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PubMed2004

Polyamine analysis for chemotaxonomy of thermophilic eubacteria: Polyamine distribution profiles within the orders Aquificales, Thermotogales, Thermodesulfobacteriales, Thermales, Thermoanaerobacteriales, Clostridiales and Bacillales.

Cellular polyamines of 45 thermophilic and 8 related mesophilic eubacteria were investigated by HPLC and GC analyses for the thermophilic and chemotaxonomic significance of polyamine distribution profiles. Spermidine and a quaternary branched penta-amine, N4-bis(aminopropyl)norspermidine, were the major polyamine in Thermocrinis, Hydrogenobacter, Hydrogenobaculum, Aquifex, Persephonella, Sulfurihydrogenibium, Hydrogenothermus, Balnearium and Thermovibrio, located in the order Aquificales. Thermodesulfobacterium and Thermodesulfatator belonging to the order Thermodesulfobacteriales contained another quaternary penta-amine, N4-bis(aminopropyl)spermidine. In the order Thermotogales, Thermotoga contained spermidine, norspermidine, caldopentamine and homocaldopentamine. The latter two linear penta-amines were not found in Marinitoga and Petrotoga. In the order Thermales, Thermus and Marinithermus contained homospermidine, norspermine and the linear penta-amines. Meiothermus lacked penta-amines. Vulcanithermus contained linear penta-amines and hexa-amines but not homospermidine. Oceanithermus contained spermine alone. Within the order Thermoanaerobacteriales, the two quaternary branched penta-amines were found in Thermanaeromonas and Thermoanaerobacter. Caldanaerobacter contained N4-bis(aminopropyl)spermidine. Thermoanaerobacterium lacked penta-amines. Thermaerobacter of the order Clostridiales contained N4-bis(aminopropyl)spermidine and agmatine. Thermosyntropha, Thermanaerovibrio, Thermobrachium ( the order Clostridiales), Sulfobacillus, Alicyclobacillus, Anoxybacillus, Ureibacillus, Thermicanus ( the order Bacillales), Desulfotomaculum, Desulfitobacterium and Pelotomaculum (the family Peptococcaceae) ubiquitously contained spermine. Some thermophiles of Bacillales added linear and branched penta-amines.

Bacteria↗
PubMed1990

Effects of starvation and difluoromethylornithine (DFMO) on diamine oxidase activity in rat ileum.

Starvation and difluoromethylornithine (DFMO) administration have profound affects on intestinal proliferation, ornithine decarboxylase activity, and tissue polyamine levels. Diamine oxidase activity may play a role in the regulation of proliferation, and the activity of this enzyme may be influenced by ornithine decarboxylase activity. To determine if diamine oxidase is influenced by starvation and DFM administration, ileal diamine oxidase activities were determined on mucosal homogenates from five groups of rats: fed control, starved for 48 h, fed group receiving DFMO, a starved/refed group, and a starved/refed group receiving DMFO. The homogenates from starved rats were found to have decreased ornithine decarboxylase activity and increased diamine oxidase activity when compared to control values. The homogenates from the DFMO group also were found to have decreased ODC activity however, mucosal diamine oxidase activity was also decreased. Refeeding produced a dramatic increase in ornithine decarboxylase activity and a minimal change in diamine oxidase activity. The preservation of diamine oxidase activity during starvation implies a need for the enzyme not related to mucosal proliferation or digestion. However, in the fed state, diamine oxidase activity may be more dependent on ornithine decarboxylase activity or its reaction product putrescine.

Amine Oxidase (Copper-Containing)↗
PubMed1997

Degradation of the polyamine alkaloid aphelandrine by endophytic fungi isolated from Aphelandra tetragona.

