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Biomedical subjects

A Pusztai

Publications and source records attributed to A Pusztai.

At least 55 records · Page 3Linked to original sources
PubMed1997

Red kidney bean lectin is a potent cholecystokinin releasing stimulus in the rat inducing pancreatic growth.

BACKGROUND: Lectins are proteins capable of specific binding to carbohydrates without altering their covalent structure. As an essential part of plants they are ingested in our daily diet. By binding to glycosyl side chains of receptors lectins can mimic or inhibit the action of the ligand. Oral administration of phytohaemagglutinin (PHA) in rats dose dependently induces growth of the small intestine and the pancreas by an unknown mechanism. AIMS: To investigate the mechanism of PHA induced intestinal and pancreatic growth. METHODS: Thirty day old male rats were pairfed for 10 days with lactalbumin as a control diet or lactalbumin plus PHA or purified soybean trypsin inhibitor (STI) as a positive control (42 mg/rat/day) with or without 20 micrograms of the cholecystokinin A (CCK-A) antagonist MK 329. To investigate further the effect of PHA on CCK release intestinal mucosal cells were isolated from rats which were continuously perfused in a perfusion apparatus. CCK release into the medium was assayed. RESULTS: PHA and STI significantly stimulated growth of the pancreas and the small intestine. MK 329 blocked this growth effect in the pancreas but not in the small intestine. In vivo, PHA significantly increased CCK plasma levels from 0.75 to 6.67 (SEM 2.23) compared with 2.3 (0.35) pM in the control group. In addition, in vitro PHA dose dependently stimulated CCK release with a maximal effect at 100 ng/ml. CONCLUSION: In vivo and in vitro PHA is a potent stimulus for CCK release in the rat, thereby inducing pancreatic growth, whereas intestinal growth is stimulated by a CCK independent mechanism.

Animals↗
PubMed1997

Both free and complexed trypsin inhibitors stimulate pancreatic secretion and change duodenal enzyme levels.

Secretion of pancreatic digestive enzymes was measured in pancreatic cannulated rats after duodenal stimulation with Kunitz or Bowman-Birk protease inhibitors or their complexes with trypsin and/or chymotrypsin. Free and complexed inhibitors were bound by the duodenal epithelium, stimulated the discharge of cholecystokinin, and significantly increased secretion rates of alpha-amylase, trypsinogen, and chymotrypsinogen. Inasmuch as secretion rates returned to basal levels with cholecystokinin-A receptor antagonists, the stimulation was likely to be mediated by cholecystokinin. Soya factors also influenced the duodenal concentration of pancreatic enzymes under simulated feeding conditions. Thus the level of alpha-amylase increased while the trypsin concentration decreased in rats gavaged with free or complexed inhibitors. The same was true for chymotrypsin when the Bowman-Birk inhibitor was used, but the Kunitz inhibitor and its trypsin complex actually raised the luminal concentration of chymotrypsin. Accordingly, because soya inhibitors remained effective in stimulating pancreatic secretion after elimination of their inhibitory activity by complex formation, it is questionable whether the signal for cholecystokinin secretion was solely due to lowering of duodenal protease levels.

Animals↗
PubMed1996

Response of intestinal transglutaminase activity to dietary phytohaemagglutinin.

The behaviour of the activity of tissue transglutaminase, a calcium-dependent enzyme, and the levels of polyamines which are physiological substrates for the enzyme, were studied in rat small intestine induced to grow by lectin phytohaemagglutinin. Transglutaminase activity greatly increased in the homogenates and the cytosolic fractions of the intestinal mucosa of lectin-treated rats compared to that of untreated animals. The measurement of enzyme activity in the presence of monodansylcadaverine, a competitive inhibitor of transglutaminase, testified that the assayed enzyme activity was authentic transglutaminase. As regards polyamines, the level of spermine did not change, whereas putrescine and spermidine contents were enhanced. The activation of transglutaminase, which was probably due to Ca2+ accumulation in enterocytes, could have a role in maintaining enterocyte adhesion and intestinal cell homeostasis, and/or repairing lectin-induced damages of microvilli of the gut epithelium.

