 of which increased from baseline to study end (including all branched-chain amino acids). Eight metabolites decreased after conversion to nocturnal dialysis, including l-carnitine and acetylcarnitine. By contrast, several established uremic retention solutes, including p-cresol sulfate, indoxyl sulfate, and trimethylamine N-oxide, did not change with extended dialysis. CONCLUSIONS: Across a wide array of metabolites examined, extended duration hemodialysis was associated with modest changes in the plasma metabolome, with most differences relating to metabolite increases, despite increased dialysis time. Few metabolites showed reduction with more dialysis, and no change in several established uremic toxins was observed.


Assuntos
Metaboloma , Diálise Renal/métodos , Insuficiência Renal Crônica/sangue , Acetilcarnitina/sangue , Adulto , Idoso , Aminoácidos de Cadeia Ramificada/sangue , Estudos de Casos e Controles , Cresóis/sangue , Feminino , Humanos , Indicã/sangue , Estudos Longitudinais , Masculino , Metilaminas/sangue , Pessoa de Meia-Idade , Estudos Prospectivos , Ésteres do Ácido Sulfúrico/sangue , Fatores de Tempo
11.
J Proteome Res ; 17(3): 1120-1128, 2018 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-29364680

RESUMO

Equine grass sickness (EGS) is a frequently fatal disease of horses, responsible for the death of 1 to 2% of the U.K. horse population annually. The etiology of this disease is currently uncharacterized, although there is evidence it is associated with Clostridium botulinum neurotoxin in the gut. Prevention is currently not possible, and ileal biopsy diagnosis is invasive. The aim of this study was to characterize the fecal microbiota and biofluid metabolic profiles of EGS horses, to further understand the mechanisms underlying this disease, and to identify metabolic biomarkers to aid in diagnosis. Urine, plasma, and feces were collected from horses with EGS, matched controls, and hospital controls. Sequencing the16S rRNA gene of the fecal bacterial population of the study horses found a severe dysbiosis in EGS horses, with an increase in Bacteroidetes and a decrease in Firmicutes bacteria. Metabolic profiling by 1H nuclear magnetic resonance spectroscopy found EGS to be associated with the lower urinary excretion of hippurate and 4-cresyl sulfate and higher excretion of O-acetyl carnitine and trimethylamine-N-oxide. The predictive ability of the complete urinary metabolic signature and using the four discriminatory urinary metabolites to classify horses by disease status was assessed using a second (test) set of horses. The urinary metabolome and a combination of the four candidate biomarkers showed promise in aiding the identification of horses with EGS. Characterization of the metabolic shifts associated with EGS offers the potential of a noninvasive test to aid premortem diagnosis.


Assuntos
Acetilcarnitina/urina , Cresóis/urina , Disbiose/diagnóstico , Hipuratos/urina , Doenças dos Cavalos/diagnóstico , Metilaminas/urina , Ésteres do Ácido Sulfúrico/urina , Acetilcarnitina/sangue , Animais , Bacteroidetes/classificação , Bacteroidetes/isolamento & purificação , Biomarcadores/sangue , Biomarcadores/urina , Clostridium botulinum/metabolismo , Clostridium botulinum/patogenicidade , Cresóis/sangue , Disbiose/sangue , Disbiose/microbiologia , Disbiose/urina , Fezes/microbiologia , Firmicutes/classificação , Firmicutes/isolamento & purificação , Microbioma Gastrointestinal , Hipuratos/sangue , Doenças dos Cavalos/sangue , Doenças dos Cavalos/microbiologia , Doenças dos Cavalos/urina , Cavalos , Espectroscopia de Ressonância Magnética , Metilaminas/sangue , RNA Ribossômico 16S/genética , Ésteres do Ácido Sulfúrico/sangue
12.
Epilepsia ; 58(5): 893-900, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-28369834

