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Tertiary tetraamines and quaternary pentaamines composed of aminopropyl and/or aminobutyl groups were synthesized as authentic samples for the identification of naturally occurring branched polyamines. Four tertiary tetraamines were obtained by alkylating the free secondary amine group of diphthaloyl derivatives of sym-norspermidine or sym-homospermidine with N-(3-bromopropyl)phthalimide or N-(4-bromobutyl)phthalimide in the presence of KF-Celite. Five quaternary pentaamines were obtained by fusing triphthaloyl derivatives of the tertiary tetraamines with an excess amount of N-(3-iodopropyl)phthalimide or N-(4-iodobutyl)phthalimide. The present methods are simple and achieved high yields. The 13C-NMR spectra of these branched polyamines were recorded in D2O as fully protonated forms, and all 13C chemical shifts were assigned consistently.

The synthesis and translocation of the diamine cadaverine during soybean (Glycine max L. Meer cv. Sakai) germination were studied using 15N-labelled lysine (the cadaverine precursor) and 15N-labelled cadaverine, both under light/dark (12 h/12 h) and total dark germinating conditions. 15N-cadaverine and non-labelled polyamines were simultaneously detected using ionspray ionization–mass spectrometry. Both 15N-cadaverine and 15N-lysine were taken up by soybean. 15N-lysine was transported to the shoot and root and converted into 15N-cadaverine, whereas relatively little 15N-cadaverine was formed from 15N-lysine in the cotyledon. The acropetal translocation of 15N-cadaverine from the cotyledon to the shoot seemed to predominate over basipetal transport to the root. Although no other 15N-derivatised polyamines were found, supplying exogenous 15N-lysine seemed to indirectly affect the metabolism of 14N putrescine, spermidine and spermine, while no significant effect was detected after supplying 15N-cadaverine.

Triamines such as norspermidine, spermidine, and homospermidine and tetraamines such as norspermine, spermine, thermospermine, and aminopropylhomospermidine were found to be distributed ubiquitously in the eight extremely thermophilic (growing at 70 degrees C) Thermus species tested. Three linear pentaamine (caldopentamine, homocaldopentamine, and thermopentamine), two linear hexaamines (caldohexamine and homocaldohexamine), two tertiary branched tetraamines (N4-aminopropylnorspermidine and N4-aminopropyl-spermidine), and quaternary branched pentaamines such as N4-bis(aminopropyl)norspermidine and N4-bis(aminopropyl)spermidine were detected in T. thermophilus HB8, T. filiformis Wai33 A1, T. flavus AT-62, and T. caldophilus GK24. The linear hexaamines and branched polyamines were absent in T. aquaticus YT-1, T. sp. X-1, T. sp. T2, and T. sp. T351, in which linear pentaamines were minor components. Moderately thermophilic Thermus ruber and Thermus sp. K-2 contained putrescine, spermidine, norspermidine, homospermidine, spermine, norspermine, thermospermine, and aminopropylhomospermidine. No pentaamines, hexaamines, or branched polyamines were found in these two moderately thermophilic Thermus species. On the other hand, moderately thermophilic, acidophilic Acidothermus cellulolyticus was devoid of all the polyamines.

1. 1. Polyamines were extracted from the guts and ovaries of the sea urchin Anthocidoris crassispina, and the guts and flesh of the sea cucumber Stichopus japonicus and the sea squirt Halocynthia roretzi, the oyster Crassostrea gigas and the short-necked clam Tapes philippinarum, and analyzed by ion-exchange high-performance liquid chromatography and gas chromatography-mass spectrometry. 2. 2. Norspermidine and norspermine as well as putrescine, cadaverine, spermidine, spermine and agmatine were the ubiquitous polyamines in these invertebrates. These results suggest the widespread distribution of norspermidine and norspermine in invertebrates. 3. 3. Thermopentamine, thermohexamine and homothermohexamine were found in the sea urchin. This in the first report on the occurence of thermopentamine and hexaamine in invertebrates. 4. 4. Homospermidine, canavalmine, aminopropylhomospermidine, homospermine, caldopentamine, homocaldopentamine and aminopropylcanavalmine were found in the sea cucumber. Homospermidine, aminopropylhomospermidine and homospermine were found in the squirt. This is the first report on the occurence of canavalmine, aminopropylhomospermidine, homospermine, homocaldopentamine and aminopropylcanavalmine in invertebrates.

