Cancer-protective effect of a synbiotic combination between Lactobacillus gasseri 505 and a Cudrania tricuspidata leaf extract on colitis-associated colorectal cancer.

Previously, a synbiotic combination of probiotic Lactobacillus gasseri 505 (LG) and a new prebiotic, Cudrania tricuspidata leaf extract (CT) in fermented milk, designated FCT, showed an in vitro immunomodulatory effect and antioxidant activity. Although synbiotic combination might have cancer-protective effects, these activities have not been fully validated in vivo. Ten-week treatment of LG, CT, or FCT to azoxymethane (AOM)/dextran sodium sulfate (DSS)-induced colitis-associated colorectal cancer (CAC) mouse model reduced both the incidence of colonic tumors and damage to the colonic mucosa effectively, suggesting a cancer-protective effect. To understand these, biomarkers associated with inflammation, colon barrier, apoptosis, and cancer cell proliferation were monitored in AOM/DSS group versus LG/CT/FCT groups. A synbiotic combination (FCT) down-regulated pro-inflammatory cytokines (TNF-α, IFN-γ, IL-1ß, and IL-6) and inflammation-associated enzymes (iNOS and COX-2), and up-regulated anti-inflammatory cytokines (IL-4 and IL-10). In addition, colon barrier experiment revealed that biomarkers of mucus layer (MUC-2 and TFF3) and tight junction (occludin and ZO-1) were up-regulated. Subsequent apoptosis experiment showed that pro-apoptotic factors (p53, p21, and Bax) were up-regulated and anti-apoptotic factors (Bcl-2 and Bcl-xL) were down-regulated. Furthermore, comparative metagenome analysis of gut microbiota revealed that Staphylococcus decreased but Lactobacillus, Bifidobacterium, and Akkermansia increased, supporting their protective effects, accompanied by increased short-chain fatty acids (SCFAs). Taken together, the FCT administration showed cancer-protective effects by reducing the risk of colitis-associated colon cancer via regulation of inflammation, carcinogenesis, and compositional change of gut microbiota. Consequently, the synbiotic combination (FCT) could be a novel potential health-protective natural agent against CAC.

Identification of bioactive compounds in Lactobacillus paracasei subsp. paracasei NTU 101-fermented reconstituted skimmed milk and their anti-cancer effect in combination with 5-fluorouracil on colorectal cancer cells.

Chemotherapy is currently used to treat colorectal cancer (CRC), the most common cancer worldwide. However, chemotherapeutic drugs are limited by severe side effects or drug resistance. In this study, bioactive compound(s), a mixture of palmitic acid, stearic acid, and glyceryl 1,3-dipalmitate (PSG), in Lactobacillus paracasei subsp. paracasei NTU 101-fermented reconstituted skimmed milk ethanol extract (NTU 101-FMEE) were isolated and identified. PSG (1 : 1.5 : 6.3) at 125 µg mL-1 could significantly decrease CRC cell viability at dosages that were not cytotoxic to healthy colon epithelial cells or macrophages. Moreover, the inhibitory effect of the combination of 62.5 µg mL-1 PSG (1 : 1.5 : 6.3) and 5-fluorouracil (5-FU) was significantly higher than that of 5-FU alone (p < 0.05). PSG up-regulated the activities of apoptosis-related proteins and down-regulated the nuclear factor-κB signaling pathway compared to the levels in the control group. Overall, PSG purified from NTU 101-FMEE possesses the potential to ameliorate CRC by improving the effects of adjuvant chemotherapy drugs and reducing side effects.

Impact of Buckwheat Fermented Milk Combined with High-Fat Diet on Rats' Gut Microbiota and Short-Chain Fatty Acids.

