Uptake and bioavailability of dietary polyamines.
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Animal Feed↗
Biomedical subjects
D S Brown
Publications and source records attributed to D S Brown.
The fate of absorbed and exogenous ammonia as influenced by forage or forage-concentrate diets in growing sheep.
Changes in splanchnic energy and N metabolism were studied in sheep, prepared with vascular catheters across the portal-drained viscera (PDV) and the liver, and maintained on supramaintenance intakes of either gross or grass + barley pellets. The animals were challenged, on both diets, with 4 d intramesenteric vein infusions of NH4Cl (25 mumol/min) plus NH4HCO3 (at either 0 or 125 mumol/min). On the final day of each treatment the natural abundance NH4Cl was replaced with 15NH4Cl over a 10 h infusion while over the same period [1-13C]leucine was infused via a jugular vein. Measurements were made of blood flow plus mass transfers of NH3, urea, free amino acids and O2 across the PDV and liver. Enrichments of [14N15N]urea and [15N15N]urea plus [15N]glutamine, aspartate and glutamate were also monitored. Whole-body urea flux was determined by infusion of [14C]urea. At the end of the study the animals were infused for 3 h with 15nH4Cl, killed and liver samples assayed for intracellular free amino acid enrichments and concentrations. Blood flows across the splanchnic region were unaffected by either diet or level of ammonium salt infusion. At the lower ammonium salt infusion there was a trend for greater absorption of NH3 across the PDV (P < 0.10) with grass + barley than with the grass diet, while removal of urea was unaltered. At the higher ammonium salt infusions there was a significantly greater appearance of NH3 across the PDV and this exceeded the extra infused. Urea-N removal, however, was also elevated and by more than that required to account for the additional NH3. The PDV contributed 19-28% to whole-body O2 consumption and the liver 23-32%. Hepatic extraction of absorbed NH3 was complete on all treatments and systemic pH remained constant. The fractions of urea-N apparently derived from NH3 were similar on the grass (0.50-0.64) and grass + barley (0.64-0.67) diets. Hepatic production of urea agreed well with urea flux measurements. Between the two levels of ammonium salt infusion and within diets the additional NH3 removed across the PDV was accounted for by the increased urea-N production. The [14N15N]:[15N15N] ratio of the urea produced was 97:3, while the enrichment of hepatic intracellular free aspartate was lower than that of [14N15N]urea. Glutamine enrichments were 0.23-0.37 those of [14N15N]urea, indicating a minor role for those hepatocytes (probably perivenous) which contain glutamine synthetase (EC 6.3.1.2). Leucine kinetics, either for the whole body or splanchnic tissues, were not different between diets or level of ammonium salt infusion, except for oxidation which was less on the grass + barley ration. Amino acid concentrations were lower on the grass + barley diet but net PDV absorptions were similar. The pattern of essential amino acids absorbed into the PDV showed good agreement with the published composition of mixed rumen microbial protein. Fractional disappearances of absorbed free essential amino acids across the liver varied from 0.4 (branched chains) to near unity (histidine, phenylalanine).
Amino Acids↗
Splanchnic-bed transfers of amino acids in sheep blood and plasma, as monitored through use of a multiple U-13C-labelled amino acid mixture.
The response in whole-body and splanchnic tissue mass and isotope amino acid transfers in both plasma and blood has been studied in sheep offered 800 g lucerne (Medicago sativa) pellets/d. Amino acid mass transfers were quantified over a 4 h period, by arterio-venous procedures, across the portal-drained viscera (PDV) and liver on day 5 of an intravenous infusion of either vehicle or the methylated products, choline (0.5 g/d) plus creatine (1.0 g/d). Isotopic movements were monitored over the same period during a 10 h infusion of a mixture of U-13C-labelled amino acids obtained from hydrolysis of labelled algal cells. Sixteen amino acids were monitored by gas chromatography-mass spectrometry, with thirteen of these analysed within a single chromatographic analysis. Except for methionine, which is discussed in a previous paper, no significant effects of choline plus creatine infusion were observed on any of the variables reported. Whole-body protein irreversible-loss rates ranged from 158 to 245 g/d for the essential amino acids, based on the relative enrichments (dilution of the U-13C molecules by those unlabelled) of free amino acids in arterial plasma, and 206-519 g/d, when blood free amino acid relative enrichments were used for the calculations. Closer agreement was obtained between lysine, threonine, phenylalanine and the branched-chain amino acids. Plasma relative enrichments always exceeded those in blood (P < 0.001), possibly due to hydrolysis of peptides or degradation of protein within the erythrocyte or slow equilibration between plasma and the erythrocyte. Net absorbed amino acids across the PDV were carried predominantly in the plasma. Little evidence was obtained of any major and general involvement of the erythrocytes in the transport of free amino acids from the liver. Net isotope movements also supported these findings. Estimates of protein synthesis rates across the PDV tissues from [U-13C] leucine kinetics showed good agreement with previous values obtained with single-labelled leucine. Variable rates were obtained between the essential amino acids, probably due to different intracellular dilutions. Isotope dilution across the liver was small and could be attributed predominantly to uni-directional transfer from extracellular sources into the hepatocytes and this probably dominates the turnover of the intracellular hepatic amino acid pools.
