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Bacteroides ruminicola, pure or combined with Selenomonas ruminantium, and Lachnospira multiparus, pure or combined with Succinivibrio dextrinosolvens, were grown on a medium with pectin as energy source. There was a difference in fermentation products between the pure and combined cultures and efficiency of substrate utilization was better with the combined cultures.
In rats fed a semisynthetic diet, streptozotocin-induced diabetes [45 mg/kg, 17 days] led to hypertriglyceridaemia [6.4 mmol/l], to a marked increase in the proportion of plasma cholesterol present in the very low density lipoproteins [VLDL] [to 40 %] and to a decrease in the amount present in the high density lipoproteins [HDL] [to 34 %]. The addition of 0.25 % cholesterol to the above diet led in healthy rats to hypercholesterolaemia [4.3 mmol/l] and to similar changes in the distribution of cholesterol in the lipoproteins. In diabetic rats, the same diet led to pronounced hypertriglyceridaemia [13.8 mmol/l] and hypercholesterolaemia [18.9 mmol/l], while the proportion of HDL-borne plasma cholesterol fell still further to 6 % and rose in the VLDL to 70 %. The addition of pectin to the diet in 6 % concentration markedly inhibited triglyceridaemia [3.3 mmol/l] and cholesterolaemia [4.4 mmol/l] and raised the proportion of HDL plasma cholesterol to 47 %.
The effect of four kinds dietetic fruit products, sweetened by Sorbitol (pectin nectar, apricot nectar, apple and strawberry jam) on blood sugar and insulin needs of the patients was studied in 40 patients with insulin-dependent diabetes. The results were compared with those, obtained with the consumption of similar fruit products, sweetened by industrial sugar. The study was performed by an apparatus for automatic regulation of blood sugar (artificial endocrine pancreas--Biostator), which supplies the organism of the patients, overloaded with the respective product, with insulin, depending on the degree of glycemia and guarantees "normal" utilization of the product. The results obtained reveal that the blood sugar is slightly increased with the dietetic products, the peak of the blood sugar curve is lower and less insulin is supplied automatically by the biostator during the loading with the product. All values are lower, with a statistical significance, for the dietetic products as compared with the non-dietetic ones (p less than 0.05 to 0.001). Those data provide grounds to recommend the products studied as proper to be included in the diet of the diabetic patients.
1. In the course of 34 days the detectable methanol concentration in freshly squeezed grapefruit and organ juices as well as in grapefruit and orange juice mixes increased to a maximum level of approximately 600 mg/kg. This increase could already be detected at the beginning of the examination i.e. as early as 1 or 2 days after squeezing the fruit juices. At the beginning of the experiments the methanol concentrations were between 10 and 270 mg/kg. 2. After the consumption of pectin (40 g on one day and 2 x 40 g on two days) blood or serum-methanol values respectively were observed which were clearly above the level of chronic alcohol abuse (10 mg/kg). After the ingestion of ethanol (0.5 g/kg bodyweight) the serum-methanol concentration increased even further.
Effect of natural and enriched with pectin tanned fruits on radiation-induced metabolic disorders was studied in persons subjected to radiation due to Chernobyl accident. It was shown that products in question beneficially influenced blood antioxidant system as well as brought to the norm contents of triglicerides and albumins in patients with IIa and IV types of hyperlipoproteinaemia.
Five isozymes (four acidic and one basic) of polygalacturonase were separated by chromatofocusing from the culture filtrate of Botrytis cinerea grown on apple pectin. The isozymes, designated as Polygalacturonase I to V, have isoelectric points of 9.7, 4.9, 4.6, 3.7, and 2.7, respectively, with Polygalacturonase III exhibiting the highest specific activity. Polygalacturonase I appeared to function as an endo-polygalacturonase while the other four isozymes act as exo-polygalacturonases. The pH optima of the isozymes range from pH 4.5 to 5.5 with Polygalacturonase V being less sensitive to higher pH compared with the rest of the isozymes.