RESUMO

OBJECTIVE: In the absence of specific metabolic disorders, accurate predictors of response to ketogenic dietary therapies (KDTs) for treating epilepsy are largely unknown. We hypothesized that specific biochemical parameters would be associated with the effectiveness of KDT in humans with epilepsy. The parameters tested were ß-hydroxybutyrate, acetoacetate, nonesterified fatty acids, free and acylcarnitine profile, glucose, and glucose-ketone index (GKI). METHODS: Biochemical results from routine blood tests conducted at baseline prior to initiation of KDT and at 3-month follow-up were obtained from 13 adults and 215 children with KDT response data from participating centers. One hundred thirty-two (57%) of 228 participants had some data at both baseline and 3 months; 52 (23%) of 228 had data only at baseline; 22 (10%) of 228 had data only at 3 months; and 22 (10%) of 228 had no data. KDT response was defined as ≥50% seizure reduction at 3-month follow-up. RESULTS: Acetyl carnitine at baseline was significantly higher in responders (p < 0.007). It was not associated with response at 3-month follow-up. There was a trend for higher levels of free carnitine and other acylcarnitine esters at baseline and at 3-month follow-up in KDT responders. There was also a trend for greater differences in levels of propionyl carnitine and in ß-hydroxybutyrate measured at baseline and 3-month follow-up in KDT responders. No other biochemical parameters were associated with response at any time point. SIGNIFICANCE: Our finding that certain carnitine fractions, in particular baseline acetyl carnitine, are positively associated with greater efficacy of KDT is consistent with the theory that alterations in energy metabolism may play a role in the mechanisms of action of KDT.


Assuntos
Biomarcadores/sangue , Dieta Cetogênica , Epilepsia/sangue , Epilepsia/dietoterapia , Acetilcarnitina/sangue , Adolescente , Adulto , Fatores Etários , Criança , Pré-Escolar , Epilepsia/genética , Feminino , Seguimentos , Humanos , Masculino , Resultado do Tratamento , Adulto Jovem
13.
Int J Mol Sci ; 18(3)2017 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-28287417

RESUMO

Asthma is a chronic inflammatory disorder of the airway and is characterized by airway remodeling, hyperresponsiveness, and shortness of breath. Modified Kushen Gancao Formula (mKG), derived from traditional Chinese herbal medicines (TCM), has been demonstrated to have good therapeutic effects on experimental allergic asthma. However, its anti-asthma mechanism remains currently unknown. In the present work, metabolomics studies of biochemical changes in the lung tissue and plasma of ovalbumin (OVA)-induced allergic asthma mice with mKG treatment were performed using ultra high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). Partial least squares-discriminate analysis (PLS-DA) indicated that the metabolic perturbation induced by OVA was reduced after mKG treatment. A total of twenty-four metabolites involved in seven metabolic pathways were identified as potential biomarkers in the development of allergic asthma. Among them, myristic acid (L3 or P2), sphinganine (L6 or P4), and lysoPC(15:0) (L12 or P16) were detected both in lung tissue and plasma. Additionally, l-acetylcarnitine (L1), thromboxane B2 (L2), 10-HDoHE (L10), and 5-HETE (L11) were first reported to be potential biomarkers associated with allergic asthma. The treatment of mKG mediated all of those potential biomarkers except lysoPC(15:0) (P16). The anti-asthma mechanism of mKG can be achieved through the comprehensive regulation of multiple perturbed biomarkers and metabolic pathways.


Assuntos
Asma/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Hipersensibilidade/metabolismo , Pulmão/efeitos dos fármacos , Metaboloma/efeitos dos fármacos , Acetilcarnitina/sangue , Acetilcarnitina/metabolismo , Animais , Asma/etiologia , Biomarcadores/sangue , Biomarcadores/metabolismo , Feminino , Hipersensibilidade/complicações , Pulmão/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Ácido Mirístico/sangue , Ácido Mirístico/metabolismo , Esfingosina/análogos & derivados , Esfingosina/sangue , Esfingosina/metabolismo , Tromboxano B2/sangue , Tromboxano B2/metabolismo
14.
J Med Screen ; 24(2): 58-61, 2017 06.
Artigo em Inglês | MEDLINE | ID: mdl-27216769

RESUMO

Background Newborn screening for propionic acidemia and methylmalonic acidurias using the marker propionylcarnitine (C3) is neither sensitive nor specific. Using C3 to acetylcarnitine (C3/C2) ratio, together with conservative C3 cut-offs, can improve screening sensitivity, but the false positive rate remains high. Incorporating the marker 2-methylcitric acid has been suggested, to improve the positive predictive value for these disorders without compromising the sensitivity. Methods Between July 2011 and December 2012 at the Newborn Screening Ontario laboratory, all neonatal dried blood spot samples that were reported as screen positive for propionic acidemia or methylmalonic acidurias based on elevated C3 and C3/C2 ratio were analyzed for 2-methylcitric acid, using liquid chromatography tandem mass spectrometry. Results Of 222,420 samples screened, 103 were positive for methylmalonic acidurias or propionic acidemia using C3 and C3/C2 ratio as markers. There were nine true positives: propionic acidemia (n = 3), Cobalamin (Cbl) A (n=1), and Cbl C (n = 5). Among false positives there were 72 neonates not affected, 20 with maternal B12 deficiency, and two incidental finding (transcobalamin II and unclassified Cbl defect). 2-Methylcitric acid was analyzed in all 103 samples and ranged between 0.1 and 89.4 µmol/l (reference range 0.04-0.36). Only 14 samples exceeded the set 2-methylcitric acid cut-off of 1.0 µmol/l, including the samples from all nine true positives. Conclusion By including 2-methylcitric acid in the screening algorithm, the positive predictive value of our primary and secondary screening targets improved from 8.7 to 64.3%. This would have eliminated 89 unnecessary referrals while maintaining 100% sensitivity.