Amino Acids↗
Monitoring of median nerve somatosensory evoked potentials during cervical spinal cord decompression.
We evaluated the intraoperative utility of monitoring median nerve somatosensory evoked potentials (SEPs) in 31 consecutively hospitalized neurosurgical patients (mean age 55.3 +/- 2.1 years) who underwent spinal cord decompression for cervical herniated disc, spondylosis, or tumor. Pre- and postoperative standard neurological examinations included evaluation of motor function, sensory responses, gait, tone, and reflexes. Evoked potentials were recorded from clavicular Erb's (N9) and contralateral cortical (N20) points. Intraoperatively, SEP measurements were obtained after the onset of anesthesia (baseline control) and were repeated throughout the operative procedures. N20 and N9-N20 conduction latencies were measured for each SEP recording; disappearance of the SEP waveform was interpreted as a nonquantifiable increase in latency. Follow-up neurological examinations were made immediately after and up to 6 months following surgery. Postoperatively, the 31 study subjects were assigned to one of two groups based on neurological evaluation: 27 group I subjects had either no change or improvement (good outcome) whereas four group II subjects had postoperative neurological deterioration (poor outcome). Intraoperative N9-N20 interpeak latency was found to increase during cervical decompression in six of 27 group I and in two of four group II subjects. Only two of the Group I subjects exhibited increases > 10% (14 and 19%, respectively). Intraoperative communication to the surgeon of a marked increase of N9-N20 latency during positioning for cervical traction clearly obviated a poor outcome in one group I subject; Upon removal of traction, latency decreased and significant changes in neurological function were not detected postoperatively. The SEP waveform disappeared in two of the group II and in none of the group I subjects. In the two group II subjects exhibiting increases of N9-N20 latencies, increments were > 20%. These findings indicate that in patients undergoing cervical spinal cord decompression, disappearance of SEPs or increases > 20% in the N9-N20 interpeak latency are suggestive of operative conditions that place patients at risk for poor neurological outcome.
Adult↗
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↗
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↗
Rapid high-performance liquid chromatographic method for the quantitation of polyamines as their dansyl derivatives: application to plant and animal tissues.
A rapid high-performance liquid chromatographic method for the separation of polyamines as their dansyl derivative has been developed. The chromatographic system used consisted of a reversed-phase column and a mobile phase of acetonitrile and water. The separation of 1,3-diaminopropane, putrescine, cadaverine, spermidine and spermine takes only 9 min. This method provides a good resolution between 1,3-diaminopropane and putrescine. It has been applied to quantify polyamines from seeds of wheat, petals of Phalaenopsis hybrids and various rat tissues.
Animals↗
Hepatic detoxification of ammonia in the ovine liver: possible consequences for amino acid catabolism.