The lipid-lowering effect of a new mixture of fiber consisting in guar and apple-pectin in combination with apple-pomaces has been estimated in 15 female type 2-diabetics [age mean = 62 (52-70) yr] with hypercholesterolemia (total-chol > 240 mg/dl and LDL-chol > 130 mg/dl). After a dietetic run-in-phase of 3 weeks the patients received the fiber mixture (1 package of 17 g with about 5.9 g water-soluble fiber) dissolved in 250 ml water for the next 9 weeks: during the first 3 weeks 2 portions per day, the next 3 weeks twice 1/2 portion and the last 3 weeks one 1/2 portion daily. The fiber mixture had to been consumed 30 minutes before taking a main meal. Regular intake of the fiber product led to a significant decrement in blood lipids, and this improvement under 2 portions daily has been maintained for the most part with the reduced dosage. The total-chol levels decreased by 11.3% during the first 3 weeks of fiber intake (p < 0.05) and by -12.6% during the next 3 weeks (p < 0.05), the decrease during the last 3 weeks under the reduced fiber intake was -9.6% (ns). The HDL-chol levels remained approximately the same during the whole period of observation. The triglyceride concentrations in serum could be lowered by -15.5% during the first 3 weeks of fiber intake, and by -19.2% during the next 3 weeks (p < 0.05), finally the decrease was -12.3% (ns). The atherogenic index (Chol-/HDL-chol) could be reduced on an average of 14.3% in relation to the initial value.(ABSTRACT TRUNCATED AT 250 WORDS)
Dietary soluble fiber significantly lowers plasma low density lipoprotein (LDL) cholesterol concentrations in humans and animals. In male guinea pigs, alterations in hepatic cholesterol homeostasis induced by dietary fiber in part account for the decrease in plasma LDL levels (Fernandez et al. 1994. Am. J. Clin. Nutr. 59: 869-878; 1995. 61: 127-134, and J. Lipid Res. 1995. 36: 1128-1138). To test whether dietary fiber elicited similar hypocholesterolemic responses in both genders, female guinea pigs were fed diets containing 12.5% pectin (PE), 12.5% guar gum (GG), 7.5% psyllium (PSY), or 12.5% cellulose (control diet). In addition, physiological (0.04%) (LC) or pharmacological (0.25%) (HC) amounts of cholesterol were tested with the fibers to determine whether dietary cholesterol altered the plasma cholesterol response. Significant reductions in plasma cholesterol were observed in females fed LC diets with PE, GG, or PSY (P < 0.01) while the responses to fiber with high cholesterol intake were more moderate. Hepatic cholesterol concentrations were reduced in the LC group (P < 0.001) with increased HMG-CoA reductase and cholesterol 7 alpha-hydroxylase and decreased acyl CoA:cholesterol acyltransferase (ACAT) activities accompanied by a reduction in hepatic cholesterol pools induced by fiber intake. In addition, plasma LDL lowering in animals fed the LC diets was associated with increases in hepatic LDL receptor Bmax values. Effects of fiber on hepatic cholesterol in animals fed HC diets were moderate and hepatic enzymes were not altered to the same extent as in the LC groups. For the LC groups there was no gender effect on the magnitude of plasma LDL lowering, depletion of hepatic cholesterol, or alterations in hepatic cholesterol metabolism, although hepatic HMG-CoA reductase and ACAT activities were lower in females compared to males (P < 0.01). In contrast, females fed the control HC diet had higher plasma LDL levels than males and dietary fiber did not reduce hepatic cholesterol concentrations nor alter hepatic enzyme activities as effectively as in males. These studies demonstrate that female, compared to male, guinea pigs are more responsive to a dietary cholesterol challenge and, that with this pharmacological perturbation, fiber effects are moderate compared to males. In contrast, with low cholesterol intakes, the cholesterol lowering effects of fiber are similar in both genders.
Modified citrus pectin (MCP) added to the media of cultured androgen-independent human prostatic JCA-1 cells reduced cell growth and correspondingly [3H]thymidine incorporation into DNA, which was correlated with the down-regulation of cyclin B and p34cdc2 MCP also induced distinct increases in specific endogenous phosphoproteins, including a cAMP-stimulated 52,000 (52-kDa) protein. Since metastatis has been inversely correlated with nm23 gene expression in some cancer cells and was reportedly inhibited by MCP in a rat prostate model, we investigated steady state level changes in the nm23 protein in JCA-1 cells and found it to be unexpectedly suppressed as a result of exposure to MCP.
