RESUMEN
ß-Hydroxybutyrate (BHB) is a substantial physiological ketone body. Its elevated concentration causes ketoacidosis, which is a disorder with a high mortality rate. Therefore, there is an urgent need to develop a simple method for the in-situ monitoring of BHB in urine. In this study, a photonic crystal hydrogel (PCH) sensing material for the detection of urinary ketones was prepared by embedding a two-dimensional polystyrene photonic crystal array (PCA) in a hydrogel functionalized with ß-hydroxybutyrate dehydrogenase (BHBDH). BHBDH catalyzes the interconversion between ß-hydroxybutyrate and acetoacetic acid and relies on the cofactor nicotinamide adenine dinucleotide (NAD+) to participate in the reaction process. The catalytic cycle of converting ß-hydroxybutyrate to acetoacetate generates H+, which reduces the electrostatic repulsion between the carboxyl groups in the hydrogel network, ultimately leading to the shrinkage of the hydrogel volume. The hydrogel volume change was detected by measuring the diameter of the Debye diffraction ring, thus reflecting the concentration of BHB. When the concentration of BHB was increased from 0 to 10 mM, the reflection spectrum of PCH shifted for 117 nm within 60 min, consequently, the structural color of PCH changed from red to green and finally to blue. The material was used for quantitative detection of BHB with a detection limit of 48.94 µM. Then it was used for detection in artificial urine samples. While, this smart and reusable sensing material could provide a more convenient and efficient strategy for the ketone body detection in clinical diagnosis and point-of-care monitoring.
Asunto(s)
Ácido 3-Hidroxibutírico , Técnicas Biosensibles , Hidroxibutirato Deshidrogenasa , Hidroxibutirato Deshidrogenasa/química , Ácido 3-Hidroxibutírico/química , Ácido 3-Hidroxibutírico/orina , Ácido 3-Hidroxibutírico/análisis , Humanos , Cuerpos Cetónicos/orina , Cuerpos Cetónicos/química , Hidrogeles/química , Enzimas Inmovilizadas/química , Límite de Detección , FotonesRESUMEN
Ketone bodies are essential energy substrates in the absence of exogenous nutrients, and more recently, they have been suggested to prevent disease and improve longevity. ß-hydroxybutyrate (ßHB) is the most abundant ketone body. The secondary alcohol, 1,3-butanediol (1,3-BD), is commonly administered to raise ßHB bioavailability in vivo and in the absence of nutrient deprivation. However, the concentration of 1,3-BD that yields a systemic concentration of ßHB similar to that observed after a 24-hour fast has yet to be determined. To evaluate this knowledge gap, we administered 5%, 10%, or 20% 1,3-BD via the drinking water to adult, male Wistar-Kyoto rats for four weeks. In addition to systemic and excreted ßHB concentration, physiologic, metabolic, and toxicologic parameters were measured. We report that only 20% 1,3-BD significantly elevates the systemic and urinary concentrations of ßHB. Rats treated with 20% 1,3-BD had a rapid and sustained reduction in body mass. All concentrations of 1,3-BD decreased food consumption, but only the 20% concentration decreased fluid consumption. Urine volume, red blood cell count, and hematocrit suggested dehydration in the 10% and 20% 1,3-BD-treated rats. Finally, 20% 1,3-BD-treated rats presented with indicators of metabolic acidosis and sinusoidal dilation, but no evidence of fatty liver or hepatotoxicity. In summary, we report that 20% 1,3-BD, but not 5% or 10%, produces a systemic concentration of ßHB similar to that observed after a 24-hour fast. However, this concentration is associated with deleterious side effects such as body mass loss, dehydration, metabolic acidosis, and sinusoidal dilation. SIGNIFICANCE STATEMENT: 1,3-Butanediol (1,3-BD) is often administered to stimulate the biosynthesis of the most abundant ketone body, ß-hydroxybutyrate (ßHB), and its purported salubrious effects. This article reports that suprapharmacological concentrations of 1,3-BD are necessary to yield a systemic concentration of ßHB similar to that observed after a 24-hour fast, and this is associated with undesirable side effects. On the other hand, low concentrations of 1,3-BD were better tolerated and may improve health independent of its conversion into ßHB.
Asunto(s)
Ácido 3-Hidroxibutírico/sangre , Ácido 3-Hidroxibutírico/orina , Butileno Glicoles/metabolismo , Butileno Glicoles/toxicidad , Animales , Butileno Glicoles/farmacología , Ingestión de Alimentos/efectos de los fármacos , Ingestión de Alimentos/fisiología , Masculino , Ratas , Ratas Endogámicas WKYRESUMEN
Ketone bodies (acetoacetate, beta-hydroxybutyrate (ßHB), acetone) are generated as a result of fatty acid oxidation in the liver and exist at low concentrations in urine and blood. Elevated concentrations can indicate health problems such as diabetes, childhood hypoglycemia, alcohol, or salicylate poisoning. Development of portable and cost-effective bedside point-of-care (POC) tests to detect such compounds can help to reduce the risk of disease progression. In this study, ßHB was chosen as a model molecule for developing an optical sensor-integrated microplatform. Prior to sensor optimization, ßHB levels were measured at a concentration range of 0.02 and 0.1 mM spectrophotometrically, which is far below the reported elevated ranges of 1-2 mM and resulting absorbance changes were converted into an Arduino microcontroller code for the correlation. Measurements performed with the designed integrated microplatform were found significant. Integrated microplatform was verified with the benchtop spectrophotometer. Measurements between 0.02 and 0.1 mM substrate concentration were found highly sensitive with "y = 0.7347x + 0.00184" with R2 value of 0.9796, and the limit of detection was determined as 0.02 mM. Based on these results, the proposed system will allow on-site and early intervention.
