Short communication: Effect of milk and milk containing Lactobacillus casei on the intestinal microbiota of mice

BALB/c mice were fed milk or Lactobacillus casei BL23 in milk for 14 d and fecal samples were collected at d 0, 4, and 7 as well as 1 and 8 d after the last administration. According to high-throughput DNA sequencing of the 16S rRNA genes extracted from the fecal microbiota, the bacterial diversity in the fecal samples of all mice increased over time. After 14 d of administration, the consumption of milk and milk containing L. casei BL23 resulted in distinct effects on the microbial composition in the intestine. Specifically, the proportions of bacteria in the Lactobacillaceae, Porphyromonadaceae, and Comamonadaceae were significantly higher in mice fed the L. casei BL23-milk culture compared with one or more of the other groups of mice. The relative amounts of Lachnospiraceae were higher and Streptococcaceae were lower in mice fed milk alone. The changes were not found at d 4 and 7 during milk and L. casei feeding and were no longer detected 8 d after administration was stopped. This study shows that consumption of milk or probiotic L. casei-containing milk results in non-overlapping, taxa-specific effects on the bacteria in the distal murine intestine.     

Carbohydrate metabolic conversions to lactic acid and volatile derivatives, as influenced by Lactobacillus plantarum ATCC 8014 and Lactobacillus casei ATCC 393 efficiency during in vitro and sourdough fermentation

The use of sourdough returned to be popular due to its beneficial nutritional, sensorial properties and physical features. In this study, two types of experiments aimed to evaluate comparatively the efficiency of two Lactobacillus (Lb.) strains, Lactobacillus plantarum ATCC 8014 and Lactobacilus casei ATCC 393, to metabolize different carbohydrates (glucose, fructose, maltose and sucrose) in vitro and to compare their efficiency, combined with/without Saccharomyces cerevisiae on sourdough fermentation. Their efficiency was monitored by cellular growth dynamics, pH values and metabolite production (lactic and acetic acid, volatile derivatives) determined by HPLC and GC–MS analysis. Lb. plantarum ATCC 8014 was, generally, more efficient than Lb. casei for carbohydrate consumption in vitro and release of lactic acid. Unexpected, Lb. casei was able to produce trehalose in vitro, especially when glucose and sucrose were added in media. During sourdough fermentation, Lb. plantarum had a superior growth dynamic and a dominant nonaerobic fermentation, opposite to Lb. casei. By the addition of yeast to lactobacilli after a short-time lactic fermentation, a combined nonaerobic and aerobic fermentation was noticed, reflected by a more complex mixture of volatile derivatives. In vitro, both Lb. strains produced seven volatile compounds (carbonyl derivatives), but during sourdough fermentation, especially after yeast addition, a complex mixture of alcohols, carbonyl derivatives and acetate esters were determined. Lb. strains (especially Lb. plantarum) determined a moderate volatiles production dominated by alcohols, while addition of yeast to Lb. intensified the volatiles’ production and increase the alcohols proportion against carbonyl derivatives.     

Two strains of nonstarter lactobacilli increased the production of flavor compounds in soft cheeses

The contribution to flavor generation and secondary proteolysis of 2 strains of mesophilic lactobacilli isolated from cheese was studied. Miniature soft cheeses (200 g) were produced with or without the inclusion of a culture of Lactobacillus plantarum I91 or Lactobacillus casei I90 in the starter composed of Streptococcus thermophilus. During ripening, cheeses containing the added lactobacilli showed an increased content of total free amino acids, but this increase was only significant in cheeses with Lb. plantarum I91. In addition, free amino acid profiles were modified by selective increases of some amino acids, such as Asp, Ser, Arg, Leu, and Phe. Cheeses inoculated with Lb. plantarum I91 or Lb. casei I90 were also characterized by a significantly higher concentration of diacetyl, a key flavor compound, and an increased content of acetoin. Results suggest an increase in the catabolism of either citrate or aspartate, with the production of the derived aroma compounds. Overall, aspartate content increased in both lactobacilli-added cheeses, whereas citrate was more or less constant, suggesting that aspartate could be the source of increased diacetyl and acetoin. A triangle aroma test showed that the addition of the lactobacilli strains significantly changed the sensory attributes of cheeses. At least 11 of 12 panelists commented that the aroma of cheeses with adjuncts was more buttery than that of control cheeses, which is desirable in most soft cheeses. Both Lb. plantarum I91 and Lb. casei I90 performed well as adjunct cultures by influencing cheese aroma development and cheese proteolysis.     

