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PubMed1990

Differential effects of sex steroids on uterine and renal ODC activity in ovariectomized rats.

Many hormones are known to induce the activity of ornithine decarboxylase (ODC), the first and rate-limiting enzyme for polyamine biosynthesis, in their target tissues. Using ovariectomized rats, we have compared the effects of sex hormones on ODC activity in the uterus and the kidney which contain estrogen and androgen receptors. The results show that 1) both estrogen and androgen stimulate renal ODC activity, 2) estrogen but not androgen effectively increases ODC activity in the uterus, 3) estrogen at higher dosage can stimulate renal ODC activity to an extent similar to that in the uterus, 4) daily treatment with estradiol for 5 days results in the desensitization of uterine ODC activity, but not that of renal ODC activity to the hormonal stimuli. Although both uterus and kidney are targets of sex hormones, our results indicate that estrogen and androgen have differential influences on the ODC activity in these two organs.

Androgens↗
PubMedDate not supplied

Effects of dietary arginine supplementation on protein turnover and tissue protein synthesis in scald-burn rats.

We assessed the effects of dietary arginine supplementation on protein turnover and organ protein synthesis in burned rats. Male Wistar rats weighing about 200 g underwent catheter jejunostomy and received scald burns covering 30% of the whole-body surface area. Animals were divided into a control group (n = 9) and an arginine group (n = 9) and continuously received total enteral nutrition for 7 d (250 kcal.kg-1.d-1, 1.72 gN.kg-1.d-1). Changes in body weight, plasma total protein, plasma albumin, urinary excretion of polyamines, nitrogen balance, whole-body protein kinetics, and tissue protein synthesis rates were determined. Whole-body protein kinetics and tissue fractional protein synthetic rates (Ks, percent/d) were estimated using a 24-h constant enteral infusion of 15N glycine on the last day. The changes in body weight were not different between the control and arginine groups. The urinary excretion of polyamines was higher in the arginine group than in the control group (P < 0.01). Burned rats enterally fed arginine-supplemented diet yielded significantly greater cumulative and daily nitrogen balance on days 3 and 5 than those fed a control diet (cumulative, P < 0.05; day 3, P < 0.01; day 5, P < 0.01). Whole-body protein turnover rate was significantly elevated in the arginine group as compared to that in the control group (P < 0.05). The Ks of rectus abdominis muscles were significantly increased in the arginine group in comparison to the control group (P < 0.01). We have shown that dietary arginine supplementation improved protein anabolism and attenuated muscle protein catabolism after thermal injury.

Animals↗
PubMed1995

A photoaffinity probe for the polyamine site regulating the NMDA receptor.

Polyamines produce on the NMDA-receptor channel activity a regulatory effect subsequent to their binding to specific sites on the receptor-complex. The photoactivatable polyamine derivative L-azidophenylspermine shows properties which suggest its potential as a photoaffinity probe to investigate the nature and topology of these sites. In the dark, its effect on the binding of tritiated N-(1-[thienyl]cyclohexyl)piperidine ([3H]TCP) to synaptosomal plasma membranes is similar to that of diaminodecane. Arcaine antagonizes both the effects of L-azidophenylspermine and diaminodecane on [3H]TCP binding. L-Azidophenylspermine competes in post-synaptic densities with tritiated spermidine for a unique binding site with an EC50 similar to that of spermine. Upon irradiation, L-azidophenyl-spermine incorporates into this material with a high efficiency to a level consistent with both the Bmax for tritiated spermidine and the estimated density of NMDA receptors in this fraction.

Affinity Labels↗
PubMed2006

Herminiimonas aquatilis sp. nov., a new species from drinking water.

A bacterial strain (CCUG 36956T) isolated from drinking water was taxonomically studied in detail. Phylogenetic analyses using the 16S rRNA gene sequence of the isolate indicated that it belongs to family Oxalobacteraceae of the beta-subclass of the Proteobacteria, with the highest sequence similarity of 99.3% to the type strain of Herminiimonas fonticola. In the polyamine pattern putrescine and 2-hydroxyputrescine were the predominant compounds. In the polar lipid profile major compounds were phosphatidyl ethanolamine and diphosphatidyl glycerol. Phosphatidyl glycerol and an unknown phospholipid were detected in moderate proportions. The major respiratory quinone was a ubiquinone Q-8 and the major whole cell fatty acids were 16:1 omega7c, 17:1 omega6c, and 16:0. The strain also contained 10:0 3-OH and other fatty acids typical for members of the genus Herminiimonas. The results of DNA-DNA hybridizations and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain CCUG 36956T from H. fonticola. For this reason, we propose that strain CCUG 36956T represents a new species of the genus Herminiimonas for which we propose the name Herminiimonas aquatilis sp. nov.

