In this study, the Autokit Total Ketone Bodies kit (Wako Pure Chemical, Japan), a total ketone measurement assay using an enzymatic method, was evaluated using a Roche Cobas e702 instrument (Roche Diagnostics, Germany). Precision, linearity, carryover, and reference range verification were evaluated with reference to Clinical Laboratory Standards Institute guidelines. Standard materials provided by the manufacturer and patient samples were used for the evaluation. The precision and carryover of the evaluation result satisfied the acceptance criteria. Linearity was also acceptable at more than 0.99. The quantitative Autokit Total Ketone Bodies kit is precise, and can be widely used in clinical laboratories.
3-Hydroxybutyric Acid ; Evaluation Studies as Topic ; Humans ; Ketone Bodies* ; Methods ; Reference Values

PURPOSE: Ketogenic diet (KD) remains a therapy in search of explanation although it is an established treatment for patients with intractable epilepsy. It has been clinically proven more efficacious at younger ages, presumably because of the enhanced ability of the immature brain to extract and utilize ketone bodies. The study was designed to investigate whether ketosis induced by the KD is age-dependent. METHODS: A KD ([fat]:[protein+carbohydrate] ratio of 4.3:1) was administered to male Sprague-Dawley rats for 3 weeks, while control animals were fed a standard rodent chow. Dietary treatment was initiated at either postnatal 3 or 12 weeks. Blood beta-hydroxybutyrate (BHB) levels were assayed from blood obtained via the tail vein with the Keto-SiteTM reflectance meter and test cards on treatment day 21. RESULTS: Blood BHB levels in the KD-treated group were significantly higher than those in the control group in 3 week-old rats (4.18+/-0.62 [n=30] vs. 0.27+/-0.02 [n=30] mM, respectively; p<0.0001) and 12 week-old rats (0.86+/-0.06 [n=30] vs. 0.22+/-0.02 [n=30] mM, respectively; p<0.0001). In the KD-treated groups, blood BHB levels of 3 week-old animals were significantly higher than those of 12 week-old ones (p<0.0001), whereas in the control groups, no significant differences in blood BHB levels between the two age groups (p>0.05). CONCLUSIONS: The present study demonstrates that the KD induces more severe ketosis in younger rats. Age-dependent differences in the degree of ketosis induced by the KD may explain that the diet is clinically more efficacious at younger ages.
3-Hydroxybutyric Acid ; Animals ; Blood Group Antigens ; Brain ; Diet ; Epilepsy ; Humans ; Ketogenic Diet* ; Ketone Bodies ; Ketosis* ; Male ; Rats ; Rats, Sprague-Dawley ; Rodentia ; Veins

BACKGROUND: Diabetes mellitus and alcohol consumption are the most common causes of ketoacidosis in adults. Recently, beta-hydroxybutyric acid (betaHBA) was reported to be a potential serum biomarker in the diagnosis and monitoring of ketoacidosis. We evaluated the performance of T-KB-H and 3-HB kits for the measurement of ketone bodies [acetoacetate (AcAc)+betaHBA] and betaHBA, respectively. METHODS: Quantitative enzymatic assays were performed using the T-KB-H and 3-HB kits (Nittobo Medical Co., Japan) and the Architect ci16200 Integrated System (Abbott Laboratories, USA). Simultaneously, the ketone body levels in these serum samples were determined by gas chromatography-mas spectrometry (GC-MS). We evaluated precision and linearity of these kits and correlation with GC-MS, and established reference intervals in children and adults. RESULTS: The coefficients of variation for the T-KB-H and 3-HB kits were less than 4.0% at analyte levels of 50, 100, and 400 micromol/L. Linearity was observed for AcAc and betaHBA over a 0-1,000 micromol/L range (R2<0.99). Results from the T-KB-H and 3-HB kits were in good agreement with those from the GC-MS analysis, with correlation coefficients of 0.94 for AcAc and 0.96 for betaHBA. Reference intervals determined for the T-KB-H kit were 9.8-270.1 micromol/L and 18.5-531.8 micromol/L in children and adults, respectively. For the 3-HB kit, the reference intervals were 6.4-234.0 micromol/L and 16.0-437.2 micromol/L in children and adults, respectively. CONCLUSIONS: The T-KB-H and 3-HB kits displayed good precision, clinically acceptable linearity, and reliable correlation with an established assay. This indicates that the kits can be used clinically for measuring serum ketone bodies.
3-Hydroxybutyric Acid* ; Adult ; Alcohol Drinking ; Child ; Diabetes Mellitus ; Diabetic Ketoacidosis ; Diagnosis ; Enzyme Assays ; Gas Chromatography-Mass Spectrometry ; Humans ; Ketone Bodies ; Ketosis ; Spectrum Analysis