Selection of potential probiotic lactic acid bacteria from fermented olives by in vitro tests

The present study aims to evaluate the probiotic potential of lactic acid bacteria (LAB) isolated from naturally fermented olives and select candidates to be used as probiotic starters for the improvement of the traditional fermentation process and the production of newly added value functional foods. Seventy one (71) lactic acid bacterial strains (17 Leuconostoc mesenteroides, 1 Ln. pseudomesenteroides, 13 Lactobacillus plantarum, 37 Lb. pentosus, 1 Lb. paraplantarum, and 2 Lb. paracasei subsp. paracasei) isolated from table olives were screened for their probiotic potential. Lb. rhamnosus GG and Lb. casei Shirota were used as reference strains. The in vitro tests included survival in simulated gastrointestinal tract conditions, antimicrobial activity (against Listeria monocytogenes, Salmonella Enteritidis, Escherichia coli O157:H7), Caco-2 surface adhesion, resistance to 9 antibiotics and haemolytic activity. Three (3) Lb. pentosus, 4 Lb. plantarum and 2 Lb. paracasei subsp. paracasei strains demonstrated the highest final population (>8 log cfu/ml) after 3 h of exposure at low pH. The majority of the tested strains were resistant to bile salts even after 4 h of exposure, while 5 Lb. plantarum and 7 Lb. pentosus strains exhibited partial bile salt hydrolase activity. None of the strains inhibited the growth of the pathogens tested. Variable efficiency to adhere to Caco-2 cells was observed. This was the same regarding strains' susceptibility towards different antibiotics. None of the strains exhibited β-haemolytic activity. As a whole, 4 strains of Lb. pentosus, 3 strains of Lb. plantarum and 2 strains of Lb. paracasei subsp. paracasei were found to possess desirable in vitro probiotic properties similar to or even better than the reference probiotic strains Lb. casei Shirota and Lb. rhamnosus GG. These strains are good candidates for further investigation both with in vivo studies to elucidate their potential health benefits and in olive fermentation processes to assess their technological performance as novel probiotic starters.     

The microbial diversity of water kefir

The microbial diversity of water kefir, made from a mixture of water, dried figs, a slice of lemon and sucrose was studied. The microbial consortia residing in the granules of three water kefirs of different origins were analyzed. A collection of 453 bacterial isolates was obtained on different selective/differential media. Bacterial isolates were grouped with randomly amplified polymorphic DNA (RAPD)-PCR analyses. One representative of each RAPD genotype was identified by comparative 16S rDNA gene sequencing. The predominant genus in water kefirs I and II was Lactobacillus, which accounted for 82.1% in water kefir I and 72.1% in water kefir II of the bacterial isolates. The most abundant species in water kefirs I and II were Lactobacillus hordei and Lb. nagelii followed by considerably lower numbers of Lb. casei. Other lactic acid bacteria (LAB) were identified as Leuconostoc mesenteroides and Lc. citreum in all three water kefirs. The most abundant species in water kefir III was Lc. mesenteroides (28%) and Lc. citreum (24.3%). A total of 57 LAB belonging to the species of Lb. casei, Lb. hordei, Lb. nagelii, Lb. hilgardii and Lc. mesenteroides were able to produce exopolysacchrides from sucrose. Non LABs were identified as Acetobacter fabarum and Ac. orientalis. The Acetobacter species were more prevalent in consortium III. Cluster analyses of RAPD-PCR patterns revealed an interspecies diversity among the Lactobacillus and Acetobacter strains. Aditionally, Saccharomyces cerevisiae, Lachancea fermentati, Hanseniaospora valbyensis and Zygotorulaspora florentina were isolated and identified by comparison of partial 26S rDNA sequences and FTIR spectroscopy.     