A slightly thermophilic hydrogen bacterium, "Pseudomonas hydrogenothermophila, " was found to contain 2-hydroxyputrescine, 2-hydroxyspermidine, diaminopropane, putrescine, cadaverine, spermidine, aminopropylcadaverine and spermine. A mesophilic hydrogen-oxidizing pseudomonad, "Pseudomonas hydrogenovora" contained putrescine, 2-hydroxyputrescine, and spermidine. The polyamine distribution pattern of the latter is rather common in the beta subclass of Proteobacteria. The occurrence of 2-hydroxyspermidine, aminopropylcadaverine and spermine is unique in the thermophile.

1. 1. Diaminopropane, putrescine, norspermidine, spesrmidine, norspermine and spermine were commonly found in the larval silk gland and head of Bombyx mori, Antheraea yamamai and Galleria mellonella . 2. 2. The cockroach Periplaneta americana contained thermospermine, caldopentamine, caldohexamine, homospermidine, aminopropylhomospermidine and aminobutylhomospermidine (homospermine) in addition to the common polyamines. This is the first report to show the occurrence of the homospermidine derivatives and a hexamine in insects. 3. 3. In addition to thermospermine, caldopentamine, agmatine, histamine, cadaverine and other usual polyamines, three cadaverine-derivatives, aminopropylcadaverine, bis(aminopropyl)cadaverine and aminopentylnorspermidine, were detected in the silk gland and head of the spiders, Nephila clavata and Araneus ventricosus . The occurrence of aminopropyl derivatives of of cadaverine has never been reported in animals.

Loss-of-function mutants of the ACAULIS5 (ACL5) gene in Arabidopsis thaliana have severe defects in stem elongation. ACL5 was previously reported as encoding a spermine synthase. A more recent study, however, showed that the bacterial expressed recombinant ACL5 protein catalyzes the conversion of spermidine to thermospermine, a structural isomer of spermine, rather than to spermine. In the present study, we found that thermospermine was detected in wild-type seedlings but was not detectable in the acl5-1 mutant. We further examined the effect of exogenous application of these isomers on the growth of acl5-1. Daily application of 0.1 mM thermospermine onto the shoot apex partially rescued the dwarf phenotype of acl5-1, while that of spermine had no effects on the morphology of the mutant. The acl5-1 transcript level in acl5-1 seedlings, which is much higher than the ACL5 transcript level in wild-type seedlings, was reduced by exogenous thermospermine. Thus we conclude that thermospermine is indeed produced through the action of ACL5 and required for stem elongation in Arabidopsis.

Four unusual polyamines were detected in the Gram-negative, aerobic, nonsporulating eubacterium, Thermomicrobium roseum (ATCC 27502), an obligate for thermophily, in which no peptidoglycan dianmino acid occurs. They were identified as caldopentamine [NH2(CH2)3NH(CH2)3 NH(CH2)3NH(CH2)3NH2], homocaldopentamine [NH2(CH2)3NH(CH2)3NH(CH2)3 NH(CH2)4NH2], thermopentamine [NH2(CH2)3 NH(CH2)3NH(CH2)4NH(CH2)3NH2] and caldohexamine [NH2(CH2)3NH(CH2)3NH (CH2)3NH(CH2)3NH(CH2)3NH2] by high-performance liquid chromatography and gas chromatography-mass spectrometry. These unusual pentaamines and hexaamine were major polyamines in the organism grown at 60–70°C.

An unknown highly basic polyamine peak was observed in Thermus thermophilus and it was identified to be a pentaamine [NH2(CH2)3NH (CH2)3NH(CH2)4NH(CH2)NH2] by gas chromotography-mass spectrometry and named as thermopentamine. The new pentaamine coexisted with another isomer, homocaldopentamine, at a molar ratio of 1:1, as a major polyamine of the organism. This is the first report to show the occurrence of thermopentamine in nature.

