Blood Ketone Bodies Research Articles

Effect of empagliflozin on ectopic fat stores and myocardial energetics in type 2 diabetes: the EMPACEF study

BackgroundEmpagliflozin is a sodium-glucose cotransporter 2 (SGLT2) inhibitor that has demonstrated cardiovascular and renal protection in patients with type 2 diabetes (T2D). We hypothesized that empaglifozin (EMPA) could modulate ectopic fat stores and myocardial energetics in high-fat-high-sucrose (HFHS) diet mice and in type 2 diabetics (T2D).MethodsC57BL/6 HFHS mice (n = 24) and T2D subjects (n = 56) were randomly assigned to 12 weeks of treatment with EMPA (30 mg/kg in mice, 10 mg/day in humans) or with placebo. A 4.7 T or 3 T MRI with 1H-MRS evaluation–myocardial fat (primary endpoint) and liver fat content (LFC)–were performed at baseline and at 12 weeks. In humans, standard cardiac MRI was coupled with myocardial energetics (PCr/ATP) measured with 31P-MRS. Subcutaneous (SAT) abdominal, visceral (VAT), epicardial and pancreatic fat were also evaluated.The primary efficacy endpoint was the change in epicardial fat volume between EMPA and placebo from baseline to 12 weeks. Secondary endpoints were the differences in PCr/ATP ratio, myocardial, liver and pancreatic fat content, SAT and VAT between groups at 12 weeks.ResultsIn mice fed HFHS, EMPA significantly improved glucose tolerance and increased blood ketone bodies (KB) and β-hydroxybutyrate levels (p < 0.05) compared to placebo. Mice fed HFHS had increased myocardial and liver fat content compared to standard diet mice. EMPA significantly attenuated liver fat content by 55%, (p < 0.001) but had no effect on myocardial fat.In the human study, all the 56 patients had normal LV function with mean LVEF = 63.4 ± 7.9%. Compared to placebo, T2D patients treated with EMPA significantly lost weight (− 2.6 kg [− 1.2; − 3.7]) and improved their HbA1c by 0.88 ± 0.74%. Hematocrit and EPO levels were significantly increased in the EMPA group compared to placebo (p < 0.0001, p = 0.041). EMPA significantly increased glycosuria and plasma KB levels compared to placebo (p < 0.0001, p = 0.012, respectively), and significantly reduced liver fat content (− 27 ± 23 vs. − 2 ± 24%, p = 0.0005) and visceral fat (− 7.8% [− 15.3; − 5.6] vs. − 0.1% [− 1.1;6.5], p = 0.043), but had no effect on myocardial or epicardial fat. At 12 weeks, no significant change was observed in the myocardial PCr/ATP (p = 0.57 between groups).ConclusionsEMPA effectively reduced liver fat in mice and humans without changing epicardial, myocardial fat or myocardial energetics, rebutting the thrifty substrate hypothesis for cardiovascular protection of SGLT2 inhibitors.Trial registration NCT, NCT03118336. Registered 18 April 2017, https://clinicaltrials.gov/ct2/show/NCT03118336

Age- and Sex-Dependent Modulation of Exogenous Ketone Supplement-Evoked Effects on Blood Glucose and Ketone Body Levels in Wistar Albino Glaxo Rijswijk Rats.

In certain disease states, such as epilepsy, the elevation of blood ketone levels with ketogenic diets (KDs) has beneficial effects, while exogenous ketone supplements (EKSs) were shown to increase the level of blood ketone bodies (such as β-hydroxybutyrate, βHB) and decrease blood glucose levels without dietary restrictions. It has been suggested that ketone body and glucose utilization of the body may be modified by age and gender resulting in changes in blood βHB and glucose levels, but it was not investigated through several months yet. Thus, we investigated whether the effect of an EKS on blood βHB and glucose level is modulated by age and sex in Wistar Albino Glaxo Rijswijk (WAG/Rij) rats, a model animal of human absence epilepsy. We used KEMCT (1:1 mix of ketone ester/KE and medium-chain triglyceride/MCT oil) by oral gavage in female and male WAG/Rij rats. Animals were fed with standard diet, which was supplemented by KEMCT (2.5 g/kg) once per month by oral gavage for 17 months. One hour after KEMCT treatment, changes in blood R-beta-hydroxybutyrate (R-βHB) and glucose levels were measured. KEMCT gavage significantly increased blood R-βHB and decreased blood glucose levels, in both male and female rats, compared with the controls. In male rats, the KEMCT-induced increase in blood R-βHB levels was lower at the 7th and 8th months and higher at the 16th and 17th months, compared with the results at the 1st month. KEMCT-generated increase in R-βHB levels was higher in female rats, compared with male rats between the 2nd and 11th months, but older (between the 14th and 17th months) female rats showed lower levels than males. KEMCT gavage induced significantly lower glucose levels at the 4th, 9th, 10th, 12th, and 13th months in both sexes, but between the 14th and 17th months, only males showed significantly lower levels, compared with the results at the 1st month. KEMCT treatment induced lower blood glucose levels in female than in male rats between the 1st and 8th months, but higher glucose levels were measured in female rats at the 17th month than in males. These findings suggest that age and sex can modify the EKS-evoked effects on blood R-βHB and glucose concentrations.

