Bicarbonate Unlocks the Ergogenic Action of Ketone Monoester Intake in Endurance Exercise.

PURPOSE: We recently reported that oral ketone ester (KE) intake before and during the initial 30 min of a 3 h 15 min simulated cycling race (RACE) transiently decreased blood pH and bicarbonate without affecting maximal performance in the final quarter of the event. We hypothesized that acid-base disturbances due to KE overrules the ergogenic potential of exogenous ketosis in endurance exercise. METHODS: Nine well-trained male cyclists participated in a similar RACE consisting of 3 h submaximal intermittent cycling (IMT180') followed by a 15-min time trial (TT15') preceding an all-out sprint at 175% of lactate threshold (SPRINT). In a randomized crossover design, participants received (i) 65 g KE, (ii) 300 mg·kg-1 body weight NaHCO3 (BIC), (iii) KE + BIC, or (iv) a control drink (CON), together with consistent 60 g·h-1 carbohydrate intake. RESULTS: KE ingestion transiently elevated blood D-ß-hydroxybutyrate to ~2-3 mM during the initial 2 h of RACE (P < 0.001 vs CON). In KE, blood pH concomitantly dropped from 7.43 to 7.36 whereas bicarbonate decreased from 25.5 to 20.5 mM (both P < 0.001 vs CON). Additional BIC resulted in 0.5 to 0.8 mM higher blood D-ß-hydroxybutyrate during the first half of IMT180' (P < 0.05 vs KE) and increased blood bicarbonate to 31.1 ± 1.8 mM and blood pH to 7.51 ± 0.03 by the end of IMT180' (P < 0.001 vs KE). Mean power output during TT15' was similar between KE, BIC, and CON at ~255 W but was 5% higher in KE + BIC (P = 0.02 vs CON). Time to exhaustion in the sprint was similar between all conditions at ~60 s (P = 0.88). Gastrointestinal symptoms were similar between groups. DISCUSSION: The coingestion of oral bicarbonate and KE enhances high-intensity performance at the end of an endurance exercise event without causing gastrointestinal distress.

Exogenous Ketosis Impairs 30-min Time-Trial Performance Independent of Bicarbonate Supplementation.

PURPOSE: We recently demonstrated that coingestion of NaHCO3 to counteract ketoacidosis resulting from oral ketone ester (KE) intake improves mean power output during a 15-min time trial (TT) at the end of a 3-h cycling race by ~5%. This ergogenic effect occurred at a time when blood ketone levels were low, as ketosis was only induced during the initial ~2 h of the race. Therefore, in the current study, we investigated whether performance also increases if blood ketone levels are increased in the absence of ketoacidosis during high-intensity exercise. METHODS: In a double-blind crossover design, 14 well-trained male cyclists completed a 30-min TT (TT30') followed by an all-out sprint at 175% of lactate threshold (SPRINT). Subjects were randomized to receive (i) 50 g KE, (ii) 180 mg·kg-1 body weight NaHCO3 (BIC), (iii) KE + BIC, or (iv) a control drink (CON). RESULTS: KE ingestion increased blood d-ß-hydroxybutyrate to ~3-4 mM during the TT30' and SPRINT (P < 0.001 vs CON). In KE, blood pH and bicarbonate concomitantly dropped, causing 0.05 units lower pH and 2.6 mM lower bicarbonate in KE compared with CON during the TT30' and SPRINT (P < 0.001 vs CON). BIC coingestion resulted in 0.9 mM higher blood d-ß-hydroxybutyrate (P < 0.001 vs KE) and completely counteracted ketoacidosis during exercise (P > 0.05 vs CON). Mean power output during TT30' was similar between CON and BIC at 281 W, but was 1.5% lower in the KE conditions (main effect of KE: P = 0.03). Time to exhaustion in the SPRINT was ~64 s in CON and KE and increased by ~8% in the BIC conditions (main effect of BIC: P < 0.01). DISCUSSION: Neutralization of acid-base disturbance by BIC coingestion is insufficient to counteract the slightly negative effect of KE intake during high-intensity exercise.

