OBJECTIVETo establish of blood beta hydroxybutyrate (βOHB) threshold for diagnosing type 2 diabetes ketoacidosis (DKA) and explore the relationship between βOHB levels and the severity of DKA.

METHODSCorrelation analysis was performed between serum βOHB and [HCO(3)] in type 2 diabetic patients admitted in the emergency department in the past year. Regression equation was used to calculate the concentration of βOHB corresponding to a [HCO(3)] level of 18.0, 15, and 10.0 mmol/L, and βOHB concentration corresponding to a [HCO(3)] level of 18.0 mmol/l was used as the DKA diagnostic threshold.

RESULTSThe serum βOHB level and [HCO3] concentration showed a good correlation (R²=0.7023, P<0.001). βOHB concentrations that corresponded to a [HCO(3)] level of 18.0, 15, and 10.0 mmol/L were 3.0, 4.70, and 7.5 mmol/L, respectively, in accordance with the severity of DKA. Combined with the blood glucose concentration ≥ 13.9 mmol/L, a blood βOHB≥3.0 mmol/L showed a sensitivity of 99%, specificity of 86%, and total effectiveness of 92.81% for diagnosing DKA.

CONCLUSIONA serum βOHB level above 3.0 mmol/L can be used as the diagnostic threshold of DKA. βOHB can serve as an index for assessing the severity of DKA.

3-Hydroxybutyric Acid ; blood ; Diabetes Mellitus, Type 2 ; blood ; Diabetic Ketoacidosis ; blood ; diagnosis ; Humans ; Sensitivity and Specificity ; Severity of Illness Index

OBJECTIVE: To investigate the inhibitory effect of ketogenic diet (KD) on growth of neuroblastoma in mice. METHODS: BALB/c-nu mouse models bearing neuroblastoma xenografts were established by subcutaneous injection of human neuroblastoma cell line (SH-SY5Y). When the tumor volume reached 250 mm3, the mice were randomized into SD group with standard diet and PBS treatment, KD group with ketogenic diet and PBS treatment, and CP+KD group with ketogenic diet and cyclophosphamide (60 mg·kg·day) treatment, =8. The tumor volume, body weight, blood glucose, ketone body (β-Hydroxybutyrate) levels, and hepatic steatosis in the mice were assessed. The expressions of caspase-3 and caspase-8 were detected by Western blotting, and Ki67 expresison was detected using immunohistochemistry (IHC). Transmission electron microscopy (TEM) was employed for the autophagosomes, and the autophagic protein Beclin1, LC3A/B and P62 were detected by IHC and Western blotting. RESULTS: On day 28 post tumor cell injection, the mice in KD and CP+KD groups could prolong the overall survival rates than that in SD group ( < 0.001). On day 22 post the injection, the tumor volume in KD group was smaller than that in SD group ( < 0.05); on 16, 19, and 22 day post the injection, the tumor volume in CP+KD group was smaller than that in SD group ( < 0.01). The mice in SD group showed greater body weight on day 19 and higher blood glucose level on day 13 post the injection than those in the other two groups ( < 0.05). Blood ketone level and hepatic steatosis score were higher and glucose ketone index (GKI) was lower in KD and CP+KD groups than those in SD group (all < 0.05). The expressions of Ki67 and apoptotic proteins were detected in the tumor tissues of all groups. TEM revealed more autophagosomes in the tumor tissues of KD group than that of SD group. P62 expression was lowered ( < 0.01) and Beclin1 and LC3A/B expressions were up-regulated in the tumor tissues of KD group ( < 0.05), which is consisitent with IHC. CONCLUSIONS KD has a strong anti-tumor effect in the xenograft mouse model possibly by regulating cell autophagy.
3-Hydroxybutyric Acid ; Animals ; Blood Glucose ; Cell Line, Tumor ; Diet, Ketogenic ; Humans ; Mice ; Mice, Inbred BALB C ; Neuroblastoma

OBJECTIVETo evaluate an enzymatic method for determining serum beta-hydroxybutyrate (beta-HB) with the National Committee for Clinical Laboratory Standards (NCCLS) projects, and to discuss its clinical values in diabetic ketoacidosis (DKA).

