Putrescine and cadaverine formation in vacuum packed beef

Bacterial numbers, putrescine and cadaverine concentrations and pH were measured at regular intervals during the chill storage of vacuum packed beef. Odours on opening the packs were also assessed. Cadaverine concentration increased more rapidly than that of putrescine and measurable increases were evident before maximum bacterial numbers were attained and before any permanent off-odours were detected. Diamine concentrations correlated better with total viable count (TVC) than with counts of Gram negative organisms.     

Jasmonic acid,polyamine, and antioxidant levels in apple seedlings as affected by Ultraviolet-C irradiation

The effect of UV-C irradiation on jasmonates, abscisic acid (ABA), polyamines and antioxidant activities was investigated in apple plants (Malus domestica Borkh.). The EC₅₀ values of O₂ ^–-scavenging activity in the UV-C treated leaves were lower than those in the untreated control. In addition, total ascorbic acid and polyphenolic concentrations in the UV-C treated leaves were generally higher than those in the untreated control. Although the endogenous ABA concentrations were not significantly different between the untreated control and UV-C treatment, the endogenous jasmonic acid, putrescine, and spermine concentrations in the UV-C treated plants were higher than those in the untreated control. However, the application of PDJ before UV treatment inhibited the rise of endogenous jasmonate and putrescine concentrations. The increase of ascorbic acid was also inhibited by PDJ application before UV treatment. These facts suggest that the increase of jasmonates and polyamines may be associated with UV-C stimulation, and UV-C irradiation may be effective for increasing antioxidant activity in apple leaves.     

Tolerance to Putrescine Toxicity in Chinese Hamster Ovary Cells Is Associated with Altered Uptake and Export

When Chinese hamster ovary (CHO) cells were cultured with low concentrations of putrescine (< 5 mM) their cell cycle time increased significantly and a fraction of the cells died. A cell line tolerant to the cytotoxic and growth inhibitory effects of millimolar concentrations of putrescine was developed by growing CHO cells over many months in increasing concentrations of the polyamine. A putrescine-tolerant cell line was obtained which was capable of growing in concentrations up to 25 mMputrescine and displayed growth and cell division rates similar to the original untreated/parental CHO cells. The tolerant cells grown in putrescine displayed relatively high intracellular putrescine yet the cell-associated putrescine concentration was estimated to be 10-fold less than the culture medium level. This high concentration of cellular putrescine diminished within 60 min when the cells were changed to non-putrescine-containing media. The putrescine-tolerant phenotype was further characterized in regards to the mechanisms involved in putrescine uptake, efflux, and biosynthesis. The parental and tolerant cell lines had similar or identical levels of cellular spermidine and spermine and no differences in the acetylated polyamine pools or diamine oxidase activity. The activity of ornithine decarboxylase was also similar in the two cell lines in both the presence and the absence of ornithine. The tolerant cells, however, had a decreased uptake rate for putrescine. The tolerant cell line also showed a greatly enhanced ability to export putrescine, especially when treated with ornithine, suggesting that an upregulated polyamine export system may be present in the tolerant cells which could be responsible for the increased cell survival in high putrescine concentrations. The data are discussed in regard to the potential for identifying the transport protein(s) responsible for the maintenance of nontoxic intracellular concentrations of putrescine in a tolerant cell line grown in putrescine.     

Relationships among foliar chemistry, foliar polyamines, and soil chemistry in red spruce trees growing across the northeastern United States

Forest trees are constantly exposed to various types of natural and anthropogenic stressors. A major long-term goal of our research is to develop a set of early physiological and biochemical markers of stress in trees before the appearance of visual symptoms. Six red spruce (t Picea rubens Sarg.) stands from the northeastern United States were selected for collection of soil and foliage samples. All of the chosen sites had soil solution pH values below 4.0 in the Oa horizon but varied in their geochemistry. Some of these sites were apparently under some form of environmental stress as indicated by a large number of dead and dying red spruce trees. Samples of soil and needles (from apparently healthy red spruce trees) were collected from these sites four times during a two-year period. The needles were analyzed for perchloric acid-soluble polyamines and exchangeable inorganic ions. Soil and soil solution samples from the Oa and B horizons were analyzed for their exchange chemistry. The data showed a strong positive correlation between Ca and Mg concentrations in the needles and in the Oa horizon of the soil. However, needles from trees growing on relatively Ca-rich soils with a low exchangeable Al concentration and a low Al:Ca soil solution ratio had significantly lower concentrations of putrescine and spermidine than those growing on Ca-poor soils with a high exchangeable Al concentration and a high Al:Ca soil solution in the Oa horizon. The magnitude of this change was several fold higher for putrescine concentrations than for spermidine concentrations. Neither putrescine nor spermidine were correlated with soil solution Ca, Mg, and Al concentrations in the B horizon. The putrescine concentrations of the needles always correlated significantly with exchangeable Al (r²=0.73, t p0.05) and soil solution Al:Ca ratios (r²=0.91, t p0.01) of the Oa horizon. This suggests that in conjunction with soil chemistry, putrescine and/or spermidine may be used as a potential early indicator of Al stress before the appearance of visual symptoms in red spruce trees.     

