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PubMed1992

Faecal steroids and colorectal cancer: the effect of lactulose on faecal bacterial metabolism in a continuous culture model of the large intestine.

The effect of lactulose on intestinal bacterial metabolism in two identical single-stage chemostats has been studied. The study was designed as a single stage crossover. Both cultures were inoculated and treated in an identical manner except that whilst one fermenter was subject to pH control the other was not and vice versa. Complex bacterial populations were maintained for 23 days and many of the metabolic functions carried out by the micro-organisms in vivo were demonstrated in vitro. The predominant organisms belonged to the genera Bacteroides, Bifidobacterium and Clostridium with abundant levels of anaerobic Gram-positive rods. The redox potential in each fermenter showed considerable variation with maximum values of below -300 eV being attained. An indication of the stability of the bacterial communities was the production of short-chain volatile fatty acids; during steady-state conditions the mean ratio of the major acids acetic, propionic and butyric being 3.90:0.69:1.00 and 3.65:0.76:1.00 in each fermenter, respectively. During steady-state conditions, 7 alpha-dehydroxylation of the primary bile acid chenodeoxycholic acid was maintained at a constant rate with lithocholic acid representing over 85% of total steroid. Addition of the soluble fibre lactulose to the cultures had a profound effect on many of the parameters tested. Without pH control the culture pH dropped to below 5.0 and this coincided with a substantial decrease in total anaerobes, especially Bacteroides spp., an increase in Lactobacillus spp. (concomitant with an increase in lactic acid), a decrease in the concentration of short-chain volatile fatty acids and a substantial decrease in 7 alpha-dehydroxylation of chenodeoxycholic acid. These results show that it is possible to maintain viable intestinal bacteria in vitro using a continuous culture chemostat and that the cultures respond to changing conditions as evinced by the inhibition of 7 alpha-dehydroxylation of chenodeoxycholic acid on addition of lactulose. This indicates that the model may be of further use in studying the modulation of secondary bile acid formation, a possible risk factor in colorectal carcinogenesis.

Amino Acids↗
PubMed1994

Chrymutasins: novel-aglycone antitumor antibiotics from a mutant of Streptomyces chartreusis. I. Taxonomy, mutation, fermentation, isolation and biological activities.

Three novel antibiotics, named chrymutasins A, B and C, were isolated from the fermentation products of a mutant strain obtained by NTG (N-methyl-N'-nitro-N-nitrosoguanidine) treatment. The mutant strain produced the chrymutasins, which differed in the aglycone moiety from chartreusin, and related compounds. The production of these compounds needed a characteristically long fermentation period. The antitumor activity of chrymutasin A is better in vivo than that of chartreusin, the cytotoxic activity against cell lines in vitro is equivalent, and the antimicrobial spectrum is narrower.

Animals↗
PubMed1993

Comparison of copper lysine and copper sulfate as copper sources for ruminants using in vitro methods.

Two in vitro experiments were conducted to estimate the availability of Cu from Cu Lys compared with Cu from CuSO4. In experiment 1, a 24-h ruminal fermentation (alpha-cellulose substrate) containing either .1% added S or 2% added urea was performed with 4, 12, or 96 ppm of added Cu as either Cu Lys or CuSO4. Soluble Cu was measured at the end of the 24-h incubation, and concentrations were higher for Cu Lys with added urea (1.16 vs. .45 ppm), but no differences existed between sources of added S. In Experiment 2, 20 ppm of Cu added as either Cu Lys or CuSO4 with and without the addition of 10 ppm of Mo and .75% S were used in an vitro study designed to simulate digesta passage through the ruminant. A 24-h ruminal fermentation (orchardgrass substrate) was followed by a 2-h digestion in pepsin and HCl (pH 2.3) and then a 2-h digestion in NaHCO3 and pancreatin (pH 6.6). Soluble Cu concentrations were analyzed after each step. Molybdenum and S addition decreased soluble Cu after ruminal fermentation and tended to decrease soluble Cu concentrations after digestion of pancreatin and NaHCO3. The source of Cu did not affect soluble Cu concentrations. Results suggest that Cu from Cu Lys and CuSO4 behave similarly in the digestive tract of ruminants.

Animals↗
PubMed1999

Biochemical characteristics & secretory activity of Aeromonas species isolated from children with gastroenteritis in Chennai.