Animals↗
PubMed1996

Effect of three novel polyamine oxa-analogues (MTR-OSPD, DIP-SPN and APPO-TFA) on the growth and proliferation of Swiss 3T3 cells.

In order to investigate their biological function on cellular polyamine content, cell growth and proliferation, three novel polyamine oxa-analogues, 5-(4-methoxy-2,3,6-trimethylbenzenesulfonyl)-6-oxa-spermidine (MTR-OSPD); 6,9-dioxa-5,10-di-(2,2,5,7,8-pentamethylchroman-6-sulfonyl) spermine (DIP-SPN) and 3-aminopropyl N-(3-phthalimidopropyloxy) trifluoroacetimidate (APPO-TFA) were tested for their ability to stop or slow down the growth of Swiss 3T3 cells. Cells at 50-60% confluency were grown for 24 or 48 hr in the presence of a wide range of polyamine oxa-analogue concentrations and the number of cells counted. To determine whether the drugs were cytotoxic or cytostatic, the analogue-containing medium in some vials was replaced with fresh culture medium after 48 hr and the cells incubated for a further 24 hr. Cellular protein, RNA, DNA, polyamine contents and the activities of ornithine decarboxylase, S-adenosyl-L-methionine decarboxylase and spermidine/spermine N1-acetyltransferase were also determined at the lowest effective analogue concentration. All three inhibitors stopped cell proliferation at concentrations over 100 microM. Both MTR-OSPD and DIP-SPN were cytotoxic, since the cells could not be revived by removing the inhibitor from the medium, whereas APPO-TFA was only cytostatic. At the lowest effective concentration the analogues had little effect on protein, RNA and DNA content of the cells, but had varying effects on polyamine metabolism. The most interesting analogue was APPO-TFA. This drug showed concentration-dependent growth inhibition between concentrations of 5 nM and 5 microM. These novel analogues may be of value in elucidating the precise functions of polyamines in cellular metabolism. Their exact mode of action is now under investigation.

3T3 Cells↗
PubMed1996

The effect of phytohaemagglutinin at different dietary concentrations on the growth, body composition and plasma insulin of the rat.

Young growing rats weighing approximately 83 g were fed on diets containing kidney bean (Phaseolus vulgaris) lectin (phytohaemagglutinin, PHA) in the range of 0-0.45 g/kg body weight for 10 d to ascertain whether there was a minimum dose below which the lectin had no significant effect on body and skeletal muscle weights in comparison with pair-fed lectin-free controls. Averaged over all experiments, PHA doses of less than 10 mg/d (0.12 g/kg body weight) reduced body dry by 1.14 (SE 0.25) g when compared with controls. Between 10 and 27 mg/d (0.12-0.32 g/kg body weight) a further reduction of 0.64 (SE 0.21) g occurred, suggesting a slight but steady decline of body dry weight with increasing dose. However, above 27 mg/d the depression of growth and changes in body composition accelerated. The difference between the proportional losses of skeletal muscle and body weight was not significant at doses of PHA below 10 mg/d (0.12 g/kg body weight) but the ratio of these losses rose to 1.5-2.0 at doses above this. The proportional decrease in lipid weight exceeded that of both body and skeletal muscle weights at all lectin doses, suggesting that lipid catabolism was the first target of the PHA effect. Plasma insulin level was depressed at the PHA dose of 0.02 g/kg body weight at which growth depression and muscle atrophy were minimal but, contrary to expectations, plasma glucose levels remained stable over the whole PHA dose range. It appears that despite a PHA-induced lowering of blood insulin, glucose catabolism is elevated by an unknown, possibly hormonal, compensatory mechanism. Thus, because low insulin levels facilitate the mobilization and catabolism of lipids, it may be possible to use low doses of PHA to reduce hyperglycaemia and body fat.