The aim of this study was to evaluate the effect of buckwheat fermented milk on intestinal flora and short-chain fatty acids (SCFAs) of rats fed a high-fat diet (HFD). Buckwheat fermented milk was made with Lactobacillus plantarum ST-Ⅲ, Lactobacillus bulgaricus, and Streptococcus thermophilus. Thirty-six males C57BL/6 rats (aged 3 weeks and with 9 to 12 g weight) were subjected to a 2-week adaptive period on a normal diet. After a week of acclimatization, the rats were randomly divided into three groups with 12 rats in each group. The high-fat diet with fermented milk (HFDFM) group received HFD supplemented with fermented milk, and the high-fat diet supplemented with buckwheat fermented milk (HFDBFM) group received HFD plus buckwheat fermented milk. Besides, the gavage dose was 0.4 mL/day. After the feedings, colon levels of cytokines and antioxidant indices of the liver, colon, and duodenum tissues were measured. The composition of intestinal flora of rats and the content of SCFAs were determined by gas chromatography-mass spectrometry and Illumina high-throughput sequencing technology. Supplementation with buckwheat fermented milk significantly inhibited the increases in lipopolysaccharide levels in colon and antioxidant indexes in the HFD-fed rats. In addition, buckwheat fermented milk could significantly enhance Firmicutes and decrease Bacteroidetes compared to HFDFM. In addition, SCFA concentrations in HFDBFM were higher than other groups. In conclusion, buckwheat fermented milk can alleviate the damage of HFD to intestinal flora of rats. It modulates microbiota composition and SCFA concentrations to balance the intestinal environment, which will benefit to rats' intestinal health. PRACTICAL APPLICATION: This study explored the effect of buckwheat fermented milk on the regulation of intestinal microflora and SCFAs in rats fed a high-fat diet. It provides new ideas for the development of buckwheat fermented food.

Comparison of the nutritional composition of experimental fermented milk:wheat bulgur blends and commercially available kishk and tarhana products.

Dried, fermented blends of dairy products and cereals, such as kishk and tarhana, are foodstuffs traditionally consumed in many regions as they possess good nutritional qualities and extended storage stability. This study examined the nutritional composition of kishk or tarhana type products and compared with experimental blends of fermented milk and wheat bulgur containing 60-80% milk. The blends with higher milk contents had levels of protein (18.9%) and fat (5.8%) at the concentrations specified in fortified blended foods as outlined by the World Food Program. Higher milk contents were also associated with higher contents of calcium (323.2 mg/100 g), phosphorus (335.3 mg/100 g), vitamin A (486.7 µg/100 g) and α-tocopherol (174.5 µg/100 g). The nutritional content of the experimental fermented milk:wheat bulgur blends compared favourably with that of the commercial samples. These blends may be suitable as base products, to be fortified with micronutrients, for the development of fortified blended foods (FBFs) for humanitarian distribution.

Effects of an ethanol extract from Lactobacillus paracasei subsp. paracasei NTU 101 fermented skimmed milk on lipopolysaccharide-induced periodontal inflammation in rats.

The increased incidence of periodontal disease in recent years has garnered considerable attention. Numerous studies have confirmed that probiotics, such as lactic acid bacteria, can ameliorate periodontal inflammation. The current study aimed to assess the effect of an ethanol extract of Lactobacillus paracasei subsp. paracasei NTU 101-fermented skimmed milk (NTU101FM) on lipopolysaccharide (LPS)-induced periodontal inflammation in rats. NTU101FM ethanol extract significantly ameliorated the weight loss caused by periodontal inflammation. NTU101FM ethanol extract treatment also reduced the oral microbial levels and decreased the levels of alveolar bone loss. Finally, NTU101FM ethanol extract was found to ameliorate periodontal inflammation by decreasing the levels of pro-inflammatory cytokines and reducing oxidative stresses induced by LPS. Overall, our findings demonstrate that NTU101FM ethanol extract could be developed as a functional food that could ameliorate periodontal inflammation.

Applications and Uncertainty Estimation of Single Level Standard Addition Method ICP-MS for Elemental Analysis in Various Matrix.

The standard addition method (SAM) based on gravimetric sample preparation was investigated as an approach for the removal or cancelling of matrix effects in measurements by inductively coupled plasma mass spectrometry (ICP-MS). Deduction of the equations and experimental confirmation of the method are both given in the present work. After measuring both spiked and non-spiked samples by ICP-MS, the concentration of an element could be calculated based on the signal intensity ratio to an internal standard. A practical example was provided for the measurement of Fe in a certified reference material (CRM), i.e. NMIJ CRM 7512-a (milk powder). The validity of the method had been confirmed by the results of international comparisons with various kinds of matrix, including bioethanol, human serum, biodiesel fuel, drinking water, infant formula milk power, and seafood. The suggested method had been applied to measurements of multiple elements in three CRMs, including tap water, milk powder, and tea leave powder, respectively.