Asunto(s)
Ácido 3-Hidroxibutírico , Óptica y Fotónica , Pruebas en el Punto de Atención , Ácido 3-Hidroxibutírico/sangre , Ácido 3-Hidroxibutírico/orina , HumanosRESUMEN
INTRODUCTION: Exogenous ketones potentially provide an alternative, energetically advantageous fuel to power exercising skeletal muscle. However, there is limited evidence regarding their relative contribution to energy expenditure during exercise. Furthermore, the effect of blood ketone concentration and exercise intensity on exogenous ketone oxidation rates is unknown. METHODS: Six athletes completed cycling ergometer exercise on three occasions within a single-blind, random-order controlled, crossover design study. Exercise duration was 60 min, consisting of 20-min intervals at 25%, 50%, and 75% maximal power output (WMax). Participants consumed (i) bitter flavored water (control), (ii) a low-dose ß-hydroxybutyrate (ßHB) ketone monoester (KME; 252 mg·kg BW-1, "low ketosis"), or (iii) a high-dose ßHB KME (752 mg·kg BW-1, "high ketosis"). The KME contained a 13C isotope label, allowing for the determination of whole-body exogenous ßHB oxidation rates through sampled respiratory gases. RESULTS: Despite an approximate doubling of blood ßHB concentrations between low- and high-ketosis conditions (~2 mM vs ~4.4 mM), exogenous ßHB oxidation rates were similar at rest and throughout exercise. The contribution of exogenous ßHB oxidation to energy expenditure peaked during the 25% WMax exercise intensity but was relatively low (4.46% ± 2.71%). Delta efficiency during cycling exercise was significantly greater in the low-ketosis (25.9% ± 2.1%) versus control condition (24.1% ± 1.9%; P = 0.027). CONCLUSIONS: Regardless of exercise intensity, exogenous ßHB oxidation contributes minimally to energy expenditure and is not increased by elevating circulating concentrations greater than ~2 mM. Despite low exogenous ßHB oxidation rates, exercise efficiency was significantly improved when blood ßHB concentration was raised to ~2 mM.
Asunto(s)
Ácido 3-Hidroxibutírico/metabolismo , Atletas , Metabolismo Energético/fisiología , Ejercicio Físico/fisiología , Cetonas/sangre , Músculo Esquelético/metabolismo , Ácido 3-Hidroxibutírico/administración & dosificación , Ácido 3-Hidroxibutírico/sangre , Ácido 3-Hidroxibutírico/orina , Estudios Cruzados , Prueba de Esfuerzo , Femenino , Glucógeno/metabolismo , Humanos , Cetonas/administración & dosificación , Cetosis/metabolismo , Masculino , Fibras Musculares Esqueléticas/metabolismo , Oxidación-Reducción , Esfuerzo Físico , Método Simple Ciego , Factores de Tiempo , Adulto JovenRESUMEN
SLC5A8 is a sodium-coupled monocarboxylate and ketone transporter expressed in various epithelial cells. A putative role of SLC5A8 in neuroenergetics has been also hypothesized. To clarify this issue, we studied the cerebral phenotype of SLC5A8-deficient mice during aging. Elderly SLC5A8-deficient mice presented diffuse leukoencephalopathy characterized by intramyelinic oedema without demyelination suggesting chronic energetic crisis. Hypo-metabolism in the white matter of elderly SLC5A8-deficient mice was found using 99mTc-hexamethylpropyleneamine oxime (HMPAO) single-photon emission CT (SPECT). Since the SLC5A8 protein could not be detected in the mouse brain, it was hypothesized that the leukoencephalopathy of aging SLC5A8-deficient mice was caused by the absence of slc5a8 expression in a peripheral organ, i.e. the kidney, where SLC5A8 is strongly expressed. A hyper-excretion of the ketone ß-hydroxybutyrate (BHB) in the urine of SLC5A8-deficient mice was observed and showed that SLC5A8-deficient mice suffered a cerebral BHB insufficiency. Elderly SLC5A8-deficient mice also presented altered glucose metabolism. We propose that the continuous renal loss of BHB leads to a chronic energetic deficiency in the brain of elderly SLC5A8-deficient mice who are unable to counterbalance their glucose deficit. This study highlights the importance of alternative energetic substrates in neuroenergetics especially under conditions of restricted glucose availability.
Asunto(s)
Envejecimiento/metabolismo , Cuerpos Cetónicos/orina , Riñón/metabolismo , Leucoencefalopatías/metabolismo , Transportadores de Ácidos Monocarboxílicos/deficiencia , Sustancia Blanca/metabolismo , Ácido 3-Hidroxibutírico/orina , Envejecimiento/orina , Animales , Glucosa/metabolismo , Leucoencefalopatías/orina , Masculino , Ratones Endogámicos C57BL , Ratones Mutantes , Transportadores de Ácidos Monocarboxílicos/genética , Tomografía Computarizada de Emisión de Fotón Único , Sustancia Blanca/diagnóstico por imagenRESUMEN
With a prevalence of up to 43 % subclinical ketosis is one of the most common diseases in dairy cows in their transition period. In itself, this may cause subsequent diseases such as clinical ketosis or lameness. Therefore, monitoring of animals in this stage is of importance. In addition to the measurement of ß-hydroxybutyrate or acetoacetate in blood, milk, and urine as well as the observation of the animals, computer-assisted systems are suitable means of monitoring. Information such as animal identification and activity data are recorded on a data logger and transmitted to a computer. A change in activity may be an indication of an underlying disease days before the onset of additional clinical signs. In cases of ketosis, a decrease in activity may be observed 5 days before the clinical diagnosis is made. Thus, these data are a valuable contribution in monitoring the cattle herd's health status for both the farmer and the veterinarian. Activity measurement may also be employed for the detection of a beginning lameness. In the presence of lameness, the individual's activity decreases and periods of lying are longer. Activity measurement via transponder as a part of the herd monitoring provides important information on lameness prevalence in the herd. In the presence of a lameness a visual assessment should additionally be made. Lameness scores (Locomotion score, Gait score) have been developed for this purpose and add to determining the lameness status of the herd. This way the animals are divided into different lameness classes. Based on this classification those individuals in need of claw trimming or further treatment may be identified leading to amelioration or prevention of secondary diseases. Due to lameness and subsequent reduction of activity and feed intake, the animals may develop subclinical or clinical ketosis. Therefore, under consideration of both animal welfare and economic factors early disease detection and prophylaxis is desirable and should be a main objective of herd monitoring.