An attempt was made to delineate the phenotypic markers for the detection of enterotoxigenic strains of Aeromonas. Eighteen Aeromonas species comprising one isolate of A. hydrophila, six isolates of A. sobria and 11 isolates of A. caviae were obtained from 379 children suffering from acute diarrhoea in Chennai. Nine of these isolates inclusive of three A. sobria and six A. caviae were found to produce secretory response in vitro in the rabbit intestinal mucosa mounted in the Ussing chambers as revealed by significant increases in the short circuit current. Eleven strains hydrolysed aesculin, 8 fermented arabinose, 6 produced acetyl methyl carbinol, 14 produced lysine decarboxylase, 3 fermented salicin, 9 produced beta-haemolysin, 9 produced CAMP-like factor and only two isolates took up congo red dye. None of these phenotypic traits were found to correlate with the in vitro secretory activity.

Aeromonas↗
PubMed1994

Digestibility of carbohydrates from rice-, oat- and wheat-based ready-to-eat breakfast cereals in children.

OBJECTIVE: To study the effect of the presence and quality of dietary fibre in ready-to-eat (RTE) breakfast cereals on completeness of carbohydrate digestion in children and on starch susceptibility to alpha-amylase in vitro. DESIGN: A controlled intervention study. SUBJECTS: Eight 3-8-year-old healthy children. INTERVENTIONS: Completeness of digestion was evaluated by assessing the amount of carbohydrates apparently fermented into the colon using the breath-H2 technique after consumption in random order, of five breakfast tests containing boiled rice (either alone or supplemented with 3 g of lactulose) as reference food, or RTE cereals based on rice (low-fibre), wheat (high insoluble fibre) and oats (high-soluble fibre). The potential glycaemic impact of the products was estimated in vitro by assessing starch susceptibility to alpha-amylolysis using an enzymatic-dialysis method. RESULTS: Compared to boiled rice and to rice-based RTE cereal, wheat- and oat-based RTE cereals both significantly (P < 0.05) increased the amount of apparently fermented carbohydrates (+1.1 +/- 1.7% of total breakfast carbohydrate fermented for rice, +5.6 +/- 0.9% for wheat and +9.4 +/- 3.7% for oats; mean +/- SEM), calculated using the excess H2 in breath after lactulose as standard. All products showed similar in vitro digestibility, resulting in estimated glycaemic indexes of 117.5 (24.0) for rice, and 105.7 (14.1) for oats-based, 128.4 (17.6) for wheat-based, and 129.8 (16.6) [mean 95% CI)] for rice-based RTE cereals. CONCLUSIONS: Results suggest that the presence of fibre in RTE breakfast cereals, in particular soluble fibre, increases colonic fermentation in children whereas it seems not to affect glucose availability.

Avena↗
PubMed1997

Effect of processing high moisture ear corn on ruminal fermentation and milk yield.

Thirty-six multiparous dairy cows (8 fitted with ruminal cannulas) and 16 primiparous dairy cows were blocked by days in milk and parity and assigned to one of four diets containing 53% alfalfa silage [dry matter (DM) basis] plus 1) high moisture ear corn, 2) high moisture ear corn plus expeller soybean meal, 3) ground high moisture ear corn, or 4) ground high moisture ear corn plus expeller soybean meal. The high moisture ear corn was rolled before ensiling at 68% DM. Ground high moisture ear corn was prepared by further grinding through a 9.5-mm screen; grinding reduced the geometric mean particle size from 4.33 to 1.66 mm. Diets contained 1.69 Mcal of net energy for lactation/kg of DM. Relative to cows fed diet 1, milk yield was 4 kg/d greater for cows fed diet 2; numerically, milk yield was about 2 kg/d greater for cows fed diets 3 and 4 than for cows fed diet 1. Yield of milk components also was greater for cows fed diets 2 and 3 but not for cows fed diet 4. Intake of DM and yield of 4% FCM were greatest for cows fed diet 3 and lowest for cows fed diet 1. Digestibilities of DM, organic matter, starch, neutral detergent fiber, and acid detergent fiber were increased, and ruminal NH3 concentration was depressed, by the grinding of high moisture corn; expeller soybean meal increased ruminal NH3. Total volatile fatty acid concentration was not different among in vivo treatments, but the molar proportion of acetate decreased, and propionate increased, for cows fed diet 3. The grinding of high moisture corn significantly decreased pH, increased total volatile fatty acid concentration, and increased the rate of decline of NH3 concentration in ruminal in vitro incubations. Grinding improved the utilization of high moisture corn by lactating cows by stimulating ruminal fermentation.

