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Carcinogen binding to various types of dietary fiber.

The percent of the carcinogen 1,2-dimethylhydrazine (DMH) bound to a variety of fibers, such as wheat bran, corn bran, citrus pulp, citrus pectin, and alfalfa, was examined at pH values ranging from 1 to 12. The percent of DMH bound to wheat bran increased from 4% at PH 1 to 55% at pH 2 to 77% at pH 12. A sharp rise in carcinogen binding to corn bran occurred between pH 5% of the DMH was bound and pH 8 where 51% of the DMH was bound. The percent of DMH bound to dehydrated citrus pulp also increased as the pH increased with 10% binding observed at pH 1 and with 57% binding observed at pH 12. Between pH 2 and pH 7, the percent of DMH bound to pectin decreased from 60 to 11%. As the pH became more basic, the percent of DMH bound to pectin increased to 42% at pH 12. The sharpest rise in the percent of DMH bound to alfalfa meal occurred between pH 10.5 and pH 12.0. Results from this experiment showed that the affinity to various types of dietary fibers for the colon carcinogen DMH was differentially affected by pH. These results suggested that the protective effect of certain types of dietary fiber against chemically induced colon cancer my in part be attributed to enhanced carcinogen binding by dietary fiber in the colon.

Cellulose↗

The effect of different pectic growth substrates on beta-glucosidase in Fusarium oxysporum f. sp. radicis lycopersici: partial purification and characterization.

Fusarium oxysporum f. sp. radicis lycopersici produces beta-glucosidase activities when it grows on pectin and glucose. The pectins were better substrates than glucose. In the medium containing galacturonic acid or sucrose the activity was present in low levels and at the end of autolysis. A beta-glucosidase from the pectin medium was purified by ion exchange chromatography followed by gel filtration. The enzyme was a unique band of protein in SDS-PAGE and isoelectric focussing. It had a molecular weight of 86,000 and a pI of 4.8. This beta-glucosidase was a glycoprotein.

Chromatography, Ion Exchange↗

Regulation of pectic enzymes from the exo-1 mutant strain of Neurospora crassa: effects of glucose, galactose, and galacturonic acid.

The exo-1 mutant of Neurospora crassa produced and secreted pectolytic activities when incubated in the presence of pectin-containing biological materials. This study shows that polygalacturonase, pectate lyase and pectin lyase activities were induced in media supplemented with galactose or galacturonic acid, indicating that these sugars induced the synthesis of pectinases. Pectinesterase activity was undetectable. Polygalacturonase activity was better induced by galactose than by galacturonic acid. The reverse was true for lyase activities. The inducing effect of galactose and galacturonic acid seemed to be different: (i) a mixture of galactose and galacturonic acid synergistically increased the production of pectic enzymes, as compared to that in the presence of one of these sugars; (ii) the inducing effect of galacturonic acid was partially repressed by glucose; (iii) in contrast, the inducing effect of galactose, rather than repressed, was enhanced by the presence of glucose. Altogether, these data point out to a complex mechanism of regulation of pectolytic enzymes by pectin-containing organic substances.

Enzyme Induction↗

Dietary regulation and localization of apoptosis cascade proteins in the colonic crypt.

This study was designed primarily to assess the localization of apoptosis cascade proteins along the rat colonic crypt and secondarily to test whether the activity and/or localization of these proteins are affected by the enrichment of the diet with the soluble fiber pectin. Expression of apoptosis cascade proteins was assessed in isolated colonocytes harvested from the luminal and basal crypt colonocyte populations. Two different dietary regimens were tested: a standard diet (diet A), and a diet enriched in pectin (diet B), a soluble fiber that undergoes fermentation in the cecum and produces high concentrations of intracolonic short-chain fatty acids. Caspase-1 expression was maximal in luminal colonocytes of rats fed diet B, as evidenced by Western blot and immunohistological analyses. Expression of the cleaved poly(ADP-ribose) polymerase product was elevated in both the luminal and basal colonocytes of the pectin-fed group, whereas in rats fed diet A, the expression was lower, especially in basal crypt colonocytes. The highest expression of the antiapoptotic protein Bcl-2 was observed in the lower compartments of the colonic crypt tissue and was maximal in the rat group fed a standard diet. The apoptotic index in colonocytes of rats fed diet B was higher than that measured in rats fed diet A. Cumulatively, our results indicate that apoptosis cascade proteins are differentially localized along the lumen-crypt axis, and their expression and activity may be controlled by dietary components. These results may, at least partially, account for the documented protective effect of butyrogenic fibers on colorectal cancer.

