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Biomedical subjects

A Pusztai

Publications and source records attributed to A Pusztai.

At least 73 records · Page 4Linked to original sources

Salmonella typhimurium and Salmonella enteritidis induce gut growth and increase the polyamine content of the rat small intestine in vivo.

The effects of infection by Salmonella enteritidis and S. typhimurium on the small and large intestines, liver, spleen and mesenteric nodules of rats were studied in vivo. Both Salmonella serotypes persisted and proliferated in the gastrointestinal tract and invaded sub-epithelial tissues, mainly the ileum, leading to the systemic distribution of these pathogens. Coincidental with infection, the rate of crypt cell proliferation increased resulting in substantial growth of the small intestine. The extent of this and the accompanying accumulation of polyamines was particularly dramatic in the ileum where there was also some disruption of the villus epithelium. It is possible that these effects of the infection on the metabolism and morphology of the small bowel, which strongly resembled the changes induced by some plant lectins, may facilitate the colonisation and invasion of the gut by Salmonellae.

Animals↗

Reversible effect of phytohaemagglutinin on the growth and metabolism of rat gastrointestinal tract.

The lectin, phytohaemagglutinin, present in beans survives passage through the gastrointestinal tract in a biologically and immunologically intact form. It is known that by binding to the brush border membranes of the small intestine phytohaemagglutinin induces its hyperplastic growth. However, its effect on the other parts of the gut are not known. This study considered the dose and time dependent changes in the gastrointestinal tract exposed to phytohaemagglutinin. Lectin binding was detected by polyclonal antibodies using PAP staining to the surface and the parietal cell region of the stomach, the brush border epithelium of the small intestine and to the surface membrane of the caecum and colon. To characterise the metabolic changes in the gut organ weights, protein, RNA, DNA, and polyamine contents were measured. While phytohaemagglutinin induced a dose and time dependent growth of the small intestine by lengthening the tissue and thickening the gut wall by increasing the number of crypt cells, the lectin also changed the size and metabolism of the large intestine and pancreas, but this growth was by hypertrophy. Phytohaemagglutinin in the diet influences the size, metabolism, and function of the entire digestive tract. The lectin induced changes were fully or partially reversed within three days.

Animals↗

Consumption of diets containing raw soya beans (Glycine max), kidney beans (Phaseolus vulgaris), cowpeas (Vigna unguiculata) or lupin seeds (Lupinus angustifolius) by rats for up to 700 days: effects on body composition and organ weights.

Feeding trials have been done with rats to assess the effects of long-term (700 d) consumption of diets based on raw cowpeas (Vigna unguiculata; moderate Bowman-Birk inhibitor content, low lectin content), lupin seeds (Lupinus angustifolius; low lectin and protease inhibitor content) or soya beans (Glycine max; high Kunitz inhibitor content, moderate Bowman-Birk inhibitor content, moderate lectin content) or diets containing low levels of raw kidney bean (Phaseolus vulgaris; high lectin content, low Bowman-Birk inhibitor content) on body weight and composition and organ weights. All the legume-based diets reduced feed conversion efficiency and growth rates during the initial 250 d. However, after 250 d the weight gains by rats given legume-based diets were similar to those of controls given the same daily feed intake. Long-term consumption of diets containing low levels of kidney bean significantly altered body composition of rats. The levels of lipid in the body were significantly reduced. As a result, carcasses of these rats contained a higher proportion of muscle/protein than did controls. Small-intestine relative weight was increased by short- and long-term consumption of the kidney-bean-based diet. However, the increase in relative pancreatic weight observed at 30 d did not persist long term. None of the other legume-based diets caused any significant changes in body composition. However, long-term exposure to a soya-bean- or cowpea-based diet induced an extensive increase in the relative and absolute weights of the pancreas and caused an increase in the incidence of macroscopic pancreatic nodules and possibly pancreatic neoplasia. Long-term consumption of the cowpea-, kidney-bean-, lupin-seed- or soya-bean-based diets by rats resulted in a significant increase in the relative weight of the caecum and colon.

Animal Feed↗

[Effect of a mite-killing agent on house dust and on symptoms of house dust allergy].