The Gram-negative facultative chemolithotroph, Paracoccus denitrificans contains putrescine, cadaverine, agmatine, spermidine, aminopropylcadaverine, spermine, thermospermine and aminopentylnorspermidine. This bacterium has the ability to produce norspermidine from supplemented diaminopropane. The halophile, Paracoccus halodenitrificans is devoid of any polyamines. Neither decarboxylation of ornithine, lysine or arginine, nor triamine synthetic activity from diamines was detected in this halophile. Two Gram-negative facultative photoautotrophs, Rhodobacter sphaeroides and Rhodobacter capsulatus contain putrescine, cadaverine, agmatine and spermidine and can produce norspermidine from supplemented diaminopropane. A Gram-negative eubacterium, Micrococcus cryophilus, contains histamine and homospermidine in addition to putrescine, cadaverine and spermidine. Hence, polyamine distribution patterns and polyamine biosynthetic activities were very different among the four groups of Gram-negative eubacteria examined.

In the extreme thermophile Thermus thermophilus, a disruption mutant of a gene homologous to speB (coding for agmatinase = agmatine ureohydrolase) accumulated N1-aminopropylagmatine (N8-amidino-1,8-diamino-4-azaoctane, N8-amidinospermidine), a new compound, whereas all other polyamines produced by the wild-type strain were absent from the cells. Double disruption of speB and speE (polyamine aminopropyltransferase) resulted in the disappearance of N1-aminopropylagmatine and the accumulation of agmatine. These results suggested the following. 1) N1-Aminopropylagmatine is produced from agmatine by the action of an enzyme coded by speE. 2) N1-Aminopropylagmatine is a metabolic intermediate in the biosynthesis of unique polyamines found in the thermophile. 3) N1-Aminopropylagmatine is a substrate of the SpeB homolog. They further suggest a new biosynthetic pathway in T. thermophilus, by which polyamines are formed from agmatine via N1-aminopropylagmatine. To confirm our speculation, we purified the expression product of the speB homolog and confirmed that the enzyme hydrolyzes N1-aminopropylagmatine to spermidine but does not act on agmatine.

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.

This study reports the first identification of 4-methylspinaceamine (4-MSPA)-a Pictet Spengler condensation reaction product of histamine with acetaldehyde-in human urine. 4-MSPA was identified and quantified as follows: the target compound was partially purified by solvent extraction from a urine sample spiked with N-methylpiperazine (N-MP) as an internal standard, then derivatized to a naphthylthiourea derivative with 1-naphthylisothiocyanate (NITC) and finally analyzed by HPLC. For verification, 4-MSPA was also analyzed by ion spray-mass spectrometry (IS-MS), using 4-MSPA-d4 as an internal standard. The amount of 4-MSPA in human urine varied between individuals and from day-to-day, ranging from undetectable to 0.80 nmol/mL.

The reactivity of malondialdehyde(MDA) with spermidine at the physiological pH was considerably higher than that with putrescine, glycine, lysine, methylamine or dimethylamine, and the major reaction product of MDA-spermidine adduct was identified as N4-(2-propenal)spermidine.

Cellular polyamines of newly isolated acidophilic, thermophilic and thermoacidophilic archaebacteria were investigated for the chemotaxonomic significance of polyamine distribution profiles. In addition to spermidine, spermine and agmatine, a quaternary branched penta-amine, N 4 -bis(aminopropyl)spermidine, was found in thermophilic Thermococcus waiotapuensis, Thermococcus aegaeus and Pyrococcus glycovorans belonging to the order Thermococcales. An acidophilic euryarchaeon, Ferroplasma acidiphilum located in the order Thermoplasmatales, contained spermidine and agmatine. Norspermidine, spermidine, norspermine and spermine were found in thermoacidophilic Acidilobus aceticus and thermophilic Thermodiscus maritimus located in the order Desulfurococcales, and in thermophilic Pyrobaculum arsenaticum, Pyrobaculum oguniense, Vulcanisaeta distributa and Vulcanisaeta souniana belonging to the order Thermoproteales; however, the four genera differ on their tetra- and penta-amine levels. Thermophilic Staphylothermus hellenicus belonging to Desulfurococcales contained caldopentamine, caldohexamine and N 1 -acetylcaldopentamine in addition to norspermidine, spermidine and norspermine. This is the first report on the occurrence of acetylated penta-amine in nature.