Untargeted UPLC-MS metabolomics reveals multiple changes of urine composition in healthy adult volunteers after consumption of curcuma longa L. extract.

Curcuma longa L. is used as food supplement to prevent diseases, although limited studies have been performed on healthy subjects up to now. In the present work, an untargeted UPLC-MS metabolomics approach was applied to study the changes of 24-hours urinary composition on healthy volunteers due to a 28-days daily consumption of a dried C. longa extract containing a standardized amount of curcuminoids. Changes in the excretion of different metabolites were observed after supplementation. Curcumin and two metabolic derivatives (hexahydrocurcumin and dihydrocurcumin) were detected in urine, indicating the absorption of the main curcuminoid from the extract and its further metabolism by liver and gut microbiota. For the first time ar-turmerone, the main apolar constituent of curcuma, was detected in urine in intact form, and its presence was confirmed by a targeted GC-MS analysis. The increase of tetranor-PGJM and tetranor-PGDM, two prostaglandin-D2 metabolites, was observed, being related to the anti-inflammatory effect exerted by curcuma. The variation of the amounts of HPAG, PAG, proline-betaine and hydroxyphenyllactic acid indicate that the supplementation induced changes to the activity of gut microbiota. Finally, the reduced excretion of niacin metabolites (nicotinuric acid, trigonelline and 2PY) and medium- and short-chain acylcarnitines suggests that curcuma could induce the mitochondrial ß-oxidation of fatty acids for energy production in healthy subjects. Overall, the results indicate that a prolonged daily consumption of a dried curcuma extract exerts multiple effects on healthy subjects, furthermore they show the opportunity offered by untargeted metabolomics for the study of the bioactivity of natural extracts in healthy human volunteers.

Serum vitamin D concentrations at dry-off and close-up predict increased postpartum urine ketone concentrations in dairy cattle.

Vitamin D is commonly supplemented to dairy cows as vitamin D3 to support calcium homeostasis and in times of low sunlight exposure. Vitamin D has beneficial immunomodulatory and anti-inflammatory properties. Serum 25-hydroxyvitamin D [25(OH)D] concentrations fluctuated during lactation, with the lowest concentrations measured in healthy cows within 7 d of calving. However, it is unknown if serum 25(OH)D concentrations measured during the previous lactation are associated with transition diseases or health risk factors in dairy cattle. We collected serum samples from 279 dairy cattle from 5 commercial dairy herds in Michigan at dry-off, close-up, and 2-10 d in milk (DIM). Vitamin D concentrations were determined by measuring serum 25(OH)D by radioimmunoassay. Total serum calcium was measured by colorimetric methods. Body condition scores (BCS) were assigned at the time of blood collection. Clinical disease incidence was monitored until 30 d postparturition. Separate bivariable logistic regression analyses were used to determine if serum 25(OH)D at dry-off, close-up, and 2-10 DIM was associated with various clinical diseases including mastitis, lameness, and uterine disorders (classified as metritis, retained placenta, or both) and increased urine ketone concentrations at P < 0.05. Among all significant bivariable analyses, multivariable logistic regression analyses were built to adjust for potential confounding variables including parity, BCS, season, and calcium. Receiver operator characteristic (ROC) curve analyses were used to determine optimal concentrations of serum 25(OH)D. We found that higher serum 25(OH)D concentrations at dry-off and close-up predicted increased urine ketone concentrations in early lactation, even after adjustment for confounders. Alternatively, we found that lower serum 25(OH)D at 2-10 DIM was associated with uterine diseases. Optimal concentrations for serum 25(OH)D at dry-off and close-up for lower risk of increased urine ketone concentrations were below 103.4 and 91.1 ng/mL, respectively. The optimal concentration for serum 25(OH)D at 2-10 DIM for uterine diseases was above 71.4 ng/mL. These results indicate that serum 25(OH)D at dry-off and close-up may be a novel predictive biomarker for increased urine ketone concentrations during early lactation. Increased urine ketone concentrations are not necessarily harmful or diagnostic for ketosis but do indicate development of negative energy balance, metabolic stress, and increased risk of early lactation diseases. Predicting that dairy cattle are at increased risk of disease facilitates implementation of intervention strategies that may lower disease incidence. Future studies should confirm these findings and determine the utility of serum 25(OH)D concentrations as a predictive biomarker for clinical and subclinical ketosis.