UNLABELLED: The efficacy of acaricid benzyl-benzoate (Acarosan) has been followed in an open study for one year in patients with house dust mite allergy; the clinical signs and the mite allergen level have been considered. METHODS: the furniture (beds, upholstered pieces, carpets) of 17 house dust mite allergic patients suffering from bronchial asthma and/or allergic rhinitis has been investigated. The mite content of the dust gained from the furniture has been determined with the help of the semiquantitative Acarex test. This test has been done before the mite elimination and 3, 6, 9 and 12 months after it. The registered clinical signs of the patients: symptoms, drug consumption, expiratory peak flow values have been measured twice a day. RESULTS: 9/17 beds have become free from mites, 6/17 beds have had less mites than before, 1/17 no change, 1/17 augmentation of mite content. The proportion of days free from complaints has been 26.8% at the beginning of the trial and 47.1% after 12 months, the drug consumption has diminished meanwhile. At the beginning of the trial 6 children had pathologic lability index based on peak-flow measurements, they improved significantly. The information about the mite content of the furniture gives help for the elimination measures. The chemical mite elimination reduces the mite content of the flat and results in clinical improvement of house dust mite allergic patients.

Acari↗

The intestinal true absorption of 65Zn in rats is adversely affected by diets containing a faba bean (Vicia faba L.) nonstarch polysaccharide fraction.

The effects on the absorption of 65Zn by two varieties of raw faba bean (Vicia faba L., minor) or seed components that may interfere with mineral metabolism in the gut, have been studied in growing rats. In bean diets all protein was supplied by the meals, and the fractions were tested by incorporating them in control diet at the same levels as they occur in the seeds. Absorption of 65Zn was also measured in rats fed dephytinized bean meal produced by including phytase in the diet. Rats were pair-fed diets supplemented with amino acids and minerals to target requirements and containing 40 mg Zn/kg diet. True absorption of Zn was 50-70% lower in rats fed diets containing both cultivars of faba bean meals than in those fed the control diet. Although soluble nonstarch polysaccharides caused a significant reduction in the absorption of Zn, this effect disappeared after the removal of phytate by demineralization. In contrast, despite its negligible content of phytate, the insoluble nonstarch polysaccharides in the cell wall fraction of the cotyledon accounted for most of the reduction in Zn absorption in rats fed the faba bean diets. Addition of phytic acid to the control diet significantly reduced the absorption of 65Zn but only from 44 to 36%. Moreover, the increase in the absorption of Zn was similarly small, from 21% to 29%, with the addition of phytase to the faba bean diet.(ABSTRACT TRUNCATED AT 250 WORDS)

6-Phytase↗

Lectins as markers of rumen epithelial cell differentiation.

Lectins of different carbohydrate specificities (GNA (Galanthus nivalis), con A (Canavalia ensiformis), VFL (Vicia faba), PSL (Pisum sativum), LCA (Lens culinaris), PNA (Arachis hypogaea; with or without prior neuraminidase treatment), WGA (Triticum vulgare), SBA (Glycine max), UEA-I (Ulex europaeus), LPA (Limulus polyphemus), BS-I B4 (Bandeiraea simplicifolia, isolectin B4)) were explored for use as differentiation markers of rumen epithelial cells in vivo and in vitro. Lectins specific for mannose (GNA), mannose/glucose (con A, VFL, PSL and LCA), N-acetylglucosamine (WGA) or for N-acetylneuraminic acid (LPA) reacted generally with all types of rumen epithelial cell from both rumen tissue and cell culture. They were, therefore, not suitable markers of epithelial differentiation. SBA was unsuitable because, although it reacted with both tissue and cultured rumen epithelial cells, it was also bound to non-stratified areas of primary rumen epithelial cell cultures. Both BS-I B4 and PNA (after neuraminidase treatment) had to be ruled out because they did not react with differentiated rumen tissue epithelial cells, although they did bind to both stratified and non-stratified cultured cells. In contrast, UEA-I reacted strongly with differentiated rumen epithelial cells both from rumen tissue and cell cultures and therefore appears to be a good general marker for rumen epithelial cell differentiation.

Acetylgalactosamine↗

Cloning and characterization of the lectin cDNA clones from onion, shallot and leek.

Characterization of the lectins from onion (Allium cepa), shallot (A. ascalonicum) and leek (A. porrum) has shown that these lectins differ from previously isolated Alliaceae lectins not only in their molecular structure but also in their ability to inhibit retrovirus infection of target cells. cDNA libraries constructed from poly(A)-rich RNA isolated from young shoots of onion, shallot and leek were screened for lectin cDNA clones using colony hybridization. Sequence analysis of the lectin cDNA clones from these three species revealed a high degree of sequence similarity both at the nucleotide and at the amino acid level. Apparently the onion, shallot and leek lectins are translated from mRNAs of ca. 800 nucleotides. The primary translation products are preproproteins (ca. 19 kDa) which are converted into the mature lectin polypeptides (12.5-13 kDa) after post-translational modifications. Southern blot analysis of genomic DNA has shown that the lectins are most probably encoded by a family of closely related genes which is in good agreement with the sequence heterogeneity found between different lectin cDNA clones of one species.