Ammonia↗
PubMed1983

Asparaginase II of Saccharomyces cerevisiae: comparison of enzyme stability in vivo and in vitro.

Asparaginase II of Saccharomyces cerevisiae is a cell wall mannan containing glycoprotein. Recent studies have demonstrated that asparaginase II activity increases in exponentially growing cell cultures and then decreases as the cells enter the stationary phase. Enzyme inactivation has been attributed to a Zn2+-dependent protease which is synthesized de novo during the late exponential phase [Pauling, K.D., & Jones, G.E. (1980) J. Gen. Microbiol. 117, 423-430; Pauling, K.D., & Jones, G.E. (1980) Biochim. Biophys. Acta 616, 271-282]. We have investigated the mechanism of asparaginase II inactivation using both whole cell suspensions and highly purified enzyme. Our data indicate that the rate of asparaginase II inactivation in cell suspensions is primarily influenced by pH changes that occur as a consequence of cell growth and glucose fermentation and that enzyme inactivation is not dependent on Zn2+ or on de novo protein synthesis. Also, in vitro studies with purified enzyme show kinetics of inactivation that are similar to those observed in vivo. Consequently, involvement of a yeast protease in the inactivation process is relatively unlikely.

Asparaginase↗
PubMed1984

Effect of pH on in vitro ruminal conversion of L-tryptophan to 3-methylindole and indole.

Three in vitro experiments were conducted to determine the effects of pH on ruminal conversion of L-tryptophan (TRP) to 3-methylindole (3-MI) and indole (IND). Experiment 1 involved 2 closed-system incubations, each with triplicate replications of buffered ruminal fluid at initial pH of 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, and 8.0. Highest molar conversions of TRP to 3-MI were 61.5 +/- 2.7% and 64.3 +/- 0.8% at initial pH treatments of 7.5 and 7.0 (final pH 7.3 and 6.9) for the 2 incubations, respectively. Experiment 2 used duplicate continuous cultures at each of 4 pH treatments. Following a common 5-day pretreatment period, effluent pH averaged 5.6, 6.1, 6.5, and 6.9 for the respective treatments. Data for 4 subsequent 5-day time periods showed production of 3-MI was affected by pH treatment (P less than 0.01), time (P greater than 0.01), and treatment X time (P less than 0.01); conversion of TRP to 3-MI reached 78% at pH 6.9 and decreased to less than 1% at pH 5.6. Production of IND was not related to pH treatment (P greater than 0.10). Total moles of volatile fatty acid (VFA) carbon produced showed a pH treatment X time interaction (P less than 0.01) which reflected a trend toward decreasing VFA production at lower pH and increasing VFA production at higher pH. Experiment 3 consisted of 2 pH treatments with additional continuous culture fermenters.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2000

Inhibition of enzymic digestion of amylose by free fatty acids in vitro contributes to resistant starch formation.

The effect of lipids on the enzymic breakdown of starch was investigated using an in vitro assay system. Mixtures of potato amylose, amylopectin and starch and various lipids were incubated at 37 degrees C for 10 min and subjected to digestion by alpha-amylase (EC 3.2.1.1) and amyloglucosidase (EC 3.2.1.33). Lauric, myristic, palmitic and oleic acids and lysolecithin inhibited enzymic hydrolysis of amylose by approximately 35% (P < 0.05). Stearic acid and cholesterol had no effect on the enzymic breakdown of amylose. Retrograded amylose was hydrolyzed less readily (P < 0.05) than solubilized amylose, but the breakdown was not further inhibited in the presence of lauric acid. Fatty acids had no effect on the enzymic hydrolysis of amylopectin, whereas inhibition by fatty acids of the breakdown of whole starch was consistent with only the amylose fraction being affected. The possibility that interactions between starch and fatty acids in the digestive tract could contribute to the formation of resistant starch is considered.

Amylose↗
PubMed1988

Nutrition and biochemistry of anaerobic Chytridiomycetes.