Members of the genus Aphelandra (Acanthaceae) produce rare macrocyclic polyamine alkaloids which consist of spermine acylated with two units of 3-(4-hydroxyphenyl)prop-2-enoic acid. Endophytic fungi were isolated from roots and shoots of Aphelandra tetragona and tested for their ability to metabolize the main alkaloid aphelandrine, which accumulates exclusively in the roots of the plants. Several endophytes were able to metabolize aphelandrine but only root endophytes belonging to the Nectriaceae were good metabolizers. In addition, the endophytes were grown on an agar medium containing putrescine, spermidine, or spermine as the sole nitrogen source. All fungi were able to grow on putrescine, but only the good aphelandrine metabolizers grew well on spermidine or spermine. Acremonium sp. 15, one of the most active metabolizers, grew also on a medium containing aphelandrine as sole nitrogen source. A number of strains thought to be conspecific with Acremonium sp. 15 were also tested for their ability to metabolize aphelandrine. The ability of the endophytes to metabolize aphelandrine suggests an ecological adaptation of the symbionts to their host. The possibility of using the aphelandrine metabolism as a taxonomic character is briefly discussed.

Alkaloids↗
PubMed2002

Plant C-N hydrolases and the identification of a plant N-carbamoylputrescine amidohydrolase involved in polyamine biosynthesis.

A nitrilase-like protein from Arabidopsis thaliana (NLP1) was expressed in Escherichia coli as a His(6)-tagged protein and purified to apparent homogeneity by Ni(2+)-chelate affinity chromatography. The purified enzyme showed N-carbamoylputrescine amidohydrolase activity, an enzyme involved in the biosynthesis of polyamines in plants and bacteria. N-carbamoylputrescine amidohydrolase activity was confirmed by identification of two of the three occurring products, namely putrescine and ammonia. In contrast, no enzymatic activity could be detected when applying various compounds including nitriles, amines, and amides as well as other N-carbamoyl compounds, indicating the specificity of the enzyme for N-carbamoylputrescine. Like the homologous beta-alanine synthases, NLP1 showed positive cooperativity toward its substrate. The native enzyme had a molecular mass of 279 kDa as shown by blue-native polyacrylamide gel electrophoresis, indicating a complex of eight monomers. Expression of the NLP1 gene was found in all organs investigated, but it was not induced upon osmotic stress, which is known to induce biosynthesis of putrescine. This is the first report of cloning and expression of a plant N-carbamoylputrescine amidohydrolase and the first time that N-carbamoylputrescine amidohydrolase activity of a recombinant protein could be shown in vitro. NLP1 is one of the two missing links in the arginine decarboxylase pathway of putrescine biosynthesis in higher plants.

Arabidopsis↗
PubMed1989

[Antitumor effect of alpha-difluoromethylornithine (DFMO) changes in ornithine decarboxylase (ODC) activity and polyamine (PA) levels in human tumor transplanted into nude mice].

Three human carcinoma xenografts serially transplanted into nude mice were used for the study of antitumor effect of DFMO. One breast carcinoma (MX-1) and two colon carcinomas (Exp-42, Co-3) were inoculated into the subcutaneous tissue of BALB/c nude mice. When the tumor weights reached 100 -300mg, 0.5%, 1% and 2% DFMO was administrated ad libitum in tap water for 21 days, MMC was administrated ip q7d x 3(DAY 0, 7,14) at 0.5mg/kg or 2.0mg/kg. Combination of 1% DFMO and MMC 1mg/kg x 3 using a similar schedule was also made. After 21 days, the tumors were weighed and assays of ODC activity and analysis of PA in the tumors were done. The antitumor effects (the lowest value of TRW/CRW) of 2% DFMO against MX-1, Exp-42 and Co-3 were 4.83%, 35.8% and 73.4% respectively. Synergic effects of MMC were observed in all three tumors. With 2% DFMO, ODC and PUT levels were decreased to 16.4%, 2.89% of control in MX-1, 14.8%, 6.62% in Exp-42, 4.18%, 11.2% in Co-3 respectively. Of MX-1 and Exp-42, the dose of DFMO administrated, the antitumor effect and the PUT levels were correlated (p less than 0.05). DFMO is one of the potent chemotherapeutic agents and may be useful in combination chemotherapy.