Allium↗

Analysis of lectin binding sites in the gut of hooded Lister rats with special emphasis on recently detected lectins.

Seven recently isolated lectins were tested for their ability to bind to tissue sections of rat gut. Binding sites for N-Acetylgalactosamine specific lectins were found in mucins, in the brush border membrane and in goblet cells. Non-reducing terminal mannose residues were absent from cell surface membranes but were detected in the supranuclear region of goblet cells and enterocytes. The results of lectin binding obtained in this study were generally similar to lectin-gut interactions observed in vivo.

Acetylgalactosamine↗

Antinutritive effects of wheat-germ agglutinin and other N-acetylglucosamine-specific lectins.

Incorporation of N-acetylglucosamine-specific agglutinins from wheat germ (Triticum aestivum; WGA), thorn apple (Datura stramonium) or nettle (Urtica dioica) rhizomes in the diet at the level of 7 g/kg reduced the apparent digestibility and utilization of dietary proteins and the growth of rats, with WGA being the most damaging. As a result of their binding and endocytosis by the epithelial cells of the small intestine, all three lectins were growth factors for the gut and interfered with its metabolism and function to varying degrees. WGA was particularly effective; it induced extensive polyamine-dependent hyperplastic and hypertrophic growth of the small bowel by increasing its content of proteins, RNA and DNA. Furthermore, an appreciable portion of the endocytosed WGA was transported across the gut wall into the systemic circulation, where it was deposited in the walls of the blood and lymphatic vessels. WGA also induced the hypertrophic growth of the pancreas and caused thymus atrophy. Although the transfer of the gene of WGA into crop plants has been advocated to increase their insect resistance, as the presence of this lectin in the diet may harm higher animals at the concentrations required to be effective against most pests, its use in plants as natural insecticide is not without health risks for man.

Acetylglucosamine↗

Pancreatic enlargement is evident in rats fed diets containing raw soybeans (Glycine max) or cowpeas (Vigna unguiculata) for 800 days but not in those fed diets based on kidney beans (Phaseolus vulgaris) or lupinseed (Lupinus angustifolius).

Pancreatic weights and composition were studied with rats fed diets containing raw legume seeds for up to 800 d. Rapid pancreatic enlargement was induced by dietary soybeans (Glycine max) (high Kunitz and Bowman-Birk trypsin inhibitor contents, moderate lectin content) during the initial 150 d. Over the next 200 d the rate of pancreatic growth was similar to that in controls. After 350 d a second period of rapid pancreatic growth occurred. Macroscopic pancreatic nodules were evident in a number of rats fed soybeans for 500 d or more. A similar pattern of pancreatic growth was observed in rats fed dietary cowpeas (Vigna unguiculata) (high Bowman-Birk inhibitor content, low lectin content). Extensive pancreatic growth was also found in young rats fed moderate dietary levels of kidney beans (Phaseolus vulgaris) (low Bowman-Birk inhibitor content, high lectin content). However, the trophic effects diminished with time, and from 100 d onwards, little enlargement was evident. Consumption of a lupinseed (Lupinus angustifolius) diet (low trypsin inhibitor, low lectin content) did not cause pancreatic enlargement. The initial pancreatic growth induced by dietary soybeans seemed to be due to the lectins and trypsin inhibitors, whereas the second period of pancreatic growth was possibly due primarily to the trypsin inhibitors.

Analysis of Variance↗

Kidney bean lectin-induced Escherichia coli overgrowth in the small intestine is blocked by GNA, a mannose-specific lectin.

The reversible and dose-dependent hyperplastic growth of the small intestine and accelerated epithelial cell turnover caused by feeding rats with diets containing kidney bean lectin (PHA) increased the proportion of immature cells on the villi whose membrane and/or cytoplasm contained mainly simple, polymannosylated glycans. These new alpha-linked mannosyl terminals, particularly of the damaged epithelium, facilitated the preferential adherence of opportunistic Escherichia coli with mannose-sensitive Type 1 fimbriae, and other coliforms, to the glycocalyx. Accordingly, the growth of the gut was accompanied by a reversible and PHA dose-dependent overgrowth with E. coli. As expected from their common carbohydrate specificity, the inclusion in the diet of the mannose-specific agglutinin from snowdrop (Galanthus nivalis) bulbs (GNA) significantly reduced the extent of E. coli overgrowth, but abolished neither the growth nor the damage caused by PHA to the small intestine. Thus, GNA and perhaps other mannose-specific lectins, especially when used in a preventive mode, can be used to specifically block the proliferation of Type 1 E. coli in the small intestine.

