Effects of Milk Fat Globule Membrane Supplementation Following Exercise Training on Physical Performance in Healthy Young Adults: A Randomized Double-Blind, Placebo-Controlled Pilot Trial.

The purpose of this study was to examine whether 4 wk of daily ingestion of milk fat globule membrane (MFGM) combined with exercise training improves physical performance-muscle strength, agility and muscle power-in healthy young adults. The study was designed as a randomized, double-blind, and placebo-controlled trial. Twenty healthy young adults received either an MFGM powder containing 1.6 g of fat and 160 mg of sphingomyelin or an isocaloric placebo powder daily throughout 4 wk of power or agility training. Physical performance tests and body composition measurements were conducted before and after the 4-wk intervention. Ingestion of MFGM did not affect isometric or isokinetic muscle strength, but it was associated with a greater increase in vertical jump peak power compared with placebo. There were no significant changes in body weight or lean body mass during the intervention period in either group, and no significant differences between groups. We conclude that daily MFGM supplementation combined with exercise training has the potential to improve physical performance in young adults; however, further studies with larger sample sizes should be conducted to obtain more evidence supporting achievement of improved physical performance through MFGM supplementation.

Milk-based culture of Penicillium camemberti and its component oleamide affect cognitive function in healthy elderly Japanese individuals: a multi-arm randomized, double-blind, placebo-controlled study.

Dairy products and fermented foods have a reported association with maintained cognitive function. Camembert cheese, a dairy product fermented by the white mold Penicillium camemberti, has also been shown to enhance cognitive function in vivo. Oleamide, derived from the fermentation of the white mold, is a candidate for an active component, and expected to improve both cognitive function and sleep conditions. Thus, this study investigated whether the milk-based culture of white mold (MCW), and oleamide, could improve cognitive function and sleep state clinically. A multi-arm randomized, double-blind, placebo-controlled trial was conducted in Tokyo, Japan. 60 healthy Japanese individuals aged 50-75 who were aware of their cognitive decline were randomly and equally divided into three groups of 20 participants using computer-generated random numbers. Participants took either MCW (equivalent to 60 µg/day of oleamide), 60 µg/day of oleamide, or placebo capsules for 12 weeks. Serum BDNF, cognitive function by Cognitrax as primary and MCI Screen as secondary outcome, and sleep status using the Japanese version of the Pittsburgh Sleep Quality Index (PSQI-J) were assessed before and after intervention. The participants, outcome assessors and analysts, and research assistants were blinded to the group assignment. Of the 60 participants, 58 completed the study and were analyzed. No adverse events related to test foods were observed. The placebo group showed a negative rate of change in serum BDNF (-10.5% ± 19.7%), whereas the MCW and oleamide groups showed positive changes (2.0% ± 27.1% and 1.3% ± 13.5%, respectively). Cognitrax scores increased after 12 weeks in all groups. Conversely, the MPI score of the MCI Screen demonstrated a significant improvement in the MCW and oleamide groups compared to the placebo group (p = 0.013 and p < 0.001, respectively). The subscales, immediate free recall and delayed free recall, also significantly increased in them compared to the placebo group. Although PSQI-J revealed no significant differences among groups, the MCW and oleamide groups showed significant improvement after intervention in overall score, subjective sleep quality, and sleep latency. Our results suggest that MCW and its component, oleamide, are safe and contribute to maintaining cognitive functions, particularly short-term and working memory, and improving sleep state. Clinical trial registration: https://center6.umin.ac.jp/cgi-open-bin/ctr/ctr_view.cgi?recptno=R000054792, identifier UMIN-CTR UMIN000048084.

Ingesting Yogurt Containing Lactobacillus plantarum OLL2712 Reduces Abdominal Fat Accumulation and Chronic Inflammation in Overweight Adults in a Randomized Placebo-Controlled Trial.

BACKGROUND: Chronic inflammation and insulin resistance are factors that are related to obesity. We have suggested that the administration of heat-treated Lactobacillus plantarum OLL2712 (OLL2712) cells can improve glucose and lipid metabolism by suppressing chronic inflammation in mouse models and a preliminary clinical study. OBJECTIVE: The aim of this study was to investigate whether ingesting OLL2712 cells can reduce body fat accumulation and improve metabolic risk factors, in overweight, healthy adults. METHODS: This study was a randomized, double-blind, placebo-controlled, parallel-group trial conducted at a single center in Japan. The study participants included 100 overweight (BMI range, ≥25 to <30 kg/m2) adults aged 20-64 y. They were randomly assigned to either the placebo or OLL2712 group (n = 50 each) and were administered conventional yogurt or yogurt containing >5 × 109 heat-treated OLL2712 cells, respectively, daily for 12 wk. The primary outcome was the 12-wk change in the abdominal fat area, as assessed by computed tomography, and the secondary outcomes were glucose and lipid metabolism-related parameters and chronic inflammation markers, which were analyzed using a linear mixed model. RESULTS: The 12-wk change of abdominal fat area (difference: 8.5 cm2; 95% CI: 0.3, 16.6 cm2; P = 0.040) and fasting plasma glucose (difference: 3.2 mg/dL; 95% CI: 0.8, 5.6 mg/dL; P = 0.021) were significantly less in the OLL2712 group than the placebo group. The overall trend of serum IL-6 was significantly decreased in the OLL2712 group compared with baseline and the placebo group. CONCLUSIONS: The ingestion of heat-treated OLL2712 cells reduces body fat accumulation and the deterioration of glycemic control and chronic inflammation, in overweight, healthy adults. We hypothesize that OLL2712 cells may prevent obesity by regulating chronic inflammation. This trial was registered at the University Hospital Medical Information Network Clinical Trials Registry as UMIN000027709.