Pectin (PE), guar gum (GG), and psyllium (PSY) lower plasma low density lipoprotein (LDL) cholesterol concentrations in guinea pigs with different orders of magnitude by inducing defined alterations in hepatic cholesterol homeostasis (Fernandez et al. 1994. Am. J. Clin. Nutr. 59: 869-879; 61: 127-134 and 1995. J. Lipid Res. 36: 1128-1138). To further explore specific mechanisms responsible for the differences in plasma and hepatic cholesterol lowering, the effects of these fibers were evaluated on cholesterol absorption, hepatic cholesterol 7 alpha-hydroxylase activity, the rate-limiting enzyme of bile acid synthesis, and in vivo LDL transport to target specific primary and secondary mechanisms accounting for the observed responses. Fibers were fed with physiological (0.04%), low cholesterol (LC), or pharmacological high cholesterol (HC) (0.25%) levels to assess whether cholesterol intake influences plasma LDL lowering mechanisms. Intake of PE, GG, or PSY with LC or HC diets lowered plasma and hepatic cholesterol concentrations (P < 0.001). PE and PSY up-regulated 7 alpha-hydroxylase activity 3-fold with LC and PE by 5-fold with HC diets. In contrast, GG intake had no effect on 7 alpha-hydroxylase activity. Cholesterol absorption was reduced 30% by PE intake while no differences were found between control and PSY groups. GG reduced cholesterol absorption only with HC diets. Intake of PE, GG, or PSY with HC diets resulted in faster plasma LDL fractional catabolic rates (FCR) (P < 0.01) with no effect on LDL apoB flux rates (FR) or pool size, suggesting that fiber reduced LDL cholesterol concentration without decreasing the number of LDL particles. In addition to reducing LDL apoB FR, PE and PSY increased LDL FCR with HC diets while GG effects were limited to lowering LDL apoB FR. These results indicate that the distinctive reductions in hepatic cholesterol induced by PE, GG, and PSY associated with plasma cholesterol lowering result from different mechanisms specific to each fiber and that the levels of dietary cholesterol contribute to the different metabolic responses.
In a prospective, double-blind, randomised, multicentre, parallel group study, children (6 months to 5.5 years of age) with acute, non-complicated diarrhea received either a preparation containing apple pectin and chamomile extract (Diarrhoesan, n = 39) or placebo (n = 40) in addition to the usual rehydration and realimentation diet. At the end of three days of treatment, the diarrhea had ended significantly (p < 0.05) more frequently in the pectin/chamomile (33/39) than in the placebo group (23/40). Pectin/chamomile reduced the duration of diarrhea significantly (p < 0.05) by at least 5.2 h. The parents documented the well-being in a diary twice daily; in contrast to placebo, a trend of continuous improvement was observed in the pectin/chamomile group. The parents expressed their contentment more frequently (82%) with pectin/chamomile than with placebo (60%, not significant). There were no further differences between the treatment groups.
The production of polygalacturonases from Fusarium oxysporum f. sp. radicis lycopersici (FORL) occurred sequentially, and endo- and exo-polygalacturonases (PG) predominated at different times during growth. At first FORL produced a PG which did not release monomers from pectins and was unable to hydrolyse dimers and trimers. Later a complex was produced in which exo-PG seemed to be the predominant activity; the monomer was released rapidly and this activity was able to degrade dimers and trimers.
Experiments conducted with albino rats have evidenced that under the effect of an atherogenic ration the level of lipids and cholesterol in the blood plasma and hepatic tissue and of sillac acids in the blood plasma increased. In the aortic intima the content of acid mucopolysaccharides was rising, this being attended by a concurrent swelling of the main interstitial substance and of the collagen fibers in the subendothelial layer. An addition of apple pectin or of cellulose to the atherogenic ration deferred the development of the mentioned changes.