NMR-based urinalysis for beta-ketothiolase deficiency.

BACKGROUND: Beta-ketothiolase deficiency is a rare inborn errors of metabolism (IEM) affecting the catabolism of isoleucine, characterized by severe ketoacidosis in children of 6 to 24months old. A prompt diagnosis is of paramount importance as the metabolic decompensation can be effectively reverted by glucose infusion and health outcomes are improved on a protein-restricted diet. Currently, majority of the laboratory diagnosis were made based on mass-spectrometry and molecular genetics while little is mentioned on the advancement of nuclear magnetic resonance (NMR) spectroscopy for the diagnosis of this condition. CASE: We report a case of beta-ketothiolase deficiency in a 1-y-old Chinese boy who presented with repeated vomiting, impaired consciousness and severe ketoacidosis. NMR urinalysis detected excessive amount of butanone (a disease specific marker of beta-ketothiolase deficiency), tiglylglycine, (intermediate of isoleucine catabolism) and ketones. Diagnosis of beta-ketothiolase deficiency was further established by molecular genetic studies of ACAT1 gene of the proband. CONCLUSIONS: This case illustrated that NMR-based urinalysis is complementary to organic acid analysis for diagnosis of beta-ketothiolase deficiency. The operation of NMR is simple and fast; sample preparation is a two-step procedure while the NMR acquisition is automatic and usually takes <15min. We envisage that NMR analysis will become more available in clinical laboratories and will play an important role in acute pediatric care.

Screening study for repeated dose and reproductive/developmental toxicity of rubber accelerator, N,N-dicyclohexyl-2-benzothiazolesulfenamide, in rats.

A screening study for a vulcanization accelerator N,N-dicyclohexyl-2-benzothiazole-sulfenamide (DCBS) was performed in rats. Rats were given DCBS by gavage daily at 0, 6, 25, 100, or 400 mg/kg. Males were dosed for a total of 44 days beginning 14 days before mating. Females were dosed for a total of 40-51 days beginning 14 days before mating to day 3 of lactation. Toxicologic changes were significantly noted only at 400 mg/kg. Three females died. An increased incidence of females showing decreased locomotor activity, soil of the lower abdominal fur, and reddish tears was observed. A lowered body weight was found in males and females. Increased urinary ketones and serum inorganic phosphorus and decreased serum glutamate pyruvate transaminase in males were found. Increased absolute and relative weights of the kidneys in males and decreased absolute weight of the thymus in both sexes were noted. Significant fatty degeneration of the renal tubular epithelia, vacuolation of the adrenocortical cells, and atrophy of the spleen were observed in females. Significant decreases in the gestation index, numbers of corpura lutea, implantations, pups born and pups born alive, live birth index, and viability index were detected. It is concluded that the No Observed Adverse Effect Levels (NOAELs) for repeat dose and reproductive/developmental toxicity are 100 mg kg-1 day-1 in this screening study.

[Pharmacodynamic study of Relinqing granules on bacterial pyelonephritis model in rats].

OBJECTIVE: To observe the effects of Relinqing granules (powder of Polygonum capitatum extract) on the bacterial pyelonephritis model in rats. METHOD: The rat bacterial pyelonephritis model was induced by injecting the escherichia coli ATCC-25922 into kidney parenchyma. The rats were divided ramdamly into Relinqing groups(52.32, 26.16 g x kg(-1)), norflorin group (0.03 g x kg(-1)), model group and normal control group, and were given experimental drugs by gastrogavage. The contents of leucocytes (WBC), occult bloo (BLD), glucose (GLU), protein (PRO), ketones, bilirubin and urobilinagen in urine were determined. RESULT: As compared with the model group, Relinqing granules 6.0 g x kg(-1) (crude drug 52.32 g x kg(-1)) could decrease significantly the contents of WBC and BLD in urine and, however, had no markedly effects on the other biochemical parameters of urine. CONCLUSION: Relinqqing granule has significant effects of decreasing urine WBC and BLD on the bacterial pyolonephritis in rats.