Animals↗

The muscular network of the sheep right atrium and frequency-dependent breakdown of wave propagation.

The complex branching structure of the right atrium (RA) muscular network may provide the substrate for complex patterns of propagation during atrial fibrillation (AF). As AF results in some cases from stable sources in the left atrium (LA) with fibrillatory conduction toward the RA, we hypothesize that periodic input to the RA at an exceedingly high frequency results in disorganized wave propagation associated with the complex structure of the RA. Optical mapping was performed in isolated coronary-perfused sheep RA. Rhythmic pacing of Bachmann's bundle allowed well-controlled and realistic conditions for LA-driven RA. Pacing at increasingly higher frequencies led to increasing delays in activation distal to major branching sites of the Crista terminalis and pectinate bundles, culminating in spatially distributed intermittent blockade at and above approximately 6.5 Hz. At this breakdown frequency, the dominant frequencies of the RA response activity became spatially nonuniform. Such frequency-dependent changes were independent of action potential duration. Rather, the spatial boundaries between proximal and distal frequencies correlated well with branch sites of the pectinate musculature. Thus, there exists a breakdown frequency in the sheep RA below which activity is periodic throughout the atrium and above which it is fibrillation-like, consistent with the ideas that during AF, high-frequency activation initiated in the LA undergoes fibrillatory conduction toward the RA, and that sink-to-source mismatch effect at branch points of the Crista terminalis and pectinate muscles is important in determining the complexity of the arrhythmia.

Action Potentials↗

Production and properties of three pectinolytic activities produced by Aspergillus niger in submerged and solid-state fermentation.

Three extracellular pectinases were produced by Aspergillus niger CH4 by submerged and solid-state fermentation, and their physicochemical and kinetic properties were studied. The highest productivities of endo- and exo-pectinase and pectin lyase were obtained with solid-state fermentation. The kinetic and physicochemical properties of these enzymes were influenced by the type of culture method used. All activities were very different in terms of pH and temperature optima, stability at different pH and temperature values and affinity for the substrate (Km values). In solid-state fermentation, all pectinase activities were more stable at extreme pH and temperature values but the Km values of endo-pectinase and pectin lyase were higher with respect to those activities obtained by the submerged-culture technique. The pectin lyase activity obtained by the submerged-culture technique showed substrate inhibition but the enzyme obtained by solid-state fermentation did not. Electrophoresis, using sodium dodecyl sulphate/polyacrylamide gel with enzymatic extracts obtained for both culture methods, showed the same number of protein bands but some differences were found in their electrophoretic position. The results obtained in this work suggest that the culture method (submerged or solid-state) may be responsible for inducing changes in some of the pectinolytic enzymes produced by A. niger.

Aspergillus niger↗

Cholesterol binding capacity of fiber from tropical fruits and vegetables.