Animals↗
PubMed2004

[Effects of exogenous spermidine on resistance of Nymphoides peltatum to Hg2+ stress].

Effects of different exogenous spermidine concentrations on the activities of some enzymes (SOD, CAT, POD, APX), O(-)(2) generation rate, the MDA, chlorophyll, soluble protein and polyamine contents in leaves of Nymphoides peltatum without Hg(2+) stress or under Hg(2+) stress were studied. The results showed that: (1) The spermidine (Spd) and spermine (Spm) content dropped significantly under Hg(2+) (3 mg/L) stress 4 d, while that of putrescine (Put) was increased markedly. Exogenous Spd could increase the Spd and Spm content in leaves, while that of Put lowered at first and rose later. (2) Hg(2+) 3 mg/L decreased drastically the activities of SOD, CAT and APX, increased the O(-)(2) generation rate, led to the accumulation of a large amount of MDA and loss of chlorophyll and soluble protein. Exogenous Spd could enhance the activities of SOD, CAT and APX, decrease O(-)(2) generation rate and reduce the MDA content. In addition, Spd could also decrease the loss of chlorophyll and soluble protein content induced by Hg(2+) stress, enhance the ability of Nymphoides peltatum to resist Hg(2+) stress. The most suitable concentration of Spd was in the range 0.1-0.5 mmol/L.

Biogenic Polyamines↗
PubMed1989

The effects of graded doses of a cholecystokinin-like peptide with and without secretin on pancreatic growth and synthesis of RNA and polyamines in rats.

The dose dependence of a cholecystokinin-like peptide (CCK-LP) on the trophic response in the rat pancreas was studied. Graded doses of Thr28Nle31CCK25-33 (0.02, 0.1, 0.5, 2.5, and 12.5 micrograms/kg/h) or saline were given as a continuous intravenous infusion to conscious and fed rats for 8 and 48 h. Secretin (5.0 micrograms/h) was given alone or combined with the three highest doses of CCK-LP for 48 h. CCK-LP showed a dose-dependent stimulating effect on pancreatic growth and synthesis of RNA and polyamines. The threshold dose ranged from 0.02 to 0.5 micrograms/kg/h and was lowest for stimulation of ornithine decarboxylase (ODC). The maximal effects on protein, RNA, and DNA contents were achieved with 2.5 micrograms/kg/h. These same variables markedly decreased with 12.5 micrograms/kg/h, whereas marked further increases were found for the activities of RNA polymerase, DNA polymerase, and thymidine kinase. This same dose of CCK-LP caused after 8 h of treatment a marked and transient increase in pancreatic weight, activity of ODC, and concentration of putrescine. When secretin was added to 0.5 and 2.5 micrograms/kg/h of CCK-LP, no additional effect (except for ODC) was found. When secretin was added to the highest dose of CCK-LP, the decreased contents of protein and RNA were significantly increased, and the markedly increased activities of RNA- and DNA-synthesizing enzymes were significantly decreased. The present study shows a clear dose-response relationship for the trophic effect of CCK-LP on the rat pancreas and indicates that the growth effect of a supramaximal dose includes components of regeneration secondary to damage.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2005

Ornithine decarboxylase activity is inhibited by the polyamine precursor amino acids at the protein stability level in Caco-2 cells.