Cycloheximide elicits in human fibroblasts a response characteristic for initiation of cell proliferation

Earlier I found that a variety of stimuli to proliferation of cultured human fibroblasts caused an increase in the rate of putrescine transport into the cells. This paper reports the effects of cycloheximide on putrescine transport in stationary and growing cultures. Cycloheximide in concentrations that inhibited protein synthesis caused increased putrescine transport in serumstarved and density-inhibited cultures. Similar effects were found with pactamycin, also an inhibitor of protein synthesis. Actinomycin D in concentrations that suppressed messenger RNA (mRNA) synthesis, did not cause increased putrescine transport. When both serum and cycloheximide were added to serum-starved cultures, the increase in putrescine transport was greater than when serum alone was added. However, cycloheximide had an inhibitory effect when added 1–2 h after addition of serum. These results suggest that one or more rapidly metabolizing proteins may be important in the regulation of putrescine transport and initiation of cell growth.     

ACETYL-COENZYME A: 1,4-DIAMINOBUTANE N-ACETYLTRANSFERASE: ACTIVITY IN RAT BRAIN DURING DEVELOPMENT, IN EXPERIMENTAL BRAIN TUMOURS AND IN BRAINS OF FISH OF DIFFERENT METABOLIC ACTIVITY

—Acetyl-CoA: 1,4-diaminobutane N-acetyltransferase catalyses the first step of putrescine catabolism in mammalian brain. It may be important in putrescine degradation of other tissues as well. Its specific activity is higher in homogenates of immature than of mature rat brains. A steady decline of putrescine acetylase activity is observed from birth until approx adult levels are reached at day 30. Microsomes and purified nuclei from brains of 2-day-old rats show considerably higher putrescine acetylase activities than the corresponding subcellular organelles from adult brains. Increased putrescine acetylase activities were found in nitroso-ethylurea-induced gliomas, together with a dramatic increase of putrescine concentration. High tissue concentrations of putrescine are, however, not necessarily correlated with enhanced putrescine acetylase activities. In trout brains a linear increase of acetyl-CoA: 1,4-diaminobutane N-acetyltransferase activity was observed together with a decrease of putrescine concentration after adaptation of the animals to increased water temperature.     

REGIONAL DISTRIBUTION OF PUTRESCINE, SPERMIDINE AND SPERMINE IN RELATION TO THE DISTRIBUTION OF RNA AND DNA IN THE RAT NERVOUS SYSTEM

—Putrescine, spermidine, spermine, RNA, DNA and protein concentrations were determined in 14 parts of the rat nervous system. If the concentrations are expressed in DNA units, putrescine and spermidine concentrations change concomitantly in the different brain parts, with the exception of hypothalamus, where relatively higher putrescine than spermidine concentrations are observed. The constancy of putrescine/spermidine ratios indicates the value of putrescine concentration as an index of spermidine biosynthesis. Spermidine correlates with RNA, except in medulla, spinal cord and peripheral nerves. It is assumed that the relative excess of spermidine in these structures indicates an additional functional role. Spermine/DNA ratios are remarkably constant in the diencephalic and telencephalic regions; they are also nearly constant, but significantly lower in midbrain, medulla, spinal cord and cerebellum. This observation gives additional support for the preferential interrelation of spermidine with RNA and spermine with DNA; i.e. for different functional roles of these two narrowly related polycations.     

Putrescine uptake by the cellular slime mould dictyostelium discoideum.