Animals↗
PubMed1996

A rat model of infection by Salmonella typhimurium or Salm. enteritidis.

Salmonellosis in the rat has many similarities with the disease in humans, with the ileum thought to be the main site of colonization/invasion in both species. Thus, the rat may be a useful way to study the mechanism of infection by these pathogenic bacteria. A series of infection trials carried out with Hooded Lister rats showed that a salmonella infection persisted for an extended period of time and that salmonellae bind to the small intestinal epithelium as early as 4 h after intragastric intubation. Reinfection from the large intestine may not therefore initially play a significant role in the salmonella infection process. The rat model may therefore provide a means to test in vivo interventionist strategies, designed to block binding of the pathogens in the gastrointestinal tract.

Animals↗
PubMed1996

Polyamine metabolism of enterocyte-like Caco-2 cells after exposure to Phaseolus vulgaris lectin.

The effect of Phaseolus vulgaris isolectin E4 on polyamine concentrations and ornithine decarboxylase activity of proliferating and differentiating Caco-2 cells was investigated. Values of putrescine, spermidine, and spermine in control cells were highest during the early phase of proliferative cell growth and lowest in the stationary phase. Phytohaemagglutinin E4 significantly increased cellular polyamine values during the late proliferative phase of cell growth. Ornithine decarboxylase activity was high during intensive proliferation and growth, but was lower when proliferation slowed down or ceased. Exposure of Caco-2 cells in the early proliferative phase of cell growth to increasing concentrations of the potent intestinal growth factor phytohaemagglutinin E4 greatly stimulated enzyme activity. In contrast, the activity of ornithine decarboxylase was not stimulated in Caco-2 cells of the late proliferative phase nor was there any increase in the enzyme activity in differentiating and fully differentiated cells of the stationary phase. Accordingly, when proliferating Caco-2 cells possessed the highest ornithine decarboxylase activity, the polyamine values were also at their highest. During differentiation, as the ornithine decarboxylase activity fell close to zero, polyamine values also decreased. In the early proliferative phase of cell growth ornithine decarboxylase activity coincided with DNA synthesis in cells exposed to Phaseolus vulgaris isolectin E4. These findings with Caco-2 cells were similar to those found in brush border cells of the rat small intestine.

Biogenic Polyamines↗
PubMed1996

Characteristics and consequences of interactions of lectins with the intestinal mucosa.

Lectins are essential and omnipresent plant (glyco)protein constituents and are ingested daily in appreciable amounts by both humans and animals. As they are biologically highly active, their consumption may have serious consequences for metabolism and health. Lectins, by virtue of their stability and specific recognition and binding by gut brush border epithelial cells, are potent exogenous metabolic growth signals for the gut and the body. As a result of their binding to surface glycans they may affect the turnover and loss of epithelial cells, damage the luminal membranes of the epithelium, interfere with their digestive/absorptive activities, stimulate shifts in the bacterial flora and modulate the immune state of the digestive tract. When eaten in relatively large quantities, these lectins have appreciable antinutritive effects for the consumers. In contrast, lectins which are not bound by the mucosa usually induce little or no harmful effects. From recent studies it is now realized that in addition to the major and sometimes dramatic effects of lectins on the gut which are mediated through their binding to pre-existing membrane glycosyl groups, lectins as metabolic signals, can also radically alter the state of glycosylation of the gut epithelium and thus further amplify their potent physiological effects. Accordingly, with the judicious use of dietary lectins it is now possible "to engineer' the digestive tract for improved physiological performance and bacterial ecology.

Animals↗
PubMed1995

Switching between control and phytohaemagglutinin-containing diets affects growth of Krebs II ascites cells and produces differences in the levels of putrescine, spermidine and spermine.