High concentrations of certain amino acids are known to affect hormonal secretion, immune function, electrolyte balance or metabolic functions. However, there is a lack of knowledge regarding the molecular mechanisms responsible for these effects. We showed that, as well as spermidine transport, the activity of ornithine decarboxylase (ODC), the first and rate-limiting enzyme in polyamine biosynthesis, is decreased in human colon adenocarcinoma cells, Caco-2, following a 4-h supplementation with one of the two polyamine precursor amino acids, L-arginine or L-methionine. Dose-response assays indicated that the inhibitory effect of supplemental L-methionine was stronger than that of supplemental L-arginine. However, it was transient, being even replaced by ODC induction after 8 h, whereas the inhibitory effect of L-arginine lasted for at least 8 h. Unlike L-cysteine, neither L-methionine nor L-arginine could inhibit ODC activity in a crude acellular preparation of the enzyme. The inhibition of ODC activity in cells exposed to L-methionine or L-arginine was due to a decreased abundance of ODC protein without change at the mRNA level and each of these amino acids could counteract ODC induction by a glycine supplement. Contrary to the latter, supplemental L-methionine or L-arginine induced a marked decrease in ODC half-life, concomitantly with an increase in the activity of antizyme, an ODC inhibitory protein. Thus, depending on their nature, amino acids can up- or downregulate ODC activity at the protein stability level.

Arginine↗
PubMed1988

Abnormal polyamine metabolism in hypertensive cardiac hypertrophy.

In order to assess myocardial hypertrophic activity during the process of hypertensive cardiac hypertrophy in the presence and absence of treatment with anti-hypertensive agents, we analyzed myocardial polyamine concentrations in spontaneous hypertensive (SHR) rats and control rats of Wistar Kyoto (WKY) strain. The anti-hypertensive agents studied were diltiazem, hydralazine and captopril, each of which was administered for 5 weeks. In comparison with WKY rats, SHR rats showed elevated blood pressure and enlarged hearts with higher myocardial spermidine concentration. Although blood pressure was lowered in the diltiazem-treated SHR rats, heart weight and myocardial spermidine concentration increased as in untreated SHR rats. In the hydralazine-treated group increases in both blood pressure and myocardial spermidine concentration were suppressed, while an increase in heart weight was not. In the captopril-treated group, increases in blood pressure, heart weight and spermidine concentration were all suppressed. Since spermidine level appears to be a sensitive indicator of hypertrophic activity in the heart, this study suggests that captopril exerts an inhibitory effect on hypertensive cardiac hypertrophy whereas diltiazem does not. It also suggests that hypertrophy may reach a certain plateau level earlier in the hydralazine-treated animals than in others.

Animals↗
PubMed2003

Onset of DNA aggregation in presence of monovalent and multivalent counterions.

We address theoretically aggregation of DNA segments by multivalent polyamines such as spermine and spermidine. In experiments, the aggregation occurs above a certain threshold concentration of multivalent ions. We demonstrate that the dependence of this threshold on the concentration of DNA has a simple form. When the DNA concentration c(DNA) is smaller than the monovalent salt concentration, the threshold multivalent ion concentration depends linearly on c(DNA), having the form alphac(DNA) + beta. The coefficients alpha and beta are related to the density profile of multivalent counterions around isolated DNA chains, at the onset of their aggregation. This analysis agrees extremely well with recent detailed measurements on DNA aggregation in the presence of spermine. From the fit to the experimental data, the number of condensed multivalent counterions per DNA chain can be deduced. A few other conclusions can then be reached: 1), the number of condensed spermine ions at the onset of aggregation decreases with the addition of monovalent salt; 2), the Poisson-Boltzmann theory overestimates the number of condensed multivalent ions at high monovalent salt concentrations; and 3), our analysis of the data indicates that the DNA charge is not overcompensated by spermine at the onset of aggregation.

Adsorption↗
PubMed2003

Biomarkers and their use in cervical cancer chemoprevention.