The effects of either low (25 mumol/min) or high (235 mumol/min) infusion of NH4Cl into the mesenteric vein for 5 d were determined on O2 consumption plus urea and amino acid transfers across the portal-drained viscera (PDV) and liver of young sheep. Kinetic transfers were followed by use of 15NH4Cl for 10 h on the fifth day with simultaneous infusion of [1-13C]leucine to monitor amino acid oxidation. Neither PDV nor liver blood flow were affected by the additional NH3 loading, although at the higher rate there was a trend for increased liver O2 consumption. NH3-N extraction by the liver accounted for 64-70% of urea-N synthesis and at the lower infusion rate the additional N required could be more than accounted for by hepatic removal of free amino acids. At the higher rate of NH3 administration additional sources of N were apparently required to account fully for urea synthesis. Protein synthesis rates in the PDV and liver were unaffected by NH3 infusion but both whole-body (P < 0.05) and splanchnic tissue leucine oxidation were elevated at the higher rate of administration. Substantial synthesis of [15N]glutamine occurred across the liver, particularly with the greater NH3 supply, and enrichments exceeded considerably those of glutamate. The [15N]urea synthesized was predominantly as the single labelled, i.e. [14N15N], species. These various kinetic data are compatible with the action of ovine hepatic glutamate dehydrogenase (EC 1.4.1.2) in periportal hepatocytes in the direction favouring glutamate deamination. Glutamate synthesis and uptake is probably confined to the perivenous cells which do not synthesize urea.(ABSTRACT TRUNCATED AT 250 WORDS)
Amino Acids↗
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↗
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↗
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↗
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↗
Hospital discharge preparation for homeward bound elderly.
The purpose of this study was to explore hospital discharge preparation for elderly patients returning to their homes, and to examine knowledge and satisfaction outcomes. A descriptive correlational design was used in an urban health maintenance organization medical center. The convenience sample included 140 English-speaking patients with medical diagnoses. Knowledge scores were obtained for those patients who went home with a regime change, and the majority were discharged with changes in diet, medications, treatments, or activities. Medication knowledge was identified as an area that needed improvement; low scorers for medication knowledge tended to be older, less ambulatory, and had more secondary diagnoses. Patients were satisfied with the instruction that they received and perceived themselves to be prepared to continue their care at home. The ability of elderly patients to learn instruction for their continued health care is still unclear. Patients may have been too ill to learn, or length of stay may have been too short, or patients may not have had the need to learn what health care providers considered essential.
Aged↗
Tumour and host tissue responses to branched-chain amino acid supplementation of patients with cancer.
1. Rates of protein synthesis have been measured from the incorporation of 57 mg of L-[1-13C]leucine/kg for 90 min into muscle tissue and colorectal tumours removed at surgery from cancer patients. 2. For the 20 h preceding surgery and during the measurement of protein synthesis, the patients received intravenous saline, conventional intravenous nutrition (0.2 g of N and 103 non-protein kJ/kg body weight) or intravenous nutrition enriched with the branched-chain amino acids leucine, isoleucine and valine (0.2 g of N with 30% from branched-chain amino acids and 103 non-protein kJ/kg body weight). 3. Conventional intravenous nutrition resulted in a significant stimulation of the rate of protein synthesis in both muscle tissue (2.64 +/- 0.75%/day versus 1.78 +/- 0.51%/day in saline control, means +/- SD) and tumour tissue (43.9 +/- 10.3%/day versus 22.6 +/- 5.6%/day in saline control). 4. Pre-operative nutrition enriched with branched-chain amino acids was less effective than conventional intravenous nutrition in stimulating protein synthesis in both muscle and tumour. The rates of protein synthesis were 2.12 +/- 0.41%/day in muscle and 33.7 +/- 5.3%/day in the tumours. 5. The expression of proliferating cell nuclear antigen in sections of the tumours showed changes with intravenous feeding of the two different amino acid mixtures that were similar to the changes in protein synthesis, and these two variables were significantly correlated. This is evidence that feeding with conventional mixtures and mixtures enriched with branched-chain amino acids stimulates tumour growth.(ABSTRACT TRUNCATED AT 250 WORDS)
Aged↗
Determination of L-methionine-dl-sulphoxide in tissue extracts.
The amino acid fraction from rat liver, heart and skeletal muscle was prepared by the separation of sulphosalicylic acid extract on Dowex 50 H+ form. The presence of L-methionine-dl-sulphoxide in these extracts was identified and compared by three independent chromatographic methods: ion-exchange, Pico-Tag and reversed-phase high-performance liquid chromatography after precolumn derivatisation with diethylethoxymethylenemalonate. Quantitative data indicate that L-methionine-dl-sulphoxide is present in the intracellular pool at the levels of free methionine.
Animals↗
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↗
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↗