Almost twice as many ascites tumour cells were recovered from mice pre-fed for 3 days on a lactalbumin (La)-based control diet, injected with Krebs II ascites cells and then maintained on the same diet for a further 8 days, when compared with mice fed on a phytohaemagglutinin-containing (PHA) diet for the whole period. A dietary switch on the day of injection of tumour cells produced an intermediate effect; mice switched to the La diet after pre-feeding on PHA for 3 days developed somewhat more tumour cells than when the opposite dietary switch was performed. The polyamine content in the tumour cells was lowest in the mice fed on La, and highest in mice fed PHA for the duration of the experiment, respectively. Since large amounts of extraneous polyamines are required in order to sustain tumour growth, and the hyperplastic growth of the gut which occurs in response to injesting the lectin is a polyamine-dependent process, it is evident that the two growth signals compete with one another for important nutrients/growth factors, including polyamines.

Animals↗
PubMed1995

Diamine and polyamine oxidase activities in phytohaemagglutinin-induced growth of rat small intestine.

The activities of diamine and polyamine oxidases, two enzymes of polyamine catabolism, were studied in hyperplastic growth of rat small intestine induced by phytohaemagglutinin. This growth, evaluated by the elongation of Lieberkühn's crypts, was more extensive in the proximal than in the distal parts of the gut. The activity of diamine oxidase was significantly reduced in the proximal (70%), medial (45%) and the distal (25%) parts. The activity of polyamine oxidase was doubled. The concentrations of putrescine, cadaverine and spermidine were significantly elevated in the three intestinal parts studied, whereas those of histamine and spermine were unchanged. It appears that changes in the activities of diamine and polyamine oxidases may contribute to the increased putrescine content, which is necessary to maintain active polyamine turnover for sustaining growth of the gut.

Amine Oxidase (Copper-Containing)↗
PubMed1995

Lectins and also bacteria modify the glycosylation of gut surface receptors in the rat.

Oral exposure to lectins or the presence or absence of bacteria in the rat small intestine were shown by histological methods using anti-lectin antibodies or digoxigenin-labelled lectins to have major effects on the state of glycosylation of lumenal membranes and cytoplasmic glycoconjugates of epithelial cells. Taken together with the dramatic effects of exposure to lectins on gut function, metabolism and bacterial ecology, this can be used as a basis for new perspectives of biomedical manipulations to improve health.

Administration, Oral↗
PubMed1995

The importance of dietary polyamines in cell regeneration and growth.

The polyamines putrescine, spermidine and spermine are essential for cell renewal and, therefore, are needed to keep the body healthy. It was previously believed that polyamines are synthesized by every cell in the body when required. However, in the present paper evidence is provided to show that, as in the case of the essential amino acids, the diet can supply sufficient amounts of polyamines to support cell renewal and growth. Systematic analysis of different foods was carried out and from the data obtained, the average daily polyamine consumption of British adults was calculated to be in the range 350-500 mumol/person per d. The major sources of putrescine were fruit, cheese and non-green vegetables. All foods contributed similar amounts of spermidine to the diet, although levels were generally higher in green vegetables. Meat was the richest source of spermine. However, only a part of the polyamines supplied by the diet is available for use by the body. Based on experiments with rats it was established that polyamines were readily taken up from the gut lumen, probably by passive diffusion, and were partly metabolized during the process of absorption. More than 80% of the putrescine was converted to other polyamines and non-polyamine metabolites, mostly to amino acids. The enzyme responsible for controlling the bioavailability of putrescine was diamine oxidase (EC 1.4.3.6). For spermidine and spermine, however, about 70-80% of the intragastrically intubated dose remained in the original form. Considering the limitations on bioavailability (metabolism and conversion), the amounts of polyamines supplied by the average daily diet in Britain should satisfy metabolic requirements.

Adult↗
PubMed1995

Inhibition of starch digestion by alpha-amylase inhibitor reduces the efficiency of utilization of dietary proteins and lipids and retards the growth of rats.

