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In a screening for proteolytically active lactic acid bacteria, three strains, Lactobacillus delbrueckii ssp. lactis 92202, Lactobacillus helveticus 92201, and Lactobacillus delbrueckii ssp. bulgaricus 92059, showed the highest activities following growth in milk. All three strains degraded α- and β-casein, but did not hydrolyse κ-casein. HPLC analysis of skim milk fermentation revealed increasing amounts of peptides after 5 and 10 h with Lb. d. ssp. bulgaricus 92059. Hydrolysates obtained with Lb. d. ssp. lactis 92202 and Lb. d. ssp. bulgaricus 92059 revealed the highest angiotensin-converting enzyme-inhibitory effect. The effect was dose dependent. Almost no effect (<10%) was seen for Lb. helveticus 92201. For Lb. d. ssp. bulgaricus 92059, maximal inhibition of approx. 65% was reached after 25 h of fermentation. In an in vitro assay measuring potential immunomodulation, hydrolysates of the three strains yielded anti-inflammatory activities in the presence of TNF-α. However, the effects were more pronounced at lower hydrolysate concentrations. In the absence of TNF-α, slight pro-inflammatory effects were observed. The hydrolysate of Lb. d. ssp. bulgaricus 92059, when purified by means of solid-phase extraction, exhibited pro-inflammatory activity. Sour whey containing Lb. d. ssp. bulgaricus 92059 cells showed pro-inflammatory activity while cell-free sour whey was clearly anti-inflammatory. In the purified hydrolysate, 20 different α- and β-casein (CN)-derived peptides could be identified by LC-MS. Most peptides originated from the central and C-terminal regions of β-casein. Peptide length was between 9 (β-CN(f 59–67)) and 22 amino acids (β-CN(f 117–138)).

Background/Objectives: Exacerbated postprandial lipid responses are associated with an increased cardiovascular risk. Dietary proteins influence postprandial lipemia differently, and whey protein has a preferential lipid-lowering effect. We compared the effects of different whey protein fractions on postprandial lipid and hormone responses added to a high-fat meal in type 2 diabetic subjects. Subjects/Methods: A total of 12 type 2 diabetic subjects ingested four isocaloric test meals in randomized order. The test meals contained 100 g of butter and 45 g of carbohydrate in combination with 45 g of whey isolate (iso-meal), whey hydrolysate (hydro-meal), α-lactalbumin enhanced whey (lac-meal) or caseinoglycomacropeptide enhanced whey (CGMP-meal). Plasma concentrations of triglyceride, retinyl palmitate, free fatty acid, insulin, glucose, glucagon, glucagon-like peptide 1 and glucose-dependent insulinotropic peptide were measured before and at regular intervals until 8-h postprandially. Results: We found no statistical significant differences between meals on our primary variable triglyceride. The retinyl palmitate response was higher after the hydro-meal than after the iso- and lac-meal in the chylomicron-rich fraction ( P =0.008) while no significant differences were found in the chylomicron-poor fraction. The hydro- and iso-meal produced a higher insulin response compared with the lac- and CGMP-meal ( P <0.001). Otherwise no significant differences in the hormone responses were found in the incremental area under the curve over the 480-min period. Conclusions: A supplement of four different whey protein fractions to a fat-rich meal had similar effects on postprandial triglyceride responses in type 2 diabetic subjects. Whey isolate and whey hydrolysate caused a higher insulin response.

Orally ingested probiotic bacteria may modulate the immune response and increase antibody titers against enteric infections by bacteria or viruses. Even though positive effects of probiotics on respiratory tract infections have been reported, overall only few studies have examined effects on virus infections concerning organs other than the gastrointestinal tract. It was the aim of the study to investigate whether and how probiotics affect the immune response to a standardized enterovirus challenge (polio) and infections not limited to the gastrointestinal tract in healthy adults. In a randomized, controlled and double-blind study 64 volunteers consumed for 5 weeks chemically acidified clotted milk without bacteria or with 10(10)/serving (Lactobacillus rhamnosus ) GG or Lactobacillus acidophilus CRL431 added. In the second week subjects were vaccinated orally against polio 1, 2 and 3. Polio virus neutralizing serum activity, the primary parameter, was determined by the standard neutralization test (WHO) before and three times after vaccination. Polio-specific IgA, IgG and IgM were detected by ELISAs. Probiotics increased poliovirus neutralizing antibody titers (NT) and affected the formation of poliovirus-specific IgA and IgG in serum. The maximum increase after immunization was about 2, 2.2, or 4-fold higher, respectively, for NT, IgG or, IgA, in volunteers consuming probiotics instead of placebo. No consistent difference was noted between bacterial strains. Probiotics induce an immunologic response that may provide enhanced systemic protection of cells from virus infections by increasing production of virus neutralizing antibodies.