The cholesterol binding capacity of 28 fiber samples from a variety of the more common tropical fruits and vegetables was determined. The binding capacity of cholestyramine, cellulose, lignin, guar gum and citrus pectin were also determined. Capacities were evaluated by an in vitro method that simulates the effect of the human digestive system on fiber using a series of enzymatic treatments before the binding was determined. Binding values varied from 3% for a soluble fraction of cassava to 84% for cholestyramine. Values for most fruit and vegetable fiber samples were less than or ca. equal to cellulose or lignin (20% and 16%, respectively). Apart from cholestyramine, sweet potato was the most effective binder (30%). Citrus pectin, at 8%, was a relatively poor binder. The capacity of guar gum (17%) was slightly less than cellulose. These data do not support the conclusion from in vivo studies that the hypocholesteremic effects observed for citrus pectin and guar gum are the result of the direct binding of cholesterol or bile acids in the large intestine.

Cellulose↗

Dietary fiber supplements: effects on serum and liver lipids and on liver phospholipid composition in rats.

Rats (6 per group) were fed semipurified diets containing either particulate fibers (alfalfa, 10%; cellulose, 10%; bran, 10%), a soluble ionic fiber (pectin 5%), soluble, nonionic fibers (guar gum, 5%; Metamucil, 10%), a mixed fiber preparation (Fibyrax, 10%, or an insoluble, ionic bile acid-binding resin (cholestyramine, 2%). The control group was fed the unsupplemented diet. The feeding period, during which diet and water were provided ad libitum, was 28 days. Compared with the control group, serum total cholesterol levels were increased by more than 10% in rats fed alfalfa and decreased by more than 10% in rats fed cellulose, guar gum, Fibyrax and cholestyramine. There were no significant differences in percentage of plasma HDL cholesterol. Serum triglycerides were elevated in the groups fed alfalfa, pectin, guar gum or Fibyrax and reduced in the group fed Metamucil. Plasma phospholipids were elevated in rats fed alfalfa or bran, unaffected in rats fed pectin or Metamucil and reduced in the other groups. Liver total cholesterol was elevated in all groups but those fed wheat bran and cholestyramine. The percentage of liver cholesterol present as ester was elevated in every group except that fed cholestyramine. Liver triglycerides were reduced in rats fed guar gum or Metamucil and elevated in those fed alfalfa. Liver phospholipids were lowered in the group fed cellulose. Liver phospholipids were fractionated by thin layer chromatography to give phosphatidylcholine (PC), phosphatidylethanolamine (PE), sphingomyelin (Sph), lysophosphatidylcholine (LPC) and phosphatidylinositol plus phosphatidylserine (PI + PS).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Estimation of available energy of dietary fibres by indirect calorimetry in rats.

BACKGROUND: Knowledge of energetic availability of dietary fibres is important for human nutrition. But up to now results are often different and depend on the methods used. Estimation of metabolisable energy of dietary fibres (mainly by balance technique) is a time-consuming procedure and needs special technical effort. AIM OF THE STUDY: Validation of the experimental design for short-term studies by using indirect calorimetry with feeding below maintenance requirement to evaluate the energetic availability of dietary fibres and their influence on absorption velocity of carbohydrates (CHO). METHODS: Energy expenditure and CHO oxidation (including short-chain fatty acids as fermentation products) were estimated in Wistar rats over 23 h after being fed a basal diet for the first day (300 KJ/kg0.75, 20% protein, 3% fat, 77% CHO) followed by supplementation with either microcrystalline cellulose, the soluble rye fibre arabinoxylan, apple pectin, amylomaize starch (with 48% of resistant starch) or gelatinised wheat starch (200 KJ/kg0.75 each) as control for the following days. Energetic availability was determined by comparing the increase of CHO oxidation after addition of gelatinised wheat starch with that of the dietary fibres tested. RESULTS: In comparison to wheat starch (100%), the following energetic availability of the dietary fibres was found: microcrystalline cellulose 14%, arabinoxylan 33%, pectin 39%, amylomaize starch 62%. The time-course of CHO oxidation indicated that microcrystalline cellulose enhances, whereas the soluble rye fibre slows down the velocity of CHO absorption due to the different consistency of the intestinal contents modified by the kind and properties of the used dietary fibres. After intake of arabinoxylan or pectin, CHO oxidation remained at a higher level during the experimental period elucidating an increased activity of fermentation to short-chain fatty acids. CONCLUSIONS: Short-term experiments in rats using indirect calorimetry are a suitable method for comparative estimation of the energetic availability of dietary fibres. Results are partly in agreement with values estimated by long-term in vivo methods.