Development of novel Meju starter culture using plant extracts with reduced Bacillus cereus counts and enhanced functional properties.

We developed a novel type of Meju starter culture using single and combined extracts of Allium sativum (garlic clove), Nelumbo nucifera (lotus leaves), and Ginkgo biloba (ginkgo leaves) to improve the quality and functionality of Meju-based fermented products. Meju samples fermented with plant extracts (10 mg/ml) showed phenolic contents of 11.4-31.6 mg/g (gallic acid equivalents). Samples of extracts (garlic clove, lotus leaves, ginkgo leaves and their combination) fermented with Meju strongly inhibited tyrosinase, α-glucosidase, and elastase activities by 36.43-64.34%, 45.08-48.02%, and 4.52-10.90%, respectively. Specifically, ginkgo leaves extract added to fermented Meju samples at different concentrations (1% and 10%) strongly inhibited tyrosinase, α-glucosidase, and elastase activities and exhibited a potent antibacterial effect against Bacillus cereus with a significant reduction in bacterial counts compared with the effects observed for garlic clove and lotus leaf added to Meju samples. Scanning electron microscopy revealed severe morphological alterations of the B. cereus cell wall in response to ginkgo leaf extracts. Gas chromatographic mass spectroscopic analysis of plant extract-supplemented Meju samples and control Meju samples identified 113 bioactive compounds representing 98.44-99.98% total extract. The proposed approach may be useful for the development of various fermented functional foods at traditional and commercial levels.

Specific Signatures of the Gut Microbiota and Increased Levels of Butyrate in Children Treated with Fermented Cow's Milk Containing Heat-Killed Lactobacillus paracasei CBA L74.

We recently demonstrated that cow's milk fermented with the probiotic Lactobacillus paracasei CBA L74 (FM-CBAL74) reduces the incidence of respiratory and gastrointestinal tract infections in young children attending school. This effect apparently derives from a complex regulation of non-immune and immune protective mechanisms. We investigated whether FM-CBAL74 could regulate gut microbiota composition and butyrate production. We randomly selected 20 healthy children (12 to 48 months) from the previous randomized controlled trial, before (t0) and after 3 months (t3) of dietary treatment with FM-CBAL74 (FM) or placebo (PL). Fecal microbiota was profiled using 16S rRNA gene amplicon sequencing, and the fecal butyrate concentration was also measured. Microbial alpha and beta diversities were not significantly different between groups prior to treatment. FM-CBAL74 but not PL treatment increased the relative abundance of Lactobacillus Individual Blautia, Roseburia, and Faecalibacterium oligotypes were associated with FM-CBAL74 treatment and demonstrated correlative associations with immune biomarkers. Accordingly, PICRUSt analysis predicted an increase in the proportion of genes involved in butyrate production pathways, consistent with an increase in fecal butyrate observed only in the FM group. Dietary supplementation with FM-CBAL74 induces specific signatures in gut microbiota composition and stimulates butyrate production. These effects are associated with changes in innate and acquired immunity.IMPORTANCE The use of a fermented milk product containing the heat-killed probiotic strain Lactobacillus paracasei CBAL74 induces changes in the gut microbiota, promoting the development of butyrate producers. These changes in the gut microbiota composition correlate with increased levels of innate and acquired immunity biomarkers.

Changes in Adiposity and Body Composition during Anemia Recovery with Goat or Cow Fermented Milks.