Assuntos
Erros Inatos do Metabolismo dos Aminoácidos/sangue , Citratos/sangue , Teste em Amostras de Sangue Seco , Triagem Neonatal/métodos , Acetilcarnitina/sangue , Algoritmos , Erros Inatos do Metabolismo dos Aminoácidos/diagnóstico , Biomarcadores/sangue , Carnitina/análogos & derivados , Carnitina/sangue , Cromatografia Líquida , Reações Falso-Positivas , Humanos , Recém-Nascido , Ácido Metilmalônico/sangue , Valor Preditivo dos Testes , Valores de Referência , Encaminhamento e Consulta , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem
15.
Sci Rep ; 6: 24642, 2016 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-27089826

RESUMO

The failure to adapt metabolism to the homeorhetic demands of lactation is considered as a main factor in reducing the productive life span of dairy cows. The so far defined markers of production performance and metabolic health in dairy cows do not predict the length of productive life span satisfyingly. This study aimed to identify novel pathways and biomarkers related to productive life in dairy cows by means of (targeted) metabolomics. In a longitudinal study from 42 days before up to 100 days after parturition, we identified metabolites such as long-chain acylcarnitines and biogenic amines associated with extended productive life spans. These metabolites are mainly secreted by the liver and depend on the functionality of hepatic mitochondria. The concentrations of biogenic amines and some acylcarnitines differed already before the onset of lactation thus indicating their predictive potential for continuation or early ending of productive life.


Assuntos
Bovinos/metabolismo , Lactação , Longevidade , Metaboloma , Acetilcarnitina/sangue , Animais , Aminas Biogênicas/sangue , Biomarcadores/sangue , Bovinos/crescimento & desenvolvimento , Bovinos/fisiologia , Feminino , Mitocôndrias Hepáticas/metabolismo
16.
Biomed Res Int ; 2016: 2920108, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26998483

RESUMO

Blood carnitine and/or acetylcarnitine deficiencies are postulated in the literature as possible causes of higher ammonia levels. The aim of this study was to investigate if the use of valproic acid, the age of the patients, or certain central nervous system pathologies can cause carnitine and/or acetylcarnitine deficiency leading to increased ammonia levels. Three groups of patients were studied: (A) epileptic under phenytoin monotherapy (n = 31); (B) with bipolar disorder under valproic acid treatment (n = 28); (C) elderly (n = 41). Plasma valproic acid and blood carnitine and acyl carnitine profiles were determined using a validated HPLC and LC-MS/MS method, respectively. Blood ammonia concentration was determined using an enzymatic automated assay. Higher ammonia levels were encountered in patients under valproic acid treatment and in the elderly. This may be due to the lower carnitine and/or acetylcarnitine found in these patients. Patients with controlled seizures had normal carnitine and acetylcarnitine levels. Further studies are necessary in order to conclude if the uncontrolled bipolar disorder could be the cause of higher carnitine and/or acetylcarnitine levels.


Assuntos
Acetilcarnitina , Amônia/sangue , Transtorno Bipolar , Carnitina , Epilepsia , Fenitoína/administração & dosagem , Ácido Valproico/administração & dosagem , Acetilcarnitina/sangue , Acetilcarnitina/deficiência , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Transtorno Bipolar/sangue , Transtorno Bipolar/tratamento farmacológico , Carnitina/sangue , Carnitina/deficiência , Epilepsia/sangue , Epilepsia/tratamento farmacológico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
17.
Cancer Res ; 76(10): 2912-20, 2016 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-26976432

RESUMO

The identification of serum biomarkers to improve the diagnosis and prognosis of hepatocellular carcinoma has been elusive to date. In this study, we took a mass spectroscopic approach to characterize metabolic features of the liver in hepatocellular carcinoma patients to discover more sensitive and specific biomarkers for diagnosis and progression. Global metabolic profiling of 50 pairs of matched liver tissue samples from hepatocellular carcinoma patients was performed. A series of 62 metabolites were found to be altered significantly in liver tumors; however, levels of acetylcarnitine correlated most strongly with tumor grade and could discriminate between hepatocellular carcinoma tumors and matched normal tissues. Post hoc analysis to evaluate serum diagnosis and progression potential further confirmed the diagnostic capability of serum acetylcarnitine. Finally, an external validation in an independent batch of 58 serum samples (18 hepatocellular carcinoma patients, 20 liver cirrhosis patients, and 20 healthy individuals) verified that serum acetylcarnitine was a meaningful biomarker reflecting hepatocellular carcinoma diagnosis and progression. These findings present a strong new candidate biomarker for hepatocellular carcinoma with potentially significant diagnostic and prognostic capabilities. Cancer Res; 76(10); 2912-20. ©2016 AACR.