Oleic acid and ergosterol supplementation mitigates oxidative stress in wine strains of Saccharomyces cerevisiae

During fermentation of high-sugar-containing medium lacking lipid nutrients, wine yeasts undergo oxidative stress and oxidative damage to cell membranes and proteins. Considering that cell membranes are important stress sensors, and that under hypoxic conditions wine yeasts modulate cell membranes composition by incorporating lipids available in the growth medium, in the present work, the effects of lipid nutrition on wine yeast oxidative stress response were evaluated on two strains of Saccharomyces cerevisiae. Biomarkers of oxidative stress, oxidative damage and antioxidant response were evaluated together with viability and acetic acid production during fermentation of a synthetic must lacking lipid nutrients as compared to added oleic acid and ergosterol. The results show that the availability of lipid nutrients causes a significant reduction in the intracellular content of reactive oxygen species and in the oxidative damage to membranes and proteins, as indicated by flow cytometry of cells stained with dihydroethidum (DHE) and propidium iodide (PI) and by Western blot of protein carbonyls. Accordingly, lipid nutrients feeding results in the increase in cell viability and superoxide activity, and the reduction in trehalose accumulation, proteinase A activity and production of acetic acid. In summary, these results are compatible with the hypothesis that the supplementation of lipid nutrients mitigates oxidative stress and oxidative damage in wine strains of S. cerevisiae during growth under unfavourable conditions.     

Textural properties of alginate–pectin beads and survivability of entrapped Lb. casei in simulated gastrointestinal conditions and in yoghurt

Lactobacillus casei was entrapped in beads made with sodium alginate (A), amidated low-methoxyl pectin (P), and blends of A–P (1:2, 1:4, 1:6 ratios) by the extrusion technique. Diameter, sphericity and textural properties of the beads and the survivability of entrapped Lb. casei in yoghurt and in simulated gastrointestinal conditions were determined. Entrapment efficiency of Lb. casei and diameter increased as the proportion of P and the total biopolymers concentration increased. Principal Component Analysis showed that the survivability of Lb. casei in yoghurt, and in simulated gastric juice, was positively correlated with the diameter and all the textural properties of the beads, while survivability after exposure to simulated gastric juice and bile salts was positively correlated with the sphericity of the beads. The beads made with A–P blends in 1:4 and 1:6 ratios provided a significant better protection to the entrapped Lb. casei under all conditions studied.     

Proteolytic pattern and organic acid profiles of probiotic Cheddar cheese as influenced by probiotic strains of Lactobacillus acidophilus,Lb. paracasei,Lb. casei or Bifidobacterium sp.

Cheddar cheeses were produced with starter lactococci and Bifidobacterium longum 1941, B. lactis LAFTI^® B94, Lactobacillus casei 279, Lb. paracasei LAFTI^® L26, Lb. acidophilus 4962 or Lb. acidophilus LAFTI^® L10 to study the survival of the probiotic bacteria and the influence of these organisms on proteolytic patterns and production of organic acid during ripening period of 6 months at 4 °C. All probiotic adjuncts survived the manufacturing process of Cheddar cheese at high levels without alteration to the cheese-making process. After 6 months of ripening, cheeses maintained the level of probiotic organisms at >8.0 log₁₀ cfu g⁻¹ with minimal effect on moisture, fat, protein and salt content. Acetic acid concentration was higher in cheeses with B. longum 1941, B. lactis LAFTI^® B94, Lb. casei 279 and Lb. paracasei LAFTI^® L26. Each probiotic organism influenced the proteolytic pattern of Cheddar cheese in different ways. Lb. casei 279 and Lb. paracasei LAFTI^® L26 showed higher hydrolysis of casein. Higher concentrations of free amino acids (FAAs) were found in all probiotic cheeses. Although Bifidobacterium sp. was found to be weakly proteolytic, cheeses with the addition of those strains had highest concentration of FAAs. These data thus suggested that Lb. acidophilus 4962, Lb. casei 279, B. longum 1941, Lb. acidophilus LAFTI^® L10, Lb. paracasei LAFTI^® L26 and B. lactis LAFTI^® B94 can be applied successfully in Cheddar cheese.     