PURPOSE: The ketogenic diet(KD) has been felt to be clinically more efficacious at younger ages, presumably because of the enhanced ability of the immature brain to extract and utilize ketone bodies. The present study was designed to investigate age-dependent effects of the KD on pentylenetetrazole(PTZ)-seizure severity in rats. METHODS: A KD([fat]:[protein+carbohydrate] ratio of 4.3:1) was administered to male Sprague-Dawley rats for 3 weeks, while control animals were fed a standard rodent chow. Dietary treatment was initiated at either postnatal 9 or 12 weeks. Seizures were chemically induced by intraperitoneal injection of PTZ(60 mg/kg) and blood beta-hydroxybutyrate (BHB) levels were assayed on treatment day 21. Seizure severity was evaluated by using a scoring system of seizure behaviors:0, no seizure; 0.5, abnormal behavior; 1, myoclonic jerk; 2, myoclonic jerk with jumping; 3, forelimb clonus with preserving righting reflex; 4, generalized clonic seizure with brief loss of righting reflex; 5, generalized tonic clonic seizure; 6, expire. A greater score represents a more severe seizure. RESULTS: In 9 weeks old rats, the mean(+/-SEM) seizure behavior scores were 3.5+/-1.2 [n=19] and 4.4+/-0.9[n=17] for the KD-treated and control groups, respectively(P<0.05), whereas in 12 weeks old animals, no significant differences in seizure behavior scores between the two groups(3.9+/-0.3[n=17] vs. 4.1+/-0.3[n=16], respectively). Blood BHB levels in the KD-treated group were significantly higher than those of the control group in 9 (1.21+/-0.14[n=19] vs. 0.14+/-0.12[n=17] mM, respectively; P<0.001) and 12(0.64+/-0.08[n=17] vs. 0.18+/-0.02[n=16] mM, respectively; P<0.001) weeks old animals. CONCLUSION: The KD was previously reported to decrease PTZ-seizure severity in 3 weeks old rats. In this study, the KD decreases PTZ-seizure severity in 9 weeks old rats, but is ineffective in 12 weeks old rats. These results parallel clinical experience, where the beneficial effects of the KD are felt to be age-dependent.
3-Hydroxybutyric Acid ; Animals* ; Brain ; Forelimb ; Humans ; Injections, Intraperitoneal ; Ketogenic Diet* ; Ketone Bodies ; Male ; Models, Animal* ; Myoclonus ; Pentylenetetrazole ; Rats ; Rats, Sprague-Dawley ; Reflex, Righting ; Rodentia ; Seizures

3-Hydroxybutyric Acid ; blood ; Animals ; Dietary Carbohydrates ; administration & dosage ; Dietary Fats ; administration & dosage ; Dietary Proteins ; administration & dosage ; Disease Models, Animal ; Epilepsy ; diet therapy ; metabolism ; pathology ; Hippocampus ; metabolism ; Kainic Acid ; Ketone Bodies ; metabolism ; Male ; Mossy Fibers, Hippocampal ; pathology ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, Kainic Acid ; analysis ; genetics

BACKGROUND: Urine ketone test is commonly used to screen for diabetic ketoacidosis (DKA). Ketonuria also develops in patients with disease conditions other than DKA. However, the prevalence of DKA in patients with ketonuria is not known. We investigated the prevalence of ketonuria and characteristics of patients with ketonuria and estimated the prevalence of DKA among them to study the clinical significance of ketonuria as an indicator of DKA. METHODS: We studied 1,314 adult and 1,027 pediatric patients who underwent urinalysis. The prevalence of ketonuria in the different groups of patients, classified according to the types of their visits to the institution, was investigated, and the relationships between ketonuria and albuminuria, glycosuria, and bilirubinuria were evaluated. RESULTS: The overall prevalence of ketonuria was 9.1%. The prevalences of ketonuria in adult and pediatric patients were 4.3% and 15.2%, respectively. The prevalences of ketonuria were the highest in the adult (9.7%) and pediatric (28%) patients in the group that had visited the emergency department. Among patients with ketonuria, 7% adult and 3.8% pediatric patients showed glycosuria. CONCLUSIONS: This study showed that the prevalence of DKA in patients with ketonuria, defined as the simultaneous presence of ketone bodies and glucose in urine, was only 7%. Therefore, we concluded that ketonuria might be clinically significant as an indicator of acute or severe disease status rather than of DKA.
Adult ; Albuminuria ; Diabetic Ketoacidosis ; Emergencies ; Glucose ; Glycosuria ; Humans ; Ketone Bodies ; Ketosis ; Prevalence ; Urinalysis