Animals↗

Binding and degradation of lectins by components of rumen liquor.

The binding of 15 different plant lectins to feed particles and microbes in rumen liquor, and their degradation were studied in vitro. The rate of degradation assessed from the label released when radioactive iodine-labelled lectins were incubated with rumen liquor conflicted with the rates calculated from measurements of the survival of the antigenic structure (immuno-rocket electrophoresis) or the biological function (haemagglutination) of the lectins. Thus solubilization of the radioactive label indicated that Concanavalin A (Con A), but not the soyabean agglutinin, SBA, or kidney bean phytohaemagglutinin, PHA-E3L, was stable to rumen proteolysis. In contrast, both SBA and PHA-E3L were shown by immuno-rocket electrophoresis or haemagglutination tests to be highly resistant to breakdown, while the degradation of Con A proceeded at a constant slow rate under the same conditions. This was in accord with the previously established general stability of lectins in the gut of single-stomach animals. Of the 15 lectins, SBA, favin (Vicia faba lectin) and Con A were bound by hay and the particle fraction of rumen liquor. This was, in part, specific and reversible in the presence of appropriate sugars. Most pure bacterial strains preferentially bound lectins with specificity for glucose/mannose (favin and Con A), while rumen fungi reacted with SBA. The level of binding was low with other lectins. However, inter-strain differences of lectin-binding were found in Selenomonas ruminantium and Ruminococcus flavefaciens. Clearly, as some lectins were not fully degraded in the rumen, they could be expected to depress the utilization of the diet not only in single-stomach animals but, possibly, also in ruminants.

Animals↗

Fate of the antinutritive proteins of soyabean in the ovine gut.

Defatted raw soyabean flour was administered intraruminally at a level of 10 g/kg body weight to a sheep fitted with ruminal, duodenal, and ileal cannulas. Soyabean lectin and trypsin inhibitor appeared in the duodenum within one hour, both reached the terminal ileum and disappeared within 24 hours. The passage of the lectin along the gut progressively lagged behind that of the inhibitor, which indicates that the lectin binds to the intestinal surface. The effect of the inhibitors was neutralized by a marked pancreatic hypersecretion. Contrary to earlier assumptions, the antinutritional proteins were not effectively degraded in the forestomachs.

Animal Feed↗

Dietary lectins are metabolic signals for the gut and modulate immune and hormone functions.

Although it is common knowledge that some dietary lectins can adversely affect the growth and health of young animals and that, therefore, lectins are implicated in nutritional disorders of the digestive system, it has not been rigorously established that findings with animals are also directly applicable to humans. However, because the glycosylation state of the human gut is basically similar to that of higher animals, it may be confidently predicted that the effects of dietary lectins will have similarities in both humans and animals. The more recent but not generally appreciated realization that lectins also have many beneficial effects on the gut and metabolism of animals makes the exploration of these for possible use in medical-clinical practice even more attractive. Most lectins in our diet are resistant to breakdown during gut passage and are bound and endocytosed by epithelial cells. These lectins are powerful exogenous growth factors for the small intestine, can induce dramatic shifts in its bacterial flora and interfere with its hormone secretion. In addition, lectins which are transported across the gut wall into the systemic circulation can modulate the body's hormone balance, metabolism and health. Although these physiological effects are mediated or reinforced by immune responses, they are primarily the result of the specific chemical reactivity of lectins with cell surface receptors of the gut. Clearly, as the interactions between lectins and the gut are predictable and may be manipulated to our advantage, the exciting scientific challenge is now to explore the possible transfer of the existing knowledge gained from animal experimentation to medical-clinical practice.

Animals↗

Alterations in the level of insulin receptor and GLUT-4 mRNA in skeletal muscle from rats fed a kidney bean (Phaseolus vulgaris) diet.

1. A decline in the level of circulating insulin was observed in rats fed a diet containing kidney bean. 2. Consumption of a diet containing kidney bean caused an increase in the level of mRNAs for the insulin receptor (327%) and GLUT-4 (185%) in the gastrocnemius muscle. In contrast there was only a small increase in the amount of actin mRNA (125%). Since the kidney bean-fed rats are euglycaemic the results suggest that insulin receptor and GLUT-4 mRNA levels are regulated in response to circulating insulin concentrations rather than glucose. 3. No increases in the level of insulin receptor and actin mRNA were evident in the soleus muscle of rats fed the diet containing kidney bean; however a decline was observed in the level of GLUT-4 mRNA. 4. It is proposed that a component of kidney beans, most likely the lectin phytohaemagglutinin, has systemic effects which lead to changes in expression of the insulin receptor and GLUT-4 genes and to the sensitivity of muscle to insulin.