Probiotic Cheddar cheese: Influence of ripening temperatures on survival of probiotic microorganisms, cheese composition and organic acid profiles

Bifidobacterium longum 1941, Bifidobacterium animalis subsp. lactis LAFTI^®B94 (B94), Lactobacillus casei 279, Lb. casei LAFTI^®L26 (L26), Lactobacillus acidophilus 4962 or Lb. acidophilus LAFTI^®L10 (L10) were used as an adjunct in the production of Cheddar cheeses which were ripened for 24 wk at 4 and 8 °C. Effects of ripening temperatures on survival of starter lactococci and probiotic microorganisms, pH and composition of cheeses and production of organic acids were examined. The counts of starter lactococci in cheeses produced with B. animalis B94, Lb. casei L26 or Lb. acidophilus 4962 ripened at 8 °C were significantly lower than those ripened at 4 °C (P < 0.05) at 24 wk. Probiotic microorganisms remained viable (>7.50 log₁₀ CFU/g) at the end of 24 wk and their viability was not affected by the ripening temperatures. There were significant effects of the type of probiotic microorganisms used, ripening time, ripening temperatures and their interactions on the concentration of lactic and acetic acids in the cheeses (P < 0.05). The acetic acid concentration in cheeses made with Bifidobacterium sp. or Lb. casei sp. was significantly higher than that of the control cheese (P < 0.05). Citric, propionic and succinic acids contents of the cheeses were not significantly affected by the type of probiotic microorganisms or ripening temperatures (P > 0.05).     

Production of organic acids by Lactobacillus strains in three different media

Ten strains of Lactobacillus (Lb). casei, Lb. rhamnosus, Lb. plantarum, Lb. paracasei and Lb. curvatus species were chosen to determine the production of organic acids after cultivation in skimmed milk, MRS broth and Jerusalem artichoke (JA) medium. The highest acidity was obtained in MRS broth and the weakest acidification was found in skimmed milk. Lb. casei Shirota produced the highest amount and Lb. rhamnosus VT1 the lowest amount of substances being estimated as titratable acidity. All strains produced lactic acid in the investigated broth and most of the strains produced acetic acid in MRS broth except Lb. curvatus 2768 and Lb. casei Shirota, in JA broth except Lb. paracasei SF1 and in skimmed milk except Lb. casei 2750, Lb. curvatus 2768, Lb. curvatus 2775 and Lb. casei Shirota. All strains, except Lb. plantarum 01, produced butyric acid in MRS broth. Beside the lactic and acetic acids, formic, citric, succinic and glutamic acids were also produced in MRS broth; formic and succinic acids were produced in skimmed milk and succinic acid in JA broth. Some strains showed change in their fermentation profile from homofermentative to mix-acid fermentation in milk. The antifungal efficiency of the lactic and acetic acid in the amount produced by lactobacilli was investigated. None of the investigated aspergilli were inhibited. The inhibitory effect of acids against Fusarium increased unequivocally with the increasing concentration. The study pointed at the dissimilarity of organic acid production of Lactobacillus strains, which was considerably influenced by the media.     