1. Rapid labelling occurs when myxamoebae of Dictyostelium discoideum strain AX2 are incubated with [1,4-14C]putrescine. Labelling is energy-dependent. 2. The label enters a pool from which rapid exchange with extracellular putrescine does not occur, and labelling is believed to represent uptake into the cells. 3. The concentration-dependence of putrescine uptake indicates that a number of systems are involved, at least one of which is saturable, with a Km of 9.1 micro M-putrescine. At high putrescine concentrations the overall uptake process is non-saturable. 4. Significant metabolism of putrescine and loss of intracellular putrescine to the medium only occurred when cells were incubated with millimolar concentrations of extracellular putrescine. 5. Putrescine uptake was inhibited by diamines, polyamines, bivalent metal ions and omega-aminocarboxylic acids. 6. The ability to take up putrescine at low concentrations decreased during starvation of myxamoebae. 7. The results are interpreted in terms of a model for putrescine uptake involving adsorptive pinocytosis at low concentrations and fluid-phase pinocytosis at high concentrations.     

Involvement of Putrescine in the Development of Kindled Seizure in Rats

During the development of kindling by daily electrical stimulations applied to the left amygdala of rats, concentrations of the polyamines putrescine, spermidine, and spermine were measured in the left amygdala and the remainder of the cerebrum. A significant increase of putrescine concentration appeared first at the left amygdala in prekindled rats and then propagated to the remainder of the cerebrum with the development of kindling. This increase in putrescine concentration in the left amygdala was higher in prekindled rats than in fully kindled rats and lasted for at least 24 h after the final kindling stimulation. The concentrations of spermidine and spermine were slightly increased in a fully kindled state. To clarify the role of putrescine in kindling, the development of amygdaloid kindling was examined in rats after microinjections of alpha-difluoromethylornithine, a specific inhibitor of polyamine synthesis, and putrescine into the ipsilateral amygdala. Pretreatment with alpha-difluoromethylornithine (50 nmol) for 10 days accelerated both the development of behavioral kindling and the propagation of the afterdischarge from the left amygdala to the frontal cortex. In contrast, pretreatment with putrescine (200 nmol) for 10 days retarded the development of kindling. These results suggest that the increase in putrescine concentration in the kindled brain has an inhibitory effect on the development of kindling.     

POLYAMINES OF THE HUMAN BRAIN DURING NORMAL FETAL AND POSTNATAL GROWTH AND DURING POSTNATAL MALNUTRITION

Abstract— The concentrations of the polyamines and putrescine were measured in the brains of human infants during fetal and early postnatal development. The concentrations of the amines were also measured in the brains of children who were malnourished during the first 2 years of life. In the brains of the adequately-nourished infants there were differences in the developmental profiles of the amines between different regions of the brain, and the changes in polyamine concentrations paralleled changes in nucleic acid accumulation. The concentration of putrescine was much higher than that of the polyamines in all regions of the brain, and in the brain stem there were marked increases in putrescine concentration at the time of most rapid rate of myelination. Putrescine also increased markedly in the forebrain at the time of neuroblast multiplication in the fetus. In children malnourished during the first year of life there were reductions in the concentrations of spermidine and putrescine in the forebrain and brainstem, but spermine was unaffected. Malnutrition had no effect on the concentration of any of the amines in the cerebellum.     

Androgenic control of polyamine concentrations in rat epididymis.

Unilateral orchidectomy resulted in a significant decrease in tissue content of putrescine and polyamines. However, no differences were detected when the results were expressed in terms of ng g-1 tissue. At 48 h after bilateral orchidectomy, a significant decrease in putrescine content was observed, but spermidine and spermine content were unaffected. The observed decrease in putrescine was prevented by treatment with testosterone propionate, but neither spermidine nor spermine were affected. Bilateral orchidectomy resulted in a significant decrease in the tissue content of putrescine, spermidine and spermine after 7 days. Treatment with testosterone propionate increased the content of putrescine, spermidine and spermine in the epididymis by about 200%, 92% and 34%, respectively. When results were expressed as nmol g-1, a significant decrease after castration in putrescine and spermidine, but not in spermine, was observed. Treatment with testosterone propionate restored putrescine concentration, but had no effect on spermidine and spermine concentrations. In castrated rats treated with testosterone propionate, the anti-androgen flutamide abolished the effect of the androgen on putrescine and spermidine content, but there was no effect on spermine. Acetylputrescine was not detected in the epididymis, while acetylpolyamines were detected at much lower concentrations than polyamines. After bilateral orchidectomy there was a decrease in the tissue content of all acetylpolyamines and an increase in their tissue concentration. The effect of castration on acetylpolyamine content was reversed by testosterone propionate treatment. We conclude that an active synthesis of polyamines occurs in the rat epididymis, and that this process depends upon the androgen environment. Regulation of ornithine decarboxylase activity appears to be the main step that is controlled by androgens.     