Asunto(s)
Enfermedades de los Bovinos/diagnóstico , Cetosis/veterinaria , Cojera Animal/diagnóstico , Ácido 3-Hidroxibutírico/análisis , Ácido 3-Hidroxibutírico/sangre , Ácido 3-Hidroxibutírico/orina , Acetoacetatos/análisis , Acetoacetatos/sangre , Acetoacetatos/orina , Animales , Conducta Animal , Bovinos , Enfermedades de los Bovinos/etiología , Enfermedades de los Bovinos/prevención & control , Diagnóstico por Computador/veterinaria , Endometritis/diagnóstico , Endometritis/etiología , Endometritis/prevención & control , Endometritis/veterinaria , Femenino , Enfermedades del Pie/etiología , Enfermedades del Pie/veterinaria , Pezuñas y Garras/patología , Cetosis/diagnóstico , Cetosis/prevención & control , Cojera Animal/etiología , Cojera Animal/prevención & control , Leche/química , Oportunidad Relativa , ProbabilidadRESUMEN
AIM: To verify the possible advantages of 3- ß-hydroxybutyrate (3HB) measurement compared to urinary assay of ketones during an intercurrent disease managed at home. METHODS: Twelve Pediatricians were asked to enroll at least 4 patients aged 3 to 5 years, affected by an intercurrent illness and showing at least one of symptoms reliable to ketosis. Recruited patients were submitted to the simultaneous assay of 3HB in capillary blood and ketones in urine at 3 (T3) and 6 hours (T6) from the first measurement (T0). For urinary and blood ketone detection commercial tests were used. RESULTS: Thirty-eight children (4.36±2.60 years old; 25 boys) were enrolled into the study. At T0 all children showed 3HB levels (1.2-3.2 mmol/L), but only 10 of them (26.3%) associated also urinary ketone bodies (2 to 4+). In response to 3 hour treatment (T3) with a glucose solution, 3HB values decreased in 19 (0,8-1,8 mmol/L) and normalized in 13 children (<0.2 mmol/L); while ketonuria disappeared in only 2 patients, it was confirmed in 8 and appeared (4+) the first time in the remaining 28 children. At T6 3HB levels fell definitively within the normal range in all children, while ketonuria was still present (2+) in 9 patients (23%). The pediatricians reported two limitations about blood 3HB dosage compared to the urinary test: the invasiveness of capillary blood collection, and the cost of supplies for finger pricking, reagent strips and reflectance meter. CONCLUSIONS: 3HB monitor in capillary blood is more effective and clinically more useful in diagnosing and managing of an ongoing ketosis in children with a mild infective disease than ketones detection in the urine. These advantages are mitigated by the cost of 3HB measurement.
Asunto(s)
Ácido 3-Hidroxibutírico/sangre , Ácido 3-Hidroxibutírico/orina , Diabetes Mellitus Tipo 1/sangre , Cetoacidosis Diabética/diagnóstico , Monitoreo Fisiológico/métodos , Capilares , Preescolar , Diabetes Mellitus Tipo 1/complicaciones , Diabetes Mellitus Tipo 1/diagnóstico , Femenino , Servicios de Atención de Salud a Domicilio , Humanos , Italia , Masculino , Pediatras , Muestreo , Sensibilidad y EspecificidadRESUMEN
AIM: To investigate sodium-glucose cotransporter 2 inhibitor (SGLT2i)-induced changes in ketogenic enzymes and transporters in normal and diabetic mice models. MATERIALS AND METHODS: Normal mice were randomly assigned to receive either vehicle or SGLT2i (25 mg/kg/d by oral gavage) for 7 days. Diabetic mice were treated with vehicle, insulin (4.5 units/kg/d by subcutaneous injection) or SGLT2i (25 mg/kg/d by intra-peritoneal injection) for 5 weeks. Serum and tissues of ketogenic organs were analysed. RESULTS: In both normal and diabetic mice, SGLT2i increased beta-hydroxybutyrate (BHB) content in liver, kidney and colon tissue, as well as in serum and urine. In these organs, SGLT2i upregulated mRNA expression of ketogenic enzymes, 3-hydroxy-3-methylglutaryl-coenzyme A synthase 2 and 3-hydroxy-3-methylglutaryl-coenzyme A lyase. Similar patterns were observed in the kidney, ileum and colon for mRNA and protein expression of sodium-dependent monocarboxylate transporters (SMCTs), which mediate the cellular uptake of BHB and butyrate, an important substrate for intestinal ketogenesis. In diabetic mice under euglycaemic conditions, SGLT2i increased major ketogenic enzymes and SMCTs, while insulin suppressed ketogenesis. CONCLUSIONS: SGLT2i increased systemic and tissue BHB levels by upregulating ketogenic enzymes and transporters in the liver, kidney and intestine, suggesting the integrated physiological consequences for ketone body metabolism of SGLT2i administration.