L-苹果酸抗氧化作用机理研究

目的:研究L-苹果酸对老年大鼠不同组织脂质过氧化水平和抗氧化能力的影响,探索L-苹果酸增强老年机体抗氧化能力的作用机理。方法:以24月龄大鼠作为老年动物模型,通过给大鼠灌胃L-苹果酸30d后,评价L-苹果酸对老年大鼠不同组织中脂质过氧化产物丙二醛(MDA)含量及抗氧化酶活性的影响。结果:补充L-苹果酸能分别提高老年大鼠肝脏总超氧化物歧化酶(TSOD)、铜锌-超氧化物歧化酶(CuZnSOD)、谷胱苷肽过氧化酶(GPx)的活性34.4%、34.2%和39.9%。结论:L-苹果酸可以增强机体的抗氧化能力,减少脂质过氧化的发生,起到抗氧化作用。     

Protective activity of probiotic bacteria against 2-amino-3-methyl-3H-imidazo[4,5-f]quinoline (IQ) and 2-amino-1-methyl-6-phenyl-1H-imidazo[4,5-b]pyridine (PhIP) – an in vitro study

Heterocyclic aromatic amines (HAAs) are carcinogenic compounds present in a typical Western diet rich in thermally processed meat. These nutritional factors can modulate the cytotoxicity of faecal water (FW) and induce tumours in the human gastrointestinal tract. Supplementation with probiotics is promising in terms of reducing the harmful effects of HAAs in the human body. The aim of the study was in vitro assessment of the protective activity of the probiotic strains Lb. rhamnosus 0900, Lb. rhamnosus 0908, Lb. casei 0919 and Lb. casei DN 114001 against IQ (2-amino-3-methyl-3H-imidazo[4,5-f]quinoline) and PhIP (2-amino-1-methyl-6-phenyl-1H-imidazo[4,5-b]pyridine) after incubation with faeces from 15 persons aged 4 months to 82 years (children, adults and the elderly). The highest mean cytotoxicity of FW was observed for the elderly (63.2% ± 3.7%) and the lowest for children (28.0% ± 9.5%), as estimated by a neutral red uptake assay. The probiotics lowered the average cytotoxicity of FW exposed to IQ or PhIP. The concentration of IQ and PhIP in FW was most effectively reduced by Lb. rhamnosus 0900 (47.5%) and Lb. casei 0919 (45.8%), respectively, as determined by high -performance liquid chromatography. All the tested strains bound PhIP to a higher extent than IQ. In an alkaline comet assay, Lb. casei 0919 and Lb. rhamnosus 0908 displayed the strongest protective effect against IQ and PhIP (up to 80% reduction of DNA damage). Also in a comet assay, Lb. rhamnosus 0908 exhibited antioxidative activity toward H₂O₂ and PhIP (up to 63% and 69.5% reduction of oxidative DNA damage, respectively). The protective activity of the probiotic strains was specific to a given person’s FW, which implies the involvement of intestinal microbiota in the process.     

Survival of probiotic adjunct cultures in cheese and challenges in their enumeration using selective media