Animals↗

Volatilization of selenium from agricultural evaporation pond sediments.

Microbial volatilization of Se was evaluated as a means of detoxifying Se-contaminated sediments. Sediment samples containing 60.7 (Kesterson Reservoir) and 9.0 mg Se kg-1 (Peck ponds) were incubated for 273 days in closed systems located in the greenhouse. Volatile Se was collected from a continuous air-exchange stream using activated carbon. Various economical and readily available organic and inorganic amendments were tested for their capacity to enhance the microbial process, including Citrus (orange) peel, Vitis (grape) pomace, feedlot manure, barley straw, chitin, pectin, ZnSO4, (NH4)2SO4, and an inoculum of Acremonium falciforme (an active Se methylating fungus). With the Kesterson sediment, the highest Se removal (44.0%) resulted from the combined application of citrus peel and ZnSO4, followed by citrus peal alone (39.6%), and citrus peel combined with ZnSO4, (NH4)2SO4 and A. falciforme (30.1%). Manure (19.5%), pectin (16.4%), chitin (9.8%) and straw plus N (8.8%) had less pronounced effects. Without the amendments, cumulative Se volatilization was 6.1% of the initial inventory. Grape pomace (3.0%) inhibited the process. With the Peck sediment, the highest amount of Se removed was observed with chitin (28.6%), manure (28.5%), and citrus peel alone (27.3%). Without amendments, 14.0% of the native Se was volatilized in 273 days. Cumulative Se volatilization was 24.7% with citrus plus Zn and N, 17.2% with citrus plus Zn, and 18.8% with citrus plus Zn, N and A. falciforme. Pectin (15.2%), straw plus N (16.4%), and grape pomace (7.3%) were among the less effective amendments for the Peck sediment. The differences in the effectiveness of each treatment between the two seleniferous soils may be a result of the residual N content of the sediments. With the Kesterson sediment, which was high in organic C and N, added N inhibited volatilization of Se, while with Peck sediments (low in organic C and N) N-rich materials tended to accelerate Se volatilization. Inoculation with A. falciforme did not enhance Se evolution from either sediment, indicating that there was a sufficient population of microflora capable of producing gaseous Se.

Acremonium↗

Accelerated ion fluxes during differentiation in zoospores of Phytophthora palmivora.

Zoospores of Phytophthora palmivora show increased fluxes of Na+ and Ca2+ 3-5 min after they have been stimulated to differentiate with pectin. Both spontaneous and pectin-induced encystment are reduced below pH 6 and accelerated above pH 7. The ionophores monensin and A23187 induce slow differentiation when added together, but not when added separately. Ethanol (0.5%) also induces slow differentiation. Amiloride and verapamil inhibit pectin-induced differentiation and also reduce the onset of the Na+ and Ca2+ flux. A requirement for Ca2+ for differentiation is confirmed, but a requirement for Na+ could not be demonstrated.

Amiloride↗

Pectinolytic enzymes secreted by yeasts from tropical fruits.

Three hundred yeasts isolated from tropical fruits were screened in relation to secretion of pectinases. Twenty-one isolates were able to produce polygalacturonase and among them seven isolates could secrete pectin lyase. None of the isolates was able to secrete pectin methylesterase. The pectinolytic yeasts identified belonged to six different genera. Kluyveromyces wickerhamii isolated from the fruit mangaba (Hancornia speciosa) secreted the highest amount of polygalacturonase, followed by K. marxianus and Stephanoascus smithiae. The yeast Debaryomyces hansenii produced the greatest decrease in viscosity while only 3% of the glycosidic linkages were hydrolysed, indicating that the enzyme secreted was an endo-polygalacturonase. The hydrolysis of pectin by polygalacturonase secreted by S. smithiae suggested an exo-splitting mechanism. The other yeast species studied showed low polygalacturonase activity.

