Investigation of the actions and antagonist activity of some polyamine analogues in vivo.

1. The ability of three putative polyamine antagonists to antagonize behavioural changes induced by spermine was assessed. 2. Injection of an excitotoxic dose of spermine (100 microg, i.c.v.) in mice results in the development of a characteristic behavioural profile, which has two temporally distinct phases. The early events include clonic convulsions, and the later, more general excitation, includes tremor and culminates in the development of a fatal tonic convulsion. 3. Co-administration of arcaine (25 microg, i.c.v.) potentiated the early phase effects after spermine injection, but antagonized the development of spermine-induced tonic convulsions. A larger dose of arcaine (50 microg, i.c.v.) given alone resulted in the development of spermine-like body tremor and convulsions. It therefore appears that arcaine is not a pure polyamine antagonist in vivo, but may be a partial agonist. 4. Similarly, 1,10-diaminodecane appeared to act as a partial agonist in vivo, although it was less potent than arcaine. 5. In contrast, diethylenetriamine (DET) effectively inhibited the development of the early effects of spermine, but was ineffective against the spermine-induced CNS excitation and tonic convulsions. 6. It is concluded that none of the putative polyamine antagonists tested behaved as effective polyamine antagonists in vivo, although each produced some antagonism.

Effects of herbicides on two submersed aquatic macrophytes, Potamogeton pectinatus L. and Myriophyllum sibiricum Komarov, in a prairie wetland.

Clopyralid, picloram, 2,4-D and a mixture of 2,4-D plus picloram, (Tordon 202C) were added to the water of 1 m square enclosures in a prairie wetland in Saskatchewan, Canada to produce concentrations of 0.01 and 0.1 mg active ingredient litre(-1). Effects on the submersed macrophytes, Potamogeton pectinatus and Myriophyllum sibiricum, were monitored by taking repeated measurements of plant weight, flower and tuber production and inspecting for injuries at 30 and 60 days after application. Clopyralid did not inhibit weight gain (growth) in either species, but stimulated growth and flowering by M. sibiricum at 0.01 mg litre(-1) and tuber production by P. pectinatus at both rates. The low rate of 2,4-D stimulated flowering by M. sibiricum and tuber production by P. pectinatus, whereas the high rate inhibited growth of M. sibiricum and injured both species. Picloram did not affect growth of either species, but injured M. sibiricum at both concentrations and inhibited flowering at 0.1 mg litre(-1). Tordon 202C at 0.1 mg litre(-1) caused reduced growth and flowering in M. sibiricum and injured both species; 0.01 mg litre(-1) also injured M. sibiricum. Mortality resulted only from Tordon 202C and 2,4-D. Field data are lacking to assess the extent to which submerged macrophytes in prairie ponds are exposed to harmful concentrations of herbicide from aerial spraying, drift from ground application, runoff or wind erosion of soil.

Investigation of the involvement of the N-methyl-D-aspartate receptor macrocomplex in the development of spermine-induced CNS excitation in vivo.

1. The involvement of the N-methyl-D-aspartate (NMDA) receptor macrocomplex in the development of spermine-induced CNS excitation in vivo was investigated. 2. Injection of 100 micrograms of spermine into the left lateral cerebral ventricle of female Laca mice (20-25 g) resulted in the development of two distinct phases of CNS excitatory effects which were quantified by a scoring system. 3. The first phase effects occurred within minutes of injection and generally lasted for about 1 h. Most mice showed scratching of the upper body, frequent face washing and some mice developed clonic convulsions. By about 2 h after injection, the second phase of effects began to develop in the form of body tremor which worsened with time and culminated in fatal tonic convulsions, generally within 8 h of injection. 4. Pretreatment of the mice with dizocilpine (0.3 mg kg-1, i.p.) resulted in antagonism of the first phase of spermine-induced effects, but a higher dose (0.3 mg kg-1, (x2), i.p.) was necessary to inhibit the second phase effects. 5. Whereas the glutamate antagonist, 3-((R)-2-carboxypiperazin-4-yl) propyl-1-phosphonic acid (D-CPP) (10, 20 mg kg-1, i.p.), the glycine antagonist 7-chlorokynurenate (10, 30, 50 nmol, i.c.v.), or the polyamine antagonist ifenprodil (30, 60 mg kg-1, i.p.) antagonized the first phase of effects produced by spermine, these agents given as monotherapy, were ineffective against the development of the second phase of effects. 6. Co-administration of ifenprodil with either D-CPP or 7-chlorokynurenate resulted in a dose-dependent antagonism of the development of the second phase of spermine-induced effects. 7. It is concluded that the development of the two temporally distinct phases of spermine-induced effects may be mediated by pharmacologically distinct mechanisms, although the results suggest that the NMDA receptor macrocomplex may be involved in both phases of effects. Furthermore, a moderate dose of D-CPP or 7-chlorokynurenate appears to enhance the inhibitory potential of ifenprodil in vivo.

