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[The energetic utilization of sugar beets, dried chips, cane sugar and apple pectin by growing swine. 2. Nitrogen and energy metabolism].

The energetic utilization of rations with raw and steamed sugar beets, dried sugar beet pulp (16 measured values each), sucrose and apple pectin (8 measured values each) as supplement feed to a basic ration was measured at 4 growing pigs each in the live weight range of between 40 and 130 kg. The utilization of the energy in the rations amounted to 69.1 +/- 5.5, 68.3 +/- 7.1, 60.9 +/- 5.8, 70.5 +/- 4.1 and 69.9 +/- 6.9% (same sequence as above). The utilization values for the supplemented feedstuffs were 69.2 +/- 7.4, 68.0 +/- 10.0, 55.9 +/- 9.7, 72.2 +/- 7.6 and 71.7 +/- 17.3%. The average retention value of the digestible pectin was derived from the test results with dried sugar beet pulp and apple pectin as 5.8 kJ/g. An influence of the ontogenetic development on energy utilization cannot be derived from the comparison of the retention effect of digestible nutrients measured at growing pigs and that derived from adult pigs.

Animal Feed↗

In situ gelling pectin formulations for oral sustained delivery of paracetamol.

The purpose of this study was to evaluate the potential of a pectin formulation with in situ gelling properties for the oral sustained delivery of paracetamol (acetaminophen). The formulations consisted of dilute aqueous solutions (1% to 2% w/v) of low methoxy pectin containing calcium ions in complexed form, which on release in the acidic environment of the stomach caused gelation of the pectin. In vitro studies demonstrated diffusion-controlled release of paracetamol from the gels over a period of 6 h. A bioavailability of approximately 96% of that of a paracetamol solution could be achieved from gels containing an identical dose of drug formed in situ in the stomachs of rats, with appreciably lower peak plasma levels and a sustained release of drug over a period of at least 6 h.

Acetaminophen↗

Novel modulation of drug delivery using binary zinc-alginate-pectinate polyspheres for zero-order kinetics over several days: experimental design strategy to elucidate the crosslinking mechanism.

A Box-Behnken design was applied to mathematically establish whether different degrees of crosslinking were induced by Zn2+ and Ca2+ ions in polyspheres composed of alginate and/or pectin, and the model drug ibuprofen. Based on their different crystal structures and coordination numbers, a theoretical model was proposed demonstrating that Zn2+ ions preferentially crosslink alginate and pectin. In addition, the lower coordination number of Zn2+ (4-6) would significantly retard hydration of both polymers, as opposed to Ca2+ (7-9). The responses studied for 28 statistically derived polyspheres included drug encapsulation efficiency, physicomechanical behavior, and in vitro drug release potential. Single-tailed Student's t-tests on data generated for the encapsulation efficiencies, primary facture values, and rupture energies indicated that Zn2+ was statistically superior (p<0.05) in crosslinking alginate and pectin. Further textural analysis revealed a good correlation between the Brinell hardness number and fracture load, while an inverse relationship was found for matrix tensile strength. Viscosity studies demonstrated different in situ crosslinking thresholds for Zn2+. The Durbin-Watson statistic and correlation coefficient revealed that the quadratic regression function was highly accurate in predicting the responses. Using a generalized reduced gradient algorithm on dissolution values obtained after 2 hours (t2h) provided optimized solutions for achieving zero-order release extending from 2 hours to 7 days. Mathematical simulations projected drug release from 25 to 50 days.

Alginates↗

Multiunit controlled-release diclofenac sodium capsules using complex of chitosan with sodium alginate or pectin.