Cervical cancer chemoprevention agents under study include diet and micronutrients (particularly beta-carotene, folate, and vitamins A, C, and E); medications such as retinoids (retinyl acetate gel, all-trans-retinoic acid, and 4-hydroxyphenylretinamide) that are chemically related to micronutrients; and other chemopreventives meant to affect the carcinogenic process at the cellular level, including such polyamine synthesis inhibitors as alpha-difluoromethylornithine. Agents become reasonable candidates for study when they have a biologic rationale, they are of low toxicity, and they can be taken for a long period of time. Since the human papillomavirus (HPV) is the major etiologic agent, the medication should show activity against HPV-positive preinvasive and invasive cell lines. The medication needs to be of low toxicity because it may be taken for long periods of time and less toxicity is tolerated in the precancerous setting. Until 1995, none of the studies used surrogate end point biomarkers (SEBs), relying instead on histologic and colposcopic regression as end points. All studies typically included subjects with cervical intraepithelial neoplasia. Conclusions to be drawn from these studies include the following: Though micronutrients are logical candidates for chemoprevention, they haven't worked consistently, and the reasons remain unclear. Furthermore, SEBs need to be validated in phase I trials. Finally, a better understanding of the role of HPV needs elucidation, including an understanding of the relationship of the medication to HPV status and of the immunobiology of HPV throughout the trial.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed2003

[Relationship between floral differentiation and endogenous hormones and polyamines contents in cucumber cotyledonary nodes cultured in vitro].

Changes of endogenous hormones and polyamines(PAs) contents during floral differentiation (0-6 day) in cucumber cotyledonary nodes cultured in vitro were determined by using HPLC. The results showed that all four endogenous hormones decreased obviously within 0-2 day and increased slightly within 4-5 day after culture. This indicated that low levels of indole-3-acetic acid (IAA), gibberelline(GA3) and abscisin acid(ABA) during 0-2 day were favorable to floral primordia differentiation, while high level of zeatin(ZT) during 3-5 day was favorable to floral organ primordia differentiation. Contents of spermine(Spm), spermidine(Spd) and cadavarine(Cad) were decreased within 0-1 day, but increased within 1-4 day, and then decreased again within 4-5 day after culture. On the contrary, content of putrescine(Put) was increased significantly on the first day and then declined during 1-6 day. Changes of PAs contents indicated that high levels of PAs and Put were favorable to floral primordia differentiation, the increasing of Spm after the second day was favorable to floral organ primordia differentiation, and change of Cad content may be one of the characteristic symbols between floral and vegetative bud differentiation.

Biogenic Polyamines↗
PubMed1988

Effect of estradiol on the activities of ornithine decarboxylase and S-adenosyl-methionine decarboxylase in tissues of ovariectomized rats.

Polyamines play an important role in cellular growth and differentiation. Ornithine decarboxylase (ODC) and S-adenosyl-methionine decarboxylase (SAMD) catalyze the rate-limiting steps of polyamine formation. Many hormones have been known to affect these decarboxylase activities in their target tissues. This study investigated the effect of estradiol (E2) on the activities of ODC and SAMD and on the growth of three estrogen receptor-containing organs: uterus, kidney and liver. Two weeks after ovariectomy, Sprague-Dawley rats received a subcutaneous injection of E2 (10 micrograms/rat). The ODC activity and organ weight were determined at various intervals after E2 administration. The E2 treatment increased the weight of uterus but not that of liver and kidney. In the uterus, ODC activity showed biphasic increases at 6 h and 16-24 h respectively after E2 injection. Renal ODC activity increased gradually and peaked at 18-24 h whereas hepatic ODC activity remained unchanged. Daily injection of E2 over a 3-day period provoked an increase of uterine weight, uterine ODC and SAMD activity. The same treatment also increased renal ODC activity, however it had no effect on the renal SAMD activity and the kidney Our results indicate that E2 has different effects on the activities of ODC and SAMD in three organs tested, and the increase of ODC activity is not necessarily correlated with the tissue growth.

Adenosylmethionine Decarboxylase↗
PubMed2001

Polyamine metabolism and glutamate receptor agonists-mediated excitotoxicity in the rat brain.