Relative reactivities of histamine and indoleamines such as tryptamine, 5-hydroxytryptamine and 5-methoxytryptamine with acetaldehyde (AA) under physiological conditions were investigated. AA was found to have much higher reactivity towards histamine than towards indoleamines. For example, when a reaction mixture of AA (1 mM) and histamine or tryptamine (5 mM) in 0.1 M phosphate buffer (pH 7.4) was incubated at 37 degrees C for 24 h, AA decreased by 11% in the case of tryptamine, while in the case of histamine, it decreased 88%. In addition, the reaction product of AA with histamine was investigated. Mixtures of a fixed amount of histamine (5 mM) and various amounts of AA (1-20 mM) in phosphate buffer (pH 7.4) were incubated for 5 h at 37 degrees C. In all cases, only one product, 4-methylspinaceamine (4-MSPA), was observed. The yield of 4-MSPA was in approximate agreement with the losses of histamine and AA, indicating that the loss of histamine caused by the reaction of AA was quantatively converted to 4-MSPA. These results show that the reaction of AA with histamine easily takes place to produce 4-MSPA in an aqueous medium close to physiological conditions.

We studied absorption, distribution, metabolism, and excretion of polyamines (putrescine, spermidine, and spermine) in the gastrointestinal tract using (15)N-labeled polyamines as tracers and ionspray ionization mass spectrometry (IS-MS). The relatively simple protocol using rats bearing solid tumors provided useful information. Three (15)N-labeled polyamines that were simultaneously administered were absorbed equally from gastrointestinal tract, and distributed within tissues at various concentrations. The uptake of (15)N-spermidine seemed preferential to that of (15)N-spermine since the concentrations of (15)N-spermidine in the liver and tumors were higher, whereas those of (15)N-spermine were higher in the kidney, probably due to the excretion of excess extracellular spermine. Most of the absorbed (15)N-putrescine seemed to be lost, suggesting blood and tissue diamine oxidase degradation. Concentrations of (15)N-spermidine and (15)N-spermine in the tumor were low. We also describe the findings from two rats that were administered with (15)N-spermine. The tissue concentrations of (15)N-spermine were unusually high, and significant levels of (15)N-spermidine were derived from (15)N-spermine in these animals.

The major cytotoxic activity of Moxa was extracted with CH2Cl2 and partially purified by three cycles of silica gel column chromatography. The active fractions showed higher cytotoxicity against six human tumor cell lines (two oral squamous cell carcinoma, one salivary gland tumor, one melanoma, two leukemia) than three normal oral human cells (gingival fibroblast, periodontal ligament fibroblast, pulp cell). All fractions failed to protect the cells from the cytopathic effect induced by HIV infection. ESR spectroscopy showed that all fractions produced little or no radical under alkaline conditions, while showing much lower O2- scavenging activity, generated by hypoxanthine-xanthine oxidase reaction, than antioxidants and polyphenols. Active fractions induced DNA fragmentation in HL-60 cells, but failed to modify the mobility and activity of mitochondrial Mn-containing superoxide dismutase (MnSOD), in contrast to Moxa smoke. These data suggest that the active principles in the Moxa extract might be different from that in Moxa smoke, which produced carbon radical and modified MnSOD mobility and activity.

Cellular polyamine fraction of alkaliphilic Bacillus species was analyzed by HPLC. 2-Phenylethylamine was found selectively and ubiquitously in the five strains belonging to Bacillus cohnii within 27 alkaliphilic Bacillus strains. A large amount of this aromatic amine was produced by the decarboxylation of L-phenylalanine in the bacteria and secreted into the culture medium. The production of 2-phenylethylamine may serve for the chemotaxonomy of alkaliphilic Bacillus.