Asunto(s)
Ácido 3-Hidroxibutírico/metabolismo , Colon/efectos de los fármacos , Hidroximetilglutaril-CoA Sintasa/efectos de los fármacos , Riñón/efectos de los fármacos , Hígado/efectos de los fármacos , Transportadores de Ácidos Monocarboxílicos/efectos de los fármacos , Oxo-Ácido-Liasas/efectos de los fármacos , Inhibidores del Cotransportador de Sodio-Glucosa 2/farmacología , Ácido 3-Hidroxibutírico/sangre , Ácido 3-Hidroxibutírico/orina , Animales , Compuestos de Bencidrilo/farmacología , Colon/metabolismo , Estrés del Retículo Endoplásmico/efectos de los fármacos , Glucósidos/farmacología , Humanos , Hidroximetilglutaril-CoA Sintasa/genética , Hipoglucemiantes/farmacología , Insulina/farmacología , Mucosa Intestinal/citología , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/metabolismo , Cuerpos Cetónicos/metabolismo , Riñón/metabolismo , Túbulos Renales Proximales/citología , Túbulos Renales Proximales/efectos de los fármacos , Túbulos Renales Proximales/metabolismo , Hígado/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Obesos , Transportadores de Ácidos Monocarboxílicos/genética , Oxo-Ácido-Liasas/genética , ARN Mensajero/efectos de los fármacos , ARN Mensajero/metabolismo , Distribución Aleatoria , RatasRESUMEN
Single large-scale mitochondrial DNA deletions disorders are classified into three main phenotypes with frequent clinical overlap: Pearson marrow-pancreas syndrome (PMS), Kearns-Sayre syndrome (KSS) and chronic progressive external ophtalmoplegia (PEO). So far, only few anecdotal studies have reported on the urinary organic acids profile in this disease class. In this single-center retrospective study, we performed quantitative evaluation of urinary organic acids in a series of 15 pediatric patients, 7 with PMS and 8 with KSS. PMS patients showed an organic acids profile almost constantly altered, whereas KSS patients frequently presented with normal profiles. Lactate, 3-hydroxybutyrate, 3-hydroxyisobutyrate, fumarate, pyruvate, 2-hydroxybutyrate, 2-ethyl-3-hydroxypropionate, and 3-methylglutaconate represented the most frequent metabolites observed in PMS urine. We also found novel metabolites, 3-methylglutarate, tiglylglycine and 2-methyl-2,3-dihydroxybutyrate, so far never reported in this disease. Interestingly, patients with a disease onset as PMS evolving overtime into KSS phenotype, presented persistent and more pronounced alterations of organic acid signature than in patients with a pure KSS phenotype. Our study shows that the quantitative analysis of urinary organic acid profile represents a helpful tool for the diagnosis of PMS and for the differential diagnosis with other inherited diseases causing abnormal organic acidurias.
Asunto(s)
Acil-CoA Deshidrogenasa de Cadena Larga/deficiencia , ADN Mitocondrial/genética , Síndrome de Kearns-Sayre/orina , Errores Innatos del Metabolismo Lipídico/orina , Enfermedades Mitocondriales/orina , Enfermedades Musculares/orina , Ácido 3-Hidroxibutírico/orina , Acil-CoA Deshidrogenasa de Cadena Larga/genética , Acil-CoA Deshidrogenasa de Cadena Larga/orina , Adolescente , Niño , Preescolar , Síndromes Congénitos de Insuficiencia de la Médula Ósea , Fumaratos/orina , Glutaratos/orina , Humanos , Hidroxibutiratos/orina , Lactante , Síndrome de Kearns-Sayre/diagnóstico , Síndrome de Kearns-Sayre/genética , Ácido Láctico/orina , Errores Innatos del Metabolismo Lipídico/diagnóstico , Errores Innatos del Metabolismo Lipídico/genética , Enfermedades Mitocondriales/diagnóstico , Enfermedades Mitocondriales/genética , Enfermedades Musculares/diagnóstico , Enfermedades Musculares/genética , Ácido Pirúvico/orina , Estudios Retrospectivos , Valeratos/orinaRESUMEN
For the enantioselective and simultaneous analysis of lactate and 3-hydroxybutyrate, a validated online two-dimensional high-performance liquid chromatography system using 4-nitro-7-piperazino-2,1,3-benzoxadiazole as a fluorescent derivatization reagent has been developed. For the reversed-phase separation in the first dimension, a Capcell Pak C18 ACR column (1.5 × 250 mm, particle size 3 µm) was used, and the target fractions were isolated by their hydrophobicity. In the second dimension, a polysaccharide-coated enantioselective column, Chiralpak AD-H (2.0 × 250 mm, 5 µm), was used. The system was validated by the calibration curve, intraday precision, interday precision, and accuracy using standards and real human samples, and satisfactory results were obtained. The present method was applied to human plasma and urine, and in the plasma, trace amounts of d-lactate (8.4 µM) and l-3-hydroxybutyrate (1.0 µM), besides high levels of l-lactate (860.9 µM) and d-3-hydroxybutyrate (59.4 µM), were successfully determined. In urine, trace levels of d-lactate (3.7 µM), d-3-hydroxybutyrate (2.3 µM), and l-3-hydroxybutyrate (3.3 µM) in addition to a relatively large amount of l-lactate (15.4 µM) were observed. The present online two-dimensional high-performance liquid chromatography system is useful for the simultaneous determination of all the lactate and 3-hydroxybutyrate enantiomers in human physiological fluids, and further clinical applications are ongoing.