Bacterial Typing Techniques↗
PubMed1998

Polyamine profiles in the urine of patients with leukemia.

Eleven urinary polyamine levels were determined in controls (32 cases) and 43 patients with varying stages of leukemia including initial, relapse and complete remission, using gas chromatography nitrogen-phosphorus detection. Also, to indirectly evaluate the possible involvement of enzymes, precursor to product concentration ratios were compared between controls and patients with each stage of leukemia. As a result, it is confirmed that the ratio of N1-acSpd/N8-acSpd could be used as a diagnostic marker and the level of N1,N12-diacetylspermine could be used for determining disease stage and as a malignancy marker for leukemia. An altered metabolic pathway related to leukemia is also proposed in which N1,N12-diacetylspermine can be produced directly from spermine and N1,N12-diacetylspermine is a major source of N1-acetylspermidine.

Adolescent↗
PubMed2005

The involvement of polyamines as substrates of transglutaminase in zonal different hepatocyte proliferation after partial hepatectomy.

We have recently demonstrated the inverse correlation between transglutaminase (TGase) activity and DNA synthesis in periportal hepatocytes (PPH) and perivenous hepatocytes (PVH) at 1 d after partial hepatectomy. In order to elucidate a role of polyamines as substrates of TGase in the differential growth capacities between PPH and PVH from regenerating liver, we investigated the zonal differences in alteration of ornithine decarboxylase (ODC) activity and polyamines. In two subpopulations, the inverse correlation between DNA synthesis and epsilon-(gamma-glutamyl) lysine (Gln-Lys) cross-linking catalyzed by TGase was demonstrated at 1 d after partial hepatectomy. ODC activity in PPH significantly increased with a peak at 1 d after partial hepatectomy, whereas did not in PVH. Protein-binding SPD in PPH also transiently increased with a peak at 1 d after partial hepatectomy, but did not in PVH. These results suggest that at 1 d after partial hepatectomy, in PPH, the inhibition of Gln-Lys cross-linking by the formation of N-gamma-glutamyl SPD leads to the increase of DNA synthesis, whereas in PVH, enhanced formation of Gln-Lys cross-linking leads to the lower DNA synthesis.

Animals↗
PubMed1992

Developmental differences in antagonism of NMDA toxicity by the polyamine site antagonist ifenprodil.

Antagonists of 4 distinct regulatory sites on the N-methyl-D-aspartate (NMDA) receptor were tested for their ability to attenuate NMDA-mediated acute excitotoxicity in isolated chick retina of various embryonic ages between days 11 and 19 in ovo. Acute excitotoxicity was monitored by histology and by release of endogenous gamma-aminobutyric acid (GABA) into the medium during 30 min of incubation with 50 microM NMDA. The uncompetitive PCP channel site antagonist, MK-801, the competitive antagonist, CGS 19755, and the strychnine-insensitive glycine site antagonist, 7-chlorokynurenate, completely blocked NMDA-induced cell swelling and increased GABA release at all ages tested. Potencies versus NMDA were MK-801 greater than CGS 19755 greater than 7-chlorokynurenate with IC50S of 0.02, 0.62, and 15 microM, respectively. NMDA antagonism by the polyamine site antagonist, ifenprodil, differed from other classes of antagonists in several respects. At the earlier embryonic ages tested (E12-13) ifenprodil provided differential protection; completely blocking somal and neuritic swelling in most but not all inner nuclear layer neurons and inner plexiform processes. In dose-response studies, ifenprodil attenuated the NMDA-induced increase in medium GABA at all ages tested with an Imax of 10 microM. Ifenprodil, however, showed a decreased ability to completely protect some NMDA-sensitive neurons. This was reflected both histologically and by GABA release. Maximal attenuation of NMDA evoked GABA release was 83, 80, 62 and 50% at days E12, 13, 15 and 19, respectively. Histologically, differential protection was seen at E12 and 13, in limited areas at E15, and was no longer present at E19.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗
PubMed1995

Effects of polyamines on voltage-activated calcium channels in guinea-pig intestinal smooth muscle.