This study explored the application of chitosan-alginate (CA) and chitosan-pectin (CP) complex films as drug release regulator for the preparation of multiunit controlled-release diclofenac sodium capsules. Pellets containing drug and microcrystalline cellulose, in a ratio of 3:5, were prepared in a fluidized rotary granulator. The pellets were coated with CA, CP, sodium alginate, pectin, and chitosan solutions. The pellets, equivalent to 75 mg drug, were filled into capsules. After 2 h of dissolution test in acidic medium, the amount of the drug released from any preparation was negligible. The pellets were further subject to pH 6.8 phosphate buffer More than 80% drug release at 12 h was observed with the uncoated pellets and those coated with sodium alginate, pectin or chitosan. Both 1% CA and 3% CP coated pellets exhibited drug release profiles similar to that of Voltaren SR75. It was found that approximately 60% and 85% of the drug were released at 12 and 24 h, respectively. Both Differential thermal analysis (DTA) and Fourier transform infrared spectroscopy (FTIR) analyses revealed complex formation between chitosan and these anionic polymers. It could be concluded that CA and CP complex film could be easily applied to diclofenac sodium pellets to control the release of the drug.

Alginates↗

Sustained pectin ingestion: effect on gastric emptying and glucose tolerance in non-insulin-dependent diabetic patients.

The effects of sustained pectin ingestion on gastric emptying, glucose tolerance, and hormone responses were studied in 12 stable, non-insulin-dependent (type 2) diabetic patients. Patients were placed on a 2400 kcal, low-fiber (3 g) diet for 2 wk, followed by 4 wk of an isocaloric diet supplemented with 20 g apple pectin/d. Gastric-emptying half-time, plasma glucose, glucagon, and human pancreatic polypeptide levels were determined. Gastric-emptying half-time was prolonged 43% (p less than 0.025) by pectin supplementation and returned to normal 3 d after its discontinuation. Fiber supplementation decreased the incremental area under the glucose tolerance curve from 34.8 +/- 3.0 to 27.9 +/- 3.2 mmol/L (p less than 0.01) but did not affect hormonal responses to a meal. Sustained pectin ingestion slowed the gastric-emptying rate and improved glucose tolerance; however, a direct relationship could not be demonstrated between changes in gastric emptying and changes in the incremental area under the glucose curve (r = 0.22).

Adult↗

Evaluation of a pectin agar medium for isolation of Yersinia enterocolitica within 48 hours.

A modified pectin agar medium was evaluated for the rapid isolation and presumptive identification of Yersinia enterocolitica. Of 118 isolates of Enterobacteriaceae tested, only the 13Y. enterocolitica and the three Klebsiella oxytoca strains produced colonies that depressed and sank into the agar. Yersinia enterocolitica was also easily identified in mixed cultures, even from inocula containing three times as many other Enterobacteriaceae organisms as Y. enterocolitica. The recovery of Y. enterocolitica was evaluated on Mueller-Hinton, pectin, Hektoen enteric, xylose lysine desoxycholate, Salmonella-Shigella, and MacConkey agars. Compared with Mueller-Hinton agar, the pectin agar showed a 100% recovery of Y. enterocolitica, with all strains having depressed colonies, while the other media showed lesser recoveries of only 5 to 25%, with no other discriminating colonial characteristic.

Culture Media↗

Dietary supplementation with pectin and guar gum on 1,2-dimethylhydrazine-induced colon carcinogenesis in rats.

The effect of dietary supplementation with pectin and/or guar gum on 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis was studied using 120 male Sprague-Dawley rats. The rats were given a weekly injection of DMH for 8 weeks and were maintained on a basal fiber-free diet supplemented with 5% cellulose. The rats were then subdivided into four groups and kept on the basal fiber-free diet supplemented with either no fiber, 10% pectin, 10% guar gum or a combination of 5% pectin/5% guar gum for a period of 24 weeks. The 8 weeks of DMH administration were defined as the initiation stage of carcinogenesis and the next 24 weeks were defined as the promotional stage of carcinogenesis. Food and water were available ad libitum. The rats were killed 32 weeks after the start of the experiment and tumor incidence, location and frequency in the colon were determined. Other parameters measured were body weight and caloric intake. Dietary fiber supplementation with 10% pectin or with 10% guar gum but not with the combination of 5% pectin/5% guar gum (fed during the promotional stage of carcinogenesis), was found to suppress colon cancer incidence to a significant extent.

1,2-Dimethylhydrazine↗

Interaction of dietary protein differing in sulfur amino acid content and pectin on bile acid conjugation in immature and mature rats.