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Ácido 3-Hidroxibutírico/sangre , Ácido 3-Hidroxibutírico/orina , Internet , Ácido Láctico/sangre , Ácido Láctico/orina , Adulto , Cromatografía Líquida de Alta Presión , Femenino , Voluntarios Sanos , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Masculino , Estructura Molecular , Estereoisomerismo , Adulto JovenRESUMEN
Urinary tract infection is one of the most common bacterial infections worldwide. Current diagnosis of urinary tract infection chiefly relies on its clinical presentation, urine dipstick tests and urine culture. Small molecules found in bio-fluids related with both infection and recovery would facilitate diagnosis and management of UTI. Mass spectrometry-based fingerprinting of plasma and urine at 3 time points, pre-infection (t = -24h), infection (t = 24h) and post 3-day treatment (t = 112h), were acquired in the following four groups: mice which were healthy, infected but not treated, infected and treated with ciprofloxacin, and infected and treated with Relinqing® granules (n = 6 per group). A metabolomics workflow including multivariate analysis and ROC regression was employed to select metabolic features that correlated with UTI and its treatment. Circa 4,000 molecular features were acquired for each sample. The small acid 3-hydroxybutyrate in plasma was found to be differentiated for urinary tract infection, with an area under the curve = 0.97 (95% confidence interval: 0.93-1.00, accuracy = 0.91, sensitivity = 0.92 and specificity = 0.91). The level of 3-hydroxybutyrate in plasma was depleted after infection with a fold change of -22 (q < 0.0001). Correlation between plasma 3-hydroxybutyrate and urine bacterial number in all groups and time points was r = -0.753 (p < 0.0001). The findings show that 3-hydroxybutyrate is depleted in blood and strongly associated with UTI at both infection and post-treatment stage in a UTI mouse model. Further work is envisaged to assess the clinical potential of blood tests to assist with UTI management.
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Ácido 3-Hidroxibutírico/sangre , Metabolómica , Fenotipo , Infecciones Urinarias/sangre , Ácido 3-Hidroxibutírico/orina , Animales , Modelos Animales de Enfermedad , Perros , Femenino , Ratones , Factores de Tiempo , Infecciones Urinarias/orinaRESUMEN
OBJECTIVE: Pharmacologically induced glycosuria elicits adaptive responses in glucose homeostasis and hormone release, including decrements in plasma glucose and insulin levels, increments in glucagon release, enhanced lipolysis, and stimulation of ketogenesis, resulting in an increase in ketonemia. We aimed at assessing the renal response to these changes. RESEARCH DESIGN AND METHODS: We measured fasting and postmeal urinary excretion of glucose, ß-hydroxybutyrate (ß-HB), lactate, and sodium in 66 previously reported patients with type 2 diabetes and preserved renal function (estimated glomerular filtration rate ≥60 mL · min-1 · 1.73 m-2) and in control subjects without diabetes at baseline and following empagliflozin treatment. RESULTS: With chronic (4 weeks) sodium-glucose cotransporter 2 inhibition, baseline fractional glucose excretion (<2%) rose to 38 ± 12% and 46 ± 11% (fasting vs. postmeal, respectively; P < 0.0001) over a range of BMIs (range 23-41 kg/m2) and creatinine clearance (65-168 mL · min-1 · m-2). Excretion of ß-HB (median [interquartile range]: 0.08 [0.10] to 0.31 [0.43] µmol · min-1), lactate (0.06 [0.06] to 0.28 [0.25] µmol · min-1), and sodium (0.27 [0.22] to 0.36 [0.16] mEq · min-1) all increased (P ≤ 0.001 for all) and were each positively related to glycosuria (P ≤ 0.001). These parameters changed in the same direction in subjects without diabetes, but changes were smaller than in the patients with diabetes. Although plasma N-terminal pro-B-type natriuretic peptide levels were unaltered, plasma erythropoietin concentrations increased by 31 (64)% (P = 0.0078). CONCLUSIONS: We conclude that the sodium-glucose cotransporter 2 inhibitor-induced increase in ß-HB is not because of reduced renal clearance but because of overproduction. The increased lactate excretion contributes to lower plasma lactate levels, whereas the increased natriuresis may help in normalizing the exchangeable sodium pool. Taken together, glucose loss through joint inhibition of glucose and sodium reabsorption in the proximal tubule induces multiple changes in renal metabolism.