Some factors affecting the release of imipramine from gel-precipitated aluminium hydroxide spheres.

Changing the pH of the dissolution medium has been found to affect the release of imipramine from gel-precipitated aluminium hydroxide spheres. Release from unwashed, unheated spheres into solutions of pH 1.2 was controlled by dissolution of the gel matrix, whereas that into solutions of pH 3 and pH 5 appeared to be under diffusion control. The liberation of drug from unwashed, heated spheres into the media of higher pH exhibited more complex kinetics. Washed spheres failed to release significant amounts of imipramine into the solutions of pH 3 and 5. Changing the ionic strength of the media had little effect on drug release. These phenomena have been explained with reference to model theories of the precipitation and ageing of aluminum hydroxide gels and their pH-solubility profiles.

The production and properties of gel precipitated aluminium hydroxide spheres containing imipramine.

The production of aluminium hydroxide spheres containing imipramine by gel precipitation of liquid feed solutions has been described. Washing and heating procedures applied to the precipitated spheres markedly affected the rate of drug release from the spheres into 0.1M hydrochloric acid. Increasing the imipramine content of the spheres also altered the drug release rate. The effects described have been explained with reference to model theories of the precipitation and ageing of aluminium hydroxide gels in the presence and absence of 'foreign' anions.

High-affinity uptake of spermine by slices of rat cerebral cortex.

The accumulation of the polyamine spermine into 0.1-mm prisms of rat cerebral cortex has been investigated at both 37 percent C and at 4 percent C. Kinetic analysis of the temperature-sensitive portion of uptake indicates two high-affinity saturable components together with an unsaturable component at high concentrations. The "very high'-affinity saturable system (K(m)= 3.8 nM) was temperature- and sodium-dependent, and significantly reduced by metabolic inhibitors, finding that are consistent with an active transport system for spermine into brain tissue. The "high'-affinity saturable component (K(m)= 0.44 micron) was sodium-dependent and inhibited by ouabain, but only partially susceptible to inhibition by 2,4-dinitrophenol and sodium cyanide. The significance of these results with respect to the function of spermine in the central nervous system is discussed.

The spontaneous and evoked release of spermine from rat brain in vitro.

1 The efflux of previously accumulated [3H]-spermine from brain slices was measured using a continuous perfusion system. The spontaneous efflux was biphasic, consisting of an initial rapid efflux followed by a much slower release. 2 The slices were depolarized by the addition to the medium of high potassium concentrations, ouabain or veratrine. 3 At concentrations greater than 30 mM, potassium evoked a striking increase in the release of [3H]-spermine. Following uptake in the presence of 5.7 x 10(-9)M [3H]-spermine, K+-evoked release was dependent on the presence of calcium ions. Release of spermine after uptake at 5.6 x 10(-8)M or 5.0 x 10(-7)M was not calcium-dependent. 4 The calcium-dependent, K+-stimulated release of spermine was inhibited in the presence of diphenylhydantoin (5 x 10(-5)M) or ruthenium red (10(-5)M). 5 Following uptake of 5.7 x 10(-9)M [3H]-spermine in a sodium-free medium, the calcium-dependent, K+-stimulated release was significantly inhibited. 5 Ouabain (10(-4)M) caused a large but calcium-independent increase in the efflux of [3H]-spermine. 7 Veratrine-induced release was less substantial but was increased in a calcium-free medium. Release evoked by veratrine was abolished in the absence of sodium. 8 These results are discussed with respect to a possible 'neurotransmitter' or 'neuromodulator' role for spermine.

The synthesis and turnover of spermidine and spermine in mouse brain.

Following the administration to mice of radiolabeled putrescine by intraventricular injection, changes in the specific radioactivity of putrescine, spermidine, and spermine have been measured. Putrescine decline was biphasic, being more rapid over the first 12 hr(t 1/2 = 5 hr) than over the remainder of the 48-hr period (t 1/2 = 11 hr) that significant labeling was detected. Spermidine was rapidly labeled during the decline in putrescine radioactivity and maximum incorporation of label occurred at 18 hr. Subsequently, spermidine specific activity declined with a half-life of 22 days. Spermine synthesis was slower, with maximum labeling occurring after 4 days. Spermine turnover, measured at a time when spermidine radioactivity had substantially declined, was extremely slow (t 1/2 = 92 days). The data supports the view that putrescine is a precursor of spermidine which in turn is required for spermine synthesis.