Rapidly growing immature (4-wk-old) and slowly growing mature (15-wk-old) rats were fed fiber-free or 10 g/100 g pectin diets containing various proteins differing in the sulfur amino acid content for 30-32 d. Soybean protein, casein, whole egg protein and egg albumen were used at the nitrogen level of 2.7 g/100 g diet. These experimental diets contained 0.354, 0.540, 0.945 and 1.22 g sulfur amino acids/100 g, respectively. In the rats fed fiber-free diets, a substantial quantity of glycine-conjugated bile acids was detected in the bile of immature rats fed soybean protein and casein (73 and 25% of total bile acids, respectively), but not in the other groups (less than 13%). Dietary pectin increased bile acid excretion both in immature (48-77%) and mature (34-114%) rats irrespective of the protein source, except in immature rats fed egg albumen and mature rats fed whole egg protein. Because a pectin-dependent increase in bile acid excretion was essentially attributed to the increase in glycine-conjugates, this dietary fiber significantly increased the ratio of glycine-conjugates to taurine-conjugates (2.4- to 6.5-fold). This increase was accompanied by a 40-50% decrease in the concentration of liver taurine, except in immature rats fed soybean protein and egg albumen. However, there was no consistent relationship between the extent of taurine conjugation and the activity of liver cysteine dioxygenase, one of the rate-limiting enzymes in taurine synthesis.

Aging↗

Prune fiber or pectin compared with cellulose lowers plasma and liver lipids in rats with diet-induced hyperlipidemia.

Previous research indicated that prunes in the diet of men with hypercholesterolemia lowered plasma and LDL cholesterol concentrations. To further study lipid metabolism in response to ingesting prunes, we conducted an animal study to test the hypotheses that fiber extracted from prunes, compared with purified cellulose, lowers plasma and liver cholesterol in rats with diet-induced hyperlipidemia and that the response is dose dependent. Rats were randomly assigned to one of five experimental diet groups. Four of the diets contained cholesterol and cholic acid to induce hyperlipidemia. The fiber source in the hyperlipidemic diets was 6% cellulose, 3% prune fiber, 6% prune fiber or 3% pectin. The fifth group, the nonhyperlipidemic control, was fed a diet containing 6% cellulose without cholesterol or cholic acid. Rats consumed one of the five diets ad libitum for 28 d, then were killed after 16 h without food. Plasma, LDL and liver cholesterol concentrations were higher in the hyperlipidemic control than the nonhyperlipidemic control and lower in the groups fed diets containing pectin or prune fiber than in the hyperlipidemic control group. No differences in plasma or liver cholesterol concentrations were detected between groups fed either dose level of prune fiber or between groups fed 6% prune fiber and pectin. These results indicate that fiber extracted from prunes lowers plasma and liver cholesterol in hyperlipidemic rats, although a dose-dependent response was not detected.

Analysis of Variance↗

Cholesterol 7 alpha-hydroxylase activity is increased by dietary modification with psyllium hydrocolloid, pectin, cholesterol and cholestyramine in rats.

Sources of dietary fiber known to alter cholesterol metabolism and/or bile acid pool size were fed to rats, and activity of the rate-limiting step in bile acid synthesis, cholesterol 7 alpha-hydroxylase, was measured. In the first experiment, semipurified diets containing 5% cellulose, psyllium hydrocolloid, pectin or oat bran as dietary fiber sources or 2% cholestyramine were fed to groups of 10 male Wistar rats for 4 wk. In the second experiment, groups of six rats were fed diets containing 5% cellulose, rice bran, oat bran or psyllium with and without 0.25% cholesterol. In the first experiment, the activity of cholesterol 7 alpha-hydroxylase (pmol.min-1.mg protein-1) was highest in the cholestyramine-treated group (95.6 +/- 3.6), followed by groups fed psyllium (35.5 +/- 3.5) or pectin (36.0 +/- 4.5), which exhibited more than twice the enzyme activity of groups fed cellulose (16.9 +/- 1.9) or oat bran (12.3 +/- 2.0). In the second experiment, feeding cholesterol resulted in significantly higher enzyme activity when cellulose (65%), oat bran (118%) and rice bran (60%) were fed, but no difference in activity was observed when cholesterol was added to the psyllium-containing diet. Higher activity of cholesterol 7 alpha-hydroxylase when pectin or psyllium rather than cellulose was fed may explain the almost twofold higher bile acid pool sizes previously reported in response to feeding either of these fibers. These data support the hypothesis that the hypocholesterolemic effect of soluble fibers is modulated through increased synthesis and therefore pool size of bile acids.