Chronic Ketosis Modulates HIF1α-Mediated Inflammatory Response in Rat Brain.

Hypoxia inducible factor alpha (HIF1α) is associated with neuroprotection conferred by diet-induced ketosis, but the underlying mechanism remains unclear. In this study, we use a ketogenic diet in rodents to induce a metabolic state of chronic ketosis, as measured by elevated blood ketone bodies. Chronic ketosis correlates with neuroprotection in both aged and following focal cerebral ischemia and reperfusion (via middle cerebral artery occlusion, MCAO) in mouse and rat models. Ketone bodies are known to be used efficiently by the brain, and metabolism of ketone bodies is associated with increased cytosolic succinate levels that inhibits prolyl hydroxylases allowing HIF1α to accumulate. Ketosis also regulates inflammatory pathways, and HIF1α is reported to be essential for gene expression of interleukin 10 (IL10). Therefore, we hypothesized that ketosis-stabilized HIF1α modulates the expression of inflammatory cytokines orchestrating neuroprotection. To test changes in cytokine levels in rodent brain, 8-week-rats were fed either the standard chow diet (SD) or the KG diet for 4weeks before ischemia experiments (MCAO) were performed and the brain tissues were collected. Consistent with our hypothesis, immunoblotting analysis shows IL10 levels were significantly higher in KG diet rat brain compared to SD, whereas the TNFα and IL6 levels were significantly lower in the brains of KG diet-fed group.

The Ketogenic Diet Increases In Vivo Glutathione Levels in Patients with Epilepsy.

The Ketogenic Diet (KD) is a high-fat, low-carbohydrate diet that has been utilized as the first line treatment for contrasting intractable epilepsy. It is responsible for the presence of ketone bodies in blood, whose neuroprotective effect has been widely shown in recent years but remains unclear. Since glutathione (GSH) is implicated in oxidation-reduction reactions, our aim was to monitor the effects of KD on GSH brain levels by means of magnetic resonance spectroscopy (MRS). MRS was acquired from 16 KD patients and seven age-matched Healthy Controls (HC). We estimated metabolite concentrations with linear combination model (LCModel), assessing differences between KD and HC with t-test. Pearson was used to investigate GHS correlations with blood serum 3-B-Hydroxybutyrate (3HB) concentrations and with number of weekly epileptic seizures. The results have shown higher levels of brain GSH for KD patients (2.5 ± 0.5 mM) compared to HC (2.0 ± 0.5 mM). Both blood serum 3HB and number of seizures did not correlate with GSH concentration. The present study showed a significant increase in GSH in the brain of epileptic children treated with KD, reproducing for the first time in humans what was previously observed in animal studies. Our results may suggest a pivotal role of GSH in the antioxidant neuroprotective effect of KD in the human brain.

Ketone bodies as an energy source: regular-grade, premium, or super-fuel to power the mitochondrial engine?

Ketone bodies as an energy source: regular-grade, premium, or super-fuel to power the mitochondrial engine?

CR reprograms acetyl-CoA metabolism and induces long-chain acyl-CoA dehydrogenase and CrAT expression.

Calorie restriction (CR), an age delaying diet, affects fat oxidation through poorly understood mechanisms. We investigated the effect of CR on fat metabolism gene expression and intermediate metabolites of fatty acid oxidation in the liver. We found that CR changed the liver acylcarnitine profile: acetylcarnitine, short‐chain acylcarnitines, and long‐chain 3‐hydroxy‐acylcarnitines increased, and several long‐chain acylcarnitines decreased. Acetyl‐CoA and short‐chain acyl‐CoAs were also increased in CR. CR did not affect the expression of CPT1 and upregulated the expression of long‐chain and very‐long‐chain Acyl‐CoA dehydrogenases (LCAD and VLCAD, respectively). The expression of downstream enzymes such as mitochondrial trifunctional protein and enzymes in medium‐ and short‐chain acyl‐CoAs oxidation was not affected in CR. CR shifted the balance of fatty acid oxidation enzymes and fatty acid metabolites in the liver. Acetyl‐CoA generated through beta‐oxidation can be used for ketogenesis or energy production. In agreement, blood ketone bodies increased under CR in a time of the day‐dependent manner. Carnitine acetyltransferase (CrAT) is a bidirectional enzyme that interconverts short‐chain acyl‐CoAs and their corresponding acylcarnitines. CrAT expression was induced in CR liver supporting the increased acetylcarnitine and short‐chain acylcarnitine production. Acetylcarnitine can freely travel between cellular sub‐compartments. Supporting this CR increased protein acetylation in the mitochondria, cytoplasm, and nucleus. We hypothesize that changes in acyl‐CoA and acylcarnitine levels help to control energy metabolism and contribute to metabolic flexibility under CR.