Effects of 12-wk Lactobacillus plantarum OLL2712 treatment on glucose metabolism and chronic inflammation in prediabetic individuals: A single-arm pilot study.

OBJECTIVE: Previously, we demonstrated that the administration of heat-killed OLL2712 cells suppressed chronic inflammation and improved hyperglycemia in a mouse model of obesity and diabetes. The aim of this study was to preliminarily examine the effect of OLL2712 supplementation on glucose metabolism and chronic inflammation in prediabetic subjects. METHODS: This study was a prospective, 12-wk, single-arm, open trial, followed by a 4-wk posttreatment period. Inclusion criteria were fasting plasma glucose levels of 105 to 130 mg/dL in an age range of 35 to 65 y. Thirty individuals consumed a dairy beverage containing ∼1 × 1010 heat-killed OLL2712 cells for 12 wk. RESULTS: The ingestion of the OLL2712 beverage significantly improved fasting plasma glucose levels, serum glycoalbumin levels, and insulin resistance indexes compared with baseline levels. The intervention also suppressed serum monocyte chemotactic protein-1 and interleukin-6 levels, which are proinflammatory cytokines involved in the development of insulin resistance and hyperglycemia. Furthermore, stratified analysis by these proinflammatory cytokine levels revealed that the beneficial effects of OLL2712 beverage were observed particularly in individuals with chronic inflammation at baseline. CONCLUSION: The results of this study suggested that heat-killed OLL2712 cells have the potential to improve insulin resistance and glucose metabolism by suppressing chronic inflammation.

Enhanced antibody production following intermediate addition based on flux analysis in mammalian cell continuous culture.

Generally, mammalian cells utilize glucose and glutamine as primary energy sources. To investigate the effect of energy sources on metabolic fluxes and antibody production, glucose- or glutamine-limited serum-free continuous culture of hybridoma 3A21 cells, which produce anti-ribonuclease A antibody, was carried out. The cell volume and dry cell weight were evaluated under various steady-state conditions. The specific consumption and production rates were evaluated on the basis of dry cell weight. On the basis of these results, the fluxes of the metabolic pathway were calculated. It was found that increasing the specific growth rate causes the specific ATP and antibody production rates to decrease. The fluxes between malate and pyruvate also decreased with the increase in specific growth rate. To increase the ATP production rate under steady-state conditions by the enhancement of fluxes between malate and pyruvate, the reduced metabolic fluxes were increased by an intermediate (pyruvate, malate, and citrate) addition. As a result, higher specific ATP and antibody production rates were achieved following the intermediate addition at a constant dilution rate.

Effect of growth conditions of Lactobacillus gasseri OLL2809 on the immunostimulatory activity for production of interleukin-12 (p70) by murine splenocytes.

Lactobacillus gasseri OLL2809, a probiotic lactic acid bacterium, strongly stimulates interleukin (IL)-12 (p70) production by murine splenocytes; therefore, it is expected to ameliorate allergic diseases. Although many studies have investigated characteristics of the immunostimulatory activity of probiotics, little is known about how bacterial growth conditions affect the activity. In this study, we investigated the effects of the growth conditions of L. gasseri OLL2809 on the stimulation of IL-12 (p70) production. L. gasseri OLL2809 was grown under various culture conditions including different cultivation periods, media, and culture pH, and IL-12 (p70) production by murine splenocytes stimulated with these bacterial cells was determined. The results revealed that IL-12 (p70) production (i) increased with the growth of the bacterial cells and was higher in stationary-phase cells than in logarithmic-phase cells; (ii) it was higher in the cells grown in acidic pH; and (iii) it decreased when the cells were incubated in a buffer at neutral pH prior to heat treatment. These observations indicated that stimulation of IL-12 (p70) production is affected by culture medium pH. In addition, the observations of a difference in the stimulation of IL-12 (p70) production by L. gasseri OLL2809 grown under various conditions are consistent with the characteristics of autolysis. Therefore, it was deduced that the integrity of the bacterial cells is necessary for the stimulatory effect on IL-12 (p70) production and that acidic pH and heat treatment contributed to the stimulation by inhibiting the activity of autolysins indigenous to the bacteria. Our result suggests that cultivation until the stationary phase under acidic pH is required for the effective production of probiotics with immunostimulatory activity.