METHODSThe precision, accuracy, specificity, linearity and interference of the enzymatic method were analyzed. This method was used to determine serum beta-HB in 60 cases of normals, 50 cases of diabetes, and 34 cases of DKA by autochemistry analyzer.

RESULTSEnzymatic beta-HB assay was precise (within-run CV, day-to-day CV, and total CV < 5%). The linearity studies showed the method was linear up to 4 mmol/L. Recovery rate was 98.5%-104.1%. Hemolysis (Hemoglobin up to 18.2 g/L), icteric samples with total bilirubin up to 224 mumol/L, and lipemia up to triglyceride concentration of 2.28 mmol/L did not interfere with the beta-HB results in this method. Serum beta-HB levels were significantly elevated in DKA patients compared with DM patients and controls (P < 0.01). Positive rate of serum beta-HB in DKA patients was significantly higher than that of urinary ketone (P < 0.05).

CONCLUSIONSEnzymatic method is convenient and reliable, allows full automation, and is rapid enough to be used for both routine and urgent determinations of serum beta-HB. It can be used in diagnosing and monitoring treatment of DKA.

3-Hydroxybutyric Acid ; blood ; Adolescent ; Adult ; Autoanalysis ; Diabetes Mellitus ; blood ; Diabetic Ketoacidosis ; blood ; Evaluation Studies as Topic ; Female ; Humans ; Male ; Middle Aged

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

In human medicine, diagnosis of diabetic ketoacidosis (DKA) is usually based on measurement of capillary 3-beta-hydroxybutyrate (3-HB) with a hand held ketone sensor. This study was conducted to determine if measurement of capillary 3-HB could be useful for the diagnosis and monitoring of canine DKA. Fifteen dogs with diabetic ketosis and 10 with DKA were evaluated. Paired measurements of 3-HB of capillary and venous blood samples were analysed by the electrochemical sensor and reference method. Use of capillary 3-HB measurement during DKA management was then evaluated through simultaneous measurements of capillary 3-HB, urinary AcAc and venous blood gas analysis. Good agreement between capillary and venous 3-HB measurement was detected by the electrochemical sensor and reference method. Monitoring treatment of DKA revealed a significant correlation between capillary 3-HB and acidosis markers, while no significant correlation was observed between AcAc and acidosis markers. A cut-off value of capillary blood 3-HB >3.8 mmol/L for diagnosis of DKA resulted in 70% and 92% sensitivity and specificity. The electrochemical sensor accurately measures 3-HB concentration in both capillary and venous blood samples, is accurate in diagnosing canine DKA, and appears to reflect the patient's metabolic status during DKA treatment.
3-Hydroxybutyric Acid/blood/*diagnostic use ; Animals ; Blood Chemical Analysis/standards/*veterinary ; Blood Specimen Collection/instrumentation/*veterinary ; Capillaries/chemistry ; Diabetic Ketoacidosis/diagnosis/therapy/*veterinary ; Dog Diseases/*diagnosis/therapy ; Dogs ; Electrochemical Techniques/instrumentation/*veterinary

BACKGROUNDA major shortcoming in tissue engineered blood vessels (TEBVs) is the lack of healthy and easily attainable smooth muscle cells (SMCs). Smooth muscle progenitor cells (SPCs), especially from peripheral blood, may offer an alternative cell source for tissue engineering involving a less invasive harvesting technique.