Pectin methylesterase (PME) was extracted from bananas (cv Cavendish) and purified by affinity chromatography on a CNBr-Sepharose-PME inhibitor (PMEI) column. A single protein and PME activity peak was obtained. For banana PME, a biochemical characterization in terms of molar mass (MM), pI, and kinetic parameters was performed. In a second step, the thermal and high-pressure stability of the enzyme was studied. Isothermal inactivation of purified banana PME could be described by a first-order kinetic model in a temperature range of 65 degrees to 72.5 degrees C, whereas its isobaric-isothermal inactivation followed a fractional-conversion model. Banana PME was found to be more thermally stable compared with PMEs extracted from orange, tomato, and apple.
Determination of the composition of the oral microflora has traditionally been based on cultivation. Treponemes are prevalent in many oral infections but, unfortunately, are not regularly cultured. In this study a new method was established for routine isolation of oral treponemes from clinical samples. Bacterial samples from 47 periodontal pockets and 4 endodontic infections were incubated anaerobically under nitrogen atmosphere at 37 degrees C in U-tubes containing pectin medium. In the U-tube a 'bacterial sample side' and a 'sterile medium side' were established on separate sides of a membrane filter and an agar plug. Using this method we were able to isolate viable treponemes from all bacterial samples. This was in contrast to previously established methods such as the agar dilution technique, the technique involving the membrane filter placed on the surface of solid agar media and the well in agar plate technique. We believe that the 'U-tube method' is a valuable supplement to previously described techniques in routine isolation of treponemes from clinical samples.
The effects of extended heat stress on polygalacturonase (PG; EC 3.2.1.15) and pectin methylesterase (PME; EC 3.1.1.11) gene expression at mRNA, protein and activity levels in ripening tomato fruits were investigated. Steady state levels of PG mRNA declined at temperatures of 27 degrees C and above, and a marked reduction in PG protein and activity was observed at temperatures of 32 degrees C and above. Exogenous ethylene treatment did not reverse heat stress-induced inhibition of PG gene expression. Transfer of heat-stressed fruits to 20 degrees C partly restored PG mRNA accumulation, but the rate of PG mRNA accumulation declined exponentially with duration of heat stress. Heat stress-induced inhibition of PME mRNA accumulation was recoverable even after 14 days of heat stress. In fruits held at 34 degrees C, both PG and PME protein and activity continued to accumulate for about 4 days, but thereafter PG protein and activity declined while little change was observed in PME protein and activity. In spite of increases in mRNA levels of both PG and PME during the recovery of heat-stressed fruit at 20 degrees C, levels of PG protein and activity declined in fruits heat-stressed for four or more days while PME protein and activity levels remained unchanged. Collectively, these data suggest that PG gene expression is being gradually and irreversibly shut off during heat stress, while PME gene expression is much less sensitive to heat stress.
Two pectin esterase cDNA clones representing different isozymes with ca. 95% homology were isolated from an early ripening tomato fruit cDNA library. Both clones were longer than previously published sequences, and the encoded proteins possessed extended (229-233 amino acid) putative N-terminal extensions. In addition, the mRNA species corresponding to the two clones showed differential levels of expression in fruit.
Differential screening of a Brassica napus genomic library led to the isolation of the clone named Bp 19 containing a gene which is highly expressed during microspore development. The accumulation of Bp19 mRNA starts in uninucleate microspores, increases during development reaching a peak in the late stages but declines considerably in mature pollen. The nucleotide sequence of the entire coding region and of extended portions of the 5' and 3' flanking regions was determined. Several homologous cDNA clones were also isolated and sequenced. The Bp 19 gene contains a single intron of 137 bp and gives origin to a mRNA of ca. 1.9 kb which codes for a polypeptide of 584 amino acids. Bp 19 protein has an estimated molecular weight of 63 kilodaltons and has a highly hydrophobic amino terminal region which shows features of a signal peptide. The carboxy half of the Bp 19 protein, starting at amino acid 269, has striking sequence similarity to the pectin esterases of tomato and of the plant pathogen Erwinia chrysanthemi. Four short domains are extremely well conserved in all the three proteins and therefore could represent catalytic sites responsible for enzyme activity. Comparison of the 5' flanking region of the Bp 19 gene with the sequence of other pollen-specific promoters revealed the presence of several conserved regions. These short promoter sequences could correspond to regulatory elements responsible for pollen-specific gene expression.