Enhancement of 1,4-dihydroxy-2-naphthoic acid production by Propionibacterium freudenreichii ET-3 fed-batch culture.

The production of 1,4-dihydroxy-2-naphthoic acid (DHNA) was investigated using a fed-batch culture of Propionibacterium freudenreichii ET-3. DHNA is a precursor of menaquinone (MK) and is transformed to MK by combination with an isoprenoid unit. We found that ET-3 stopped MK production and increased DHNA production in an anaerobic fed-batch culture by maintaining the lactose concentration at approximately zero. The maximum DHNA concentration observed in the anaerobic fed-batch culture was markedly higher than the maximum DHNA concentration observed in an anaerobic batch culture. Moreover, MK or DHNA production was affected by the lactose feeding rate; this suggests that lactose metabolism participates in the syntheses of these products. On the other hand, accumulation of propionate was found to inhibit DHNA production in the fed-batch culture. Based on the fact that ET-3 increases DHNA production in an aerobic culture by consuming propionate, we carried out a cultivation experiment in which an anaerobic fed-batch culture was switched to an anaerobic batch culture and found that the DHNA production was increased to a greater extent than the DHNA production in an anaerobic fed-batch culture. These results suggest that DHNA production by ET-3 is markedly influenced by carbon source limitation and the oxygen supply.

Optimal aerobic cultivation method for 1,4-dihydroxy-2-naphthoic acid production by Propionibacterium freudenreichii ET-3.

To investigate the effects of oxygen supply on Propionibacterium freudenreichii ET-3 metabolism and 1,4-dihydroxy-2-naphthoic acid (DHNA) production in detail, the strain was cultured by switching from anaerobic condition to aerobic condition at 72 h (termed anaerobic-aerobic switching culture hereafter) employing different oxygen transfer rates (OTRs) in the range of 0.08-0.90 mg/(l.h). It was found that a 0.08 mg/(l.h) OTR could not change the metabolism or improve the DHNA production of P. freudenreichii ET-3. When the OTR was in the range of 0.23-0.66 mg/(l.h), propionate, which inhibits DHNA production significantly, was consumed during the aerobic phase. Final DHNA concentration increased to 0.22 mM, irrespective of OTR. When the OTR was 0.90 mg/(l.h), a sudden increase in dissolved oxygen (DO) concentration during the aerobic phase resulted in a sudden decrease in DHNA concentration. To attenuate the stresses caused by propionate and oxygen exposure, we designed an optimal cultivation in which the anaerobic and aerobic phases were repeated three times alternately. As a result, propionate concentration was maintained below the level that inhibits DHNA production, and no DO concentration was detected throughout the culture. The final DHNA concentration in this culture was 0.33 mM, which is 2.7-fold that in the anaerobic culture and 1.5-fold that in the anaerobic-aerobic switching culture.

Aerobic culture of Propionibacterium freudenreichii ET-3 can increase production ratio of 1,4-dihydroxy-2-naphthoic acid to menaquinone.

This is the first report on the production of both 1,4-dihydroxy-2-naphthoic acid (DHNA) and menaquinone by Propionibacterium freudenreichii ET-3. DHNA can be a stimulator of bifidogenic growth, and menaquinone has important roles in blood coagulation and bone metabolism. During anaerobic culture, DHNA and menaquinone concentrations reached 0.18 mM and 0.12 mM, respectively. The molar ratio between these products was approximately 3:2, which was not affected by culture pH and temperature over the ranges of 6.0-7.0 and 31-35 degrees C, respectively. As for organic acid, propionate and acetate accumulated at concentrations of 0.3 M and 0.15 M, respectively, and the propionate accumulation particularly inhibited further production of DHNA. To improve DHNA production, we switched from anaerobic condition to aerobic condition during the culture when lactose was depleted. DHNA concentration continued to increase even after lactose exhaustion, reaching 0.24 mM. In contrast to DHNA production, menaquinone production stopped after the switch to aerobic condition. The total molar production of DHNA and menaquinone was 0.3 mM irrespective of aerobic culture and anaerobic-aerobic switching culture. Therefore, the anaerobic-aerobic switching culture could increase the production ratio of DHNA to menaquinone. The DHNA concentration obtained from the anaerobic-aerobic switching culture was 1.3-fold higher than that in the anaerobic culture, because P. freudenreichii ET-3 utilized propionate accumulated in the medium via the reversed methylmalonyl CoA pathway under aerobic condition. The culture method proposed in this study could be applicable to industrial-scale fermentation using 1000 l of media, by which 0.23 mM DHNA was produced.