Objective: To examine the effects of a s1 -casein hydrolysate on females with stress-related symptoms. Design: Double-blind, randomized, crossover, placebo-controlled trial. Setting: The a s1 -casein hydrolysate was manufactured by INGREDIA (Arras, France) and the placebo was manufactured by DIETAROMA (Bourg, France). Study was designed and performed at PROCLAIM (Rennes, France), and the statistical analyses were performed by D Desor (Nancy, France). Subjects: A total of 63 female volunteers suffering from at least one disorder that may be related to stress such as anxiety, sleep problems and general fatigue. Interventions: A total of 63 volunteers participated in a double-blind, randomized, crossover, placebo-controlled study. Subjects were randomly allocated to receive either tablets containing a s1 -casein hydrolysate or placebo at the dose of 150 mg/ day for 30 days. After a 3 weeks washout period, they were crossed over for a new 30-day period of tablets intake. The outcome measure was a questionnaire including 44 items of symptoms that may be related stress in which the severity of each sign was evaluated using a 10-degree scale. These measures were studied repeatedly at the day of 0, 15 and 30 after the start of each interventional period. Results: The 30-day treatment by a s1 -casein hydrolysate in females with stress-related symptoms reduced their symptoms, particularly in digestion (Po0.01), cardiovascular (Po0.05), intellectual (Po0.01), emotional (Po0.05) and social problems (Po0.05). Conclusion: This study showed that a 30-day ingestion of a s1 -casein hydrolysate decreased the stress-related symptoms in females suggesting that this product may be used as an effective functional ingredient alleviating such symptoms.

Background: Various extracts of Hovenia dulcis have been commonly used in Asia for cases of alcohol-related disorders. Fermentation is reported to enhance the level and biological activities of various bio-constituents of plant extracts. Therefore, this study was undertaken to evaluate the effects of fermented H. dulcis extract (FHDE) on ethanol-induced liver injury in mice. Methods: FHDE was prepared using Bacillus subtilis and Lactobacillus plantarum. The effects of FHDE on ethanolinduced liver injury were evaluated in C57BL/6 N CrSlc mice. A mixed feed preparation containing the fermented extract with and without ethanol was given to mice for 29 days, according to its group. At the end of the experiment, blood and liver samples were collected from all mice in the group. Plasma biochemical analysis and histopathological investigation were performed to evaluate the impacts of treatment on the biomarkers of hepatic damage and inflammatory changes. Besides, the expression of genes that regulate the activities of enzymes associated with alcohol metabolism, antioxidant activity, and fatty acid oxidation was assessed using a quantitative real-time polymerase chain reaction. Moreover, the amino acid contents and the active ingredients of the extract were evaluated before and after fermentation.

Hovenia dulcis Thunb. extract (HDE) was co-fermented by Bacillus subtilis HA and Lactobacillus plantarum EJ2014. The co-fermentation was optimized by adding glucose (3% for the first fermentation, 1.5% for the second fermentation), MSG (5%), and skim milk (5%). The HDE broth fermented by B. subtilis HA showed pH 7.3, 0.09% acidity, 0.52 mg/g tyrosine equivalents, and viable bacterial counts of 1.4×10 9 CFU/mL. The viscous broth obtained indicated a consistency index of 3.90 Pa.s n and 32.70 mg/mL mucilage after 2 days. Subsequently, after the second fermentation, the broth indicated pH 4.5, 1.08% acidity, 0.87 mg/g tyrosine equivalents and viable bacterial counts of 2.0×10 6 and 3.0×10 9 CFU/mL for B. subtilis HA and L. plantarum EJ2014, respectively. The MSG substrate was effectively converted into GABA, which showed a final concentration of 16.08 mg/mL in the broth. Therefore, the co-fermented HDE was effectively fortified with mucilage, GABA, peptides, and probiotics, and could be used as a functional ingredient for nutraceuticals.

Milk was co-fermented with Bacillus subtilis HA and Lactiplantibacillus plantarum EJ2014 to produce a dairy ingredient enriched with poly-γ-glutamic acid (γ-PGA) and γ-aminobutyric acid (GABA). The first fermentation of milk with B. subtilis HA resulted in a viscous broth with pH 6.56, 0.26% acidity, 1.40 mg/g tyrosine equivalent, and 17.21 U/g protease activity. The viable cell counts of B. subtilis indicated 8.74 log CFU/mL, and the consistency index of the alkaline fermented milk was 1.82 Pa·sn. In addition, 4.65% mucilage was produced with 35.93% γ-PGA content. The milk co-fermented by L. plantarum indicated 1.34% acidity and pH 4.91. The viable bacterial counts of B. subtilis decreased to 4.44 log CFU/mL, whereas those of L. plantarum increased to 9.42 log CFU/mL. Monosodium glutamate (MSG) as a precursor was effectively converted into γ-PGA by B. subtilis, and then residual MSG was completely converted into GABA by L. plantarum with a yield of 26.15 mg/g. Furthermore, the co-fermented milk produced volatiles, including hexanoic acid, 2,3-butanediol, and acetoin, which may be responsible for its aged cheese-like aroma.

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