Either the volunteer or a relative gave their written informed consent, and the study was approved by the ethical committee of Hospital District of Southwest Finland. Exclusion criteria were the consumption of antibiotics in the last selleck month and use of medication expected to either affect the immune function and/or the intestinal microbiota of the subject. Another exclusion criterion was the habitual use of pro- and/or prebiotic-containing products. The study protocol consisted of three consecutive phases. In phase 1, the subjects
consumed a control cheese during breakfast for 2 weeks (run-in). In phase 2, the subjects consumed a probiotic cheese for 4 weeks (intervention). In phase 3, the subjects consumed the same control cheese Selumetinib manufacturer again for 4 weeks (wash-out). The products were blinded to the volunteers and were identical in taste and appearance. The total duration of the study was 10 weeks, and during the time, the food at the elderly home remained stable. Heparinized peripheral blood (9 mL) was drawn by a venipuncture from each subject at baseline (T0), after run-in (T1), after intervention (T2), and after wash-out (T3) for immunological analysis. On the same occasion, a blood sample was collected for general health monitoring tests carried out at the University of Turku Hospital. The probiotic and the control Gouda cheese were commercial products (Mills DA, Oslo, Norway). Identical slices
of both types of cheese (15 g) were prepared and packed before the commencement of the study. The probiotic cheese slice contained approximately 109 CFU of L. rhamnosus HN001 (AGAL NM97/09514) and L. acidophilus NCFM (ATCC 700396). The viability of the strains was assessed throughout the study and was observed to remain stable. Both probiotic and control cheese contained proprietary starter strains (Choozit 712™, Danisco, Paris). The volunteers consumed one slice of cheese per day during breakfast. The probiotic cheese had been on the Norwegian
market for approximately 1 year. The probiotic strains Enzalutamide cost have been in commercial use for approximately 7 years (L. rhamnosus HN001) and 30 years (L. acidophilus NCFM) and have substantial safety and efficacy data (Shu et al., 1999; Zhou et al., 2000; Gill & Rutherfurd, 2001; Sanders & Klaenhammer, 2001; Sheih et al., 2001). The same probiotic cheese was tested for bacterial survival using a human gastrointestinal tract-simulating model, and it was shown that the strains (L. acidophilus NCFM and L. rhamnosus HN001) survived the simulated upper gastrointestinal tract (Makelainen et al., 2009). The cytotoxicity of the peripheral blood mononuclear cells (PBMCs), proportions of CD3−CD56+ cells (NK cells), CD3+CD56+ cells (NKT cells), CD3+CD56− cells, and CD3−CD56− cells in the total PBMCs, and phagocytic activity were assessed using flow cytometry (FACScan flow cytometer, BD biosciences). The data were analyzed using cellquest pro software.