The Hyperglycemia and Hyperketonemia Impaired Bone Microstructures: A Pilot Study in Rats.

Though diabetes mellitus (DM) is one of the known causes of osteoporosis, it is also realized that ketogenic diet (KD), an effective regimen for epilepsy, impairs bone microstructures. However, the similarities and differences of effects between these two factors are still unknown. The purpose of this study is to identify different effects between hyperglycemia and hyperketonemia, which are manifestations of DM and KD, on bone in rats. Thirty male Sprague-Dawley rats were randomly divided into three groups: the sham, DM, and KD groups. Hyperglycemia was achieved by intravenous injection of streptozotocin in DM group, while hyperketonemia was induced by application of ketogenic diet (carbohydrates-to-fat as 1:3) in KD group. The body weight, blood ketone body, and blood glucose were recorded, and the bone turnover markers, bone length, bone microstructures, bone biomechanics and histomorphology were measured after 12 weeks intervention. Compared with the control and KD groups, a significant body weight loss was found in the DM group, and the bone lengths of tibia and femur of the group were shortened. The blood glucose and blood ketone were noticeably increased in the DM and KD rats, respectively. Microstructures and properties of cancellous bone were significantly deteriorated in both the DM and KD groups compared with the sham group, as the bone volumes were decreased and the bone trabecula structures were disturbed. Meanwhile, the thickness and strength of cortical bone was reduced more in the DM group than those in the sham and KD groups. The HE staining showed that bone trabecula was significantly decreased in both the DM and KD groups, and more adipose tissue was observed in the KD rats. The activity of osteoblasts was decreased more in both the KD and DM groups than that in the sham group, while the activity of osteoclasts of the two groups was remarkably increased. The present study indicates that both hyperglycemia and hyperketonemia have adverse effects on bone. Therefore, it is worth paying more attention to the bone status of patients with hyperglycemia and hyperketonemia in clinic.

Placebo-Controlled, Double-Blind Study of Empagliflozin (EMPA) and Implantable Cardioverter-Defibrillator (EMPA-ICD) in Patients with Type2 Diabetes (T2DM): Rationale and Design.

IntroductionType 2 diabetes (T2DM) is associated with cardiovascular death, including sudden cardiac death due to arrhythmias. Patients with an implantable cardioverter-defibrillator (ICD) are also at high risk of developing a clinically significant ventricular arrhythmia. It has been reported that sodium–glucose cotransporter 2 (SGLT2) inhibitors can reduce cardiovascular deaths; however, the physiological mechanisms of this remain unclear. It is, however, well known that SGLT2 inhibitors increase blood ketone bodies, which have been suggested to have sympatho-suppressive effects. Empagliflozin (EMPA) is an SGLT2 inhibitor. The current clinical trial titled “Placebo-controlled, double-blind study of empagliflozin (EMPA) and implantable cardioverter-defibrillator (EMPA-ICD) in patients with type 2 diabetes (T2DM)” was designed to investigate the antiarrhythmic effects of EMPA.MethodsThe EMPA-ICD study is a prospective, multicenter, placebo-controlled, double-blind, randomized, investigator-initiated clinical trial currently in progress. A total of 210 patients with T2DM (hemoglobin A1c 6.5–10.0%) will be randomized (1:1) to receive once-daily placebo or EMPA, 10 mg, for 24 weeks. The primary endpoint is the number of clinically significant ventricular arrhythmias for 24 weeks before and 24 weeks after study drug administration, as documented by the ICD. The secondary endpoints of the study are the change from baseline concentrations in blood ketone and catecholamine 24 weeks after drug treatment.ConclusionThe EMPA-ICD study is the first clinical trial to assess the effect of an SGLT2 inhibitor on clinically significant ventricular arrhythmias in patients with T2DM and an ICD.Trial registrationUnique trial number, jRCTs031180120 (https://jrct.niph.go.jp/latest-detail/jRCTs031180120).Electronic Supplementary MaterialThe online version of this article (10.1007/s13300-020-00924-9) contains supplementary material, which is available to authorized users.

Urinary glucose and ketone bodies as indicators of acute caffeine poisoning.

AimIn various countries, many fatal health problems have been reported due to high intake of caffeine‐rich energy drinks, tablets, and powders. In patients with acute caffeine poisoning, determination of blood caffeine concentration is an important yet difficult task. We aimed to assess whether the presence of glucose and ketone bodies in urine reflected the blood caffeine concentration in patients with acute caffeine poisoning.MethodsFrom April 2010 to March 2018, 25 patients with an overdose of only caffeine‐rich tablets were admitted to our hospital. Their clinical features were investigated. In addition, we investigated whether the glucose and ketone bodies in the urine reflected blood caffeine concentration in 23 patients who underwent the urine qualitative test at admission.ResultsThe majority of the patients were young healthy women, whose average caffeine ingestion was 15.6 ± 8.1 g. Initial urine examinations showed glucose in 60% (14/23) of patients and ketone bodies in 57% (13/23) of patients. Ketone bodies or glucose were found in 78% (18/23) of the patients. The correlation between blood caffeine concentration and urinary glucose was R = 0.625, blood caffeine concentration and ketone bodies was R = 0.596, and blood caffeine and both was R = 0.76.ConclusionUrine qualitative test is effective for differential diagnosis and severity assessment of acute caffeine poisoning in patients.

Transition period and subclinical ketosis in dairy cattle: association with milk production, metabolic and reproductive disorders and economic aspects

Many dairy cows experience a high incidence of health problems during the transition period (TP). The TP is an intermediate stage of various digestive, metabolic and reproductive functions which determine the general health status at the time of calving and during the first weeks postpartum. Negative energy balance due to increased energy demand at parturition and significantly reduced dry matter intake relative to demand is an important determinant. Consequently, substantial lipid mobilization from adipose tissue, increased oxidative stress and impaired immunity are associated with higher incidences of periparturient health problems including ketosis or subclinical ketosis (SCK), which have tremendous economic impact on dairy productivity. SCK is defined as the presence of increased blood ketone bodies (BHBA: betahydroxybutyric acid, acetone, acetoacetic acid) without clinical ketosis signs. Varying blood and milk cut-off values have been reported for BHBA concentrations defining SCK, but the most commonly accepted values are ≥ 1.2 mmol/L and ≥ 200 μmol/L respectively. This underestimated disease can impact dairy cow productivity through decreased milk production in the order of roughly 300 kg/lactation and increases the risk of metabolic and reproductive diseases such as displaced abomasum, retained placenta, metritis, mastitis, prolong oestrus interval and reduces conception rates. SCK also referred to as ‘profit robber or killer’ can cause productivity and economic losses of between $200-290 per dairy cow annually. Options for the control and prevention of SCK include controlled-release monensin capsules, and the injectable combination butaphosphan and cyanocobalamin and oral propylene glycol. SCK is easy to detect in early lactation using cow-side validated BHBA analysers with high specificity and sensitivity.

Exogenous Ketone Supplementation and Keto-Adaptation for Endurance Performance: Disentangling the Effects of Two Distinct Metabolic States.

Ketone bodies (KB) provide an alternative energy source and uniquely modulate substrate metabolism during endurance exercise. Nutritional ketosis (blood KBs > 0.5mM) can be achieved within minutes via exogenous ketone supplementation or days-to-weeks via conforming to a very low-carbohydrate, ketogenic diet (KD). In contrast to short-term (< 2weeks) KD ingestion, chronic adherence (> 3weeks) leads to a state of keto-adaptation. However, despite elevating blood KBs to similar concentrations, exogenous ketone supplementation and keto-adaptation are not similar metabolic states as they elicit diverse and distinct effects on substrate availability and metabolism during exercise; meaning that their influence on endurance exercise performance is different. In contrast to contemporary, high(er)-carbohydrate fuelling strategies, inducing nutritional ketosis is rarely ergogenic irrespective of origin and, in fact, can impair endurance performance. Nonetheless, exogenous ketone supplementation and keto-adaptation possess utility for select endurance events and individuals, thus warranting further research into their performance effects and potential strategies for their optimisation. It is critical, however, that future research considers the limitations of measuring blood KB concentrations and their utilisation, and assess the effect of nutritional ketosis on performance using exercise protocols reflective of real-world competition. Furthermore, to reliably assess the effects of keto-adaptation, rigorous dietary-training controls of sufficient duration should be prioritised.

Ketone body supplement label claims: what supplement has been supplemented?

There is a keen interest in performance-enhancing supplementation and the associated benefits, despite reports of incorrect label claims made by manufacturers and the questionable efficacy of the supplements. The use of ketone body supplements as a source of fuel during exercise and sporting performance, in particular, is of interest to sportspeople. By increasing blood ketone body levels, with an accompanying decrease in blood glucose, may indicate a state of nutritional ketosis, whereby the body no longer relies on glucose metabolism but rather the metabolism of ketone bodies. This could be beneficial for long, slow steady-state endurance exercise. There are numerous ketone body supplements on the market manufactured in South Africa and internationally. However, unlike medicines, the sports supplementation industry is poorly regulated. Furthermore, ketone body supplementation with regard to its effects on improving exercise and athletic performance is still unconvincing. Within the ever-changing sports supplementation industry, ketone body supplements are being used despite controversies regarding the accuracy and scientific merit of label claims. The ingredients and their quantities, as well as the performance benefits, need to be objectively validated.

Liver Glycogen Phosphorylase Deficiency Leads to Profibrogenic Phenotype in a Murine Model of Glycogen Storage Disease Type VI

Mutations in the liver glycogen phosphorylase (Pygl) gene are associated with the diagnosis of glycogen storage disease type VI (GSD‐VI). To understand the pathogenesis of GSD‐VI, we generated a mouse model with Pygl deficiency (Pygl −/−). Pygl −/− mice exhibit hepatomegaly, excessive hepatic glycogen accumulation, and low hepatic free glucose along with lower fasting blood glucose levels and elevated blood ketone bodies. Hepatic glycogen accumulation in Pygl −/− mice increases with age. Masson's trichrome and picrosirius red staining revealed minimal to mild collagen deposition in periportal, subcapsular, and/or perisinusoidal areas in the livers of old Pygl −/− mice (>40 weeks). Consistently, immunohistochemical analysis showed the number of cells positive for alpha smooth muscle actin (α‐SMA), a marker of activated hepatic stellate cells, was increased in the livers of old Pygl −/− mice compared with those of age‐matched wild‐type (WT) mice. Furthermore, old Pygl −/− mice had inflammatory infiltrates associated with hepatic vessels in their livers along with up‐regulated hepatic messenger RNA levels of C‐C chemokine ligand 5 (Ccl5/Rantes) and monocyte chemoattractant protein 1 (Mcp‐1), indicating inflammation, while age‐matched WT mice did not. Serum levels of aspartate aminotransferase and alanine aminotransferase were elevated in old Pygl −/− mice, indicating liver damage. Conclusion: Pygl deficiency results in progressive accumulation of hepatic glycogen with age and liver damage, inflammation, and collagen deposition, which can increase the risk of liver fibrosis. Collectively, the Pygl‐deficient mouse recapitulates clinical features in patients with GSD‐VI and provides a model to elucidate the mechanisms underlying hepatic complications associated with defective glycogen metabolism.

Effect of Annona muricata Leaves Extract on Colon Antioxidant Status and Ketone Bodies in Blood and Urine of Cycas-treated Rats

The effect of Annona muricata methanol leaf extract (AMELE) on colon antioxidant status and ketone bodies in blood and urine of cycas treated rats was investigated. Seventy male Wistar albino rats were randomly assigned into seven groups of ten experimental animals in a study that lasted for twenty-eight days. After 28 days, using standard methods, serum and urinary ketone bodies were assayed. Malondialdehyde (MDA), antioxidants enzymes in the colon and histopathological analysis were assayed. The results showed ketogenic diet had significant (p&lt;0.05) increase in serum and urinary ketone body levels compared to other groups on normal feed. Annona muricata extract enhanced cellular consumption and uptake of ketone bodies whereas cycas reduced cellular consumption of ketone bodies. Annona muricata given either with ketogenic diet or cycas or with both ketogenic diet and cycas led to higher tissue levels of antioxidant enzymes. Tissue MDA levels were significantly (p&lt;0.05) increased in all cycas treated groups compared to the control group with the highest levels seen in the two groups placed on normal feed and cycas. Lactate dehydrogenase activities were significantly (p&lt;0.05) increases in all the groups placed on ketogenic diet compared to the groups placed on normal feed. Histopathological analysis revealed normal rat colon tissues in all the groups except 43% of the group 2 animals. This therefore indicates that cycas- induced oxidative stress and tissue injury may be mitigated by the combined administration of Annona muricata extract and ketogenic diet.

Effect of Empagliflozin on Free Fatty Acids and Ketone Bodies in Japanese Patients with Type 2 Diabetes Mellitus: A Randomized Controlled Trial.

We report a randomized, double-blind, placebo-controlled, 4-week study to investigate the effect of empagliflozin on free fatty acids and blood ketone bodies in Japanese patients with type 2 diabetes mellitus. Patients (baseline mean [standard deviation] glycated hemoglobin 7.91% [0.80%]; body mass index 24.3 [3.2] kg/m2) were randomized to empagliflozin 10mg (n = 20), empagliflozin 25mg (n = 19), or placebo (n = 21) daily as monotherapy for 28days. Meal tolerance tests (MTTs; breakfast, lunch, dinner) were performed on day - 1, day 1 (first day of treatment), and day 28. On day 1 and day 28, study drug was administered 1h before breakfast. Free fatty acids and blood ketone bodies were measured before and 1, 2, and 3h after each MTT, and the next morning (overnight fast). Empagliflozin significantly reduced plasma glucose and insulin and reduced body weight vs. placebo. Empagliflozin increased free fatty acids and total ketones bodies at day 1 and day 28. At day 28, the adjusted mean (95% confidence interval) difference vs. placebo in the time-corrected area under curve over 24h for total ketone bodies was 67.1 (12.3, 121.8) µmol·h/L·h (P = 0.017) with empagliflozin 10mg and 178.1 (123.9, 232.2) µmol·h/L·h (P < 0.001) with empagliflozin 25mg. Increases in ketones with empagliflozin vs. placebo peaked just before and declined after meals, with the highest peak before breakfast. Changes in total ketone bodies appeared to be associated with changes in plasma glucose, insulin, and free fatty acids. Empagliflozin modestly increased free fatty acids and blood ketone bodies after a single dose and 28days' treatment. Increases in ketones appeared to be related to the duration of fasting and were most pronounced before breakfast. Increases in ketones appeared to be associated with changes in well-known metabolic determinants of ketone production. ClinicalTrials.gov identifier, NCT01947855. Boehringer Ingelheim & Eli Lilly and Company.

Changes of serum procalcitonin and lactate levels in patients with diabetic ketoacidosis complicated with infection and its clinical significance

Objective To investigate the changes and clinical significance of serum procalcitonin (PCT) and lactate levels in patients with diabetic ketoacidosis (DKA). Methods A total of 112 patients with DKA admitted to Wenling Hospital of Traditional Chinese Medicine from June 2016 to June 2018 were enrolled.Thirty-five healthy controls in our hospital were selected as the control group.White blood cell counts (WBC), ketone bodies, creatinine (Scr), urea nitrogen (BUN), alanine aminotransferase (ALT), aspartate aminotransferase (AST), PCT and lactic acid were measured in each group. Results Compared with the control group, the WBC [(10.26±4.19)×109+/L vs.(14.81±3.79)×109/L], ketone bodies [(2.36±0.79) mmol/L vs.(6.25±1.02)mmol/L], Scr [(116.51±41.47)μmol/L vs.(113.15±37.68)μmol/L], BUN [(76.15±17.33)ng/L vs.(143.64±16.59)ng/L], ALT [(81.65±22.56)U/L vs.(79.36±25.34)U/L], AST [(61.38±12.52)U/L vs.(78.37±11.74)U/L], PCT [(1.53±0.59)ng/mL vs.(5.75±1.21)ng/mL] and lactic acid [(3.67±0.56)mmol/L vs.(10.31±2.51)mmol/L] levels in the uninfected group and the infected group were significantly increased, the differences were statistically significant (t=3.706, 10.841, 13.484, 32.826, 8.195, 8.465, 23.546, 48.493, 13.622, 11.715, 13.115, 21.148, 11.979, 26.426, 34.766, 23.538, all P<0.05). Compared with the uninfected group, the WBC [(14.81±3.79)×109+/L], ketone body [(6.25±1.02)mmol/L], BUN [(143.64±16.59)ng/L], AST [(8.37±11.74)U/L], PCT [(5.75±1.21)ng/mL] and lactic acid [(10.31±2.51)mmol/L] levels of the infected group were significantly increased (t=5.349, 19.230, 18.559, 6.547, 19.225, 15.383, all P<0.05). Multivariate logistic regression analysis showed that serum lactate and PCT levels were stronger factors in patients with DKA.The ROC analysis showed that the areas under the ACU of PCT, lactic acid and combined detection were 0.849, 0.805 and 0.930, respectively.The sensitivity (90.91%) and specificity (89.47%) of the combined detection were better than the separate tests (PCT: 78.33% and 82.69%, lactic acid: 70.18% and 70.91%) (sensitivity χ2=3.398, 7.092, specificity χ2=3.896, 0.048, all P<0.05). Conclusion The serum PCT and lactate levels of DKA patients are significantly higher than healthy people, and those in the infected patients are higher than the uninfected patients.The combined detection of these two indicators has higher value in the diagnosis of patients with DKA co-infection. Key words: Diabetic ketoacidosis; Calcitonin; Lactic acid; Infection; Diagnostic value