Effects of polyamines on the spontaneous mechanical and electrical activity of guinea-pig intestinal smooth muscle were studied. Spermine and spermidine inhibited action potential generation and contractions, while putrescine had no effect. Single smooth muscle cells were isolated from the longitudinal muscle layer of the guinea-pig ileum. Whole-cell voltage-clamp experiments were carried out to investigate the effects of polyamines on current through voltage-activated Ca2+ channels. Spermine and spermidine (0.1-1 mM) reduced the inward current in a concentration-dependent manner. Spermine blocked current activated by the dihydropyridine agonist BAY K 8644 (1 microM), whereas no additional inhibition by spermine was seen after blockage of dihydropyridine-sensitive channels by nifedipine (0.1 microM). Inhibition by spermine or spermidine did not shift the peak of the current voltage relation of the inward current. Steady-state activation and inactivation relationships were not affected and thus the amplitude, but not the voltage dependence, of the window current responsible for Ca2+ inflow during sustained depolarization was affected. Putrescine (1 mM) had no significant effect on the inward current. These results suggest that spermine and spermidine inhibit contraction in spontaneously active intestinal smooth muscle by inhibiting Ca2+ current responsible for generation of action potentials.

Action Potentials↗
PubMed1991

Regulation of polyamine and magnesium inhibition of the N-methyl-D-aspartate (NMDA) receptor ionophore complex.

The inhibitory effects of spermidine, putrescine and magnesium on [3H]TCP binding in crude synaptosomes and well washed buffy coat membranes were studied. We report that the IC50 values for the drugs tested varied depending on the state of activation of the channel, being lowest when the channel is maximally activated and highest when the channel is least activated in the control buffy coat preparation. Comparison of the inhibitory characteristics of these 3 agents when activated by glutamate, glycine, and low concentrations of spermidine or magnesium strongly suggest that putrescine and magnesium share a common inhibitory mechanism, perhaps mediated by the voltage-dependent channel site for magnesium. Inhibition by spermidine, however, is clearly mediated by a distinct site, non-identical to the phencyclidine receptor. Glutamate appears to uncouple this inhibition in about one-half of the N-methyl-D-aspartate (NMDA) complexes, suggesting the possibility of distinct subpopulations of NMDA-operated ion channels. The potential physiological relevance of these findings is discussed.

Animals↗
PubMed2002

Heterologous expression of a bacterial homospermidine synthase gene in transgenic tobacco: effects on the polyamine pathway.

Homospermidine synthase (HSS) is a branch-point enzyme that links the secondary pathway (pyrrolizidine alkaloids) to primary metabolism (polyamines). Since the diamine putrescine is a precursor of homospermidine and nicotine in tobacco, we performed heterologous expression of a bacterial homospermidine synthase gene (hss)in Nicotiana tabacum and determined the effect on free and conjugated polyamine levels. The hss gene from Rhodopseudomonas viridis was placed under the control of the cauliflower mosaic virus (CaMV) 35S promoter in Agrobacterium tumefaciens Ti-plasmid in sense and antisense orientation and both hss constructs were transformed into tobacco plants. Expression of the hss gene was verified by "Northern" and "Southern Blot" analysis. 2 transgenic sense lines were generated from 1000 calli which showed weak expression of homospermidine synthase, i.e. 50 pktal/mg protein and 45 pktal/mg protein. These transgenic sense plants showed a significantly decreased content of free spermidine while the pool of conjugated spermidine was not affected. The 2 sense plants exhibited a range of abnormal phenotypes such as dwarfness and stunted growth. Homospermidine was sporadically detectable in wild type tobacco. To our knowledge, this is the first biotechnological approach to express a prokaryotic homospermidine synthase gene in tobacco plants.

Alkyl and Aryl Transferases↗
PubMedDate not supplied

[Effect of polyamines on actomyosine-ATP-ase activity in smooth muscles of the uterus].