During the analysis of the cellular polyamines by high-performance liquid chromatography and gas chromatography within the Cytophaga-Flavobacterium Bacteroides phylum, an usual amine corresponding to 2-phenylethylamine was detected in Psychroflexus torqu s belonging to the family Flavobacteriaceae. Homospermidine, agmatine and 2-phenylethylamine were detected as the major cellular polyamine components of the organism. A part of 2-phenylethylamine produced in the cells was secreted into the medium whereas agmatine and homospermidine were scarcely secreted. i

POLYAMINE OXIDASE 1 (OsPAO1), from rice (Oryza sativa), and POLYAMINE OXIDASE 5 (AtPAO5), from Arabidopsis (Arabidopsis thaliana), are enzymes sharing high identity at the amino acid level and with similar characteristics, such as polyamine specificity and pH preference; furthermore, both proteins localize to the cytosol. A loss-of-function Arabidopsis mutant, Atpao5-2, was hypersensitive to low doses of exogenous thermospermine but this phenotype could be rescued by introduction of the wild-type AtPAO5 gene. Introduction of OsPAO1, under the control of a constitutive promoter, into Atpao5-2 mutants also restored normal thermospermine sensitivity, allowing growth in the presence of low levels of thermospermine, along with a concomitant decrease in thermospermine content in plants. By contrast, introduction of OsPAO3, which encodes a peroxisome-localized polyamine oxidase, into Atpao5-2 plants could not rescue any of the mutant phenotypes in the presence of thermospermine. These results suggest that OsPAO1 is the functional ortholog of AtPAO5.

We found a tertiary methylated tetraamine, N 4 -methylthermospermine [NH 2 (CH 2 ) 3 N (CH 3 ) (CH 2 ) 3 NH (CH 2 ) 4 NH 2 ] [PA 3(Me) 34] in addition to spermidine, homospermidine, spermine, thermospermine and canavalmine in extracts of mature seeds of leguminous plants, Wistaria floribunda , Canavalia gladiata and Astragalus sinicus by HPLC and GC-mass spectrometry. This is the first report to show the presence of a tertiary branched methyl derivative of any polyamine.

Ultra-filtrable and macromolecule-bound polyamines in rat liver homogenates, made without buffer, were determined, using Potter-Elvehjem homogenizer and commercially available, pressure-aided ultrafiltration device with a membrane pore size that allows passage of particles of molecular weight no larger than 5000. About 90% of polyamines in the liver were shown to be equilibrated with externally added 15N-labeled polyamines, based on the difference in the ratio of the natural to 15N-labeled polyamine in the liver homogenate and the ultrafiltrate. The entire amount of ultrafiltrate in the homogenized liver, required for calculation of the amounts of ultra-filtrable and macromolecule-bound polyamines, was estimated to be about 0.25 g in one gram of the homogenate, using a limited dilution curve of spermine in the ultrafiltrate with phosphate buffered saline and distilled water. With this value, ultra-filtrable polyamines in normal rat liver homogenate were calculated as about 25%, 8%, and 2% of the total amount of putrescine, spermidine, and spermine, respectively. The method was then used to measure ultra-filtrable and macromolecule-bound polyamines in regenerating rat liver homogenates, to examine possible changes of polyamines during cell growth. The method was also applied to measure other ultra-filtrable compounds such as amino acids and inorganic ions in rat liver homogenate.

The analytical method for various 15N-enriched spermidines was first applied for studies on polyamine metabolism of the yeast, Candida boidinii CBS 5777. Analyses were carried out during growth of the yeast in two defined media containing 15N-enriched ammonium sulfate as a sole nitrogen source, and newly prepared α-15N-enriched L-ornithine with ammonium sulfate as nitrogen sources. Observation in the former culture indicated that the ammonia pool was negligible, and the putrescine pool was larger than the decarboxylated S-adenosylmethionine pool in this organism.The latter culture suggested that α, δ-nitrogens of ornithine behaved equally throughout polyamine metabolism.