Background/Objectives: Postprandial lipaemia is an established risk factor for atherosclerosis. To investigate the acute effect of four milk-derived dietary proteins (alpha-lactalbumin, whey isolate, caseinoglycomacropeptide and whey hydrolysate) on postprandial lipaemia, we have conducted a randomized, acute, single-blinded clinical intervention study with crossover design. Subjects/Methods: A total of 11 obese non-diabetic subjects (age: 44–74, BMI: 30–41.4 kg m -2 ) were included. On 4 different days the subjects ingested a high-fat meal with the following energy distribution: 66% energy from fat (100 g of butter), 15% of energy from carbohydrate (90 g of white wheat bread) and 19% of energy from protein (45 g of pure protein). Our primary variable was plasma triglyceride measured in the 8-h postprandial period. Secondarily, retinyl palmitate, non-esterified free fatty acids, glucose, insulin, glucagon, GLP-1 and GIP, active and total grehlin and cholecystokinin were measured. Results: We observed no statistically significant ( P =0.8) differences between meals on our primary variable that is, triglycerides. Whey hydrolysate was associated with a significantly ( P =0.02) smaller postprandial suppression of non-esterified free fatty acids compared with the other dietary proteins. Conclusion: We did not observe significant differences in postprandial lipaemia to the four milk-derived dietary proteins. Whey hydrolysate caused less postprandial suppression of free fatty acids.

This study assessed the protective potential of salivary pellicles formed in situ over periods ranging from 2 to 24 h. Pellicles were produced on enamel slabs mounted on the palatal aspect of removable acrylic splints and exposed to the oral environment in three subjects for 2, 6, 12 and 24 h. Enamel specimens with and without pellicles were immersed in citric acid (1%) for 60 s, and the amount of dissolved calcium was measured by atomic absorption spectroscopy. In addition, specimens were processed for transmission electron microscopy (TEM). Mean values (standard deviations) for calcium release (mg/l related to the specimen's surface area of 5 x 5 mm(2)) were: 2-h pellicle 6.94 (1.55); 6-h pellicle 6.69 (2.05); 12-h pellicle 6.57 (2.31); 24-h pellicle 5.71 (2.46); enamel without pellicle 8.95 (1.66). There were no significant differences in calcium release that were dependent on pellicle formation time, but in comparison to enamel specimens without pellicle, significantly less (p <0.05) demineralization of the enamel was observed in pellicle-covered specimens. TEM showed that the pellicle was partly, but not completely dissolved following acid exposure. It is concluded that even a 2-h in-situ-formed pellicle layer protects the enamel surface to a certain extent against demineralization.

Orally ingested probiotic micro-organisms do not exert health effects exclusively in the intestine. Some strains can alleviate or prevent bacterial, fungal or viral infections in other organs by stimulation of the immune system. By preservation or improvement of the barrier function of the intestinal mucosa, they may inhibit translocation of potential pathogens and thus prevent infections of the blood stream and other tissues and organs. Modulation of the intestinal microflora can affect the local microflora of the urogenital tract and possibly of the oral cavity. Finally, some strains of orally ingested bacteria reach target organs like the urogenital tract in a viable state; alternatively they can be applied locally. Despite the infection-preventing properties of probiotic bacteria, lactic acid bacteria have rarely been identified in infections of the blood stream, heart valves and other organs, usually only in patients with severe disease. It is the general opinion that in most cases the source of infection was the commensal microflora of the intestine or the oral cavity. Until now only one case of infection associated with administration of a probiotic strain has been published. The most promising health-promoting effects have been seen in vaginosis, urinary tract infections, Helicobacter pylori gastritis and infections of the respiratory tract in children. More controlled clinical trials with sufficient numbers of participants are needed to determine the scientific basis for the use of probiotic bacteria in infections in locations of the body other than the intestine.

To investigate the effect of long-term consumption of probiotic bacteria on viral respiratory tract infections (common cold, influenza), a randomized, double blind, controlled intervention study was performed during two winter/spring periods (3 and 5 month). Four hundred and seventy-nine healthy adults were supplemented daily with vitamins plus minerals with or without probiotic lactobacilli and bifidobacteria. The intake of the probiotic had no effect on the incidence of common cold infections (verum = 158, control = 153 episodes, influenza was not observed), but significantly shortened duration of episodes by almost 2 days (7.0 ± 0.5 versus 8.9 ± 1.0 days, p = 0.045), reduced the severity of symptoms and led to larger increases in cytotoxic T plus T suppressor cell counts and in T helper cell counts.

