Glucose glu

Bifidobacterium longum subsp. infantis ATCC 15697 and B. breve SC95 were routinely grown for 48 h anaerobically at 37 °C in the semisynthetic de Man-Rogosa-Sharpe (MRS) broth or MRS agar (Becton Dickinson) supplemented with 1 % (wt/vol) L-cysteine. Single colony isolates were inoculated into modified MRS (mMRS) without sugar supplemented with 2 % of filter-sterilized (wt/vol) lactose (LAC) (Sigma Aldrich), 2 % glucose (GLU) (Fisher) or 2 % purified human milk oligosaccharide mixture (HMO) as the sole carbon source. HMO was kindly provided by the laboratory of Dr. Bruce German (Department of Food Science and Technology, University of California, Davis) and purified according to the method described in Gnoth et al. [53 (link)].

All chemicals were of analytical grade and de-ionized water (conductivity <0.1 μS/cm) was employed. The following reagents were obtained from Sigma-Aldrich: Breast Tumour Antigen from human fluids (CA15-3), myoglobin (Myo), o-phenylenediamine (oPDA), p-phenylenediamine (pPDA), 3-hydroxytyraminium chloride (Dopamine, DPM) and trypsin. Potassium hexacyanoferrate III (K₃[Fe(CN)₆]), L-ascorbic acid (AA) and potassium hexacyanoferrate II (K₄[Fe(CN)₆] trihydrate were obtained from Riedel-De Haën. 4-Styrenesulfonic Acid Sodium Salt (SA) was obtained from Alfa Aesar. Potassium di-hydrogenphosphaste (KH₂PO₄) and Sodium chloride (NaCl) were obtained from Panreac. Potassium chloride (KCl) was obtained from Merck. Glucose (Glu) was obtained from Fisher BioReagents. Creatine kinase isoenzyme (CK-MB) was obtained from ERM-European Reference Materials. Creatinine (Crea) and bovine serum albumin (BSA) were obtained from Amresco and Aniline from Analar Normapur.

After being enrolled in the study, participants were assigned a participant code in an ascending order. This number operated as study ID throughout the course of the study. They were then allocated to the order in which the trial visits were conducted in a randomized crossover fashion set using the software Research Randomizer^® [21 ]. A researcher not otherwise involved in the trial randomized the order of exercises (1:1:1) and the order of consumed beverages in a crossover fashion (1:1:1:1) as shown in Figure 1.
At the start of each study visit, participants received either 1 g/kg body mass (BM) of glucose (Glu) (Fisher Scientific, Lough-borough, United Kingdom), 1 g/kg BM of fructose (Fru) (Grüssing, Felsum, Germany), and 0.5 g/kg BM of a mixture of glucose and fructose (GluFru) (each) dissolved in 300 mL water. The amount of placebo, Sucralose (MyProtein, Norwich, United Kingdom), was administered as a fixed 0.2 g per dosage with the intention to mimic the taste of the other trial products. Between each visit, a minimum period of 48 h was maintained to ensure that glycogen stores were replenished. Participants were allowed to eat regularly in this period but were told to avoid strenuous exercise sessions. This procedure was conducted for any of the twelve visits (four exercise sessions consisting of including cycling, chest- and leg-press visits).