A large amount of 2-phenylethylamine was produced in cells of alkalophilic Bacillus sp. strain YN-2000. This amine is secreted in the medium during the cell growth. The amounts of 2-phenylethylamine in both cells and medium change upon changing the pH of the medium.

When polyamines in mature seeds of a leguminous plant, Dolichos lablab were analysed by HPLC, among unknown compounds N-(3-aminopropyl)aminoethanol [NH2 (CH2)3NH(CH2)2OH] and N-(3-aminopropyl)amino-propanol [NH2(CH2)3NH(CH2)3OH] were identified by GC-MS. Polyamines found in the seeds were diaminopropane (PA 3), putrescine (PA 4), spermidine (PA 34), spermine (PA 343), thermospermine (PA 334) and aminopropylhomospermidine (PA 344). This is the first report to show the occurrence of aminoalkylaminoalkylalcohols (oxyalkyldiamines), which are possibly produced by aminopropyltransfer to aminoalkylalcohol (oxyalkylamine) in nature.

We found new pentaamines in extracts of sword bean Canavalia gladiata. They are identified as aminopropyl and aminobutyl derivatives of canavalmine (PA 434), i.e. aminopropylcanavalmine [NH2(CH2)3NH (CH2)4NH(CH2)3NH(CH2)4NH2] (PA 3434) and aminobutylcanavalmine [NH2(CH2)4NH(CH2)4NH(CH2)3NH(CH2)4NH2] (PA 4434). The content of PA 3434 in the sword bean was ca 5% that of PA 434 and PA 4434 is present in much lower concentrations. The two pentaamines were identified by HPLC coupled with enzymatic cleavage method and GC-MS.

The coloration of iron(III)-thiocyanate complexes in acidic solution was dependent on the kind of coexistent polyoxyethylene nonionic surfactants, RO(CH2CH2O)nH. The surfactants with R = oleyl group allowed not to fade the red complexes for 120 min or more. The average addition number (n) of ethylene oxide groups for the surfactant affected the color intensity. When n≥ 20, e = 2.2 × 104 mol-1 dm3 cm-1 (at 480 nm).

When the yeasts Saccharomyces cerevisiae, Candida albicans and Schizosaccharomyces pombe and the thermophilic bacteria Bacillus stearothermophilus and Bacillus acidocaldarius were cultured in the presence of homospermidine, a new compound accumulated in the cells within a few days. This compound was identified as aminopropylhomospermidine [NH2(CH2)3NH (CH2)4NH(CH2)4NH2] by gas chromatographymass spectrometry (GC-MS) and by the enzymatic cleavage method developed in our laboratories. This polyamine was not produced from homospermidine in Escherichia coli, Bacillus subtilis, Bacillus alkalophilus, or a eukaryotic protozoon, Tetrahymena pyriformis, none of which usually contains appreciable amounts of spermine. These findings suggest that the synthesis of aminopropylhomospermidine from homospermidine is mediated by spermine synthase.

The equilibrium constants for the complex formation of cobalt(II) and zinc(II) with several β-diketones in 1M sodium perchlorate solution ( β n ), the two-phase distribution constants of the bis-chelates of these metals between carbon tetrachloride and the above aqueous solution ( Kdm ), and the adduct formation of the bis-chelates with trioctylphosphine oxide (TOPO) in the organic phase ( β n ( org ) ) have been determined at 25°C. The distribution constants of the bis-zinc(II) chelates were also determined when the solvent was hexane, chloroform or benzene. The weaker the acidity of the ligand the higher the β n value for these complexes. The relationship, log Kdm = 2 log Kd + C , where Kd is the distribution constant of the extractant is obeyed for each of the zinc(II) chelates when the solvent is different. The Kdm values of the metal chelates with the same ligand are always lower for cobalt(II) than for zinc(II), and among the series of bis-chelates of the same metal ion, there is a tendency for β n ( org ) of TOPO adducts to be higher when the β 2 is lower, but the stability order of the adducts of a set of cobalt(II) and zinc(II) chelates with the same β-diketone is not always the same.