Putrescine (PUT) increases have been seen in a range of models of neuropathological disturbances. The present study was designed to compare the ability of various types of glutamate receptor agonist to promote excitotoxic brain damage and to examine whether a PUT increase is a general marker of excitotoxic brain damage. To that end, we evaluated features of brain damage associated with the excitotoxicity induced by both ionotropic glutamate receptor (iGluR) and metabotropic glutamate receptor (mGluR) agonists in the conscious rat and the changes produced in the regulation of polyamine metabolism. Intracerebroventricular infusion of N-methyl-D-aspartate (NMDA; 80 nmol), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA; 15 nmol), kainic acid (KA; 2.3 nmol), (R,S)-3,5-dihydroxyphenylglycine (3,5-DHPG; 1.5 micromol), and (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD; 2 micromol) produced similar seizure incidences (76-84%) in the rat. The convulsant episodes appeared sooner after iGluR (13-22 min) than after mGluR agonists (50-179 min). Histological analysis of the hippocampus 24 hr after seizures indicated several degrees of excitotoxic injury after equiconvulsive doses of the iGluR and mGluR agonists assayed. The agonists can be placed in the following order, according to the degree of damage they produce: AMPA > 3,5-DHPG approximately KA > NMDA > 1S,3R-ACPD. In the frontal cortex, moderate to low levels of damage were observed after all GluR agonists. Both iGluR- and mGluR-induced seizures produced an overshoot in the hippocampal and cortical PUT concentration, whereas spermidine and spermine levels were similar to control. Moreover, a concurrence of increased PUT levels and brain damage was observed, indicating that PUT is a general marker of excitotoxic brain damage.

Animals↗
PubMed1990

Effect of fasting and feeding on polyamines and related enzymes along the villus: crypt axis.

Fasting and feeding have profound effects on crypt cell production and small bowel mucosal growth but the mechanism whereby food stimulates villus tip enterocytes to influence crypt cell production is unknown. We therefore measured the activities of ornithine decarboxylase (ODC), diamine oxidase (DAO) and alkaline phosphatase (ALP--a marker of enterocyte maturity) and polyamine concentrations in epithelial cells from villus tips, mid villi, lower villi and crypts of small intestine in non-fasted controls and 18-24 h fasted rats. Fasting reduced crypt cell production and caused villus hyperplasia, DAO activity (mU/g) increased in control villus tips from 9.6 +/- (SEM) 0.8 to 12.3 +/- 1.5 after fasting (NS), from 7.6 +/- 0.4 to 13.9 +/- 3.0 in mid villi (p less than 0.01), from 5.7 +/- 1.0 to 10.4 +/- 7.4 in lower villi (p less than 0.01) and from 5.4 +/- 0.9 to 12.8 +/- 1.5 in the crypts (p less than 0.001). ALP showed a similar pattern of results. In contrast, fasting lowered ODC activity (pmol/mg protein/h) dramatically from 319 +/- 82 in control villus tips to 16.7 +/- 3.0 during fasting, from 297 +/- 59 to 10.7 +/- 3.6 in mid villi, from 224 +/- 45 to 6.3 +/- 2.8 in lower villi and from 150 +/- 31 to 5.8 +/- 3.3 in the crypts. Fasting reduced putrescine concentrations in all fractions but particularly in the crypts and in general was associated with increases in spermidine and spermine concentrations. The role of DAO in the maintenance of low putrescine concentrations during fasting is unclear.

Alkaline Phosphatase↗
PubMed1995

Inhibition of oral carcinogenesis by the arotinoid mofarotene (Ro 40-8757) in male F344 rats.