Animals↗

Bifidobacterium lactis Bb-12 and Lactobacillus salivarius UCC500 modify carboxylic acid formation in the hindgut of rats given pectin, inulin, and lactitol.

The effect of Bifidobacterium lactis (Bb-12) and Lactobacillus salivarius (UCC500) on the formation of carboxylic acids (CAs) was studied in the hindgut of rats fed pectin, inulin of low solubility, and lactitol. When the pectin diet was supplemented with Bb-12, the formation of CAs was larger throughout the colon of rats, due to increased formation of acetic acid (P < 0.01) and, in the distal part of the colon, also because of propionic and butyric acids (P < 0.01). In rats fed pectin and UCC500, there was a shift in the formation of CAs from the cecum to the distal colon. Thus, the cecal pool of CAs in the rats was lower (P < 0.05), whereas the concentration of CAs in the distal part of colon was larger (P < 0.01) than without this strain. Concerning the slowly fermentable inulin, there was a greater formation of CAs in the cecum (P < 0.05) of rats, especially propionic acid, and a lower formation in the distal part of the colon (P < 0.01) when the diets were supplemented with Bb-12, whereas UCC500 had no effect except for a lower proportion of acetic acid in the distal part of the colon (P < 0.001). In rats fed lactitol and Bb-12, the concentration of CAs was lower in the distal part of colon (P < 0.001) than without this strain, whereas the cecal pool of CAs was greater in rats supplemented with UCC500 (P < 0.001). We conclude that the probiotic bacteria affect the amount, the pattern, and the site of release of CAs in the hindgut of rats, but the combination of pre- and probiotics is of great importance for the outcome.

Animals↗

Cloning of a Cicer arietinum beta-galactosidase with pectin-degrading function.

The cDNA clone (CanBGal-3) encoding a cell wall pectin-degrading beta-galactosidase (beta III-Gal) from Cicer arietinum L. cv. Castellana has been identified. The identification was carried out by comparing the deduced amino acid sequences of several isolated chickpea beta-galactosidase clones with the purified beta III-Gal protein sequence. The expression pattern of the gene corresponding to CanBGal-3 was in concordance with the fluctuations of the enzyme beta III-Gal in different seedling organs, being specific to elongating organs such as epicotyls and roots. Transformation of Solanum tuberosum plants with the chickpea CanBGal-3 clone indicated that the beta-galactosidase encoded by this clone is a pectin-degrading enzyme. The authors propose an important role for chickpea beta III-Gal in pectin degradation in cell walls of vegetative organs such as epicotyls and roots. The degradation of galactan carried out by this enzyme may determine structural changes and affect cell wall porosity. It is suggested that the increase in the size of cell wall pores could permit access of other cell wall-modifying enzymes to their substrate.

Amino Acid Sequence↗

In vivo expression of a Cicer arietinum beta-galactosidase in potato tubers leads to a reduction of the galactan side-chains in cell wall pectin.