The dual action of the non-physiological diamines 1,3 diaminopropane and cadaverine on ornithine decarboxylase of HTC cells

In rat hepatoma tumor (HTC) cells 1,3 diaminopropane and cadaverine induced the ornithine decarboxylase antizyme as well as the end product of the ornithine decarboxylase reaction putrescine. Although at equal exogenous concentrations (10^?3M) the two non-physiological diamines penetrated the cells as effectively as putrescine; they decreased cellular ornithine decarboxylase considerably less rapidly than the naturally present diamine. Cell extracts treated with high concentrations of 1,3 diaminopropane and putrescine, and which as a result had a high specific activity of ornithine decarboxylase antizyme, were chromatographed on a superfine Sephadex G-75 column in the presence of 250 mM NaCl. No ornithine decarboxylase-antizyme complex could be detected indicating the original decrease of ornithine decarboxylase in the cells was likely due to some mechanism other than antizyme. These results indicate that 1,3 diaminopropane and cadaverine probably can act on ornithine decarboxylase, like putrescine, by two distinct regulatory mechanisms.     

Effect of dietary putrescine on whole body growth and polyamine metabolism

Putrescine (1,4-diaminobutane) is the simplest of the mammalian polyamines. These are small, positively charged molecules which are essential for cell growth and are thought to play a role in regulation of anabolic events such as synthesis of DNA, RNA, and protein. Recent reports have indicated the potential for dietary precursor amino acids of putrescine to alter tissue putrescine concentrations. The current study was conducted to determine the physiologic significance of these effects by feeding up to flooding doses of putrescine to determine any influence on whole body growth and polyamine metabolism. A total of 96 chicks were fed purified crystalline amino acid diets containing 0.0, 0.2, 0.4, 0.6, 0.8, or 1.0% purified putrescine (four birds per pen, four pens per diet) for 14 days. The feeding of 0.2% putrescine increased growth rate beyond that of controls while further supplements reduced growth and were toxic when 0.8 and 1.0% putrescine were fed. Hepatic and muscle concentrations of ornithine increased with dietary putrescine while the effect in kidney was much less. Putrescine concentrations in liver, kidney, and muscle rose when 0.4% putrescine or more was fed. This effect was particularly obvious in muscle in which there were also increases in the concentrations of spermidine and spermine. In a subsequent similar experiment, putrescine was fed at 0.0, 0.1, 0.2, 0.3, 0.4, or 0.5% to determine the effect on the activities of the key enzymes regulating polyamine synthesis. The feeding of putrescine at even 0.1% caused a rapid reduction in hepatic ornithine decarboxylase activity while S-adenosylmethionine decarboxylase and arginase activities were not influenced by diet. It was concluded that excess tissue putrescine can be toxic to whole organisms but small, orally administered doses of this metabolite can promote growth.     

Changes in endogenous polyamines during flower development in two diverse species of rose

Changes in the concentrations of endogenous free, conjugated and bound polyamine were determined in petals of two different species of rose, viz. Rosa damascena and Rosa bourboniana, from small bud (stage 1) till full bloom (stage 8). High free putrescine and spermidine concentrations were associated with early stages of flower development and then decreased in R. damascena. At full bloom, the concentration of free putrescine was higher than rest of the polyamines measured. A steady increase in conjugated putrescine, spermidine and spermine was observed during entire period of flower development with predominance of conjugated putrescine at full bloom. In R. damascena the bound spermine was higher than rest of the polyamines during full bloom. In R. bourboniana, during the early stages of flower development, similar situation was observed, however, at full bloom, free spermidine concentration was higher than rest of the polyamines. In this species, the concentration of conjugated and bound spermine was higher than rest of the polyamines during full bloom. Polyamine concentrations were generally lower in the petals of R. bourboniana than R. damascena which may be due to genotypic differences. The possible roles of the observed polyamines are discussed in relation to flower development.IHBT Communication no, 0345.     

Inhibition of L-arginine iminohydrolase (EC 3.5.3.6) from Tetrahymena thermophila by putrescine and spermidine: feedback control of polyamine biosynthesis

L-Arginine iminohydrolase (arginine deiminase, ADI) from Tetrahymena thermophila was purified approx. 75-fold by means of gel permeation chromatography. The Km of the purified enzyme for L-arginine was 412 +/- 25 microM and L-ornithine inhibited the reaction competitively with a Ki of 985 +/- 105 microM. D-Ornithine was a weak inhibitor with a Ki of greater than 10mM. The polyamines putrescine and spermidine inhibited ADI incompetitively with a Kii of 2.8mM for putrescine and 4.3mM for spermidine. Since the concentrations required for inhibition were within the range of the normal intracellular polyamine concentrations in Tetrahymena (maximally 14mM putrescine and 4mM spermidine), it is suggested that the polyamine effects on ADI are of regulatory nature. Thus, polyamine biosynthesis in Tetrahymena thermophila is regulated not only on the level of ornithine decarboxylase activity, but also on an earlier step, the supply of ODC with substrates.     