In previous investigations, was shown to lack the metabolic pathway from fructose to mannitol and to produce ethanol when cultivated in the presence of fructose. Hence, we assessed the effect of oral administration of (strain NRRL-B-14171) on blood and fecal ethanol concentrations, glucose and lipid metabolism and traits of the metabolic syndrome in Wistar rats (n=27) fed diets with two different fat and fructose levels and with or without the addition of during a total intervention time of 15 weeks (105 days). From week 1 to 6, rats were given a medium fructose and fat (MFru-MF) diet containing 28% fructose and 10% fat without the addition of (control group, n=13) or mixed with 30 g per kg diet of lyophilized (10.56 ± 0.20 log CFU/g; group, n=14). From week 7 to 15, the percentage of dietary fructose and fat in the control and group was increased to 56% and 16%, respectively (high fructose-high fat (HFru-HF) diet). In HFru-HF-fed rats, was detected in feces, regardless of whether was added to the diet or not, but not in rats receiving the MFru-MF diet without added or in an additional control group (n=10) fed standard rat food without fructose, increased fat content and . This indicates that fecal may be derived from orally administered as well as - in the case of high fructose and fat intake and obesity of rats - from the intestinal microbiota. As shown by multifactorial ANOVA, blood ethanol, the relative liver weight, serum triglycerides, and serum cholesterol as well as fecal ethanol, ADH, acetate, propionate and butyrate, but not lactate, were significantly higher in the compared to the control group. This is the first in vivo trial demonstrating that heterofermentative lactic acid bacteria lacking the mannitol pathway (like ) can increase fecal and blood ethanol concentrations in mammals on a high fructose-high fat diet. This may explain why resulted in hyperlipidemia and may promote development of NAFLD in the host.

Background Milk products are good sources of calcium and their consumption may reduce bone resorption and thus contribute to prevent bone loss. Aim of the study We tested the hypothesis that bedtime consumption of fermented milk supplemented with calcium inhibits the nocturnally enhanced bone resorption more markedly than fermented milk alone, and postulated that this effect was most pronounced when calcium absorption enhancers were added. Methods In a controlled, parallel, double-blind intervention study over 2 weeks we investigated the short-term effects of two fermented milks supplemented with calcium from milk minerals (f-milk + Ca, n = 28) or calcium from milk minerals, inulin-type fructans and caseinphosphopeptides (f-milk + Ca + ITF + CPP; n  = 29) on calcium and bone metabolism in healthy, postmenopausal women, and compared them with the effect of a fermented control milk without supplements (f-milk, n  = 28). At bedtime 175 ml/d of either test milk was consumed. Fasting blood samples and 48 h-urine were collected at baseline and at the end of the intervention. Urine was divided into a pooled daytime and nighttime fraction. Multifactorial ANOVA was performed. Results Fermented milk independent of a supplement ( n  = 85) reduced the nocturnal excretion of deoxypyridinoline, a marker of bone resorption, from 11.73 ± 0.54 before to 9.57 ± 0.54 µmol/mol creatinine at the end of the intervention ( P  = 0.005). No effect was seen in the daytime fraction. Differences between the three milks ( n  = 28 resp. 29) were not significant. Fermented milk reduced bone alkaline phosphatase, a marker of bone formation, from 25.03 ± 2.08 to 18.96 ± 2.08 U/l, with no difference between these groups either. Fermented milk increased the nocturnal but not daytime urinary excretion of calcium and phosphorus. The effects on calcium and phosphorus excretion were mainly due to the group supplemented with Ca + ITF + CPP. Conclusion Bedtime consumption of fermented milk reduced the nocturnal bone resorption by decelerating its turnover. Supplemented calcium from milk mineral had no additional effect unless the absorption enhancers ITF + CPP were added. A stimulated intestinal calcium absorption may be assumed, since urinary calcium excretion increased at a constant bone resorption.

Orally ingested probiotic micro-organisms do not exert health effects exclusively in the intestine. Some strains can alleviate or prevent bacterial, fungal or viral infections in other organs by stimulation of the immune system. By preservation or improvement of the barrier function of the intestinal mucosa, they may inhibit translocation of potential pathogens and thus prevent infections of the blood stream and other tissues and organs. Modulation of the intestinal microflora can affect the local microflora of the urogenital tract and possibly of the oral cavity. Finally, some strains of orally ingested bacteria reach target organs like the urogenital tract in a viable state; alternatively they can be applied locally. Despite the infection-preventing properties of probiotic bacteria, lactic acid bacteria have rarely been identified in infections of the blood stream, heart valves and other organs, usually only in patients with severe disease. It is the general opinion that in most cases the source of infection was the commensal microflora of the intestine or the oral cavity. Until now only one case of infection associated with administration of a probiotic strain has been published. The most promising health-promoting effects have been seen in vaginosis, urinary tract infections, Helicobacter pylori gastritis and infections of the respiratory tract in children. More controlled clinical trials with sufficient numbers of participants are needed to determine the scientific basis for the use of probiotic bacteria in infections in locations of the body other than the intestine.