METHODSSPCs were isolated from 5-ml fresh rat peripheral blood by density-gradient centrifugation and cultured for 3 weeks in endothelial growth medium-2-MV (EGM-2-MV) medium containing platelet-derived growth factor-BB (PDGF BB). Before seeded on the synthesized scaffold, SPC-derived smooth muscle outgrowth cell (SOC) phenotypes were assessed by immuno-fluorescent staining, Western blot analysis, and reverse transcription polymerase chain reaction (RT-PCR). The cells were seeded onto the silk fibroin-modified poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (SF-PHBHHx) scaffolds by 6x10(4) cells/cm2 and cultured under the static condition for 3 weeks. The growth and proliferation of the seeded cells on the scaffold were analyzed by 3-(4,5-dimethylthiazol-2-yl)-diphenyltetrazolium bromide (MTT) assay, scanning electron microscope (SEM), and 4,6-diamidino-2-phenylindole (DAPI) staining.

RESULTSSOCs displayed specific "hill and valley" morphology, expressed the specific markers of the SMC lineage: smooth muscle (SM) alpha-actin, calponin and smooth muscle myosin heavy chain (SM MHC) at protein and messenger ribonucleic acid (mRNA) levels. RT-PCR results demonstrate that SOCs also expressed smooth muscle protein 22alpha (SM22alpha), a contractile protein, and extracellular matrix components elastin and matrix Gla protein (MGP), as well as vascular endothelial growth factor (VEGF). After seeded on the SF-PHBHHx scaffold, the cells showed excellent metabolic activity and proliferation.

CONCLUSIONSPCs isolated from peripheral blood can be differentiated into the SMCs in vitro and have an impressive growth potential in the biodegradable synthesized scaffold. Thus, SPCs may be a promising cell source for constructing TEBVs.

3-Hydroxybutyric Acid ; chemistry ; Animals ; Blood Vessels ; cytology ; Caproates ; chemistry ; Cell Adhesion ; Cell Differentiation ; Cell Proliferation ; Immunophenotyping ; Microscopy, Electron, Scanning ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; RNA, Messenger ; analysis ; Rats ; Tissue Engineering ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETo investigate the mechanism of beta-cell dysfunction induced by glucolipotoxicity in high fat-fed obese rats.

METHODSEighteen high-fat obese male Wistar rats were assigned into 3 groups and underwent 48-hour infusion through the jugular vein with normal saline (n=6), 20% intralipid + heparin (FFA group, n=6), or 25%glucose +20% intralipid + heparin (GS-FFA group, n=6). The plasma beta-hydroxybutyric acid (beta-HBA) was measured before and at the end of the infusion. After the infusion, the rats were sacrificed following an intravenous glucose tolerance test (IVGTT) to remove the tail of the pancreas for detection of apoptotic islet cells using TUNEL method. Immunohistochemical staining was performed to detect the expression of cytochrome c (cyt c), apoptosis-inducing factor (AIF), caspase-9 and caspase-3 in the islet cells.

RESULTSAt the end of the infusion, all the rats exhibited increased plasma beta-HBA levels, which was the highest in the GS-FFA group (P<0.05). IVGTT performed after the infusion showed a significantly lower insulinogenic index in GS-FFA group than that in NS and FFA groups. Greater number of apoptotic islet cells was found in the GS-FFA group than in the FFA and NS groups (P<0.05), and the islets had significantly higher levels of cyt c, AIF, caspase-9 and caspase-3 in the former group than in the latter two groups (P<0.05).

CONCLUSIONSHyperglycemia and high free fatty acid level synergistically impair insulin secretions to cause ketone overproduction in high fat-fed obese rats. The beta-cell dysfunction due to glucolipotoxicity is associated with increased beta-cell apoptosis and activation of mitochondrial apoptotic pathway.