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Diabetes Mellitus Tipo 2/orina , Cetonas/metabolismo , Riñón/efectos de los fármacos , Inhibidores del Cotransportador de Sodio-Glucosa 2 , Ácido 3-Hidroxibutírico/orina , Compuestos de Bencidrilo/uso terapéutico , Glucemia , Índice de Masa Corporal , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Eritropoyetina/sangre , Femenino , Tasa de Filtración Glomerular , Glucagón/metabolismo , Glucósidos/uso terapéutico , Glucosuria/sangre , Glucosuria/orina , Humanos , Hipoglucemiantes/uso terapéutico , Riñón/metabolismo , Ácido Láctico/orina , Masculino , Persona de Mediana Edad , Natriuresis , Péptido Natriurético Encefálico/sangre , Fragmentos de Péptidos/sangre , Sodio/orina , Transportador 2 de Sodio-Glucosa/metabolismoRESUMEN
OBJECTIVE: Idiopathic sudden sensorineural hearing loss is a frequent emergency, with unknown aetiology and usually treated with empiric therapy. Steroids represent the only validated treatment but prognosis is unpredictable and the possibility to select the patients who will not respond to steroids could avoid unnecessary treatments. Metabolomic profiling of the biofluids target the analysis of the final product of genic expression and enzymatic activity, defining the biochemical phenotype of a whole biologic system. METHODS: We studied the metabolomics of the urine of a cohort of patients with idiopathic sudden sensorineural hearing loss, correlating the metabolic profiles with the clinical outcomes. Metabolomic profiling of urine samples was performed by 1H Nuclear Magnetic Resonance spectroscopy in combination with multivariate statistical approaches. RESULTS: 26 patients were included in the study: 5 healthy controls, 13 patients who did not recover after treatment at 6 months while the remaining 8 patients recovered from the hearing loss. The orthogonal partial least square-discriminant analysis score plot showed a significant separation between the two groups, responders and non-responders after steroid therapy, R2Y of 0.83, Q2 of 0.38 and p value <0.05. The resulting metabolic profiles were characterized by higher levels of urinary B-Alanine, 3-hydroxybutyrate and Trimethylamine N-oxide, and lower levels of Citrate and Creatinine in patients with worst outcome. CONCLUSION: Idiopathic sudden sensorineural hearing loss is a specific disease with unclear systemic changes, but our data suggest that there are different types of this disorder or patients predisposed to effective action of steroids allowing the recover after treatment.
Asunto(s)
Ácido 3-Hidroxibutírico/orina , Alanina/orina , Pérdida Auditiva Súbita/orina , Metabolómica , Metilaminas/orina , Adolescente , Adulto , Anciano , Anticoagulantes/uso terapéutico , Audiometría de Tonos Puros , Estudios de Casos y Controles , Niño , Ácido Cítrico/orina , Estudios de Cohortes , Creatinina/orina , Enoxaparina/uso terapéutico , Femenino , Pérdida Auditiva Súbita/terapia , Humanos , Oxigenoterapia Hiperbárica/métodos , Inmunosupresores/uso terapéutico , Masculino , Persona de Mediana Edad , Pregnenodionas/uso terapéutico , Pronóstico , Estudios Prospectivos , Espectroscopía de Protones por Resonancia Magnética , Orina/química , Adulto JovenRESUMEN
Several rapid tests for use on farm have been validated for the detection of hyperketonemia (HK) in dairy cattle, however the reported sensitivity and specificity of each method varies and no single study has compared them all. Meta-analysis of diagnostic test accuracy is becoming more common in human medical literature but there are few veterinary examples. The objective of this work was to perform a systematic review and meta-analysis to determine the point-of-care testing method with the highest combined sensitivity and specificity, the optimal threshold for each method, and to identify gaps in the literature. A comprehensive literature search resulted in 5196 references. After removing duplicates and performing relevance screening, 23 studies were included for the qualitative synthesis and 18 for the meta-analysis. The three index tests evaluated in the meta-analysis were: the Precision Xtra(®) handheld device measuring beta-hydroxybutyrate (BHB) concentration in whole blood, and Ketostix(®) and KetoTest(®) semi-quantitative strips measuring the concentration of acetoacetate in urine and BHB in milk, respectively. The diagnostic accuracy of the 3 index tests relative to the reference standard measurement of BHB in serum or whole blood between 1.0-1.4mmol/L was compared using the hierarchical summary receiver operator characteristic (HSROC) method. Subgroup analysis was conducted for each index test to examine the accuracy at different thresholds. The impact of the reference standard threshold, the reference standard method, the prevalence of HK in the population, the primary study source and risk of bias of the primary study was explored using meta-regression. The Precision Xtra(®) device had the highest summary sensitivity in whole blood BHB at 1.2mmol/L, 94.8% (CI95%: 92.6-97.0), and specificity, 97.5% (CI95%: 96.9-98.1). The threshold employed (1.2-1.4mmol/L) did not impact the diagnostic accuracy of the test. The Ketostix(®) and KetoTest(®) strips had the highest summary sensitivity and specificity when the trace and weak positive thresholds were used, respectively. Controlling for the source of publication, HK prevalence and reference standard employed did not impact the estimated sensitivity and specificity of the tests. Including only peer-reviewed studies reduced the number of primary studies evaluating the Precision Xtra(®) by 43% and Ketostix(®) by 33%. Diagnosing HK with blood, urine or milk are valid options, however, the diagnostic inaccuracy of urine and milk should be considered when making economic and treatment decisions.
Asunto(s)
Enfermedades de los Bovinos/diagnóstico , Pruebas Diagnósticas de Rutina/veterinaria , Cetosis/veterinaria , Ácido 3-Hidroxibutírico/sangre , Ácido 3-Hidroxibutírico/orina , Animales , Bovinos , Enfermedades de los Bovinos/sangre , Enfermedades de los Bovinos/orina , Pruebas Diagnósticas de Rutina/métodos , Pruebas Diagnósticas de Rutina/normas , Femenino , Cetosis/diagnóstico , Leche/química , Sistemas de Atención de Punto , Sensibilidad y EspecificidadRESUMEN
Auxiliary enzymes participate in ß-oxidation of unsaturated fatty acids. The objective of the study was to investigate the impact of a moderate zinc deficiency and a high intake of polyunsaturated fat on Δ(3)Δ(2)-enoyl-CoA isomerase (ECI) in the liver and other tissues. Five groups of eight weanling rats each were fed moderately zinc-deficient (ZD) or zinc-adequate (ZA) semisynthetic diets (7 or 50 mg Zn/kg) enriched with 22 % cocoa butter (CB) or 22 % safflower oil (SO) for 4 weeks: (1) ZD-CB, fed free choice; (2) ZA-CBR, ZA-CB diet fed in equivalent amounts consumed by the ZD-CB group; (3) ZD-SO, fed free choice; (4) ZA-SOR, ZA-SO diet fed in equivalent amounts consumed by the ZD-SO group; and (5) ZA-SO, fed free choice. Growth and Zn status markers were markedly reduced in the ZD groups. ECI activity in the liver of the animals fed the ZD- and ZA-SO diets were significantly higher (approximately 2- and 3-fold, respectively) as compared with the CB-fed animals, whereas activities in extrahepatic tissues (kidneys, heart, skeletal muscle, testes, adipose tissue) were not altered by dietary treatments. Transcript levels of the mitochondrial Eci gene in the liver did not significantly differ between ZD and ZA rats, but were 1.6-fold higher in the ZA-SO- than in the ZD-CB-fed animals (P < 0.05). It is concluded that diets enriched with safflower oil as a source high in linoleic acid induce markedly increased hepatic ECI activities and that a moderate Zn deficiency does not affect transcription of the mitochondrial Eci gene in the liver.