The nutrition and biochemistry of anaerobic Chytridiomycetes is at present poorly understood. Data has been obtained principally from studies of rumen isolates of Neocallimastix spp. grown in vitro. Our knowledge of the nutrition of Neocallimastix is summarised. Current information on glycolysis and fermentation product generation via cystosolic and hydrogenosomal systems, production of enzymes involved in plant cell wall hydrolysis, lipid metabolism and the role of Chytridiomycetes in ruminal proteolysis is discussed. At present this is insufficient to provide useful phylogenetic information.

Anaerobiosis↗
PubMed2006

Fermented papaya preparation attenuates beta-amyloid precursor protein: beta-amyloid-mediated copper neurotoxicity in beta-amyloid precursor protein and beta-amyloid precursor protein Swedish mutation overexpressing SH-SY5Y cells.

Recent studies indicate that the deposition of beta-amyloid (Abeta) is related in the pathogenesis of Alzheimer's disease (AD), but the underlying mechanism is still not clear. The abnormal interactions of Abeta with metal ions such as copper are implicated in the process of Abeta deposition and oxidative stress in AD brains. In the present study, we established a new AD model, using which we found that copper triggered the Abeta neurotoxicity in SH-SY5Y cells overexpressing the Swedish mutant form of human APP (APPsw) in a concentration dependent manner. Fermented papaya preparation (FPP) has shown high free radical scavenging ability in vivo and in vitro. FPP post-treatment increased cell viability and decreased the intracellular [Ca2+]i, reactive oxygen species (ROS) generation such as hydroxyl free radical and superoxide anion and nitric oxide (NO) accumulation in the cell. Our results also show that FPP prevents the cell apoptosis through bax/bcl-2 sensitive pathway.

Amyloid beta-Peptides↗
PubMed2005

[Plague in Algeria: about five strains of Yersinia pestis isolated during the outbreak of June 2003].

In this study, we isolated and identified five strains of Yersinia pestis during an epidemic occurred in west of Algeria in June 2003. The bacteriological identification was confirmed by bacteriophage susceptibility. All these strains belonged to the biovar Orientalis (they did not ferment glycerol but did reduce nitrate to nitrite) which caused the current pandemic. The in vitro activities of antimicrobial agents used to treat plague and recommended for prophylaxis, showed that they are active against all strains. The comparison of these strains by plasmid profile analyse demonstrated that all isolates had three plasmids: 110, 70 and 9.5 kb, which are present in Y. pestis strains.

Algeria↗
PubMed2006

Beneficial effects of green tea--a review.

Tea is the most consumed drink in the world after water. Green tea is a 'non-fermented' tea, and contains more catechins, than black tea or oolong tea. Catechins are in vitro and in vivo strong antioxidants. In addition, its content of certain minerals and vitamins increases the antioxidant potential of this type of tea. Since ancient times, green tea has been considered by the traditional Chinese medicine as a healthful beverage. Recent human studies suggest that green tea may contribute to a reduction in the risk of cardiovascular disease and some forms of cancer, as well as to the promotion of oral health and other physiological functions such as anti-hypertensive effect, body weight control, antibacterial and antivirasic activity, solar ultraviolet protection, bone mineral density increase, anti-fibrotic properties, and neuroprotective power. Increasing interest in its health benefits has led to the inclusion of green tea in the group of beverages with functional properties. However, although all the evidence from research on green tea is very promising, future studies are necessary to fully understand its contributions to human health, and advise its regular consumption in Western diets, in which green tea consumption is nowadays limited and sporadic.

Anticarcinogenic Agents↗
PubMed2004

Differentiation-promoting activity of pomegranate (Punica granatum) fruit extracts in HL-60 human promyelocytic leukemia cells.

Differentiation refers to the ability of cancer cells to revert to their normal counterparts, and its induction represents an important noncytotoxic therapy for leukemia, and also breast, prostate, and other solid malignancies. Flavonoids are a group of differentiation-inducing chemicals with a potentially lower toxicology profile than retinoids. Flavonoid-rich polyphenol fractions from the pomegranate (Punica granatum) fruit exert anti-proliferative, anti-invasive, anti-eicosanoid, and pro-apoptotic actions in breast and prostate cancer cells and anti-angiogenic activities in vitro and in vivo. Here we tested flavonoid-rich fractions from fresh (J) and fermented (W) pomegranate juice and from an aqueous extraction of pomegranate pericarps (P) as potential differentiation-promoting agents of human HL-60 promyelocytic leukemia cells. Four assays were used to assess differentiation: nitro blue tetrazolium reducing activity, nonspecific esterase activity, specific esterase activity, and phagocytic activity. In addition, the effect of these extracts on HL-60 proliferation was evaluated. Extracts W and P were strong promoters of differentiation in all settings, with extract J showing only a relatively mild differentiation-promoting effect. The extracts had proportional inhibitory effects on HL-60 cell proliferation. The results highlight an important, previously unknown, mechanism of the cancer preventive and suppressive potential of pomegranate fermented juice and pericarp extracts.