3-Hydroxybutyric Acid ; blood ; Animals ; Apoptosis ; drug effects ; Fat Emulsions, Intravenous ; pharmacology ; Glucose ; pharmacology ; Glucose Tolerance Test ; Insulin-Secreting Cells ; cytology ; pathology ; Male ; Mitochondria ; drug effects ; Obesity ; physiopathology ; Rats ; Rats, Wistar

Blood indicators are used as a tool to diagnose metabolic disorders. The present work was conducted to study the relationships among blood indicators of lipomobilization and hepatic function in high-yielding dairy cows. Two groups of Holstein cows were studied: 27 early lactation cows and 14 mid lactation cows from four different herds with similar husbandry characteristics in Galicia, Spain. Blood samples were obtained to measure beta-hydroxybutyrate (BHB), non-esterified fatty acids (NEFA), triglycerides (TG), and the activity of aspartate transaminase (AST) and gamma-glutamyl transferase. Cows in early lactation had higher levels of BHB and NEFA than mid lactation cows. High lipomobilization (NEFA > 400 micromol/L) was detected in 67% and 7% of early lactation and mid lactation cows, respectively, while subclinical ketosis (BHB > 1.2 mmol/L) was detected in 41% and 28% of the early lactation and lactation cows, respectively. TG concentrations were low in all cows suffering subclinical ketosis and in 61% of the cows with high lipomobilization. During early lactation, 30% of cows suffered hepatic lipidosis as detected by levels of AST. Compromised hepatic function was observed in early lactation cows as shown by lower concentrations of glucose, total protein, and urea.
3-Hydroxybutyric Acid/blood/diagnostic use ; Animals ; Aspartate Aminotransferases/blood/diagnostic use ; Blood Glucose/analysis/metabolism ; Blood Proteins/analysis/diagnostic use ; Cattle ; Cattle Diseases/blood/*diagnosis ; Fatty Acids, Nonesterified/blood/diagnostic use ; Female ; Ketosis/blood/diagnosis/*veterinary ; Lactation ; *Lipid Mobilization ; Lipidoses/blood/diagnosis/*veterinary ; Liver Function Tests/veterinary ; Spain ; Triglycerides/blood/diagnostic use ; Urea/blood/diagnostic use ; gamma-Glutamyltransferase/blood/diagnostic use

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

PURPOSE: The ketogenic diet has long been used to treat epilepsy, but its mechanism is not yet clearly understood. To explore the potential mechanism, we analyzed the changes in gene expression induced by the ketogenic diet in the rat kainic acid (KA) epilepsy model. MATERIALS AND METHODS: KA-administered rats were fed the ketogenic diet or a normal diet for 4 weeks, and microarray analysis was performed with their brain tissues. The effects of the ketogenic diet on cathepsin E messenger ribonucleic acid (mRNA) expression were analyzed in KA-administered and normal saline-administered groups with semi-quantitative and real-time reverse transcription polymerase chain reaction (RT-PCR). Brain tissues were dissected into 8 regions to compare differential effects of the ketogenic diet on cathepsin E mRNA expression. Immunohistochemistry with an anti-cathepsin E antibody was performed on slides of hippocampus obtained from whole brain paraffin blocks. RESULTS: The microarray data and subsequent RT-PCR experiments showed that KA increased the mRNA expression of cathepsin E, known to be related to neuronal cell death, in most brain areas except the brain stem, and these increases of cathepsin E mRNA expression were suppressed by the ketogenic diet. The expression of cathepsin E mRNA in the control group, however, was not significantly affected by the ketogenic diet. The change in cathepsin E mRNA expression was greatest in the hippocampus. The protein level of cathepsin E in the hippocampus of KA-administered rat was elevated in immunohistochemistry and the ketogenic diet suppressed this increase. CONCLUSION: Our results showed that KA administration increased cathepsin E expression in the rat brain and its increase was suppressed by the ketogenic diet.
3-Hydroxybutyric Acid/blood ; Animals ; Cathepsin E/genetics/*metabolism ; Enzyme Activators/pharmacology ; *Gene Expression Regulation, Enzymologic/drug effects ; Hippocampus/*drug effects/*metabolism ; Immunohistochemistry ; Kainic Acid/*pharmacology ; *Ketogenic Diet ; Male ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction