Galacto-oligosaccharide preconditioning improves metabolic activity and engraftment of Limosilactobacillus reuteri and stimulates osteoblastogenesis ex vivo.

A probiotic-related benefit for the host is inherently linked to metabolic activity and integration in the gut ecosystem. To facilitate these, probiotics are often combined with specific prebiotics in a synbiotic formulation. Here, we propose an approach for improving probiotic metabolic activity and engraftment. By cultivating the probiotic strain in the presence of a specific prebiotic (preconditioning), the bacterial enzymatic machinery is geared towards prebiotic consumption. Today, it is not known if preconditioning constitutes an advantage for the synbiotic concept. Therefore, we assessed the effects galacto-oligosaccharide (GOS) addition and preconditioning on GOS of Limosilactobacillus reuteri DSM 17938 on ex vivo colonic metabolic profiles, microbial community dynamics, and osteoblastogenesis. We show that adding GOS and preconditioning L. reuteri DSM 17938 act on different scales, yet both increase ex vivo short-chain fatty acid (SCFA) production and engraftment within the microbial community. Furthermore, preconditioned supernatants or SCFA cocktails mirroring these profiles decrease the migration speed of MC3T3-E1 osteoblasts, increase several osteogenic differentiation markers, and stimulate bone mineralization. Thus, our results demonstrate that preconditioning of L. reuteri with GOS may represent an incremental advantage for synbiotics by optimizing metabolite production, microbial engraftment, microbiome profile, and increased osteoblastogenesis.

Body composition assessment in children with inflammatory bowel disease: A comparison of different methods.

AIM: To assess different techniques to measure body composition in paediatric patients with inflammatory bowel disease using dual energy X-ray absorptiometry as a reference method. We hypothesised that a three-compartment model may demonstrate superiority over other methods as skinfold thickness equations and bioelectrical impedance analysis. METHODS: Body composition was assessed using skinfold thickness equations, bioelectrical impedance analysis and the three-compartment model. Data obtained with these methods were compared to the results obtained by dual energy X-ray absorptiometry. Statistical analysis was performed using Spearman's correlation and Bland-Altman's limits of agreement method. RESULTS: Twenty-one paediatric patients with inflammatory bowel disease were included: 11 females and 10 males; mean age for the entire group: 14.3 years, range 12-16 years. In children with inflammatory bowel disease, skinfold thickness equations, bioelectrical impedance analysis and the three-compartment model showed reliable measurements with small differences in the percentage of total body fat and good limits of agreements. CONCLUSION: The assessment of body composition using bioelectrical impedance analysis provides a valid and accurate method in children with inflammatory bowel disease as compared to dual energy X-ray absorptiometry. In the future, superiority of 3-compartment model in research and clinical settings of nutritional intervention and disease status in children with inflammatory bowel disease remains to be demonstrated.

Total and activity-induced energy expenditure measured during a year in children with inflammatory bowel disease in clinical remission remain lower than in healthy controls.

BACKGROUND: Pediatric inflammatory bowel disease (IBD) is often associated with growth retardation due to malnutrition. However, knowledge on total energy expenditure (TEE), active-induced energy expenditure (AEE) and physical activity remains limited in children with IBD. OBJECTIVE: Assessment of TEE using the doubly labelled water (DLW) method, resting energy expenditure (REE) using indirect calorimetry, and physical activity level using the actigraph GT3X+ in children with IBD (in remission) and healthy controls. METHODS: TEE, REE, AEE and physical activity were measured in 21 children with IBD and 24 healthy controls at baseline. IBD children parameters were monitored further after 6 and 12 months. Predicted REE and TEE values (using Schoefield and the actigraph GT3X+, for REE and TEE respectively) were compared to measured values. RESULTS: Mean ages at baseline were 14.8 ± 1.5 and 13.2 ± 2 years in children with IBD and in healthy control children, respectively. Measured TEEDLW was significantly lower (P < 0.001) in children with IBD compared to the healthy control group. REE corrected by FFM0.5, REE and AEE were also significantly lower in children with IBD. Children with IBD had AEE of 17.5% of TEE and had a significantly higher sedentary behaviour as compared to healthy children. CONCLUSIONS: This study suggests that TEE and AEE are reduced in children with IBD in clinical remission which may result in a reduced moderate and vigorous physical activity level. Our result also highlights that the actigraph GT3X + might give good prediction of TEE in children with IBD at group level but it remains highly variable at individual level.

Exclusive Functional Treatment for Mandibular Condylar Fractures.

PURPOSE: Despite the frequency of condylar fractures, no consensus has been reached regarding treatment alternatives (ie, operative vs nonoperative). The purpose of the present study was to describe functional treatment without intermaxillary fixation of condylar fractures and summarize the treatment outcomes. PATIENTS AND METHODS: We designed a retrospective case series and enrolled patients with condylar fractures with malocclusion. The primary outcome was treatment success, assessed at the end of 6 weeks. We recorded 5 parameters to assess treatment success, including a stable return to the initial occlusion, lateral excursion of more than 7 mm, centered protrusion of more than 4 mm, centered mouth opening of more than 35 mm, and painless mandibular mobilization. If all 5 parameters were met, treatment was deemed a success. Treatment failure was defined as meeting less than all 5 parameters. We used numbers and percentages for these nominal qualitative variables (healing criterion absent or present). Two prognostic variables (ie, fracture type and age range), stratified by treatment success, were compared using the Fisher exact test for patients presenting with all healing criteria. RESULTS: We included 30 patients with a median age of 33 years. Of the 30 patients (34 fractures), 15 had low subcondylar fractures (44.1%), 12 had high subcondylar fractures (35.3%), and 7 had head condylar fractures (20.5%). At the last consultation, 21 patients (70%) had exhibited all the criteria defining treatment success. A significant difference was found in the success rate when stratified by age (P = .002) in favor of the younger patients. Also, high subcondylar and head fractures were associated with a better success rate. CONCLUSIONS: Exclusive functional treatment showed promising results and should be considered for cooperative patients, avoiding the risks of surgery and the discomfort with intermaxillary fixation.

Time course of fractional gluconeogenesis after meat ingestion in healthy adults: a D2O study.

In the postprandial state, glucose homeostasis is challenged by macronutrient intake, including proteins that trigger insulin secretion and provide glucose precursors. However, little is known about the postprandial response of gluconeogenesis to a protein meal. We aimed to quantify the evolution of fractional gluconeogenesis after a meat meal. Thirteen healthy subjects received oral doses of D2O. After fasting overnight, they ingested a steak (120 g). Glycemia, insulinemia, and 2H enrichments in glucose and plasma water were measured for 8 h after the meal. Fractional gluconeogenesis was assessed using the average method. Glucose was stable for 5 h and then decreased. There was a slight increase of insulin 1 h after the meal. 2H enrichment in the carbon 5 position of glucose (C5) increased after 2 h, whereas it decreased in plasma water. Consequently, fractional gluconeogenesis increased from 68.2 ± 7.2% before the meal to 75.5 ± 5.8% 8 h after the meal, the latter corresponding to 22 h without a glucose supply. These values are consistent with the exhaustion of glycogen stores after 24 h but represent the highest among values in the literature. The impact of methodological conditions is discussed.

A high-throughput method for liquid chromatography-tandem mass spectrometry determination of plasma alkylresorcinols, biomarkers of whole grain wheat and rye intake.

Plasma alkylresorcinols are increasingly analyzed in cohort studies to improve estimates of whole grain intake and their relationship with disease incidence. Current methods require large volumes of solvent (>10 ml/sample) and have relatively low daily sample throughput. We tested five different supported extraction methods for extracting alkylresorcinols from plasma and improved a normal-phase liquid chromatography coupled to a tandem mass spectrometer method to reduce sample analysis time. The method was validated and compared with gas chromatography-mass spectrometry analysis. Sample preparation with HybridSPE supported extraction was most effective for alkylresorcinol extraction, with recoveries of 77-82% from 100 µl of plasma. The use of 96-well plates allowed extraction of 160 samples per day. Using a 5-cm NH2 column and heptane reduced run times to 3 min. The new method had a limit of detection and limit of quantification equivalent to 1.1-1.8 nmol/L and 3.5-6.1 nmol/L plasma, respectively, for the different alkylresorcinol homologues. Accuracy was 93-105%, and intra- and inter-batch precision values were 4-18% across different plasma concentrations. This method makes it possible to quantify plasma alkylresorcinols in 100 µl of plasma at a rate of at least 160 samples per day without the need for large volumes of organic solvents.

A whole-grain-rich diet reduces urinary excretion of markers of protein catabolism and gut microbiota metabolism in healthy men after one week.

Epidemiological studies consistently find that diets rich in whole-grain (WG) cereals lead to decreased risk of disease compared with refined grain (RG)-based diets. Aside from a greater amount of fiber and micronutrients, possible mechanisms for why WGs may be beneficial for health remain speculative. In an exploratory, randomized, researcher-blinded, crossover trial, we measured metabolic profile differences between healthy participants eating a diet based on WGs compared with a diet based on RGs. Seventeen healthy adult participants (11 female, 6 male) consumed a controlled diet based on either WG-rich or RG-rich foods for 2 wk, followed by the other diet after a 5-wk washout period. Both diets were the same except for the use of WG (150 g/d) or RG foods. The metabolic profiles of plasma, urine, and fecal water were measured using (1)H-nuclear magnetic resonance spectroscopy and gas chromatography-mass spectrometry (plasma only). After 1 wk of intervention, the WG diet led to decreases in urinary excretion of metabolites related to protein catabolism (urea, methylguanadine), lipid (carnitine and acylcarnitines) and gut microbial (4-hydroxyphenylacetate, trimethylacetate, dimethylacetate) metabolism in men compared with the same time point during the RG intervention. There were no differences between the interventions after 2 wk. Urinary urea, carnitine, and acylcarnitine were lower at wk 1 of the WG intervention relative to the RG intervention in all participants. Fecal water short-chain fatty acids acetate and butyrate were relatively greater after the WG diet compared to the RG diet. Although based on a small population and for a short time period, these observations suggest that a WG diet may affect protein metabolism.

A protein-leucine supplement increases branched-chain amino acid and nitrogen turnover but not performance.

PURPOSE: This study aimed to determine the effect of postexercise protein-leucine coingestion with CHO-lipid on subsequent high-intensity endurance performance and to investigate candidate mechanisms using stable isotope methods and metabolomics. METHODS: In this double-blind, randomized, crossover study, 12 male cyclists ingested a leucine/protein/CHO/fat supplement (LEUPRO 7.5/20/89/22 g · h(-1), respectively) or isocaloric CHO/fat control (119/22 g · h(-1)) 1-3 h after exercise during a 6-d training block (intense intervals, recovery, repeated-sprint performance rides). Daily protein intake was clamped at 1.9 g · kg(-1) · d(-1) (LEUPRO) and 1.5 g · kg(-1) · d(-1) (control). Stable isotope infusions (1-(13)C-leucine and 6,6-(2)H2-glucose), mass spectrometry-based metabolomics, and nitrogen balance methods were used to determine the effects of LEUPRO on whole-body branched-chain amino acid (BCAA) and glucose metabolism and protein turnover. RESULTS: After exercise, LEUPRO increased BCAA levels in plasma (2.6-fold; 90% confidence limits = ×/÷ 1.1) and urine (2.8-fold; ×/÷ 1.2) and increased products of BCAA metabolism plasma acylcarnitine C5 (3.0-fold; ×/÷ 0.9) and urinary leucine (3.6-fold; ×/÷ 1.3) and ß-aminoisobutyrate (3.4-fold; ×/÷ 1.4), indicating that ingesting ~10 g leucine per hour during recovery exceeds the capacity to metabolize BCAA. Furthermore, LEUPRO increased leucine oxidation (5.6-fold; ×/÷ 1.1) and nonoxidative disposal (4.8-fold; ×/÷ 1.1) and left leucine balance positive relative to control. With the exception of day 1 (LEUPRO = 17 ± 20 mg N · kg(-1), control = -90 ± 44 mg N · kg(-1)), subsequent (days 2-5) nitrogen balance was positive for both conditions (LEUPRO = 130 ± 110 mg N · kg(-1), control = 111 ± 86 mg N · kg(-1)). Compared with control feeding, LEUPRO lowered the serum creatine kinase concentration by 21%-25% (90% confidence limits = ± 14%), but the effect on sprint power was trivial (day 4 = 0.4% ± 1.0%, day 6 = -0.3% ± 1.0%). CONCLUSIONS: Postexercise protein-leucine supplementation saturates BCAA metabolism and attenuates tissue damage, but effects on subsequent intense endurance performance may be inconsequential under conditions of positive daily nitrogen balance.

Ventilatory disorders and facial growth: benefits of early genioplasty.

AIM: The purpose of this prospective study was to determine whether combined functional genioplasty and orthodontic treatment in patients with vertical mandibular growth can be beneficial not only in achieving an esthetic outcome and orthodontic stability but also for the multi-disciplinary management of oral ventilation. METHODS: Twenty-five non-obese adolescents (mean age: 14.6 ± 1.4 years) with vertical excess of the lower third requiring genioplasty received surgery following orthodontic treatment. All were predominantly mouth-breathers despite ENT treatment and rehabilitation. The functional before-after impact of genioplasty was examined in a clinical setting using polysomnography. RESULTS: Before genioplasty, 52% of the adolescents presented an upper airways resistance syndrome (UARS) with obstructive hypopnea and poor sleep quality. Three to 6 months after genioplasty, the mode of ventilation shifted from oral to nasal (p < 0.001). Lip seal was significantly restored with no contraction of the labio-mental muscles. All nighttime symptoms improved. The proportion of patients suffering from ronchopathy (pathological snoring), dry mouth and disturbed sleep dropped significantly (p = 0.08, p = 0.001, p = 0.0009, respectively). Respiratory events and sleep pattern became normal. Below, we present two clinical reports involving obstructive sleep apnea syndrome (OSAS). CONCLUSIONS: Genioplasty performed during puberty promotes spontaneous lip closure and helps restore nasal ventilation. It improves the obstructive disorder and its manifestations during sleep.

A plasma global metabolic profiling approach applied to an exercise study monitoring the effects of glucose, galactose and fructose drinks during post-exercise recovery.

A global metabolic profiling methodology based on gas chromatography coupled to time-of-flight mass spectrometry (GC-TOFMS) for human plasma was applied to a human exercise study focused on the effects of beverages containing glucose, galactose, or fructose taken after exercise and throughout a recovery period of 6 h and 45 min. One group of 10 well trained male cyclists performed 3 experimental sessions on separate days (randomized, single center). After performing a standardized depletion protocol on a bicycle, subjects consumed one of three different beverages: maltodextrin (MD)+glucose (2:1 ratio), MD+galactose (2:1), and MD+fructose (2:1), consumed at an average of ∼1.25 g of carbohydrate (CHO) ingested per minute. Blood was taken straight after exercise and every 45 min within the recovery phase. With the resulting blood plasma, insulin, free fatty acid (FFA) profile, glucose, and GC-TOFMS global metabolic profiling measurements were performed. The resulting profiling data was able to match the results obtained from the other clinical measurements with the addition of being able to follow many different metabolites throughout the recovery period. The data quality was assessed, with all the labelled internal standards yielding values of <15% CV for all samples (n=335), apart from the labelled sucrose which gave a value of 15.19%. Differences between recovery treatments including the appearance of galactonic acid from the galactose based beverage were also highlighted.