Epilepsy ; diet therapy ; metabolism ; Food-Drug Interactions ; Humans ; Ketone Bodies ; biosynthesis

Heart failure (HF) has become a major challenge in the treatment of global cardiovascular diseases. Great progress has been made in the drug treatment of HF, however, rehospitalization rate and mortality of patients with HF are still high. Hence, there is an urgent need to explore new treatment strategy and new underlying pathogenic mechanisms. In recent years, some researchers have suggested that regulation of ketone body metabolism may become a potentially promising therapeutic approach for HF. Some studies showed that the oxidative utilization of fatty acids and glucose was decreased in the failing heart, accompanied by the increase of ketone body oxidative metabolism. The enhancement of ketone body metabolism in HF is a compensatory change during HF. The failing heart preferentially uses ketone body oxidation to provide energy, which helps to improve the body's cardiac function. This review will discuss the potential significance of ketone body metabolism in the treatment of HF from three aspects: normal myocardial ketone body metabolism, the change of ketone body metabolism in HF, the effect of ketogenic therapy on HF and its treatment.
Humans ; Heart Failure/metabolism* ; Myocardium/metabolism* ; Ketone Bodies/metabolism* ; Cardiovascular Diseases ; Fatty Acids/metabolism* ; Energy Metabolism

PURPOSE: The ketogenic diet (KD) remains a therapy in search of explanation although it is an established treatment for patients with intractable seizures. It was designed to mimic the biochemical changes seen upon fasting, specifically the formation of ketone bodies: acetoacetate (ACA), beta-hydroxybutyrate (BHB), and to a lesser extent, acetone. The present study was designed to investigate the protective effect of BHB on flurothyl-induced seizures in rats. METHODS: Thirty-four male Sprague-Dawley rats were divided into two equal groups. Experimental rats (n=17) were injected intraperitoneally with BHB (20 mmol/kg), while control animals (n=17) with normal saline. Fifteen minutes later, seizures were chemically induced by flurothyl infusion (40 mL/min). Seizure susceptibility was defined as the latency from the start of flurothyl infusion to the onset of a generalized seizure. Shorter latencies reflected greater seizure susceptibility. RESULTS: The mean (+/- SEM) latency to the onset of a generalized seizure in the experimental animals treated with BHB was 476.5 +/- 13.9 seconds, which was significantly longer (P < 0.05) than the control (438.0 +/- 10.5 seconds). CONCLUSION: This study demonstrated the significant decrease in flurothyl-induced seizure susceptibility in rats treated with BHB. Our results suggest that BHB may be directly anticonvulsant.
3-Hydroxybutyric Acid ; Acetoacetates ; Acetone ; Animals ; Fasting ; Flurothyl ; Humans ; Hydrazines ; Ketogenic Diet ; Male ; Rats ; Rats, Sprague-Dawley ; Seizures

During hemorrhagic shock, liver is susceptible to ischemia and decreased hepatic energy charge results in decreasing arterial ketone body ratio(AKBR). Reperfusion after hemorrhagic shock can greatly amplify the generation of toxic oxygen metabolites. As a result, the fluxes of these highly toxic metabolites can overwhelm the endogenous antioxident defense mechanisms and lead to tissue injury. In order to observe the effect of glutathione(GSH) on the AKBR in hemorrhagic shock, dogs(n=16) were anesthetized with 1% enflurane in 02. We pretreated glutathione (100 mg/kg) intravenously before hemorrhagic shock in glutathione (GSH) group (n=8). Shock was induced with bleeding and mean arterial pressure was maintained 50 mmHg for 30 minutes. Recovery from shock was done with transfusion of preserved blood and maintained for 30 minutes. We measured arterial ketone bodies and ketone body ratio before, during and after shock, and compared them to control group (n=8) which was not pretreated with glutathione. AKBR during and after hemorrhagic shock in GSH group (0.8 and 1.0) were higher than those in control group (0.5 and 0.8). Light microscopic examination of liver biopsy revealed less portal degeneration during and after hemorrhagic shock in GSH group than control group. Pharmacologic modulation of hepatocytic function with glutathione before hemorrhagic shock has shown some beneficial effect with protection of decreased AKBR and histological change during and after hemorrhagic shock.
Animals ; Arterial Pressure ; Biopsy ; Defense Mechanisms ; Dogs* ; Enflurane ; Glutathione* ; Hemorrhage ; Ischemia ; Ketone Bodies ; Liver ; Oxygen ; Reperfusion ; Shock ; Shock, Hemorrhagic*