Solvent extraction of chromium(VI) from 1 M (H, Na)X solutions and HX solutions at various concentrations (X − = chloride, perchlorate, or nitrate) with trioctylphosphine oxide(E) in hexane has been studied at 25°C. When the hydrogen ion concentration was higher than 10 −3 M, the chromium(VI) species in the aqueous phase were H 2 CrO 4 and HCrO 4 − in all the systems with a chloride complex also present in the chloride system. The extracted species were H 2 CrO 4 ·E 2 , H 2 CrO 4 ·HX· E n , and H 2 CrO 4 (HX) 2 · E n where n was 2 in the perchlorate and nitrate systems, and 3 in the chloride system. The equilibrium constants for the first acid dissociation and the chloride complex formation in the aqueous phase and the extraction constants for the species in the organic phase were determined. It was assumed that the chromic acid and the mineral acid in the extracted mixed acid species were combined by a hydrogen-bond.

Various polyamine analogs were examined for their ability to stimulate and to function as sparing agents for the Mg2+ requirement in polypeptide synthesis at various temperatures in Escherichia coli (37 and 47 degrees C) and the extremely thermophilic Thermus thermophilus (60 and 70 degrees C) cell-free systems. The optimal concentration of each polyamine analog increased as the incubation temperature was elevated. At a fixed temperature, the optimal concentration of polyamine analogs was in the order diamines greater than triamines greater than tetraamines greater than pentaamines. All diamines tested stimulated polypeptide synthesis almost equally but lowered the optimal Mg2+ concentration in the order diaminopropane greater than putrescine greater than cadaverine. The degree of diamine stimulation was maximal at 37 degrees C. The effects of three triamines were very similar in the E. coli system but in the T. thermophilus system spermidine was most effective in stimulation of polypeptide synthesis. From the results of experiments using tetraamines and pentaamines, it was deduced that the presence of both aminobutyl and aminopropyl groups in polyamine analogs is important for stimulation of polypeptide synthesis. In the E. coli system, triamines were the most effective polyamines for stimulation of polyphenylalanine synthesis at both 37 and 47 degrees C, while, in the T. thermophilus system, thermospermine, a tetraamine, was most effective at 60 degrees C and 3,4,4,3-pentaamine was most effective at 70 degrees C.

The kinds of linear pentaamines with various combinations of 3 or 4 methylene chain intervals were synthesized. The methods were tentatively classified into two, leading to symmetrical and unsymmetrical pentaamines, all of which were prepared by successive alkylation of secondary amino derivatives of benzylamine with N-(3-bromopropyl or 4-bromobutyl)phthalimide in the presence of KF-Celite, coupled with the purification of the phthaloyl compounds by silica gel column chromatography. Protecting benzyl and phthaloyl groups were removed by usual methods. The carbon-13 unclear magnetic resonance (13C-NMR) spectra of the ten pentaamines were recorded in D2O as fully protonated forms, and a comparative analysis of their spectra allowed the complete assignment of all 13C chemical shifts.

Polyamines were analyzed in 4 species of genus Agrobacterium . Not only putrescine, spermidine and spermine, but also homospermidine and thermospermine were found in A. tumefaciens, A. radiobacter, A. rubi and A. rhizogenes . Trace amounts of aminopropylhomospermidine were also observed. Norspermidine and norspermine were formed from diamonorpropane added to the medium. Aminopropylcadaverine and its aminopropyl derivative(s) (aminopentylnorspermidine and N , N ′-bis(3-aminopropyl) cadaverine) were produced from the supplemented cadaverine. A strain of A. rhizogenes normally contains only putrescine and homospermidine; no other diamines, triamines and tetraamines were synthesized.

Solvent extraction equilibria of phosphoric acid with trioctylphosphine oxide (TOPO) has been studied at 25°C. When the diluent was hexane, a trisolvate with TOPO was extracted but when it was benzene or carbon tetrachloride, both a trisolvate and a disolvate were extracted. Extraction into hexane was slightly enhanced by hydrochloric acid and greatly impaired by perchloric or nitric acid in the aqueous phase and the impairment was found to be due to a decrease in the free TOPO concentration caused by the extraction of the strong acid. It is concluded after the correction for this decrease in the free TOPO concentration that these strong acids enhanced the net extraction of phosphoric acid by forming a mixed acid species, H3PO4·HX·(TOPO)3 (where HX is a strong acid), which is assumed to have a hydrogen-bond between the phosphoryl oxygen of the phosphoric acid and HX, and that the order of enhancement is HClO4 > HNO3 > HCl. The constants for the above extraction equilibria were determined.