Therapeutic effect of Dongbai-Tonglin-Fang, a Chinese herbal formula, on urinary tract infection in rat model

Ethnopharmacological relevanceTraditional Chinese Medicine (TCM) has many obvious advantages in the treatment of chronic conditions such as urinary tract infection (UTI). Dongbai-Tonglin-Fang (DBTL), a Chinese herbal formula, has been used for the treatment of UTI for more than 40 years with proven efficacy. However, its mechanism of action is still unknown. Aim of the studyThe purpose of this study is to evaluate the therapeutic efficacy of DBTL and its mechanism of action in a rat UTI model. Materials and methodsE. coli solution induced UTI rat model was used to evaluate the therapeutic effect of DBTL on UTI. Biochemical indicators related to UTI were measured. The kidney tissue was stained with hematoxylin-eosin (HE) to observe pathological changes whilst the ear swelling, feet swelling, hot plate and body torsion tests were used to estimate the anti-inflammatory and analgesic effects of DBTL. ResultsAfter treatment with different doses of DBTL (1, 2, 4 g/kg), a decrease in weight of the kidney in the UTI rat model was observed. The contents of white blood cell, nitrite, urinary albumin, ketone body, bilirubin and occult blood in the urine were also reduced whilst an increase in the pH of urine was observed. HE staining showed that the pathological changes in the kidney tissue were alleviated. At the same time, ear swelling assay showed that the weight and the degree of swelling of the ear of the mice in DBTL groups were decreased remarkably. DBTL also reduced the degree of feet swelling of the rats caused by the adjuvant. Furthermore, with the DBTL treatment, the latency period of foot licking induced by thermal stimulation was increased while the number of twists was lessened. ConclusionThese results show that DBTL has an excellent therapeutic effect on UTI rats accompanying with anti-inflammation and analgesia. The data presented here lays the foundations for further investigations in the treatment of UTI.

Ketone ester supplementation blunts overreaching symptoms during endurance training overload.

Key points Overload training is required for sustained performance gain in athletes (functional overreaching). However, excess overload may result in a catabolic state which causes performance decrements for weeks (non‐functional overreaching) up to months (overtraining).Blood ketone bodies can attenuate training‐ or fasting‐induced catabolic events. Therefore, we investigated whether increasing blood ketone levels by oral ketone ester (KE) intake can protect against endurance training‐induced overreaching.We show for the first time that KE intake following exercise markedly blunts the development of physiological symptoms indicating overreaching, and at the same time significantly enhances endurance exercise performance.We provide preliminary data to indicate that growth differentiation factor 15 (GDF15) may be a relevant hormonal marker to diagnose the development of overtraining.Collectively, our data indicate that ketone ester intake is a potent nutritional strategy to prevent the development of non‐functional overreaching and to stimulate endurance exercise performance. It is well known that elevated blood ketones attenuate net muscle protein breakdown, as well as negate catabolic events, during energy deficit. Therefore, we hypothesized that oral ketones can blunt endurance training‐induced overreaching. Fit male subjects participated in two daily training sessions (3 weeks, 6 days/week) while receiving either a ketone ester (KE, n = 9) or a control drink (CON, n = 9) following each session. Sustainable training load in week 3 as well as power output in the final 30 min of a 2‐h standardized endurance session were 15% higher in KE than in CON (both P < 0.05). KE inhibited the training‐induced increase in nocturnal adrenaline (P < 0.01) and noradrenaline (P < 0.01) excretion, as well as blunted the decrease in resting (CON: −6 ± 2 bpm; KE: +2 ± 3 bpm, P < 0.05), submaximal (CON: −15 ± 3 bpm; KE: −7 ± 2 bpm, P < 0.05) and maximal (CON: −17 ± 2 bpm; KE: −10 ± 2 bpm, P < 0.01) heart rate. Energy balance during the training period spontaneously turned negative in CON (−2135 kJ/day), but not in KE (+198 kJ/day). The training consistently increased growth differentiation factor 15 (GDF15), but ∼2‐fold more in CON than in KE (P < 0.05). In addition, delta GDF15 correlated with the training‐induced drop in maximal heart rate (r = 0.60, P < 0.001) and decrease in osteocalcin (r = 0.61, P < 0.01). Other measurements such as blood ACTH, cortisol, IL‐6, leptin, ghrelin and lymphocyte count, and muscle glycogen content did not differentiate KE from CON. In conclusion, KE during strenuous endurance training attenuates the development of overreaching. We also identify GDF15 as a possible marker of overtraining.