Assuntos
Acetilcarnitina/sangue , Biomarcadores Tumorais/sangue , Carcinoma Hepatocelular/diagnóstico , Cirrose Hepática/diagnóstico , Neoplasias Hepáticas/diagnóstico , Metaboloma , Adulto , Idoso , Carcinoma Hepatocelular/sangue , Estudos de Casos e Controles , Feminino , Seguimentos , Humanos , Técnicas Imunoenzimáticas , Cirrose Hepática/sangue , Neoplasias Hepáticas/sangue , Masculino , Espectrometria de Massas , Metabolômica , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Taxa de Sobrevida
18.
PLoS One ; 11(2): e0147818, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26881790

RESUMO

Rhabdomyolysis is common in very long-chain acyl-CoA dehydrogenase deficiency (VLCADD) and other metabolic myopathies, but its pathogenic basis is poorly understood. Here, we show that prolonged bicycling exercise against a standardized moderate workload in VLCADD patients is associated with threefold bigger changes in phosphocreatine (PCr) and inorganic phosphate (Pi) concentrations in quadriceps muscle and twofold lower changes in plasma acetyl-carnitine levels than in healthy subjects. This result is consistent with the hypothesis that muscle ATP homeostasis during exercise is compromised in VLCADD. However, the measured rates of PCr and Pi recovery post-exercise showed that the mitochondrial capacity for ATP synthesis in VLCADD muscle was normal. Mathematical modeling of oxidative ATP metabolism in muscle composed of three different fiber types indicated that the observed altered energy balance during submaximal exercise in VLCADD patients may be explained by a slow-to-fast shift in quadriceps fiber-type composition corresponding to 30% of the slow-twitch fiber-type pool in healthy quadriceps muscle. This study demonstrates for the first time that quadriceps energy balance during exercise in VLCADD patients is altered but not because of failing mitochondrial function. Our findings provide new clues to understanding the risk of rhabdomyolysis following exercise in human VLCADD.


Assuntos
Acil-CoA Desidrogenase de Cadeia Longa/deficiência , Trifosfato de Adenosina/biossíntese , Exercício Físico , Erros Inatos do Metabolismo Lipídico/metabolismo , Doenças Mitocondriais/metabolismo , Modelos Estatísticos , Doenças Musculares/metabolismo , Rabdomiólise/metabolismo , Acetilcarnitina/sangue , Acil-CoA Desidrogenase de Cadeia Longa/metabolismo , Adolescente , Adulto , Estudos de Casos e Controles , Síndrome Congênita de Insuficiência da Medula Óssea , Feminino , Humanos , Erros Inatos do Metabolismo Lipídico/complicações , Erros Inatos do Metabolismo Lipídico/patologia , Erros Inatos do Metabolismo Lipídico/fisiopatologia , Masculino , Mitocôndrias/metabolismo , Doenças Mitocondriais/complicações , Doenças Mitocondriais/patologia , Doenças Mitocondriais/fisiopatologia , Fibras Musculares de Contração Rápida/metabolismo , Fibras Musculares de Contração Rápida/patologia , Fibras Musculares de Contração Lenta/metabolismo , Fibras Musculares de Contração Lenta/patologia , Doenças Musculares/complicações , Doenças Musculares/patologia , Doenças Musculares/fisiopatologia , Fosforilação Oxidativa , Fosfatos/metabolismo , Fosfocreatina/metabolismo , Rabdomiólise/complicações , Rabdomiólise/patologia , Rabdomiólise/fisiopatologia
19.
Int J Clin Pharmacol Ther ; 53(11): 980-6, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26396135