To date, no studies are available about adipose tissue modifications during anemia recovery; therefore, the aim of this study is to provide detailed information about adipose tissue homeostasis during anemia recovery with fermented milks. Forty male Wistar rats were placed on a pre-experimental period of 40 days, divided in two groups (normal-Fe diet and Fe-deficient diet). Then rats were fed fermented goat or cow milk-based diets with normal-Fe content during 30 days. Ghrelin and adiponectin decreased in both groups of animals fed fermented goat milk, whereas leptin and NEFA increased. UCP-1 decreased in anemic rats fed either fermented milk, and irisin greatly increased in both groups of animals fed fermented goat milk. Fermented goat milk reduces adiposity, inducing leptin elevation and ghrelin reduction. Conversely, plasma adiponectin concentrations decreased in animals fed fermented goat milk, showing an inverse correlation with NEFA, an important marker of lipid mobilization, indicating increased lipolysis. Irisin up-regulation in animals fed fermented goat milk contributes to a favorable metabolic profile and the browning of adipose tissue during anemia recovery.

Characteristic chromatographic fingerprint study of short-chain fatty acids in human milk, infant formula, pure milk and fermented milk by gas chromatography-mass spectrometry.

Human milk, infant formula, pure milk and fermented milk as food products or dietary supplements provide a range of nutrients required to both infants and adults. Recently, a growing body of evidence has revealed the beneficial roles of short-chain fatty acids (SCFAs), a subset of fatty acids produced from the fermentation of dietary fibers by gut microbiota. The objective of this study was to establish a chromatographic fingerprint technique to investigate SCFAs in human milk and dairy products by gas chromatography coupled with mass spectrometry. The multivariate method for principal component analysis assessed differences between milk types. Human milk, infant formula, pure milk and fermented milk were grouped independently, mainly because of differences in formic acid, acetic acid, propionic acid and hexanoic acid levels. This method will be important for the assessment of SCFAs in human milk and various dairy products.

Improved functionality of fermented milk is mediated by the synbiotic interaction between Cudrania tricuspidata leaf extract and Lactobacillus gasseri strains.

This study was designed to investigate the cooperative effect of selected Lactobacillus gasseri strains and Cudrania tricuspidata (CT) leaf extract in enhancing the health-promoting activities of fermented milk. Addition of CT increased total bacterial counts and proteolysis during fermentation of milk with L. gasseri strains. Antioxidant capacities were determined by measuring the ABTS, DPPH, and peroxyl radical scavenging activities and ferric reducing power. The antioxidant capacity of CT-supplemented milk was greater than that of milk without supplementation; moreover, the antioxidant activity of CT-supplemented milk was synergistically improved by fermentation with L. gasseri strains. In particular, CT-supplemented milk fermented by L. gasseri 505 showed the highest antioxidant activity. The phenolic compounds in CT, such as neo-chlorogenic, chlorogenic, and caffeic acid, were metabolized during fermentation with L. gasseri strains, and 3,4-dihydroxy-hydrocinnamic acid was produced as a fermentation metabolite. Moreover, the liberation of bioactive peptides of fermented milk was increased by the proteolytic activity of L. gasseri strains. In particular, six peptides, which were mainly derived from ß-casein, were newly identified in this study. These findings suggest that L. gasseri strains metabolize the phenolic acids in the CT and the bioactive peptides released through this interaction improve the antioxidant activity of the fermented milk.

Influence of a fermented protein-fortified dairy product on serum insulin-like growth factor-I in women with anorexia nervosa: A randomized controlled trial.