Various selective media for enumerating probiotic and cheese cultures were screened, with 6 media then used to study survival of probiotic bacteria in full-fat and low-fat Cheddar cheese. Commercial strains of Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus paracasei, or Bifidobacterium lactis were added as probiotic adjuncts. The selective media, designed to promote growth of certain lactic acid bacteria (LAB) over others or to differentiate between LAB, were used to detect individual LAB types during cheese storage. Commercial strains of Lactococcus, Lactobacillus, and Bifidobacterium spp. were initially screened on the 6 selective media along with nonstarter LAB (NSLAB) isolates. The microbial flora of the cheeses was analyzed during 9 mo of storage at 6°C. Many NSLAB were able to grow on media presumed selective for Lactococcus, Bifidobacterium spp., or Lb. acidophilus, which became apparent after 90 d of cheese storage, Between 90 and 120 d of storage, bacterial counts changed on media selective for Bifidobacterium spp., suggesting growth of NSLAB. Appearance of NSLAB on Lb. casei selective media [de man, Rogosa, and Sharpe (MRS) + vancomycin] occurred sooner (30 d) in low-fat cheese than in full-fat control cheeses. Differentiation between NSLAB and Lactococcus was achieved by counting after 18 to 24 h when the NSLAB colonies were only pinpoint in size. Growth of NSLAB on the various selective media during aging means that probiotic adjunct cultures added during cheesemaking can only be enumerated with confidence on selective media for up to 3 or 4 mo. After this time, growth of NSLAB obfuscates enumeration of probiotic adjuncts. When adjunct Lb. casei or Lb. paracasei cultures are added during cheesemaking, they appear to remain at high numbers for a long time (9 mo) when counted on MRS + vancomycin medium, but a reasonable probability exists that they have been overtaken by NSLAB, which also grow readily on this medium. Enumeration using multiple selective media can provide insight into whether it is the actual adjunct culture or a NSLAB strain that is being enumerated.     

Cloning,sequencing and expression analysis of a gene encoding alcohol oxidase in Paenibacillus sp. AIU 311

We have cloned a gene encoding an alcohol oxidase (AOD) specific to aldehyde alcohols from Paenibacillus sp. AIU 311. The AOD gene contains an open reading frame consisting of 618 nucleotides corresponding to 205 amino acid residues. The deduced amino acid sequence exhibits a high similarity to that of manganese superoxide dismutases (SODs). We expressed the cloned gene as an active product in Escherichia coli BL21 cells. The productivity (total units per culture broth volume) of the recombinant AOD expressed in E. coli BL21 is 26,000-fold higher than that of AOD in Paenibacillus sp. AIU 311. The recombinant AOD also exhibits aldehyde alcohol oxidase activity and SOD activity. The recombinant cells described in this study have utility for the production of glyoxal from glycolaldehyde.     

Protective effect of the fruits of Terminalia arjuna against cadmium-induced oxidant stress and hepatic cell injury via MAPK activation and mitochondria dependent pathway

Cadmium (Cd), a highly toxic environmental pollutant, induces hepatic disorders. The present study has been undertaken to investigate the protective role of the fruit extract of Terminalia arjuna (AE) against Cd-induced oxidative liver impairment using a murine model. Cadmium reduced hepatocytes viability, activated MAPKs, disturbed Bcl-2 family protein balance, increased reactive oxygen species (ROS) production and induced apoptotic cell death by mitochondria dependent caspases-3 activation. AE treatment, however, suppressed all the apoptotic actions of cadmium. Similarly, mice treated with cadmium altered a number biomarkers related to hepatic oxidative stress and other apoptotic indices. Oral administration of AE both pre and post prevented all the Cd-induced hepatic damages.     

A selective medium for isolation and accurate enumeration of Lactobacillus casei-group members in probiotic milks and dairy products

The correct identification and quantitative detection of Lactobacillus casei-group members continues to be a key issue due to the extensive use of members of this group as (probiotic) adjuncts, corrective and/or reinforcement cultures. Numerous selective/differential media use selective agents to which injured or stressed cells are sensitive, resulting in an underestimation of the actual number of target bacteria. We developed a selective medium and applied it to detect members of the Lb. casei-group reliably, regardless of their physiological condition, quantitatively detect and isolate strains of this group from probiotic milk and cheeses, and monitor the probiotic Lb. paracasei CRL 431 strain in Caciotta cheeses made either from goats' or cows' milk inoculated with this probiotic strain and a multiple strain culture. This approach could be useful for control bodies, agro-food and pharmaceutical industries to isolate Lb. casei-group strains, develop probiotic food/supplements and demonstrate their compliance with the species labelling.     