The chemopreventive effect of dietary administration of a new arotinoid, mofarotene (Ro 40-8757), which contains a morpholine structure in the polar end group, during the initiation phase of 4-nitroquinoline 1-oxide (4-NQO)-induced oral carcinogenesis was investigated in male F344 rats. Also, modulatory effects of this compound on polyamine levels (biomarkers of proliferation), the 5-bromodeoxyuridine-labeling index and the number of silver stained nucleolar organizer region proteins (AgNORs)/nucleus were assessed in the target epithelium. Rats were fed Ro 40-8757 at concentrations of 250 and 500 p.p.m. for 10 weeks. One week after the commencement of the diets, 4-NQO (20 p.p.m.) was administered in the drinking water for 8 weeks. Feeding of Ro 40-8757 at both doses caused a 78% reduction in the incidence of tongue neoplasms (squamous cell papilloma and carcinoma) by 32 weeks when compared with rats treated with 4-NQO alone (P < 0.05). Similarly, in rats treated with 4-NQO together with Ro 40-8757 the incidence of preneoplastic lesions (hyperplasia and dysplasia) was significantly less than the 4-NQO alone group (P < 0.05). Expression of three biomarkers was also decreased significantly by dietary treatment with Ro 40-8757. Thus a new arotinoid, Ro 40-8757, inhibited the oral carcinogenesis induced by 4-NQO when it was administered concurrently with the carcinogen. These results might suggest the possible application of Ro 40-8757 for cancer chemoprevention in the oral cavity, in addition to the breast.

Animals↗
PubMed1994

Ornithine decarboxylase in Pneumocystis carinii and implications for therapy.

Pneumocystis carinii pneumonia (PCP) can be treated with eflornithine (difluoromethylornithine, DFMO, Ornidyl), a competitive irreversible inhibitor of ornithine decarboxylase (ODC), a key enzyme for polyamine biosynthesis. Because ODC has been reported to be absent from P. carinii, it has been assumed that eflornithine affects P. carinii only indirectly, by affecting host polyamine biosynthesis. If this is true, then improvements in the selectivity of antipolyamine therapy for PCP would be limited. Since the presence of ODC in P. carinii is an important issue, a new search for this enzyme was made. Not only were initial assays negative, but P. carinii extract reduced the background catalytic action of pyridoxal-5'-phosphate, the coenzyme required by the enzyme. This suggested the presence of an inhibitor, which was further supported by the observation that a P. carinii extract could suppress a source of known ODC activity. The inhibitory activity could be removed by a desalting column or by dialysis, allowing detection of P. carinii ODC. Indirect evidence indicates that the inhibition is only apparent and is caused by unlabeled ornithine in the extract of P. carinii which interferes with the radiolabel-based assay system. P. carinii and host ODCs respond differently to changes in pH. P. carinii ODC is much less susceptible to inhibition by eflornithine than host ODC. The presence of ODC in P. carinii suggests that P. carinii ODC is the target of eflornithine and that P. carinii ODC may have sufficiently specific properties that inhibitors with improved selectivity against P. carinii ODC could be identified.

Animals↗
PubMed1997

Polyamine-like immunoreactivity in rat neurons.

The localization of polyamine (PA) pools in motor, sensory, and autonomic neurons and in the nerve cells of the hypothalamo-hypophysial system of rats was examined by immunocytochemical method using the monoclonal antibody ASPM-29 specific to spermine (Spm) and spermidine (Spd) fixed in situ. Strong PA immunoreactivity was found in the cytoplasm and dendrites of the large perikaryon of motor neurons in the anterior spinal column, in the Purkinje cells of the cerebellum, in the pyramidal cells of the cerebrum, in the nerve cells of the paraventricular and supraoptic nuclei in the hypothalamus, and in the nerve cells of the spinal and autonomic ganglions. No PA immunoreactivity was seen in the nucleus and nerve terminals of the neurons. The PA immunoreactivities in the motor and sensory neurons were characterized by clustered masses and blocks of immunoreactive cells. Irrespective of location, small and medium-sized neurons were weakly PA-positive. The glia cells, some stellite cells, and Schwann cells were almost completely PA-negative. These results may suggest that in neurons PAs are not transported axonally, but are located in conjunction with Nissl bodies (the rough endoplasmic reticulum), specified as sites for protein synthesis within cells.

Animals↗