A modified Atkins diet is effective for the treatment of intractable pediatric epilepsy.

PURPOSE: The Atkins diet may induce ketosis as does the ketogenic diet, without restrictions on calories, fluids, protein, or need for an inpatient fast and admission. Our objective was to evaluate the efficacy and tolerability of a modified Atkins diet for intractable childhood epilepsy. METHODS: Twenty children were treated prospectively in a hospital-based ambulatory clinic from September 2003 to May 2005. Children aged 3-18 years, with at least three seizures per week, who had been treated with at least two anticonvulsants, were enrolled and received the diet over a 6-month period. Carbohydrates were initially limited to 10 g/day, and fats were encouraged. Parents measured urinary ketones semiweekly and recorded seizures daily. All children received vitamin and calcium supplementation. RESULTS: In all children, at least moderate urinary ketosis developed within 4 days (mean, 1.9). Sixteen (80%) completed the 6-month study; 14 chose to remain on the diet afterward. At 6 months, 13 (65%) had >50% improvement, and seven (35%) had >90% improvement (four were seizure free). Mean seizure frequency after 6 months was 40 per week (p = 0.005). Over a 6-month period, mean serum blood urea nitrogen increased from 12 to 17 mg/dl (p = 0.01); creatinine was unchanged. Cholesterol increased from 192 to 221 mg/dl, (p = 0.06). Weight did not change significantly (34.0-33.7 kg); only six children lost weight. A stable body mass index over time correlated with >90% improvement (p = 0.004). CONCLUSIONS: A modified Atkins diet is an effective and well-tolerated therapy for intractable pediatric epilepsy.

Effect of a monensin controlled release capsule on metabolic parameters in transition dairy cows.

A prospective field study involving 251 Holstein cows and heifers from five dairy farms near Guelph, Ontario, Canada, was conducted to measure the effect of monensin delivered in a controlled release capsule 3 wk precalving on metabolic function in dairy cows immediately pre- and postcalving. At 3 wk before expected calving, cows were randomly assigned to receive either a controlled release capsule containing monensin or to serve as negative controls. Cows were blood sampled once per week precalving and once in the week following calving, at the same time of day and the same day of the week. Serum was evaluated for beta-hydroxybutyrate (BHBA), nonesterified fatty acids (NEFA), cholesterol, urea, glucose, calcium, and phosphorus. Monensin-treated cows had significantly decreased NEFA and BHBA and significantly increased concentrations of serum cholesterol and urea in the week immediately precalving. No effect of treatment was observed for calcium, phosphorus, or glucose in the precalving period. After calving, concentrations of phosphorus were lower and BHBA tended to be lower, and cholesterol and urea were higher in monensin-treated cows. There was no effect of treatment on NEFA, glucose, or calcium in the first week postcalving. Monensin treatment administered precalving significantly improved indicators of energy balance in both the immediate precalving and postcalving periods. The findings indicate better energy metabolism in monensin-treated cows as they approach calving. Improvement of energy balance before calving is important for the prevention of energy associated metabolic diseases, such as retained placenta, clinical ketosis, and displaced abomasum, which might occur immediately postcalving.

Pregnancy toxemia in the European ferret (Mustela putorius furo).