Distribution of polyamines in methanogenic bacteria

Members of all four families of methanogenic bacteria were analyzed for polyamine concentrations. High-performance liquid chromatography analysis of dansylated cell extracts revealed typical polyamine patterns for each family. Members of Methanobacteriaceae (family I) were characterized by very low polyamine concentrations; members of Methanococcaceae (family II) were characterized by putrescine and high spermidine concentrations; members of Methanomicrobiaceae (family III) were characterized by the presence of putrescine, spermidine, and sym-homospermidine; and members of Methanosarcinaceae (family IV) contained only high concentrations of sym-homospermidine in addition to putrescine. The highest polyamine concentration was found in Methanosarcina barkeri Jülich, with 0.35% putrescine in the dry cell material. The polyamine distribution found coincides with the dendrogram based on comparative cataloguing of 16S rRNA and offers a new, rapid chemotaxonomic method for characterizing methanogenic bacteria. Variation of the growth substrates (H2-CO2, methanol, acetate, and trimethylamine) for M. barkeri resulted in quantitative but not qualitative differences in polyamine composition.     

Polyamines and diamine oxidase activity in maternal, embryonal, and fetal tissues of rat after chronic ethanol consumption

The effects of maternal ethanol consumption for 4 weeks before and throughout gestation on polyamine content and diamine oxidase activity of maternal, embryonal and fetal tissues are reported. At the 12th day of pregnancy, a decrease of putrescine in the liver of the mother and marked increases in putrescine, cadaverine and spermidine in embryos were observed. At day 18, putrescine and cadaverine diminished in maternal liver and placenta, and no changes in amine content in fetal liver and brain were found. At day 12, diamine oxidase activity increased in maternal liver and placenta, whereas it greatly diminished in embryos. At day 18, enzyme activity decreased in maternal liver, placenta, fetal liver and brain. These results indicate that chronic ethanol ingestion induces alterations in polyamine concentrations and metabolism in growing and developing tissues during pregnancy that might contribute to the adverse effect of ethanol on conceptual development.     

Putrescine uptake in saintpaulia petals

Putrescine uptake and the kinetics of this uptake were studied in petals of Saintpaulia ionantha Wendl. Uptake experiments of [³H] or [¹⁴C] putrescine were done on single petals at room temperature at various pH values. The results show that putrescine uptake occurs against a concentration gradient at low external putrescine concentration (0.5-100 micromolar) and follows a concentration gradient at higher external putrescine concentrations (100 micromolar to 100 millimolar). 2,4-Dinitrophenol and carbonylcyanide-m-chlorophenylhydrazone, two uncouplers, had no effect on putrescine uptake. Uptake rates were constant for 2 hours, reaching a maximum after 3 to 4 hours. Putrescine uptake depended markedly on the external pH and two maxima were observed: at low external concentrations of putrescine, the optimum was at pH 5 to 5.5; at higher concentrations the optimum was at pH 8.     

Polyamine distribution and activity of their biosynthetic enzymes in the European sea bass (Dicentrarchus labrax L.) compared to the rat

1. In the liver, heart and brain of the European sea bass, putrescine concentrations are much higher than in the equivalent rat tissues; spermidine and spermine levels are smaller. 2. Ornithine decarboxylase in the bass liver is more active, but less stable than that in the rat; stability is acquired upon partial purification. Bass liver adenosylmethionine decarboxylase activity is less than that found in the rat. Both are activated and stabilized by putrescine. 4. The activating effect of putrescine decreases as the assay temperature is decreased. This may explain the high level of putrescine but low levels of spermidine and spermine in the bass liver.     

环切对苹果短枝顶芽内源多胺水平的影响及其与成花的关系

研究了环切对6年生富士、金冠苹果短枝内源多胶,包括腐胶、亚精胺和精胺的含量及其与成花的关系,发现内源多胺含量与成花作用显著相关。在花诱导阶段,Put和Spd的含量呈下降趋势,主枝环切处理在促进成花的同时减缓了多胺下降的速度,Spm的含量出现峰值;在形态分化阶段初期,Put和Spd的含量继续下降,至形态分化末期,含量迅速上升。而Spm含量呈下降趋势。表明Put有助于分生组织的活化,而Spd和Spm与花诱导的启动及促进有关。