Carboxylic Ester Hydrolases↗

Plant protein inhibitors of cell wall degrading enzymes.

Plant cell walls, which consist mainly of polysaccharides (i.e. cellulose, hemicelluloses and pectins), play an important role in defending plants against pathogens. Most phytopathogenic microorganisms secrete an array of cell wall degrading enzymes (CWDEs) capable of depolymerizing the polysaccharides in the plant host wall. In response, plants have evolved a diverse battery of defence responses including protein inhibitors of these enzymes. These include inhibitors of pectin degrading enzymes such as polygalacturonases, pectinmethyl esterases and pectin lyases, and hemicellulose degrading enzymes such as endoxylanases and xyloglucan endoglucanases. The discovery of these plant inhibitors and the recent resolution of their three-dimensional structures, free or in complex with their target enzymes, provide new lines of evidence regarding their function and evolution in plant-pathogen interactions.

Carboxylic Ester Hydrolases↗

Short-chain fatty acids induce cell cycle inhibitors in colonocytes.

BACKGROUND & AIMS: We tested the hypothesis that short-chain organic acids in the colon derived from dietary pectin, wheat bran, and oat bran are protective against the development of colon cancer because they induce transforming growth factor (TGF)-beta1, which in turn inhibits cell growth by inducing cyclin-directed kinase (cdk) inhibitors. METHODS: U4 human colon carcinoma cells differentiate into water- and salt-transporting columnar enterocytes and therefore model normal colonocytes. The composition and kinase activity of cdk/cyclin complexes were determined by immunoprecipitation and Western blotting studies in U4 cells treated in vitro with short-chain fatty acid (SCFA) mixtures that mimic the digestion products of wheat bran, oat bran, pectin, and cellulose (as control), which is largely unfermentable. RESULTS: Induction of the cdk inhibitors p21cip1 and p27kip1 by fiber-mimicking SCFA mixtures occurs much more rapidly and is many-fold greater than their induction by TGF-beta1. The SCFA mixtures most effective in causing growth inhibition and cdk inhibitor production mimicked those from wheat bran > oat bran > pectin. CONCLUSIONS: cdk inhibitor induction by SCFA mixtures is not mediated by TGF-beta1. The SCFA mixture mimicking digested wheat bran fiber was the most effective of all mixtures tested in inhibiting cell growth through induction of cdk inhibitors.

Cell Cycle↗

Polygalacturonase is the key component in enzymatic retting of flax.

Seven commercial enzyme mixtures were tested for their ability to perform retting of flax (i.e. to separate flax fibers by partly removal of middle lamella) and were assayed for hydrolysis of xylan, cellulose and four kinds of pectin. The only activity that showed correlation to the ability to perform retting was the degradation of low esterfied pectin. A purified Aspergillus niger polygalacturonase was also shown to be able to perform retting. From this data it is hypothesized that degradation of the smooth regions (i.e. non-methylated polygalacturonase) in the middle lamella pectin is the most important step in enzymatic retting.

Edetic Acid↗

Effects of polysaccharides from Silene vulgaris on phagocytes.