The solvent extraction of alkali metal(I) and cobalt(II) from 1 mol dm-3 solutions of five alkali metal thiocyanates into 1, 2-dichloroethane with Triton X-100 was examined. The extractability of cobalt (II) was closely dependent on that of the alkali metal thiocyanate. The reason for this was considered.

The solvent extraction equilibria of acetic, propionic, and butyric acid and mono-, di-, and trichloroacetic acid (HA), as well as those of oxalic, malonic, succinic, glutaric, and adipic acid (H2A), have been determined in liquid–liquid systems of hexane containing none or a certain amount of trioctylphosphine oxide (E) and an aqueous 1 mol dm3 chloride solution at 25°C. The constants, Kd=[HA]org/[HA], Kdim=[(HA)2]org/[HA]org2, Kex1′=[HAE]org/[HA][E]org, Kex2′=[H2AE2]org/[H2A][E]org2, and β1(org)=[HAE]org/[HA]org[E]org2, have then been determined from the data. It was found that, among the three fatty acids, the value of Kex1′ was higher as the chain length increased, but the values of β1(org) were nearly all the same, while among the chloroacetic acids they were higher as the number of chloride atom increased. The extraction of the dicarboxylic acids with E was not very different, although the number of the –CH2– between the two carboxyl groups changed from zero to four.

Besides putrescine, cadaverine, spermidine, spermine and thermospermine, three novel polyamines were detected in a slightly halophilic eubacterium Halococcus acetoinfaciens (IAM 12094, ATCC 25861). These novel polyamines were found to be N-3-aminopropylcadaverine [NH2(CH2)3NH(CH2)5NH2] and its aminopropyl derivatives: aminopentylnorspermidine [NH2(CH2)3NH(CH2)3NH(CH2)5NH2] and N, N′ -bis(3-aminoprophyl)cadaverine [NH2(CH2)3 NH(CH2)5NH(CH2)3NH2]. Aminopropylcadaverine was also detected in two other species, Halococcus agglomeratus (IAM 12095, ATCC 25862) and Halococcus nondenitrificans (IAM 12096, ATCC 25863).

In the phylogeny of plant polyamine oxidases (PAOs), clade III members from angiosperms, such as Arabidopsis thaliana PAO5 and Oryza sativa PAO1, prefer spermine and thermospermine as substrates and back-convert both of these substrates to spermidine in vitro. A clade III representative of lycophytes, SelPAO5 from Selaginella lepidophylla, also prefers spermine and thermospermine but instead back-converts these substrates to spermidine and norspermidine, respectively. This finding indicates that the clade III PAOs of lycophytes and angiosperms oxidize thermospermine at different carbon positions. We discuss the physiological significance of this difference.

The extraction of water from a 1 mol dm−3 sodium perchlorate solution with trioctylphosphine oxide (TOPO) in four organic solvents and the co-extraction of water with hydrochloric, hydrobromic, hydroiodic, perchloric, nitric, and thiocyanic acids from 1 mol dm−3 of the acid solution by TOPO in hexane have been studied at 25 °C. The extraction constants of water in the absence of any of the acids, K′H2O=[TOPO·H2O]org[TOPO]org−1, were determined to be 100.40, 100.00, 10−0.12, and 10−0.36 in benzene, hexane, carbon tetrachloride, and chloroform respectively. A discrepancy from the 1: 1 molar ratio of TOPO and water in the extract was observed in the higher-TOPO-concentration region in the hexane system. The co-extraction of water with the acids disolvated by TOPO was greater in the order of HCl\simeqHBr>HI>HClO4, and it was much poorer with the monosolvated acids, nitric and thiocyanic acid. The equilibria for these co-extractions of water with the acids are complicated.