BACKGROUND & AIMS: Patients with anorexia nervosa (AN) have low serum IGF-I levels that may contribute to a lower bone mineral mass. We investigated the effects of a fermented, protein-fortified, dairy product on serum IGF-I levels in patients with AN during an in-hospital refeeding program. METHODS: In this multicenter, randomized, double-blind, placebo-controlled, clinical trial conducted at 3 university hospitals and 3 private clinics in France and Switzerland, 62 women recently admitted with confirmed AN and with a baseline low serum IGF-I level were randomized to 2 daily isocaloric fresh cheese pots containing either 15 g/150 g or 3 g/150 g (controls) of protein for 4 weeks. The primary outcome was the change in IGF-I levels. RESULTS: In the primary intention-to-treat analysis, mean serum IGF-I levels increased during the intervention phase from 22.9 ± 1.5 to 28.6 ± 1.3 nmol/L (means ± SEM) (+20.2%) in the intervention group and from 20.2 ± 1.2 to 25.7 ± 1.5 nmol/L (+16.8%) in controls. In a preplanned analysis of covariance with repeated measures, the between-group difference was close to statistical significance (P = 0.071). In a post-hoc mixed-regression model analysis, the difference was statistically significant (4.9 nmol/l increase; P = 0.003), as was the change of the ratio IGF-I/IGF-BP3 (P=0.004). There was no between-group difference in biochemical markers of bone turnover (osteocalcin, P1NP, CTX) or in serum parathyroid hormone level. Serum calcium levels slightly increased during the intervention phase in the higher protein group (P = 0.02). IGF-BP2 decreased significantly more in the intervention group during the follow up period at week 4 after supplements cessation (P = 0.019). CONCLUSIONS: Intake of a fermented, protein-fortified, isocaloric dairy product during 4 weeks may slightly increase serum IGF-I levels in women with AN, without significant changes in bone turnover markers. CLINICAL TRIAL REGISTRATION NUMBER: NCT01823822 (www.clinicaltrials.gov).

Application of acid whey and set milk to marinate beef with reference to quality parameters and product safety.

BACKGROUND: The aim of the study was to evaluate the potential of acid whey and set milk as a marinade in the traditional production of fermented eye round. METHODS: Studies involved assaying pH value, water activity (aw), oxidation-reduction potential and TBARS value, colour parameters in CIE system (L*, a*, b*), assaying the number of lactic acid bacteria and certain pathogenic bacteria after ripening process and after 60-day storing in cold storage. Sensory analysis and analysis of the fatty acids profile were performed after completion of the ripening process. RESULTS: Analysis of pH value in the products revealed that application of acid whey to marinate beef resulted in increased acidity of ripening eye round (5.14). The highest value of the colour parameter a* after ripening process and during storage was observed in sample AW (12.76 and 10.07 respectively), the lowest on the other hand was observed in sample SM (10.06 and 7.88 respectively). The content of polyunsaturated fatty acids (PUFA) was higher in eye round marinated in acid whey by approx. 4% in comparison to other samples. CONCLUSIONS: Application of acid whey to marinade beef resulted in increased share of red colour in general colour tone as well as increased oxidative stability of the product during storage. It also increased the content of polyunsaturated fatty acids (PUFA) in the product. All model products had high content of lactic acid bacteria and there were no pathogenic bacteria such as: L. monocytogenes, Y. enterocolitica, S. aureus, Clostridium sp.

Estimation of the antioxidant activity of the commercially available fermented milks.

BACKGROUND: Free radicals are connected with the increased risk of certain diseases, especially cancers. There is some scientific evidence that antioxidant-rich diet may inhibit the negative impact of free radicals. The aim of the present study was to analyse the antioxidant capacity of the selected commercial natural and flavoured fermented milks offered in Poland, derived from different producers. METHODS: The following commercially available natural fermented milks: 12 yoghurts, 12 kefirs, 2 butter milks, 2 cultured milks, Turkish yoghurt drink (ayran) and the following flavoured fermented milks: 22 yoghurts, 2 acidophillus milks, 2 kefirs, butter milk and vegetable flavoured fermented milk were analysed for their antioxidant potential. The antioxidant capacity was assessed, in two replicates and twice for each product, by means of ferric reducing antioxidant power (FRAP) and DPPH radical scavenging ability (expressed as ARP - anti radical power) methods. RESULTS: Among all analysed plain products, yoghurts and kefirs were characterised by the highest antioxidant activity. The presence of probiotic Lactobacillus casei strains in the product positively affected both FRAP and ARP values. Antioxidant capacity of the flavoured fermented milks was primarily affected by the type and quality (e.g. fruit concentration) of the added flavouring preparation. The most valuable regarding the estimated parameters were chocolate, coffee, grapefruit with green tea extract as well as bilberry, forest fruits, strawberry and cherry with blackcurrant fillings. CONCLUSIONS: Protein content, inclusion of probiotic microflora as well as type and quality of flavouring preparations are the main factors affecting antioxidant properties of fermented milks.