The ability of the aliphatic polyamines to inhibit [figure: see text] the ATPase activity of smooth muscle actomyosine satisfies the succession: spermine > spermidine > putrescine that is correlated with magnitude of positive charge at physiological value of pH. The most effective inhibitor of the ATP hydrolysis process is the spermine, which highest inhibitory action is manifested at 10(-3) M concentration, in lesser concentration (10(-5) M) activates the actomyosine ATPase. While defining the kinetic parameters of the ATP hydrolysis reaction catalyzed by uterus myometrium the correlation between inhibiting the ATPase activity of myometrium contractile complex under introduction into the incubation medium of 10(-3) M spermine and decreasing the affinity of actomyosine for ATP was made; the activating effect of spermine on ATPase activity of actomyosine complex in the presence of 10(-5) M spermine correlated with the increase of actomyosine affinity for Mg2+.

Adenosine Triphosphatases↗
PubMed2007

Biological polyamines inhibit nucleic-acid-induced polymerisation of prion protein.

Nucleic-acid-induced polymerisation of prion protein, when monitored by anilino naphthalene sulfonic acid dye, shows, successively, an immediate fluorescence increase of the dye upon mixing of the reactants, followed by a lag period in which the dye fluorescence remains unchanged, and then a phase in which dye fluorescence increases with time. The biological polyamines spermine and spermidine reduce the extent of the initial fluorescence increase, increase the lag period, and reduce both the rate and the extent of increase in fluorescence intensity of the dye in the final phase of the reaction. Spermidine is less effective than spermine in all of these processes. A nearly fivefold lower concentration of spermine can inhibit polymerisation of prion protein by tRNAs compared to the same process induced by double-stranded nucleic acid. The change in the secondary structure of the globular domain of the protein induced by nucleic acid is reversed by the addition of spermine, and it prevents structural destabilization of this domain induced by nucleic acids. It is suggested that physiological event(s) that would reduce the concentrations of intracellular biological amines may make nucleic acid available to induce oligomerization and polymerisation of cellular prion protein related to prion disease.

Anilino Naphthalenesulfonates↗
PubMed1998

Two mechanisms for inward rectification of current flow through the purinoceptor P2X2 class of ATP-gated channels.

1. The ATP receptor subunit P2X2 was expressed in Xenopus oocytes and human embryonic kidney (HEK) 293 cells. ATP-activated currents were studied with two-electrode voltage clamp recordings from oocytes, whole-cell recordings from HEK 293 cells, and outside-out patch clamp recordings from both cell types. The steady-state current-voltage (I-V) relation showed profound inward rectification in all recording configurations. 2. Recordings from outside-out patches demonstrated that inward rectification does not require intracellular Mg2+ or polyamines, and that inward rectification was present when the same solution was used on both sides of the patch. 3. Voltage jump experiments were performed to evaluate the voltage dependence of channel gating. After fast voltage jumps, instantaneous current jumps were followed by substantial relaxations to the steady state. The time course of the current relaxations could be fitted by single exponential functions. The instantaneous I-V relation was less inwardly rectifying than the steady-state I-V relation; however, it was not linear. 4. Single channel recordings indicated that the single channel conductance became smaller when the membrane potential became more positive. This decrease could quantitatively account for inward rectification of the instantaneous I-V relation. 5. We conclude that inward rectification of P2X2 is due to two mechanisms: voltage-dependent gating and voltage dependence of the single channel conductance.

Adenosine Triphosphate↗
PubMed2002

Polyamine synthesis in plants. Purification and properties of amidinotransferase from soybean (Glycine max) axes.

Three-day-old soybean (Glycine max) seedlings were exposed to 0.4 M sorbitol solution for 4 h to induce amidinotransferase activity, with the corresponding enzyme being purified to homogeneity by chromatographic separation on DEAE-Sephacel, Sephacryl S-300 and L-arginine Sepharose 4B. The purified enzyme used L-arginine and L-glycine as the major donor/acceptor of the amidino group, respectively, with formation of guanidinoacetic acid and ornithine products being confirmed by ESI-MS. The enzyme is a tetrameric protein having a molecular mass of 240,000 Da, whose thiol group is needed for enzymatic activity. The K(M)s for arginine and glycine were 3.8 and 0.89 mM, respectively, with optimal temperature and pH being 37 degrees C and 9.5, respectively. The soybean amidinotransferase could be indirectly involved in nitrogen metabolism, as suggested by the observation that arginine:glycine amidinotransferase in soybean axes is indirectly involved in putrescine biosynthesis and displays feedback control at high levels of an endogenous regulator, putrescine.

Amidinotransferases↗
PubMed1990

A study on the vital reaction in wounded skin: simultaneous determination of histamine and polyamines in injured rat skin by high-performance liquid chromatography.