Multivariate analysis of proteolysis patterns differentiated the impact of six strains of probiotic bacteria on a semi-hard cheese

The individual contribution of 6 strains of probiotic bacteria (3 of Lactobacillus acidophilus and 3 of the Lactobacillus casei group) to proteolysis patterns in a semi-hard cheese was assessed. Control cheeses (without probiotics) and 2 types of experimental cheeses (with the addition of probiotics either directly to milk or by a 2-step fermentation method) were manufactured. Cheeses containing Lb. acidophilus showed the most extensive peptidolysis, which was evidenced by changes in the peptide profiles and a noticeable increase of free amino acids compared with control cheeses. The strains of the Lb. casei group showed a lower contribution to cheese peptidolysis, which consisted mainly of free amino acid increase. Two-step fermentation improved peptidolytic activity for only one of the cultures of Lb. acidophilus tested. The addition of Lb. acidophilus strains into cheese may be suitable not only for their beneficial health effect but also for their influence on secondary proteolysis, consistent with acceleration of ripening and improved flavor formation.     

Fermentation characteristics and transit tolerance of probiotic Lactobacillus casei Zhang in soymilk and bovine milk during storage

Lactobacillus casei Zhang is a novel strain that was screened out of koumiss collected in Inner Mongolia, and our previous research showed that L. casei Zhang has health benefits such as cholesterol-reducing and immunomodulating effects. The fermentation characteristics of L. casei Zhang in soymilk and bovine milk and the transit tolerance of L. casei Zhang in fermented milk products during refrigerated storage for 28 d were assessed. A faster decrease in pH and faster growth of L. casei Zhang during fermentation were observed in soymilk compared with bovine milk at various inoculation rates, probably because of the low pH buffering capacity of soymilk. The fermented bovine milk samples had much higher final titratable acidity (TA) values (between 0.80 and 0.93%) than the soymilk samples (between 0.40 and 0.46%). Dramatic increases in TA values in the fermented soymilk samples during storage were observed, and the TA values of the fermented soymilk samples changed from <0.56% to values between 0.86 and 0.98%. On the other hand, only slight increases in TA were observed in the bovine milk samples during the 28 d of storage. The survival rates of freshly prepared cultures of L. casei Zhang in simulated gastric juice at pH 2.0 and 2.5 were 31 and 69%, respectively, and the delivery of L. casei Zhang through fermented soymilk and bovine milk significantly improved the viability of L. casei Zhang in simulated gastric transit. Lactobacillus casei Zhang showed good tolerance to simulated gastric juice and intestinal juice in the fermented soymilk and bovine milk samples, and maintained high viability (>10⁸ cfu/g) during storage at 4°C for 28 d. Our results indicated that both soymilk and bovine milk could serve as vehicles for delivery of probiotic L. casei Zhang, and further research is needed to elucidate the mechanism of the change in pH and TA of L. casei Zhang in fermented milk samples during fermentation and storage and to understand the difference between soy- and milk-based systems.     

Protective effect of Cordyceps militaris against high glucose-induced oxidative stress in human umbilical vein endothelial cells

The protective effect of Cordyceps militaris against high glucose-induced oxidative stress in human umbilical vein endothelial cells (HUVECs), as compared with Cordyceps sinensis, was examined. The cytotoxicity of HUVECs induced by 40 mM glucose was ameliorated by water extracts of C. militaris (CME) and water extracts of C. sinensis (CSE). CME and CSE inhibited the increase in ROS and NO in HUVECs induced by 40 mM high glucose. Moreover, CME increased the Bcl-2/Bax ratio, modulated the mitochondrial membrane potential and reduced the caspase-3 activity in high glucose-induced HUVECs. In addition, cordycepin, a component of CME and CSE, displayed protective effects against oxidative stress, which was partly responsible for the cytoprotective effects of CME and CSE against high glucose-induced oxidative stress in HUVECs. Overall, the obtained results show C. militaris helps preventing diabetic endothelial dysfunction and related complications.