Digestion/absorption and nutritional utilization of starch, protein and lipids were studied in rats fed diets containing purified kidney bean alpha-amylase inhibitor at levels of 0, 1.6, 3.3 and 6.6 g/kg diet. At the two higher levels, the growth rate of rats and the apparent digestibilities and utilization of dietary starch and protein were significantly less than in control rats, and losses of nitrogen, lipids and carbohydrate resulted in a significant reduction in dry body weight. Some organs of the body were also affected: the relative dry weights of the intestines and the pancreas were higher, whereas liver and thymus weights were lower than in control rats. As starch digestion in the small intestine was negligible at higher inhibitor concentrations, the cecum was practically blocked by solidified digesta. This effect and the ensuing bacterial fermentation stimulated the growth of this tissue by hyperplasia and hypertrophy. However, as the distension was not always sufficient, the organ was occasionally ruptured and the rats had to be killed. Inhibitor doses in this work were comparable to those in clinical studies, implying that the use of the inhibitor is not without health risks. Moreover, diets rich in alpha-amylase inhibitor such as those containing transgenic plants with high levels of inhibitor gene expression cannot be recommended in intensive animal production.

Animals↗
PubMed1995

The utilization of lupin (Lupinus angustifolius) and faba bean globulins by rats is poorer than of soybean globulins or lactalbumin but the nutritional value of lupin seed meal is lower only than that of lactalbumin.

The effects of dietary sweet lupin (Lupinus angustifolius, Unicrop) seed meal or its insoluble fiber (nonstarch polysaccharides + lignin) on performance, digestibility and nitrogen utilization in growing rats were studied in four experiments. Globulin proteins isolated from lupin, faba bean (Vicia faba L. minor) or soybean (Glycine max) were also incorporated into purified diets as replacements for lactalbumin (control) and the nutritional effects were evaluated. Isocaloric, legume-based diets supplemented with amino acids were used. Final weight gain, gain:feed ratios, nitrogen retention and net protein utilization of the animals fed whole lupin meal-based diets for 10 d were inferior to those of controls. In contrast, adding lupin insoluble fiber to a control diet produced no adverse effects. Ileal starch and apparent nitrogen digestibilities, and fecal digestibility of starch in lupin-fed rats were higher than those of controls, but fecal true nitrogen digestibility was lower. Replacement of lactalbumin with globulin proteins from lupin or faba bean depressed food intake and protein utilization, but only performance was affected by consumption of soybean globulins. Rats consuming lupin or faba bean globulins excreted significantly more nitrogen, particularly as urea through the urine. This did not occur in rats fed soybean globulins. Urea concentration in plasma was higher in rats fed diets containing lupin meal or legume globulins. The concentrations of urea, arginine and ornithine in plasma increased significantly compared with control values after 3 to 9 h of a lupin diet. After 9 h, plasma lysine was also decreased. We concluded that the main reasons for the low nutritional value of sweet lupin seed meal are likely to be related to the chemical structure of the globulin proteins and their adverse effects on growth and nitrogen metabolism, and not to any known antinutritional factor or poor digestibility.

Amino Acids↗
PubMed1995

Purified Pseudomonas aeruginosa PA-I lectin induces gut growth when orally ingested by rats.

The effects of PA-I lectin isolated from the human pathogen Pseudomonas aeruginosa upon cellular metabolism in vivo have been studied using the rat gut as a model system. Orally ingested PA-I lectin stimulated metabolic activity and induced polyamine accumulation and growth in the small intestine, caecum and colon. The nature and extent of the changes induced by PA-I lectin were similar to those caused by dietary kidney bean lectin and were likely to lead to impaired epithelial cell function and integrity. This finding contributes to our understanding of the possible roles of these lectins in Pseudomonas aeruginosa infection.

Adhesins, Bacterial↗