Actins↗

Mineral excretion of rats fed on diets containing faba beans (Vicia faba L.) or faba bean fractions.

The effects on faecal mineral excretion of two commercial varieties (local cultivar and Troy cultivar) of raw faba beans (Vicia faba L., minor) meal (VFM) and its fractions have been studied in growing rats. Diets contained local-VFM (dark seed coat) and Troy-VFM (light seed coat) at 474-500 g/kg diet, hull (VFH) from both varieties at 65 g/kg diet, and the insoluble cotyledon residue (VFCR) obtained from the Troy variety at 237 g/kg diet. Rats were pair-fed on diets which had been supplemented with amino acids to target requirements and contained similar amounts of zinc, manganese, iron and copper. With VFM diets the apparent absorption of Zn and Mn was significantly reduced. On the other hand, with hulls the apparent absorption of Fe was reduced while that of Cu slightly increased. As the amounts of calcium, phosphorus and magnesium in VFM and VFH diets were higher than in the controls, the increased intake resulted in a significant increase in both the apparent absorption and the faecal excretion of these minerals. The inclusion of VFCR in the diet had no significant effect on the mineral content of faeces. The relatively low concentrations of phytate in the bean seeds of 7.8 and 6.7 g/kg for the local and Troy cultivars respectively, could not adequately account for the increased mineral excretion. The results suggest that other seed constituents, possibly the soluble non-starch polysaccharides, may be involved in the elevated loss of Zn and Mn in rats fed on diets containing faba bean for extended periods, while some insoluble structural hull components may interfere with the absorption of Fe from the gut.

Animals↗

Nutritional evaluation of the trypsin (EC 3.4.21.4) inhibitor from cowpea (Vigna unguiculata Walp.).

The effect of feeding rats purified cowpea (Vigna unguiculata Walp.) trypsin (EC 3.4.21.4) inhibitor in a semi-synthetic high-quality diet based on lactalbumin (10 g inhibitor/kg) for 10 d was a moderate reduction in the weight gain of rats in comparison with controls, despite an identical food intake in the two groups. The reduction in the growth rate was about 20% on a live weight basis. However, the corresponding value calculated from the weight of dry carcasses was less, only about 7%, probably because the water content of the body of the two groups of rats was different. Although most of the cowpea trypsin inhibitor (CpTI) was rapidly broken down in the digestive tract, its inclusion in the diet led to a slight, though significant, increase in the nitrogen content of faeces but not of urine. Accordingly, the net protein utilization of rats fed on inhibitor-containing diets was also slightly depressed while their energy expenditure was elevated. In agreement with results obtained for the protease inhibitors of soya bean, the slight anti-nutritional effects of CpTI were probably due mainly to the stimulation of the growth and metabolism of the pancreas. Thus, the nutritional penalty for increased insect-resistance after the transfer of the cowpea trypsin inhibitor gene into food plants is slight in the short-term.

Animals↗

Effect of the beta-adrenoceptor agonist clenbuterol and phytohaemagglutinin on growth, protein synthesis and polyamine metabolism of tissues of the rat.

1. The kidney bean lectin, phytohaemagglutinin (PHA), induced a marked atrophy of skeletal muscle which was evident from the changes in tissue composition (protein, RNA, DNA and polyamine content) and from the reduction in weight and protein synthesis of hind leg muscles of rats fed on kidney bean-diets for four days. The beta-adrenoceptor agonist, clenbuterol, induced skeletal muscle hypertrophy by transiently stimulating protein synthesis. As a consequence, the muscle loss caused by a short exposure to PHA was, in part, ameliorated by clenbuterol treatment. 2. Cardiac muscle was affected to a lesser extent than skeletal muscle by both clenbuterol and the lectin. However, there was evidence that protein synthesis in heart was reduced by PHA. 3. PHA had opposite effects on the gut, the lectin-induced hyperplasia of the jejunum was accompanied by a large increase in protein synthesis. Clenbuterol alone had no effect on the jejunum whereas a combination of PHA and clenbuterol appeared to exacerbate the effect of the lectin on gut. 4. Both the lectin-induced gut growth and the hypertrophy of skeletal muscle caused by clenbuterol were preceded by the accumulation of polyamines in the respective tissues. Of particular note was the observation that a significant increase in the proportion of the intraperitoneally injected 14C-labelled spermidine or putrescine taken up by the growing tissues could be detected by the second day. Therefore, the measurement of uptake of labelled polyamines may be used as a sensitive indicator of early alterations in tissue metabolism.

Animals↗