We report the generation of Solanum tuberosum transformants expressing Cicer arietinum betaIII-Gal. betaIII-Gal is a beta-galactosidase able to degrade cell wall pectins during cell wall loosening that occurs prior to cell elongation. cDNA corresponding to the gene encoding this protein was identified among several chickpea beta-galactosidase cDNAs, and named CanBGal-3. CanBGal-3 cDNA was expressed in potato under the control of the granule-bound starch synthase promoter. Three betaIII-Gal transformants with varying levels of expression were chosen for further analysis. The transgenic plants displayed no significant altered phenotype compared to the wild type. However, beta-galactanase and beta-galactosidase activities were increased in the transgenic tuber cell walls and this affected the potato tuber pectins. A reduction in the galactosyl content of up to 50% compared to the wild type was observed in the most extreme transformant, indicating a reduction of 1,4-beta-galactan side-chains, as revealed by analysis with LM5 specific antibodies. Our results confirm the notion that the pectin-degrading activity of chickpea betaIII-Gal reported in vitro also occurs in vivo and in other plants, and confirm the involvement of betaIII-Gal in the cell wall autolysis process. An increase in the homogalacturonan content of transgenic tuber cell walls was also observed by Fourier transform infrared spectroscopy (FTIR) analysis.

Blotting, Northern↗

Lack of kinetic interaction between valproic acid and citrus pectin.

To determine the influence of ingestion of dietary fiber (citrus pectin) in the kinetic disposition of valproic acid, two studies were conducted in adult volunteers. Twelve healthy subjects took single oral doses of 500 mg valproic acid in a form of sodium valproate as coated tablets, on two occasions: under fasting conditions (treatment A) or immediately after ingestion of 14 g citrus pectin (treatment B). In each study, a randomized, single-dose, two-way crossover design was used, and 8 days elapsed between the two trials. Assays were carried out for valproic acid in plasma samples, which were collected from zero to 48 hours after drug administration. The peak plasma concentration (60.52 vs. 55.74 micrograms x ml-1), the time to peak concentration (3.47 vs. 3.91 h), the absorption rate constant (0.12 vs. 0.12 h-1), and the area under the plasma concentration-time curve (1324.0 vs. 1268.5 micrograms x h x ml-1) did not differ significantly between treatments A and B (median, Wilcoxon test). These results suggest that the administration of citrus pectin does not alter the rate and extent of absorption of valproic acid administered as a single dose to healthy volunteers.

Adult↗

Acidic oligosaccharides from pectin hydrolysate as new component for infant formulae: effect on intestinal flora, stool characteristics, and pH.

OBJECTIVES: To come even closer to the functional composition of human milk, acidic oligosaccharides (AOS) from pectin were added to well known neutral prebiotics (galacto-oligosaccharides (GOS) and long-chain fructo-oligosaccharides (FOS)). The effect of AOS and GOS/FOS/AOS on intestinal flora, stool characteristics as well as acceptance and tolerance was investigated. METHODS: Human milk contains 75% to 85% neutral and 15% to 25% acidic oligosaccharides. In this prospective, randomized, double blind study, a mixture of 80% neutral oligosaccharides (from long-chain galacto- and long-chain fructo-oligosaccharides) with 20% acidic oligosaccharides derived from pectin hydrolysis was investigated. Forty-six term infants were fed a standard formula supplemented with either maltodextrin as control (n=15), or with 0.2 g acidic oligosaccharides (n=16), or with the latter plus 0.6 g neutral oligosaccharides (mixture of galacto- and fructo-oligosaccharides; n=15). Fecal flora using plating technique and pH were measured. Stool characteristics and possible side effects (crying, vomiting, and regurgitation) were recorded. RESULTS: There was no difference in the bifidobacteria counts between the control and the group supplemented with acidic oligosaccharides alone (8.75+/-0.50 vs. 8.58+/-0.94 log colony forming units [CFU]/g stool). In infants fed the combination of acidic and neutral oligosaccharides, bifidobacteria were increased (9.61+/-0.70 log CFU/g stool; P<0.01). The same pattern was observed with lactobacilli. Stool consistency was softest in infants fed the complete oligosaccharide mixture, but also in those fed formula supplemented with acidic oligosaccharides alone, the stool consistency was significantly softer compared with the control group. Fecal pH increased in the controls, remained constant in acidic oligosaccharides alone, and decreased in the complete mixture of oligosaccharides group. CONCLUSION: There was no difference in growth, crying, vomiting, and regurgitation patterns between the groups. In summary, acidic oligosaccharides from pectin hydrolysate are well tolerated as ingredient in infant formulae but do not affect intestinal microecology.