This paper describes a modified method of simultaneous quantitative determination of histamine and polyamines (putrescine, cadaverine, spermidine and spermine) in incised skin wounds of rats using high-performance liquid chromatography (HPLC) with a strong-cation exchange column, shim-pack ISC-05/S0504P, employing a linear gradient of 0.7-2.5 M sodium chloride. There was scarcely any change in the histamine content of the vital wounds throughout the one week experimental period, although content decreased slightly in 1-h-old wounds. On the other hand, concentrations of putrescine and spermidine increased suddenly 12 h after wounding, and spermine content increased slightly in 4-day-old wounds. There was no post-mortem change in the histamine, spermidine and spermine contents of vital wounds allowed to stand at 25 degrees C up to 24 h after death. Based on these results, this HPLC method may be useful for the determination of wound age during the period of healing.

Animals↗
PubMed1992

Polyamine levels, ornithine decarboxylase (ODC) activity, and ODC-mRNA expression in normal and cancerous human colonocytes.

Ornithine decarboxylase (ODC) and polyamines (putrescine, spermidine, and spermine) are crucial for cell proliferation. Recently, elevated ODC activity and polyamine levels have been suggested as biological markers for human colon cancer. In this study, we measured ODC activity and the levels of polyamines (putrescine, spermidine, spermine, and cadaverine) and acetyl-putrescine in human colonocytes isolated from cancerous areas compared to the adjacent normal colon tissue. In addition, ODC mRNA expression was compared between both groups. We found that colonocytes isolated from cancerous areas had significantly higher mean value of ODC activity, putrescine, spermidine, spermine, and cadaverine levels up to 1480%, 470%, 260%, 380%, and 510% respectively compared to colonocytes isolated from the adjacent normal colonic mucosa. No difference was found in acetyl-putrescine levels between cancerous and normal colonocytes. Steady-state levels of ODC mRNA were slightly elevated in cancerous colonocytes relative to normal colonocytes in two of three paired samples. However, the increase in ODC mRNA levels is not sufficient to account for the increase in ODC activity suggesting that colonocyte ODC activity is regulated post-transcriptionally.

Adult↗
PubMed2002

Cerebral distribution of polyamines in kainic acid-induced models of status epilepticus and ataxia in rats. Overproduction of putrescine and histological damage.

We study the brain regional distribution of putrescine after excitotoxic damage. After status epilepticus induced by kainic acid (9 mg/kg, i.p.) we found an increase of putrescine in all the regions analyzed. Three kind of regions can be identified according to the magnitude and persistence of the abnormal concentration of putrescine: hippocampus and cortex (9-fold 1 day, still increased 2-fold 2 weeks after injection), cerebellum and medulla oblongata+pons (2-fold 1 day after injection) and the other regions (7-8-fold 1 day, still increased 2-3-fold 1 week after injection). The histological damage was: severe, absent and moderate or low, respectively. After ataxia induced by kainic acid injection (2.34 nmols) into the cerebellum, putrescine also rises in all regions; a high concentration (9-fold) and severe damage was found in the injected cerebellar hemisphere. In conclusion, in the models studied, putrescine increases in all the regions analyzed. However, the highest concentrations and the most severe damage were found in the target regions.

Analysis of Variance↗
PubMed1990

Putrescine decreases exploration of a black and white maze.

The effect of putrescine and cyclohexylamine on rat cortical polyamine concentration and on behavior in a black and white maze was studied. The levels of polyamines in brain cortex were determined 15 min, 2, 4, and 6 hours after injection of putrescine (200 or 400 mg/kg) or cyclohexylamine (380 mg/kg). Putrescine concentration increased 6-fold 15 min after injection of putrescine followed by a decline during the next 6 hours. Cyclohexylamine increased putrescine concentration doubling it 4 hours after injection. Spermidine and spermine concentrations did not change after either putrescine or cyclohexylamine injection. Behavior was studied in the Greek cross maze which provides the choice to enter either white or black compartments. Putrescine 200 mg/kg decreased entries into white but not black compartments, while putrescine 400 mg/kg decreased entries into both. The effect of cyclohexylamine was similar to putrescine 400 mg/kg. The behavioral effect of each treatment was independent of the time between injection and testing for up to 6 hours, while the levels of putrescine changed during the same period. Therefore, behavior was not directly related to total cortical putrescine.

Animals↗