The effects of the polysaccharides isolated from the intact plant (pectic polysaccharides P1, P2 and P3) and from the callus (acidic arabinogalactan C1 and pectin C2) of Silene vulgaris on phagocytic activity were studied in relation to an uptaking capacity and a myeloperoxidase activity of the peripheral human neutrophils and monocytes and rat peritoneal macrophages in vitro. Both intact plant and callus polysaccharides were shown to increase uptaking capacity of peripheral phagocytes. The callus acidic arabinogalactan C1 was only found to stimulate lysosomal activity of the peripheral phagocytes. Some polysaccharides studied were established to effect on peritoneal resident macrophages. Pectins P1, P3 and C2 failed to enhance myeloperoxidase activity of the macrophages in calcium-free solution, whereas the effect of callus arabinogalactan C1 was established to be independent of extracellular calcium. Polysaccharides studied failed to influence neither complement receptor CR3- nor scavenger receptor SR-mediated adhesion of the macrophages. The data obtained demonstrate that the intact S. vulgaris and its callus may be used as sources of immunoactive polysaccharides and that pectins and weakly acidic arabinogalactan seemed to stimulate macrophages through different mechanisms. Complement receptor type 3 and scavenger receptor failed to mediate the cell activation induced by plant polysaccharides.

Adjuvants, Immunologic↗

Binding capacity of various fibre to pesticide residues under simulated gastrointestinal conditions.

The purpose of this study was to compare the effect of the nature and quantity of various dietary fibre (cellulose, hemicellulose, pectin, lignin) in diets on the binding capacity to pesticides azinphos-methyl (AZM), chlorpropham (CLP), chlorothalonil (CKL), permethrin (PER) as estimated by solubility under conditions of pH and temperature simulating those in the gastrointestinal tract (incubated at pH 2 for 30 min at 37 degrees C, then at pH 7 for 60 min). The ratios of fibre to pesticides were determined in omnivorous diets. In this model, the binding capacity of lignin was equal to hemicellulose for PER, AZM and CLP, but it was significantly higher for CKL. Hemicellulose bound more CKL, AZM and CLP than did cellulose. Although pectin appreciably decreased all pesticides, its effect was lower than other fibres with one exception--cellulose-CKL. In the presence of equal amounts of fibre, lignin exerted the most significant effect on pesticide solubility. Hemicellulose and cellulose bind to the same extent PER and AZM. The effect of pectin was significant only on CKL and AZM when compared to the control.

Adult↗

Progressive action potential duration shortening and the conversion from atrial flutter to atrial fibrillation in the isolated canine right atrium.

OBJECTIVES: We sought to evaluate the effects of progressive shortening of the action potential duration (APD) on atrial wave front stability. BACKGROUND: The mechanisms of conversion from atrial flutter to atrial fibrillation (AF) are unclear. METHODS: Isolated canine right atria were perfused with 1 to 5 micromol/l of acetylcholine (ACh). We mapped the endocardium by using 477 bipolar electrodes and simultaneously recorded transmembrane potentials from the epicardium. The APD(90) was measured during regular pacing (S(1)) with cycle lengths of 300 ms. Atrial arrhythmia was induced by a premature stimulus (S(2)). RESULTS: At baseline, only short runs of repetitive beats (<10 cycles) were induced. After shortening the APD(90) from 124 +/- 15 ms to 72 +/- 9 ms (p < 0.01) with 1 to 2.5 micromol/l of ACh, S(2) pacing induced single, stable and stationary re-entrant wave fronts (307 +/- 277 cycles). They either anchored to pectinate muscles (5 tissues) or used pectinate muscles as part of the re-entry (4 tissues). When ACh was raised to 2.5 to 5 micromol/l, the APD(90) was further shortened to 40 +/- 12 ms (p < 0.01); S(2) pacing induced in vitro AF by two different mechanisms. In most episodes (n = 13), AF was characterized by rapid, nonstationary re-entry and multiple wave breaks. In three episodes with APD(90) <30 ms, AF was characterized by rapid, multiple, asynchronous, but stationary wave fronts. CONCLUSIONS: Progressive APD shortening modulates atrial wave front stability and converts atrial flutter to AF by two mechanisms: 1) detachment of stationary re-entry from the pectinate muscle and the generation of multiple wave breaks; and 2) formation of multiple, isolated, stationary wave fronts with different activation cycle lengths.

Acetylcholine↗