BACKGROUND AND OBJECTIVE: Pregnancy toxemia may lead to appreciable mortality among jills and their offspring. The objective of this report was to increase awareness of the disease, its likely cause, and practical prevention and treatment measures. METHODS: Ten cases of pregnancy toxemia were evaluated. Jills were in late gestation (mean, 38 days; range, 34 to 42 days) and had large litters (mean, 11.5 kits; range, 7 to 15 kits). RESULTS: The most common clinical signs of disease were lethargy, inappetence, dehydration, and excess shedding. Hematologic and clinical biochemical abnormalities included anemia (4 of 8 jills tested), hypoproteinemia (5 of 7), azotemia (7 of 7), hypocalcemia (5 of 6), hyperbilirubinemia (3 of 3), and high liver enzyme activities (6 of 6). Two jills were found dead; two jills were euthanized, six received supportive treatment, and cesarean section was performed on five. The three jills that survived tended to have less pronounced azotemia, hypoproteinemia, and liver enzyme activity increases and were not anemic. Hepatic lipidosis was observed grossly in all jills that died and was confirmed by histologic examination in four jills. CONCLUSIONS: Pregnancy toxemia in ferrets resembles metabolic diseases in several other animal species and requires aggressive treatment, including supportive care, nutritional supplementation, and cesarean section. Maintaining adequate nutrition and avoiding stress late in gestation may prevent the disease.

Case reports of successful pregnancy in women with maple syrup urine disease and propionic acidemia.

We report on 2 women with organic acidemias, one with classical maple syrup urine disease and another with mild propionic acidemia in which protein restricted diets and carnitine supplementation were successfully employed to manage pregnancies. Healthy infants were delivered without maternal metabolic decompensation.

Hormonal and metabolic adaptation to protein-supplemented fasting in obese subjects.

Thirty hospitalized, severely obese patients (40 +/- 2 yr, 82 +/- 4 percent weight excess) were submitted to a 13-d protein-supplemented fast (PSF) with 70 g milk proteins/d (1.26 MJ or 300 kcal). The mean weight loss during PSF was 5.4 +/- 0.3 kg corresponding to 422 +/- 39 g/d. Comparison of the urinary nitrogen excretion with daily protein intake revealed that the nitrogen balance was equilibrated during PSF. Blood glucose decreased moderately but significantly during the whole PSF period whereas plasma insulin was only reduced during the first 9 d and tended to rise thereafter. Plasma FFA increased rapidly and remained elevated until the end of the study (+ 60 per cent); serum total cholesterol and plasma triglycerides showed a 26 and a 35 per cent decrease respectively. Basal plasma glucagon was slightly increased. Due to the low sodium intake (42 mmol/d) urinary sodium excretion dropped rapidly. Simultaneously both systolic (-13 mmHg) and diastolic (-7 mmHg) arterial blood pressure decreased significantly. The biological tolerance was good: metabolic acidosis was prevented with sodium bicarbonate, excessive rise in serum uric acid was corrected with allopurinol and a marked decrease in serum potassium was avoided with an appropriate dose of spironolactone. Twenty-six patients could be weighed 6 to 15 months after PSF: 12 showed a further weight reduction (6.6 +/- 1.6 kg) and seven a discrete weight gain (1.0 +/- 0.4 kg). Thus, PSF was well accepted and was profitable in 19 out of our 30 patients. It should be restricted to cases of severe and refractory obesity and performed under careful medical supervision.

A 300 kcal (1.2 MJ) diet using conventional food.

A 300 kcal (1.25 MJ) diet of conventional food is described, which has been studied under in-patient conditions for four to six weeks. It contained 22.6 g protein, 34 g CHO and 6.9 g fat but not the full RDA of vitamins and minerals since this is impossible without supplementation. Hunger disappeared after the third day. Patients developed ketonuria and hyperuricemia; serum lipids were normalised and hypertension disappeared. The diet offers advantages in that it induces better nutritional knowledge and habits.

Corticosteroid-like effects after high doses of compound (5-methoxy-2-thienyl)-thioacetic acid sodium salt in rats.