Bifidobacterium↗

Synergism between Erwinia pectate lyase isoenzymes that depolymerize both pectate and pectin.

Phytopathogenic Erwinia bacteria cause tissue maceration by secretion of pectinolytic enzymes such as pectate lyase (PL). Sequencing of overlapping genomic fragments from Erwinia carotovora subsp. atroseptica established the organization of a 7.5 kbp region encoding PL isoenzymes. Two intergenic regions of 656 and 645 bp separate three enzyme coding regions of 1125 bp exhibiting approximately 80% positional identity. The promoters of each of the three genes contain a segment with high homology to the binding sequence of the E. chrysanthemi KdgR transcription repressor, implying similar mechanisms of gene regulation in the two bacterial species. Separate expression of the pel genes in the Escherichia coli-pT7-7 system and purification of their products yielded PLs at 7-33 mg (I culture)-1 with greater than 95% purity. Availability of the recombinant enzymes allowed determination of the kinetic differences amongst the PL isoforms, PL1, PL2 and PL3. The results show that PL is not strictly confined to depolymerization of pectate since each isoenzyme more readily degrades 31% esterified pectin. Addition of isoenzyme combinations revealed no synergism with respect to degradation of pectate or 31% esterified pectin. However, addition of enzyme combinations containing PL3 enhanced the activity towards 68% esterified pectin, against which individual PL activities were low, by up to 64%. These data suggest that the combination of PL isoenzymes extends the range of pectic substrates which the bacterium can degrade.

Amino Acid Sequence↗

Pectate lyase C from Bacillus subtilis: a novel endo-cleaving enzyme with activity on highly methylated pectin.

The gene yvpA from Bacillus subtilis was cloned and expressed in Escherichia coli. It encoded a pectate lyase of 221 amino acids that was denominated PelC. The heterologously expressed enzyme was purified by His-tag affinity chromatography and characterized. PelC depolymerized polygalacturonate and pectins of methyl esterification degree from 22 % to 89 %, exhibiting maximum activity on 22 % esterified citrus pectin. It showed an absolute Ca2+ requirement and the optimum temperature and pH were 65 degrees C and pH 10, respectively. The deduced amino acid sequence of PelC showed 53 % identity to pectate lyase PelA from Paenibacillus barcinonensis, which was also characterized. Similarly to PelC, purified PelA showed activity on polygalacturonate and pectins with a high degree of methyl esterification. The two enzymes cleaved pectic polymers to a mixture of oligogalacturonates, indicating an endo mode of action. Analysis of activity on trigalacturonate showed that PelC cleaved it to galacturonic acid and unsaturated digalacturonate, whereas PelA did not show activity on this substrate. PelC and PelA showed high homology to a few recently identified pectate lyases of family 3 and form with them a cluster of small-sized pectate lyases from non-pathogenic micro-organisms.

Amino Acid Sequence↗

A novel small heat shock protein gene, vis1, contributes to pectin depolymerization and juice viscosity in tomato fruit.

We have characterized a novel small heat shock protein gene, viscosity 1 (vis1) from tomato (Lycopersicon esculentum) and provide evidence that it plays a role in pectin depolymerization and juice viscosity in ripening fruits. Expression of vis1 is negatively associated with juice viscosity in diverse tomato genotypes. vis1 exhibits DNA polymorphism among tomato genotypes, and the alleles vis1-hta (high-transcript accumulator; accession no. AY128101) and vis1-lta (low transcript accumulator; accession no. AY128102) are associated with thinner and thicker juice, respectively. Segregation of tomato lines heterogeneous for vis1 alleles indicates that vis1 influences pectin depolymerization and juice viscosity in ripening fruits. vis1 is regulated by fruit ripening and high temperature and exhibits a typical heat shock protein chaperone function when expressed in bacterial cells. We propose that VIS1 contributes to physiochemical properties of juice, including pectin depolymerization, by reducing thermal denaturation of depolymerizing enzymes during daytime elevated temperatures.

Acclimatization↗