RESUMO

BACKGROUND: Acetyl-L-carnitine (ALC) has demonstrated neuroprotective effects in several experiments and is widely prescribed to reduce cognitive impairment in Alzheimer's disease patients or manage neuropathic symptoms in diabetic patients. OBJECTIVES: This study was designed to assess the pharmacokinetic (PK) bioequivalence between a new generic (test) formulation of ALC hydrochloride 590 mg and a branded (reference) formulation of ALC hydrochloride 590 mg in healthy Korean male volunteers. METHODS: This was a randomizedsequence, single-dose, two-way crossover study. All subjects randomly received one formulation of the test or reference tablet and the other formulation with a 7-day washout period. Blood samples (7 mL) were collected immediately before dosing, and at 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, 8, and 12 hours postdose. The plasma concentrations of ALC were analyzed using liquid chromatography tandem mass spectrometry. Tolerability was assessed throughout the study. RESULTS: The PK profiles of both formulations showed similar rends. The mean (±SD) baseline (predose) concentration of ALC was 1.23±0.31 µg/mL and 1.09±0.30 µg/mL for the test and the reference formulations, respectively. The mean Cmax for the test and reference formulations were 1.74±0.43 µg/mL and 1.68±0.48 µg/mL, respectively. The mean AUClast of ALC was 12.96±1.89 µg×h/mL and 12.49±2.44 µg×h/mL for the test and reference formulations, respectively. The geometric mean ratios of test/reference (90% CI) were 1.050 (0.960-1.149) for Cmax and 1.048 (1.000-1.099) for AUClast. Both formulations were well tolerated in all treatment groups. CONCLUSION: The test and the reference formulations of ALC were bioequivalent with regard to the PK parameters.


Assuntos
Acetilcarnitina/farmacocinética , Grupo com Ancestrais do Continente Asiático , Medicamentos Genéricos/farmacocinética , Fármacos Neuroprotetores/farmacocinética , Acetilcarnitina/administração & dosagem , Acetilcarnitina/efeitos adversos , Acetilcarnitina/sangue , Acetilcarnitina/química , Administração Oral , Adulto , Área Sob a Curva , Química Farmacêutica , Cromatografia Líquida , Estudos Cross-Over , Medicamentos Genéricos/administração & dosagem , Medicamentos Genéricos/efeitos adversos , Medicamentos Genéricos/química , Voluntários Saudáveis , Humanos , Masculino , Fármacos Neuroprotetores/administração & dosagem , Fármacos Neuroprotetores/efeitos adversos , Fármacos Neuroprotetores/sangue , Fármacos Neuroprotetores/química , República da Coreia , Comprimidos , Espectrometria de Massas em Tandem , Equivalência Terapêutica , Adulto Jovem
20.
Am J Physiol Endocrinol Metab ; 309(3): E256-64, 2015 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-26037250

RESUMO

Acylcarnitines are derived from mitochondrial acyl-CoA metabolism and have been associated with diet-induced insulin resistance. However, plasma acylcarnitine profiles have been shown to poorly reflect whole body acylcarnitine metabolism. We aimed to clarify the individual role of different organ compartments in whole body acylcarnitine metabolism in a fasted and postprandial state in a porcine transorgan arteriovenous model. Twelve cross-bred pigs underwent surgery where intravascular catheters were positioned before and after the liver, gut, hindquarter muscle compartment, and kidney. Before and after a mixed meal, we measured acylcarnitine profiles at several time points and calculated net transorgan acylcarnitine fluxes. Fasting plasma acylcarnitine concentrations correlated with net hepatic transorgan fluxes of free and C2- and C16-carnitine. Transorgan acylcarnitine fluxes were small, except for a pronounced net hepatic C2-carnitine production. The peak of the postprandial acylcarnitine fluxes was between 60 and 90 min. Acylcarnitine production or release was seen in the gut and liver and consisted mostly of C2-carnitine. Acylcarnitines were extracted by the kidney. No significant net muscle acylcarnitine flux was observed. We conclude that liver has a key role in acylcarnitine metabolism, with high net fluxes of C2-carnitine both in the fasted and fed state, whereas the contribution of skeletal muscle is minor. These results further clarify the role of different organ compartments in the metabolism of different acylcarnitine species.


Assuntos
Carnitina/análogos & derivados , Metabolismo dos Lipídeos , Fígado/metabolismo , Modelos Biológicos , Acetilcarnitina/sangue , Acetilcarnitina/metabolismo , Animais , Carnitina/biossíntese , Carnitina/sangue , Carnitina/metabolismo , Cateteres de Demora , Cruzamentos Genéticos , Feminino , Mucosa Intestinal/metabolismo , Intestinos/irrigação sanguínea , Rim/irrigação sanguínea , Rim/metabolismo , Fígado/irrigação sanguínea , Azeite de Oliva , Especificidade de Órgãos , Palmitoilcarnitina/sangue , Palmitoilcarnitina/metabolismo , Óleos Vegetais/administração & dosagem , Óleos Vegetais/metabolismo , Período Pós-Prandial , Sus scrofa
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