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Grasas de la Dieta/administración & dosificación , Dodecenoil-CoA Isomerasa/metabolismo , Hígado/efectos de los fármacos , Zinc/administración & dosificación , Ácido 3-Hidroxibutírico/sangre , Ácido 3-Hidroxibutírico/metabolismo , Ácido 3-Hidroxibutírico/orina , Animales , Peso Corporal/efectos de los fármacos , Dieta , Dodecenoil-CoA Isomerasa/genética , Ingestión de Alimentos/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Hígado/enzimología , Hígado/crecimiento & desarrollo , Masculino , Tamaño de los Órganos/efectos de los fármacos , Ratas Wistar , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Aceite de Cártamo/administración & dosificación , Succinato Deshidrogenasa/metabolismo , Destete , Zinc/deficiencia , Zinc/metabolismoRESUMEN
AIMS: Alterations in organic acid biomarkers from fatty acid and carbohydrate metabolism have been documented in type 2 diabetes patients. However, their association with gestational diabetes mellitus (GDM) is largely unknown. METHODS: Participants were 25 GDM cases and 25 non-GDM controls. Biomarkers of fatty acid (adipate, suberate and ethylmalonate) and carbohydrate (pyruvate, l-lactate and ß-hydroxybutyrate) metabolism were measured in maternal urine samples collected in early pregnancy (17 weeks) using liquid chromatography-mass spectrometry methods. Logistic regression were used to calculate odds ratios (OR) and 95% confidence intervals (CI). RESULTS: GDM cases and controls differed in median urinary concentrations of ethylmalonate (3.0 vs. 2.3µg/mg creatinine), pyruvate (7.4 vs. 2.1µg/mg creatinine), and adipate (4.6 vs. 7.3µg/mg creatinine) (all p-values <0.05). Women in the highest tertile for ethylmalonate or pyruvate concentrations had 11.4-fold (95%CI 1.10-117.48) and 3.27-fold (95%CI 0.72-14.79) increased risk of GDM compared with women in the lowest tertile for ethylmalonate and pyruvate concentrations, respectively. Women in the highest tertile for adipate concentrations, compared with women in the lowest tertile, had an 86% reduction in GDM risk (95%CI 0.02-0.97). CONCLUSIONS: These preliminary findings underscore the importance of altered fatty acid and carbohydrate metabolism in the pathogenesis of GDM.
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Ácido 3-Hidroxibutírico/orina , Biomarcadores/orina , Diabetes Gestacional/fisiopatología , Ácidos Grasos/orina , Ácido Láctico/orina , Complicaciones del Embarazo/diagnóstico , Piruvatos/orina , Adulto , Femenino , Humanos , Embarazo , Complicaciones del Embarazo/etiología , Complicaciones del Embarazo/orina , Estudios Prospectivos , Factores de RiesgoRESUMEN
OBJECTIVES: The aim of this study was to determine whether a rapid, fingerstick ß-hydroxybutyrate (ß-OHB) test predicts ketonuria in pregnant women with nausea and vomiting. METHODS: A convenience sample of 82 pregnant women who presented to the Barnes-Jewish Hospital emergency department (ED) with complaints of nausea or vomiting were enrolled into an institutional review board-approved, prospective, observational study. Exclusion criteria were temperature of >38.3°C, altered mental status, prisoners, and >1 L of any intravenous (IV) fluid before screening. Subjects had fingerstick ß-OHB tests, with results reported in increments of 0.1 mmol/L. Urine ketone results were made available once reported as part of standard care and varied from 0 (trace) to +4 values. Ketonuria was defined as a urine ketone value of 3+ or 4+ and was analyzed as a dichotomous, categorical variable. A contingency table and receiver operator characteristic (ROC) curve were constructed for comparing the ß-OHB values to those urine ketone values for each patient. RESULTS: Mean (±SD) ß-OHB was 0.43 (±0.52) mmol/L, and median urine was 1+ for the cohort as a whole. The area under the ROC curve equaled 0.95. The sensitivity and specificity for a fingerstick test of >0.45 mmol/L were 89 and 94.5%, respectively. The positive likelihood ratio (LR) was 16, and the negative LR was 0.12. CONCLUSIONS: Fingerstick ß-OHB is a rapid and reliable diagnostic tool that correlates well with ketonuria and identifies ketonemia in pregnant women with nausea and vomiting. Fingerstick ß-OHB testing has the potential to increase triage efficiency, shorten length-of-stay times, and positively affect patient outcomes in an ED setting.