Cell Differentiation↗
PubMed1993

Starch bioavailability in arepas made from ordinary or high amylose corn: concentration and gastrointestinal fate of resistant starch in rats.

The purpose of the present investigation was to study the importance of the amylose/amylopectin ratio for the content and gastrointestinal fate of resistant starch in a realistic composite starchy food. Corn-based breads (arepas) from dent corn (25% amylose) and from high amylose corn (70% amylose) were used as test products. Resistant starch concentration was evaluated in vitro and in vivo using rats treated with an antibiotic drug (Nebacitin) to suppress hindgut fermentation. Experiments in rats with intact hindgut microflora allowed determination of resistant starch fermentability. The small intestinal digestibility of starch in dent corn arepas was close to 96% (total starch basis), whereas the starch in high amylose arepas was poorly digested (approximately 68%, total starch basis), as calculated from the fecal recovery of resistant starch in Nebacitin-treated animals. The main resistant starch fraction required solubilization in alkali to render it available to the analytical amylases (nonhydrated fraction). The total amount of resistant starch as well as the nonhydrated starch fraction delivered to the hindgut could be accurately predicted from analysis of starch remnants in an enzymatic gravimetric dietary fiber residue. Resistant starch present in dent corn arepas was fermented approximately 63%, whereas the fermentability of resistant starch from the high amylose product was remarkably low (< 11%).

Amylopectin↗
PubMed1994

Galactose Expulsion during Lactose Metabolism in Lactococcus lactis subsp. cremoris FD1 Due to Dephosphorylation of Intracellular Galactose 6-Phosphate.

In Lactococcus lactis subsp. cremoris FD1, galactose and lactose are both transported and phosphorylated by phosphotransferase systems. Lactose 6-phosphate (lactose-6P) is hydrolyzed intracellularly to galactose-6P and glucose. Glucose enters glycolysis as glucose-6P, whereas galactose-6P is metabolized via the tagatose-6P pathway and enters glycolysis at the tagatose diphosphate and fructose diphosphate pool. Galactose would therefore be a gluconeogenic sugar in L. lactis subsp. cremoris FD1, but since fructose 1,6-diphosphatase is not present in this strain, galactose cannot serve as an essential biomass precursor (glucose-6P or fructose-6P) but only as an energy (ATP) source. Analysis of the growth energetics shows that transition from N limitation to limitation by glucose-6P or fructose-6P gives rise to a very high growth-related ATP consumption (152 mmol of ATP per g of biomass) compared with the value in cultures which are not limited by glucose-6P or fructose-6P (15 to 50 mmol of ATP per g of biomass). During lactose metabolism, the galactose flux through the tagatose-6P pathway (r(max) = 1.2 h) is lower than the glucose flux through glycolysis (r(max) = 1.5 h) and intracellular galactose-6P is dephosphorylated; this is followed by expulsion of galactose. Expulsion of a metabolizable sugar has not been reported previously, and the specific rate of galactose expulsion is up to 0.61 g of galactose g of biomass h depending on the lactose flux and the metabolic state of the bacteria. Galactose excreted during batch fermentation on lactose is reabsorbed and metabolized when lactose is depleted from the medium. In vitro incubation of galactose-6P (50 mM) and permeabilized cells (8 g/liter) gives a supernatant containing free galactose (50 mM) but no P(i) (less than 0.5 mM). No organic compound except the liberated galactose is present in sufficient concentration to bind the phosphate. Phosphate is quantitatively recovered in the supernatant as P(i) by hydrolysis with alkaline phosphatase (EC 3.1.3.1), whereas inorganic pyrophosphatase (EC 3.6.1.1) cannot hydrolyze the compound. The results indicate that the unknown phosphate-containing compound might be polyphosphate.

Journal Article↗