Estimation Metabolic Status in High Yielding Dairy Cows During Transition Period and Full Lactation

Background: Major changes in the metabolic functions in high-yielding dairy cows occur during the transitional period and during lactation. Parturition and lactogenesis are accompanied by many physiological changes that facilitate the maintenance of homeostasis Consequently, physiological situations leading to a negative energy balance are coupled to an increased uncontrolled rate of body fat mobilisation and the increased fatty acids accumulation in hepatocytes and blood ketone bodies, resulting in disturbances of the morphological and physiological liver integrity. The objective of the present study was to estimate metabolic status in late pregnant, early lactation and full lactation Holstein dairy cows on the basis changes of blood concentrations of selected biochemical markers.Materials, Methods &amp; Results: The experiment included 36 Holstein cows. Three groups of clinically healthy cows were chosen from the herd. Group 1 consisted of late pregnant cows (n = 12) from 30 to 1 day (20 ± 15) to partus; Group 2 comprised early lactation cows (n = 12) in the first month of lactation (15 ± 12 days), and Group 3 included full lactation cows (n = 12) between 60 to 90 days of lactation (81 ± 30 days). Blood samples were collected from all cows, by punction of the jugular vein. Biochemical testing for markers in the blood serum showed significantly lower values (P &lt; 0.05) of glucose, TG, Tchol. and urea in early cows than in full lactation and late-pregnant cows. The blood concentration of NEFA and BHB was significantly increased (P &lt; 0.05) in the group of cows in early lactation compared to the other groups of cows. The mean tBIL. concentration and the serum AST, GGT and ALT activities were markedly increased (P &lt; 0.01) in the lactation cows compared to the late pregnant cows. Furthermore, the intensity of lipomobilisation (NEFA or BHB concentrations) correlated positively (P &lt; 0.05) with the markers of cell damage or liver function impairment (tBIL., serum AST, ALT and GGT activities), but negatively (P &lt; 0.05) with the circulating concentrations of compounds synthesised in liver (glucose, TG and urea).Discussion: In dairy cows, it was observed that up to 50 % of females exhibited some lipid accumulation in liver in the first 3 weeks after calving and that fatty liver occurs primarily in this period. Liver can be categorized into mild, moderate and severe fatty liver as dependent on the degree of pathology and a mild fatty infiltration of liver in dairy cows during lactation is considered to be almost physiological. The blood BHB and NEFA concentrations are markers of lipomobilisation and positively associated with the ketosis and liver steatosis intensity. The simultaneous and parallel variations observed between the extent of the fat infiltration in liver and the serum BHB and NEFA concentrations in puerperal cows clearly indicated that the intense lipomobilisation in the post-partum period has induced lipid overloading and ketogenesis in the liver. On the other hand, it was observed significant decreases in the serum biochemical markers, at least partially synthesised in the liver, such as glucose, TG, Tchol., urea, albumin and TP during the postpartum period. However, the liver steatosis has induced some cellular lesions as evidenced by significant increases in the serum tBIL. concentrations and in the AST, ALT, GGT and LDH enzyme activities in puerperal cows. All these biochemical metabolites may be used as important biochemical markers in the determination of the metabolic status in high-yielding dairy cows during the transition period and during lactation.

A Spectroelectrochemical Sweat Sensor for Improved Management of Diabetes Mellitus

Despite recent attention, wearable fitness tracking devices (e.g., Garmin®, Fitbit® and Apple Watch®) remain unable to monitor overall human and animal health in real time. Continuous measurement enables: (1) alerts of abnormal status and (2) collection of large datasets to beget better models of human health. Here, we focus on continuous measurement of diabetic status. Presently, diabetic status is assessed through invasive and expensive blood glucose measurements. As such, continuous measurement in this way is impractical. Often, insufficient measurements are collected and diabetic excursions are undetected (especially in children and animals) which can result in severe complications including death. Some noninvasive techniques exist (e.g., Breath - Ketonix® and Urine) and rely on high levels of blood ketone bodies (e.g., acetoacetate (acac), β-hydroxybutyrate (bhb), and acetone) present with dysregulated diabetes. Blood ketone bodies partition into breath, tears, and sweat and enable noninvasive measurement. However, modern noninvasive techniques are subject to significant irreproducibility and inaccuracy which limits their use by diabetic patients. Alternatively, we describe spectroelectrochemical measurement of acac and bhb in sweat through complexation with metal cations generated at the electrode surface. This concept is advantageous as it may enable continuous quantitative measurement by small portable electronics (such as a cell phone or smart watch). Furthermore, this platform may be extended to other biomarkers that partition into sweat that give insight into other aspects of human health (e.g., heart and kidney failure, exercise intensity, and weight loss).