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Ácido 3-Hidroxibutírico/sangre , Cetosis/diagnóstico , Náusea/diagnóstico , Vómitos/diagnóstico , Ácido 3-Hidroxibutírico/orina , Adulto , Femenino , Humanos , Cetosis/sangre , Cetosis/orina , Embarazo , Estudios Prospectivos , Curva ROC , Sensibilidad y EspecificidadRESUMEN
Human cytomegalovirus (HCMV) represents one of the most significant viral risks of birth defects and long-term sequelae. The severity of the infection depends on the form of the disease, which can be symptomatic or asymptomatic with or without sequelae. The aim of this study was to investigate in a population of newborns the impact of HCMV infection on the urine metabolome by using (1)H-nuclear magnetic resonance (NMR) spectroscopy combined with multivariate statistical analysis. Twenty-three children born from women with a primary HCMV infection during pregnancy were recruited. Twelve were HCMV infected at birth whereas eleven were not infected (control). The (1)H-NMR spectra were analyzed using a PLS-DA mathematical model in order to identify the discriminant metabolites between the asymptomatic and the control group. The most important metabolites characterizing the clustering of the samples were: myoinositol, glycine, 3-hydroxybutyrate, 3-aminoisobutyrate, creatine, taurine and betaine. These findings suggest the use of metabolomics as a useful new tool in the investigation of HCMV congenital infection.
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Infecciones por Citomegalovirus/congénito , Infecciones por Citomegalovirus/orina , Transmisión Vertical de Enfermedad Infecciosa , Metabolómica/métodos , Complicaciones Infecciosas del Embarazo/metabolismo , Ácido 3-Hidroxibutírico/orina , Adulto , Ácidos Aminoisobutíricos/orina , Betaína/orina , Creatina/orina , Citomegalovirus/aislamiento & purificación , Infecciones por Citomegalovirus/diagnóstico , Análisis Discriminante , Femenino , Glicina/orina , Humanos , Recién Nacido , Inositol/orina , Espectroscopía de Resonancia Magnética , Embarazo , Complicaciones Infecciosas del Embarazo/diagnóstico , Complicaciones Infecciosas del Embarazo/virología , Taurina/orinaRESUMEN
Early diagnosis of diabetic nephropathy (DN) is difficult although it is of crucial importance to prevent its development. To probe potential markers and the underlying mechanism of DN, an animal model of DN, the db/db mice, was used and serum and urine metabolites were profiled using gas chromatography/time-of-flight mass spectrometry. Metabolic patterns were evaluated based on serum and urine data. Principal component analysis of the data revealed an obvious metabonomic difference between db/db mice and controls, and db/db mice showed distinctly different metabolic patterns during the progression from diabetes to early, medium, and later DN. The identified metabolites discriminating between db/db mice and controls suggested that db/db mice have perturbations in the tricarboxylic acid cycle (TCA, citrate, malate, succinate, and aconitate), lipid metabolism, glycolysis, and amino acid turnover. The db/db mice were characterized by acidic urine, high TCA intermediates in serum at week 6 and a sharp decline thereafter, and gradual elevation of free fatty acids in the serum. The sharp drop of serum TCA intermediates from week 6 to 8 indicated the downregulated glycolysis and insulin resistance. However, urinary TCA intermediates did not decrease in parallel with those in the serum from week 6 to 10, and an increased portion of TCA intermediates in the serum was excreted into the urine at 8, 10, and 12 wk than at 6 wk, indicating kidney dysfunction occurred. The relative abundances of TCA intermediates in urine relative to those in serum were suggested as an index of renal damage.
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Ciclo del Ácido Cítrico/fisiología , Nefropatías Diabéticas/sangre , Nefropatías Diabéticas/orina , Cromatografía de Gases y Espectrometría de Masas , Ácido 3-Hidroxibutírico/sangre , Ácido 3-Hidroxibutírico/orina , Albuminuria/orina , Alantoína/orina , Aminoácidos/metabolismo , Animales , Ácido Araquidónico/sangre , Biomarcadores , Colesterol/sangre , Ácido Cítrico/sangre , Glucólisis , Concentración de Iones de Hidrógeno , Metabolismo de los Lípidos , Lisina/sangre , Malatos/orina , Masculino , Metabolómica , Ratones , Ratones Endogámicos C57BL , Ácido Succínico/orina , OrinaRESUMEN
BACKGROUND: Alcohol ketoacidosis is a frequently missed diagnosis, but is well described in the literature. We present a case of ketoacidosis, likely alcohol ketoacidosis, in a 40 y-old chronic alcoholic patient. The detection of trace serum isopropanol prompted a discussion of alcohol ketoacidosis versus toxic isopropanol ingestion or a combination of both, including comparisons with citations in current literature. METHODS: The automated instruments used to analyze the patient's urine, blood, and serum samples are described. RESULTS: The initial impression was severe metabolic acidosis with an increased anion gap and normal serum glucose and whole blood lactate. Testing for potential toxic ingestions detected only increased serum acetone and trace serum isopropanol. A urinalysis positive for ketones and an increased serum ß-hydroxybutyrate concentration clenched the diagnosis of ketoacidosis. CONCLUSION: Ketoacidosis with an increased anion gap in the absence of hyperglycemia or glycosuria in a chronic alcoholic patient should prompt the evaluation for alcohol ketoacidosis. Trace serum isopropanol may be worrisome for a toxic ingestion, but this finding in severe ketoacidosis may be explained by the reversible action of the enzyme alcohol dehydrogenase. Markedly increased serum isopropanol with a low serum acetone:isopropanol ratio would be more indicative of a toxic isopropanol ingestion.