During a systemic tolerance study in rats with compound (5-methoxy-2-thienyl)-thioacetic acid sodium salt (MTTA), which is known to decrease the lipolytic process and platelet aggregation, effects after high doses were observed which are similar to the well-known findings after corticosteroid administration. The oral application of MTTA in a dose range from 10 to 1000 mg/kg over a period of 4 weeks resulted in a decrease in total lipids and free fatty acids from 100 mg/kg onwards and an increase in total glycerin and triglycerides after 1000 mg/kg, supplemented with an increased excretion of keto-bodies in urine at all dose levels. The hematological examinations revealed a reduction of blood eosinophils and lymphocytes in males at doses from 10 mg/kg, whereas the eosinophilic granulocytopoiesis was decreased only at the 1000 mg/kg level in both sexes. Determinations of organ weights showed a decrease in thymus weight from 300 mg/kg onwards with an involution at the highest dose levels, being confirmed by histological examination, whereas the adrenal glands' weights were decreased only at the dose level of 1000 mg/kg. Since the endogenous corticosterone level remained unaltered at all doses, it is suggested that these corticosteroid-like effects are directly attributable to MTTA.

[Thioamine compound content in urine determined by using an iodine-azide reagent and selenious acid reaction]. / Soderzhanie tioaminosoedinenii v moche bol'nykh pri opredelenii s pomoshch'iu iod-azidnogo reagenta i po reaktsii s selenistoi kislotoi.

Iodine-azide reagent enabled to evaluate quantitatively content of thioamines excreted within a day. Concentration of thioketons in urine of patients was 11-15 microgram/eqv of thiourea per 1 ml, at the same time their excretion within a day accounted for 13.06 +/- 0.56 mg/eqv in women and 22.11 +/- 1.1 mg/eqv in men. Neither concentration of thioketones in urine nor their excretion in patients with various tumors studied were distinct from these patterns in urine of non-oncologic patients. Comparison of these two methods suggests that development of color on addition of selenous acid to urine is not due to reaction with thioamines present but to interaction with other unknown substances.

Carbohydrate metabolism in pregnancy. VI. Plasma fuels, insulin, liver composition, gluconeogenesis, and nitrogen metabolism during late gestation in the fed and fasted rat.

The effects of late pregnancy on metabolic fuels, liver composition, gluconeogenesis, and nitrogen metabolism have been examined in fed and fasted rats. Plasma free fatty acid (FFA) and immunoreactive insulin (IRI) are greater and glucose and ketones are lower in fed 19-day pregnant than they are in agematched virgin rats. A 48 hr fast elicits greater increases in FFA and ketones and more profound reductions in glucose in the pregnant rats and obliterates the differences in IRI. Fetal weight is not modified by such fasting but maternal weight losses exceed that of the nongravid rats. Livers from rats 19 days pregnant contain more and larger hepatocytes. Per mumole hepatic deoxyribonucleic acid (DNA)-phosphorus, water and protein are more abundant, whereas glycogen is unaffected. Livers from fed pregnant rats contain more lipid phosphorus and less neutral lipid fatty acid. After a 48 hr fast, hepatic steatosis supervenes in gravid animals due to accumulated neutral fat. The contents of hepatic acetyl-coenzyme A (CoA) and citric acid are not different in fed pregnant and virgin rats but are greater in the pregnant rats after fasting. Formation of glucose-(14)C and glycogen-(14)C from administered pyruvate-(14)C are the same in fed pregnant and virgin rats, but greater in the pregnant ones after a 24 or 48 hr fast. Pregnancy does not affect creatinine excretion, and urinary urea is not different in fed pregnant, virgin, and postpartum animals. Contrariwise, more nitrogen, potassium, and phosphorus are excreted by the pregnant animals during a 2 day fast. The increment in urinary nitrogen is due largely to urea on the 1st day, whereas heightened ammonia accounts for half the increase on the 2nd and correlates with the enhanced ketonuria. Muscle catabolism, gluconeogenesis, and diversion to fat are activated more rapidly and to a greater degree when food is withheld during late gestation in the rat. These catabolic propensities are restrained in the fed state. The capacity for "accelerated starvation" may confer survival benefit upon an intermittently eating mother in the presence of a continuously feeding fetus.